Pollen–pistil interactions in divergent wide crosses lead to spatial and temporal pre-fertilization reproductive barrier in flax (Linum usitatissimum L.)

Pollen–pistil interactions in divergent wide crosses lead to spatial and temporal pre-fertilization reproductive barrier in flax (Linum usitatissimum L.) | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Pollen–pistil interactions in divergent wide crosses lead to spatial and temporal pre-fertilization reproductive barrier in flax (Linum usitatissimum L.) Vijaykumar Kailasrao Raut, Aneeta Yadav, Vikender Kaur, Mahesh Rao, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5280537/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 25 Feb, 2025 Read the published version in Scientific Reports → Version 1 posted 11 You are reading this latest preprint version Abstract Linseed, has been a source of natural fiber for terxtile industries since its domestication. However, despite being the potential source of trait reservoir, the use of Linum wild genetic resources for the improvement of economic traits are not exploited widely. This is mainly due to the degree of genetic divergence that exists among the interspecific ecotypes causing crossability issues. Self-incompatibility due to the occurrence of heterostyly is very well reported in distantly related crop wild relatives of Linum and, the mechanism of self-incompatibility between different floral morphs is also studied. However, pollen germination and tube growth responses in the interspecific crosses are rarely studied. Thus, the present study was exclusively carried out to assess the major pre-zygotic barriers and their effect on pollen germination on foreign stigma using fluorescent microscopy of aniline blue stain-aided technology, to understand how the species barriers operate on pollen germination and pollen tube growth. The study revealed that the pollen-pistil interaction in the wide crosses among L. usitatissimum X L. grandiflorum was regulated by both temporal and spatial pre-fertilization barriers. Callose deposition within 2 hours after pollination (HAP) at the stigma surface, was the major cause inhibiting pollen germination. Various kinds of aberrations started appearing during the 2-4HAP. The complexity of interspecific hybridization was observed in terms of arrest of pollen tube (PT) growth in the ovary, ruptured, twisted and swollen pollen tube tip, tube growth in reverse direction, convoluted and terminated growth patterns. The results suggest that although, distant hybridization is usually less efficient, hybridization success can be improved by advanced techniques such as embryo rescue and in vitro culture of isolated immature embryos that will undoubtedly yield crucial information on selecting the ideal culture conditions and medium, paving the way for groundbreaking discoveries in this field. Biological sciences/Plant sciences/Plant breeding Biological sciences/Plant sciences/Plant reproduction Biological sciences/Plant sciences Pollen-pistil interaction Linum interspecific Pre-fertilization barriers fluorescent microscopy Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Introduction Flaxseed, also known as linseed has evolved as one of the predominant industrial crops during the last few decades 49 and originally domesticated for its oilseed and natural fiber usage 33 . Grown for dual purpose, the two separate main directions have been developed for utilizing the plant's products, leading to the cultivation of fiber flax and oil flax (linseed) 13 . A rich and interesting history surrounds flax. The Fertile Crescent region, saw the domestication of this plant approximately 8,000 BCE 71 first for its seeds and later for its fibres 30 . It’s fascinating that its usage existed even before domestication. Caucasus area was investigated to report flax yarn residues dated 30,000 years ago 35 . Nonetheless, the cultivation of flax as a crop for textile fiber was made possible by Ancient Egypt 28 . Vavilov, (1926) 72 highlighted the Indian subcontinent, Abyssinia, and the Mediterranean area as major regions of flax diversity. In these regions, the wild ancestor Linum bienne was cultivated, likely in isolation, leading to the domestication of L. usitatissimum . Thus, these areas could be associated with multiple domestication events of flax. These days, high-end fabrics, natural actuators 15 , and composite material reinforcements 44 all use flax fibres. Therefore, flax is a link between cultures and periods 43 . As a vital industrial resource, linseed oil is employed for multiple purposes, primarily as a fundamental component in paints, resins, printing inks, varnishes, and linoleum 16 . Advances in material science have brought fiber flax back into the spotlight, as its fibers are now being harnessed for a multitude of environmentally sustainable industrial uses, such as composites, geotextiles, insulation, and specialty papers. In India during the winter season, linseed is the second most important oilseed crop after rapeseed mustard in terms of both area and production. It holds the top position in the country for industrial oil production and holds the top spot globally in terms of acreage, representing 23.8 percent of the world total, and ranks third in production, accounting for 10.2 percent of the world total 68 . Every part of the linseed plant is commercially utilized, either directly or after processing. The seed contains 33 to 47% of oil, containing around 58% omega-3 fatty acid. Flax is cultivated on an area of about 2.31 million hectares globally. Asian countries contribute about 49.2% area in the world 18 . India, Canada, China, the USA, and Ethiopia are the main countries for linseed production. The existence of modern-day flax varieties mainly includes domestication and selection as important historical breeding phenomenon. The initial bred flax varieties to surface in Southern Europe, especially in the Danube valley, were winter oil crops 1 . Evidence from history shows that bast fiber textiles extended to the Nile Valley and even to present-day Britain by 4,000 BC. In Eastern Europe, the creation of spring-planted fiber varieties is thought to have originated from either a unique domestication event in Central Asia (Indo-Afghan region) or from the existing European varieties found in the south 77 , 12 . At the dawn of the 20th century, Russia established itself as the foremost supplier of top-quality fiber flax to Europe. The tireless efforts of local peasants in cultivation gave rise to “kryazh,” a set of heritage landraces that developed over centuries through both natural and artificial selection, focusing on the creation of tall fiber flax varieties. These kryazh varieties were considered premium commodity crops, leading to their active trade and incorporation into breeding programs worldwide. It is widely recognized that the origins of all modern fiber flax varieties can be traced back to Eastern Europe 29 . Flaxseed, being a self-pollinated crop, has faced the narrowing down of its genetic base due to selection during domestication and plant breeding just like all other cultivated crops 21 . The wild progenitor species gave rise to all the major crops around the world, mainly about 4000 to 10000 years ago through domestication 14 . New variants can be derived from wild relatives or intermediary landraces, and a portion of these can be beneficial for enhancing crops through traditional breeding techniques or biotechnology. Linum is one of the largest genus of Linaceae family, containing more than 200 species 32 growing in diverse geographical regions of world. In India, apart from cultivated flax viz., L. usitatissimum , the occurrence of other five Crop Wild Relatives (CWR’s) namely, L. strictrum , L. bienne , L. grandiflorum, L. mysorense and L. perenne 66 is reported. However, utilization of wild linseed species in crop improvement programs has been limited partly because of lack of collection and conservation of diverse species in gene banks and partly because of the degree of genetic divergence that exists among the interspecific ecotypes causing crossability issues. Large number of CWR’s of flax are available worldwide in genus Linum that could be of immense potential for their inclusion as trait donors under various flax improvement programmes. However, the genus Linum has many diploid species that exhibit a remarkable diversity in chromosome number including n = 8, 9, 10, 12, 14, 15, 16, 18, 30, and > 30 10,26 . No absolute studies have yet emerged as a reflective of phylogenetic relationships among Linum species. There are uncertainties regarding the chromosome numbers of some wild species. The differences in chromosome size and their number pose immense hurdles in hybridizing CWRs with cultivated flax. These genetic differences between wild and cultivated species have their unfavorable impact on feasibility of hybridization among flax wild species in both or at least one direction. Successful production of fertile hybrids to an extent, is assumed to occur only in species with equal chromosome numbers. A comprehensive detail on this aspect has been represented by Jhala et al. (2008) 32 . Potential usage of some wild relatives of Linum includes fiber quality and core hardness ( L. bienne ), bud fly resistance, lignan and neolignans synthesis ( L. grandiflorum ), nutritious and oily quality ( L. lewisii ), drought and rust tolerance ( L. marginale ). Despite their huge potential, the use of these CWR’s under flax improvement programme is very scarce. Studies focused on the crossability potential 63 , 32 or elaborating the reasons behind the reproductive isolation in flax are limited. Interspecific hybridization between wild and cultivated species can play a major part to add economically important characters in Linum . Pollen-pistil interaction (PPI) offers enormous potential for the manipulation of pollen screening which is obviously for the compatibility of pollen. PPI can be considered as an important contributory factor in introducing genetic diversity to the flowering plants from their wild and weedy counterparts. Assessing the compatibility of foreign pollen on cultivated species during pollination and fertilization offers enormous potential to study the PPI and understand incompatibility barriers 36 , 67 , 2 . This may broaden the scope for bringing wild species under crop breeding programs for the improvement of traits of interests or introducing genetic diversity. Self-incompatibility due to the occurrence of heterostyly is very well reported in different species related to cultivated Linum 24 , 48 , 11 . Also, the mechanism of self-incompatibility between different floral morphs has been investigated 37 , 8 , 70 . However, to the best of our knowledge, there is rarely any reference literature available pertaining to germination and tube growth of foreign pollen grain onto the stigmatic surface and stylar regions of the parent belonging to another species in Linum . Therefore, the present investigation was carried out to (i.) assess the major pre-zygotic barriers and their effect on the germination of pollen grains on foreign stigma using fluorescent microscopy of aniline blue stain aided technology, (ii.) understand how the species barriers operate on pollen germination (PG) and pollen tube growth (PTG) when different wild species are used as the female parents and (iii.) account for the impact of maternal cytoplasm on pollen rejection, PTG, and fertilization in different cross combinations. Results Pollen vitality Pollen vitality test revealed an average 69.92% vital pollen grains in L. usitatissimum in 3108 studied pollen mother cells (PMCs) ranging from 55.33–83.94%. In L. bienne , pollen vitality percentage (PV%) ranged from 77.04–92.14% with an average of 86.35% in 1368 studied PMCs. In case of L. grandiflorum , pollen vitality percentage (PV%) ranged from 82.05–96.49% in 2268 studied pollen grains with an average of 91.30% (Table 3). Pollen germination on stigmatic surface of self-pollinated buds Analysis of PG in self- and cross-pollinated stigmatic surfaces of processed pistils was done as the percent PG (%) for the respective duration. The analysis of the captured images showed more than 90% PG on the stigmas in all the three species in case of self-pollination in each time duration (Fig. 2 a, b and c). Overall PG in self-pollination for L. usitatissimum X L. usitatissimum ( Lu X Lu ) was 94.51% for all HAP, maximum 95.99% for 10HAP and minimum 92.83% for 8HAP (Table 4 , Fig. 3 a, Table S1 ). Overall PG in self-pollination for L. bienne X L. bienne ( Lb X Lb ) was 94.27% for all HAP, maximum 95.58% for 8HAP and minimum 92.17% for 24HAP. Overall PG in legitimate mating for L. grandiflorum X L. grandiflorum ( Lg X Lg ) was 94.12% for all HAP, maximum 94.82% for 2HAP and minimum 93.54% for 10HAP. Pollen germination on stigmatic surface of cross-pollinated buds For the cross combinations involving L. bienne , the PG percentage on the foreign stigmas was more than 90% in both direct and reciprocal combinations. Overall PG in cross pollination for L. usitatissimum X L. bienne ( Lu X Lb ) was 94.00% for all HAP (Table 4 ), maximum 97.00% for 4HAP and minimum 91.91% for 8HAP. For the first time duration of observation i.e., 2HAP, the PG percent was 93.71% (Fig. 2 d). Overall PG in reciprocal cross combination for L. bienne X L. usitatissimum ( Lb X Lu ) was 94.87% for all HAP, maximum 96.19% for 4HAP and minimum 93.03% for 48HAP. For the first time duration of observation i.e., 2HAP, the PG percent was 95.20% (Fig. 2 e). For the cross combinations involving L. grandiflorum , the PG percentage on the foreign stigmas was restricted to about 10% in both direct and reciprocal combinations. Overall PG in cross pollination for L. usitatissimum X L. grandiflorum ( Lu X Lg ) was 8.15% for all HAP (Table 4 ), maximum 10.92% for 4HAP and minimum 6.66% for 48HAP. While or the first-time duration of observation i.e., 2HAP, the PG percent was 7.77% (Fig. 2 f). For its reciprocal combination L. grandiflorum X L. usitatissimum ( Lg X Lu ), the overall PG was 8.82% for all HAP, ranging from a maximum of 10.33% for 6HAP to a minimum of 7.71% for 8HAP. While for the first-time duration of observation i.e., 2HAP, the PG percent was 8.65% (Fig. 2 g). Overall, in self-pollination, PG ranged from 92.17% ( Lb X Lb for 24HAP) − 95.99% ( Lu X Lu for 10HAP) and in cross combination it ranged from 6.66% ( Lu X Lg for 48HAP) − 97.00% ( Lu X Lb for 4HAP) (Table 4 ). Pollen tube growth in self-pollinated pistils In case of self-pollination in all the three species, it took only 2HAP to grow through the stylar region and reach to up-to the ovule (Fig. 3 b, c and d, Table S1 ). The maximum number of pollen tubes (PTs) observed growing in the stylar region, ovary and tubes reaching to ovules were 75, 35 and 12 in 4HAP, 2HAP and 8HAP respectively for L. usitatissimum . The germinated PTs were observed growing with different rates in pistil ranging from 4.74% (24HAP) to 23.17% (8HAP) in stylar region, 2.37% (24HAP) to 11.38% (8HAP) in attachment region of ovary and, 0.79% (24HAP) to 4.88% (8HAP) seen penetrating ovular region inside the ovary (Fig. 2 a). For L. bienne , maximum number of PTs observed growing in the stylar region, ovary and tubes reaching to ovules were 62 (4HAP), 36 (4HAP) and 14 (4 and 8HAP) respectively. The germinated PTs were observed growing with different rates in pistil ranging from 3.35% (6HAP) to 6.15% (24HAP) in stylar region, 1.32% (48HAP) to 3.28% (4HAP) in attachment region of ovary and, 0.79% (6HAP) to 1.54% (8 and 10HAP) seen penetrating ovular region inside the ovary (Fig. 2 b). In case of L. grandiflorum , the PTG was comparatively lower in style (10), ovary (7) and ovule (6) during the initial two hours of observation (2HAP). The maximum numbers could be observed only during 8HAP in the stylar region (49), ovary (43) and tubes reaching to ovules (40). The germinated PTs were observed growing with different rates in pistil ranging from 0.93% (48HAP) to 1.95% (2HAP) in stylar region, 0.87% (48HAP) to 1.67% (4 HAP) in attachment region of ovary and, 0.87% (48HAP) to 1.48% (4HAP) seen penetrating ovular region inside the ovary (Fig. 2 c). The overall extent of PTs observed growing in the stylar region, ovary and ovule were 13.36%, 6.34% and 2.10% respectively for L. usitatissimum over the germinated pollen grains. For L. bienne . overall extent of PTs observed growing in the stylar region, ovary and ovule were 4.44%, 2.46% and 1.16% respectively. For L. grandiflorum these parameters were 1.36%, 1.20% and 1.12% for the respective pistil regions (Table 4 ). Pollen tube growth in cross-pollinated pistils In case of wide cross combination, with Lu X Lb and its reciprocal Lb X Lu it took only 2HAP to reach to the stylar region (40 and 21 respectively) and ovary (18 and 10 respectively). However, the tube could be seen reaching up to the ovular region only in about 2HAP (5 and 3 respectively) (Fig. 3 b, c and d, Table S1 ). The maximum number of PTs observed growing in the stylar region, ovary and tubes reaching to ovules were 44, 18 and 8 in 4HAP, 2HAP and 8HAP respectively for Lu X Lb. The average number of PTs observed growing in the stylar region, ovary and ovule were 8.10%, 3.33% and 1.29% respectively over the germinate pollen grains for all the studied hours (Table 4 ). The germinated PTs were observed growing with different rates in pistil ranging from 4.39% (24HAP) to 12.78% (2HAP) in stylar region, 2.10% (48HAP) to 5.75% (2HAP) in attachment region of ovary and, 0.84% (48HAP) to 2.82% (8HAP) seen penetrating ovular region inside the ovary (Fig. 2 d). For its reciprocal cross combination Lb X Lu , the maximum number of pollen tubes observed growing in the stylar region (40), ovary (30) and tubes reaching to ovules (16) could be observed only in 8HAP. The average number of PTs observed growing in the stylar region, ovary and ovule were 2.80%, 1.73% and 0.82% respectively for overall time duration considered under the current study, over the germinated pollen grains ( Table 4 ). The germinated PTs were observed growing with different rates in pistil ranging from 2.11% (10HAP) to 3.47% (4 and 48HAP) in stylar region, 1.07 (2HAP) to 2.11% (4HAP) in attachment region of ovary and, 0.27% (6HAP) to 1.25% (48HAP) seen penetrating ovular region inside the ovary (Fig. 2 e). With other distant cross combination i.e., Lu X Lg and its reciprocal Lg X Lu , again it took only 2HAP to reach to the stylar region (32 and 19 respectively) and ovary (16 and 15 respectively) (Fig. 3 b, c). However, the tube could be seen reaching up to the ovular region only in about 4HAP in Lu X Lg (10) and in Lg X Lu (14) (Fig. 3 d). The maximum number of PTs observed growing in the stylar region, ovary and tubes reaching to ovules were 39 (4HAP), 22 (48HAP) and 15(48HAP) for Lu X Lg. The average number of PTs observed growing in the stylar region, ovary and ovule were 43.53%, 22.82% and 12.00% respectively over the germinate pollen grains for all the studied hours (Table 4 ). The germinated PTs were observed growing with different rates in pistil ranging from 27.91% (8HAP) to 68.09% (2HAP) in stylar region, 13.54% (6HAP) to 40.00% (24HAP) in attachment region of ovary and, none in 2HAP to 28% (24HAP) seen penetrating ovular region inside the ovary (Fig. 2 f). For its reciprocal cross combination Lg X Lu , the maximum number of PTs observed growing in the stylar region (23), ovary (21) and tubes reaching to ovules (14) was observed in 4HAP. The average number of PTs observed growing in the stylar region, ovary and ovule were 14.92%, 12.95% and 8.01%, respectively over the germinated pollen grains for all the studied hours (Table 4 ). The germinated PTs growing with different rates in pistil ranging from 11.59% (24HAP) to 22.22% (8HAP) in stylar region, 9.42% (24HAP) to 22.22% (8HAP) in attachment region of ovary and none in 2HAP to 16.05% (8HAP) were observed penetrating ovular region inside the ovary (Fig. 2 g). Discussion Fertilization between diverse species within the same genus is expected to be successful only if there is some means of compatibility between both the parents. Once there happens a successful completion of a series on sequential events following pollination, as allowed by the coordinated gene and gene complexes of both pollen and pollen parents, successful fertilization takes place 60 , 7 , 20 . As per our past experiences, in case of interspecific hybridization in L. usitatissimum and its wild progenitor i.e., L. bienne , the seed set was successful up to 81.5 to 83.3% including reciprocals (Table S2 ). However, despite many attempts for making wide crosses in L. usitatissimum with its distant relative i.e., L. grandiflorum , we had been unsuccessful getting any viable seeds under field conditions (Fig. S2 ). Earlier studies also report unsuccessful attempts for obtaining interspecific hybrids between cultivated flax and L. grandiflorum 56 and attempts to get hybrids with embryo rescue have been also done in the past 6 . It is worth mentioning here that, capsules formation was observed during most of the attempts made under field conditions to an extent of about 70% in direct crosses and 22% in reciprocal cross combinations with L. grandiflorum (Table S2 ). Since, capsules were formed without any viable seed development; we assume the occurrence of parthenocarpic development of fruits without affecting successful fertilization resulting into capsules with non-viable seeds. Wide hybridization as a driving force for the development of seedless or rudimentary fruits in higher plants have been discussed thoroughly 55 , 19 . Recently, Badiger et al. (2024) 2 reported that formation of aborted embryo with partially filled seeds suffered from severe pre-zygotic barriers, because of interspecific hybridization in the crosses in Abelmoschus species. On the other hand, pollen vitality is another important factor affecting the pollen germination percentage. Pollen represents a critical stage in the life cycle of plants, as viable pollen is crucial for efficient sexual plant reproduction. Reproductive fitness in wild plants is often determined by the quantity and quality of pollen grains produced. Both parameters can be considered as survival strategy in deciding reproductive fitness of natural populations growing spontaneously under resource limited marginal environments 57 . High pollen vitality in L. bienne (86.35%) and L. grandiflorum (91.30%) than the cultivated L. usitatissimum (69.92%) indicates towards the competitive ability of these wild species to capture more opportunities to affect efficient fertilization during sexual reproduction. Though, pollen vitality is an important factor for pollen germination and tube growth, these parameters were seemed to be drastically reduced (Table S1 ) in case of wide crosses with L. grandiflorum , despite appreciable pollen vitality percentage in L. usitatissimum (69.92%) and L. grandiflorum (91.30%) (Table 3). This emphasizes that high pollen vitality does not guarantee successful pollen germination and subsequent pollen tube growth inside the foreign style's transmitting tract when dealing with genetically distant parents. Pollen germination is a vital indicator of viability and provides valuable estimation for pollen tube growth. It is crucial to note that germination loss does not necessarily mean the pollen has died, but rather underscores the need for optimal germination conditions 47 . The genetic differences between cultivated and wild L. grandiflorum seems violating the congenial conditions required for optimal germination of pollen grains and subsequent pollen tube growth. Self-incompatibility arising from heterostyly is widely documented in various species linked to cultivated Linum , as mentioned previously. Mechanism of self-incompatibility (SI) between different floral morphs are also studied and the mechanism of SI among various floral forms in Linum grandiflorum 37 , 70 , Linum usitatissimum 8 have been investigated in the past. Based on what we know, there is a lack of available literature on the germination and tube growth of foreign pollen grain on the stigma and style of a different species in flaxseed. Pollen germination on stigmatic surface In case of self-pollination for the cultivated species L. usitatissimum (Fig. 4 a), its wild progenitor L. bienne (Fig. 4 b) and, legitimate mating with its distant relative L. grandiflorum (Fig. 4 c), the PG percentage were as high as 95% (Fig. 3 a, Fig. 2 a, b and c, Fig. 4 a-l). The occurrence of high germination percentage may be explained by the hypothesis proposed by Lewis (1960) 38 that resulted into adaptation from allogamy to autogamy in flax following two phenomena. These may include either a mutation of activation of pollen specificities, the styles maintaining their incompatibility reaction or, the development of secondary fertility genes thwarting the incompatibility reaction or partial allogamy and the lack of complete auto-sterility. PG percentages and pollen tube (PT) lengths varied in flax styles for different crosses, including reciprocals and selfings, using cultivated L. usitatissimum species 8 . Similarly, the PG percentage on the foreign stigma, when L. bienne was used either as pollen or pistil parent, were as good as in case of self-pollination of individual species (Fig. 4 m, q). This can be accounted to the genetic similarity between the cultivated Linum species and its wild progenitor which is the immediate ancestor of modern cultivated flax 22 . However, when L. grandiflorum was used as pollen or pistil parent, the PG percentage were drastically reduced during different HAP (Fig. 5 a-g, Fig. 3 a). The maximum PG on the stigmatic surface of cultivated Linum , when L. grandiflorum served as pollen parent, could reach to a maximum of 10.92% only in 4HAP. When used as pistil parent, the PG reached to a maximum of 10.33% only in 6HAP (Fig. 2 f, g). Varied response and reciprocal differences of pollen and pistil parents, on PG, PTG and compatibility parameters have been studied in wide hybridization of Vigna species 50 , pigeon pea 58 , Curcuma 75 . It is evident that there is a reproductive barrier imposed on the landed pollen grain during the pre-fertilization stage. Analysis of captured images under fluorescent microscope, indicated the formation of a thick callose layer on the stigma surface (Fig. 5 a, b). The callose deposition could be seen within the 2HAP. In the incompatible cross-combinations of Lu X Lg , and its reciprocal, larger proportions of pollen grains lying ungerminated on the foreign stigmatic surface could be observed even longer hours after (10 to 24HAP) pollen landing (Fig. 5 c, d and e) that started showing up some form of degradation. The main obstacles for PG included the callose layer forming within 2HAP, which limited the germination of foreign pollen grains on stigmatic surfaces, taking 4 to 6 hours (Fig. 5 f, g). All flowering plants rely on the crucial process of sexual reproduction. This essential process is initiated when a pollen grain reaches the stigma of a pistil and gives rise to a vital pollen tube 45 . The journey to hybridization between different species within the same genus may encounter challenges, but groundbreaking researchers such as Dumas and Knox (1983) 17 and Chen and Kim (2009) 5 have championed the use of aniline blue to unravel the callose response, shedding light on the phenomena of rejection. Researchers have stressed the importance of utilizing aniline blue to track callose response, associated with pollen rejection. The Aniline Blue Fluorescent (ABF) method and fluorescence microscopy have been employed to extensively investigate PPI in a variety of crops such as sorghum 9 , sesame 67 . However, studies for analyzing unsuccessful seed set because of pre-fertilization barrier due to incompatible PPI in Linum interspecific crosses is scarce. Callose is a crucial component of the cell wall, being dynamically deposited and degraded throughout pollen development 62 . In many angiosperms, a brief callose-rich cell wall forms around each microsporocyte in the anthers, while callose is also present on the outer pollen wall. Callose effectively separates microspores and acts as an impermeable barrier 40 . In species with callose-rich walls, mutants lacking callose production consistently produce inviable pollen 61 . Visible ungerminated pollen grains on the stigmatic surfaces in case of incompatible interspecific hybridization after longer hours of pollen landing may be explained due to deposition of the callose layer as an attempt to retain longer pollen viability (Fig. 5 a-e). Callose provides a useful phenotypic bioassay in plant breeding to determine incompatibility system, pollen competition, stigma, and ovule viability and to understand the pathogen induced defense mechanism triggered in the plant systems 74 , 17 . The role of callose in pollen-stigma interactions has many analogies with host-parasite interactions as callose deposition is viewed as an indicator of biocommunication between pollen and stigma during PPI 17 . This pathogen-induced callose deposition functions as a chemical and physical defense mechanism for reinforcing plant cell wall and plays an essential role in the defense response to invading pathogens 74 . In Linum , it is assumed that callose deposition in response to non-self-pollen grains serves as a formidable barrier to pollen germination and pollen tube growth in incompatible distant crosses under the current study. The response of stigma callose is distinctly influenced by informational molecules from self or interspecific pollen grains. Dumas and Knox's (1983) 17 proposed model establishes a compelling connection between callose, boron, and inhibitor synthesis akin to phytoalexins. In the model plant Arabidopsis thaliana two independent research groups identified twelve genes encoding putative callose synthase namely CalS (callose synthase) and GSL (glucan synthase-like) genes 69 . Callose wall plays an important role in normal pollen mother cell development. Recently, through genetic approach it is revealed that OsGSL5 gene is responsible for callose deposition in anther locules and OsGSL5 gene mutation resulted in anthers with less callose deposition, aberrant pollen mother cells and abnormal microspores 65 . Global transcriptome analysis showed that expression of OsGSL5 was downregulated in the Osspl ( OsSPOROCYTELESS ) mutant which was defective in meiosis-specific callose deposition 59 , 39 . Similarly, we posit that a mutated or malfunctioning expression of genes responsible for synthesizing callose on the stigmatic surface when encountering non-self-pollen grains may present an opportunity to overcome pre-fertilization barriers arising from callose deposition. It is recommended to conduct further studies to explore the suppression of such genes and its potential impact on enabling the overcoming of callose-related barriers. Pollen tube growth within the pistil Irrespective of self-or cross-pollination, in all the studied three Linum species, and their wide cross-combinations, the germinated pollen grains were able to grow and become visible within the various stylar regions of the pistil parent in different time intervals. The number of growing PTs kept reducing in all the cases of self and cross pollination while growing towards the ovular region from stylar region (Fig. 3 b, c and d). Just like self-pollinated pistils, wide crosses with L. bienne , were able to be seen growing PTs within various stylar regions in foreign pistils and seemed navigating ovular region within 2HAP (Fig. 4 m-p). However, the pollen grains that could successfully cross the germination barriers due to callose deposition on the stigmatic surface in Lu X Lg and its reciprocal, which were seen growing their PTs in the stylar regions of pistil parent within 2HAP were unable to reach up to the ovular region of foreign pistil parent. Appearance of PTs growing in the ovular region for distant crosses with L. grandiflorum could be noticed only during 4HAP irrespective of pistil parent (Fig. 3 d). The delayed growth of PT in the foreign style was explained due to predominant resistance force due to a high intensity of PTs in the stigmatic surface 53 . Unlike self-pollinated pistils and, crosses with its wild progenitor i.e ., L. bienne , various kinds of aberrations were observed in the developing PTs, imposing barriers in the path of further growth in wide crosses with L. grandiflorum (Figs. 6 and 7 ). The major PTG inhibition occurred in the stylar region was, due to appearance of emerging deformed growth patterns in the developing tubes. These discordant growth patterns may be attributed to cause delayed arrival of PTs to ovular region of foreign pistil in wide crosses with L. grandiflorum (Fig. 3 ). According to Hodnett et al. (2005) 31 , unfavorable conditions hindered the normal metabolism of the PT, leading to its malformation and subsequent reduction in growth towards the micropyle. Comprehensive analysis of PTG in the interspecific and intergeneric crosses as an indicator of pre-zygotic barrier is not just limited to field crops but have been extensively implied in horticultural crops too 41 , 46 . Most of the nonconductive circumstances started appearing during the 2-4HAP (Fig. 6 a-g). Partially elongated PTs along with the non-germinated pollen grains could still be seen in the upper stylar regions of L. usitatissimum during this time duration (Fig. 6 a). Molecular aspects better explain the reasons behind the rejection of incompatible PTG. Some studies mention role of putative protein O-fucosyltransferase facilitating PT penetration through the stigma-style interface 64 . Stylar region comprises various chemical signals that reside in the extracellular matrix of transmitting tract play crucial roles in the recognition of self or non-self-PT. There are various substances existing in the extracellular matrix of the transmitting tract that reject non-self-PTs and the loss-of-function of the pistil-side barrier will result in self-incompatibility transition to self-compatibility 3 . Various biomolecules involved in guiding the PT through the transmitting tract have been extensively elaborated and reviewed by Zheng et al. (2018) 76 and recently by Cheung et al. (2022) 7 . Swelling of PTs at the tip was traced through the florescent image analysis in the mid-stylar (Fig. 6 b) and attachment regions of the ovary (Fig. 6 c) in the pistil. Non-penetration of the ovular region even at 96HAP was reported in the studies of Gong et al. (2023) 27 , where they reported that PT barely entered the ovule, but remained at the base of the style and became swollen in another oilseed plant species Camellia oleifera. The control of PTG by protein factors was addressed by Mei et al. (2022) 42 in model plant Arabidopsis thaliana. Twisted PTG was witnessed in the upper (Fig. 6 d) and mid-stylar (Fig. 6 e) regions along with severe inhibition of the growing PT in the ovary (Fig. 6 f, g). The appearance of PTs growing in reverse direction (Fig. 6 h) while reaching towards the ovule, complex and convoluted PTG at the mid-stylar region (Fig. 6 i) were noticed during 6HAP. Partially germinated pollen grains with terminated PTG (Fig. 6 j) were still visible even after longer hours of pollination (10HAP). Thus, the initial 4 hours were proved critical and crucial from the PG and PTG point of view. In the interspecific crosses, there were frequent observations of delayed growth of PTs, as well as other structural abnormalities such as twisting, swelling, high branching, a bi-furcated tip, PTG before reaching the ovary and inability to navigate to micropyle and variations in callose form. These variations included reverse orientation and irregularity in callose plugs along the PT. Such abnormalities attributed for delayed pollination and delayed pollen tube growth resulting into incompatible interaction 52 . In case of distant crosses made with L. grandiflorum as pistil parent, almost similar abnormality patterns were observed during PTG of L. usitatissimum . Within 2HAP, the pre-fertilization barriers started appearing in the form of restricted and twisted PTG in the upper stylar region (Fig. 7 a). As soon as the PT arrives at the attachment region in of ovary at the lower stylar region, some PT faced twisted growth patterns (Fig. 7 b). Under-germinated pollen grains with terminated, twisted and, swollen PT was most frequent during the 4HAP (Fig. 7 c, d). Non-germinated pollen grains were quite obviously visible during different HAP from 2 to 24HAP (Fig. 7 e). Large proportions of ungerminated pollen grains along with consistently growing PTs were noticed in the stylar region (Fig. 7 f). Reduced number of PTs passing through ovular regions affecting fertilization after longer hours of pollination taking up to 18 to 24 hours have been reported in wide crosses of Vigna spp. 34 . Ruptured and bifurcated PT tip in the stylar region could also be witnessed as soon as 4 hours after pollen landed and germinated on foreign pistil (Fig. 7 g). Convoluted PTG (Fig. 7 h), partial or delayed PTG (Fig. 7 i) with swollen tip (Fig. 7 j) were visible even after longer durations following pollination. Many pollen grains germinated with swollen PT tips and twisted growth patterns showing random growth patterns could be seen in the stylar region even 10-12HAP (Fig. 7 m, n). The pre-fertilization barriers were predominant in wide crosses with L. grandiflorum , that operated in all the stages resulting into delayed PTG. These barriers not only acted on the stigmatic surfaces but also had operated gradually and mildly at various growth stages. No PTs could be noticed penetrating ovular region in wide crosses with L. grandiflorum till 4HAP as opposed in crosses attempted with L. bienne , where the ovular penetration occurred within 2HAP (Fig. 3 , Fig. 4 m-t). Low levels of PT penetration and PTG in the wide crosses due to operative pre-fertilization barriers have been advocated by Krishnasamy et al. (2008) 34 in Vigna spp., Ganesh Ram et al. (2006) 23 in Sesame spp., Badiger et al. (2024) 2 in Abelmoschus species. Based on image analysis, the non-arrival of the PT within the first four HAP could be attributed to the various kinds of pre-fertilization barriers and deformities appearing during PTG. The first two to four hours seemed critical for the PT growth in wide crosses with L. grandiflorum . Though, the PT was able to reach up to the attachment region of ovary at the lower stylar region of pistil within the first 2HAP, they remained unable to penetrate the ovular region until 4HAP. Interspecific hybridization in Linum has been advocated in the past and has been successfully used in both West European and East European cultivars. For example, Linum crepitans L. was employed to introduce early maturity traits into the L. usitatissimum L. However, the potential of these early maturing types was not fully realized in subsequent breeding efforts. While interspecies crossing generally exhibits low effectiveness, this can be improved through the application of embryo tissue culture and the cultivation of immature isolated embryos under in vitro conditions 25 , 54 . In situations where natural interspecific hybridization in Linum fails due to pollen-stigma incompatibility, the utilization of advanced techniques such as protoplast fusion and embryo rescue holds immense promise for obtaining successful interspecific hybrids. The ongoing research on protoplast culture and isolated immature embryos of distant hybridization of flax will undoubtedly yield valuable insights into the selection of optimal medium and culture conditions, paving the way for groundbreaking advancements in this field 54 . In Wang's (2008) 73 study, interspecific hybridization between L. usitatissimum and L. perenne was conducted. The results demonstrated that repeat-pollination alongside the use of plant growth regulators (such as GA 3 , NAA, and 2,4-D) proved to be an effective method for overcoming interspecific cross-incompatibility in flax. Production of haploids followed by genotype stabilization in the case of interspecific hybridization for extension of genetic variability in Linum is one of the unconventional methods for producing interspecific hybrids. For this purpose, the application of biochemical methods has been studied and suggested. Material and Methods Details of Linum species used in the study Linseed accession IC268345 represented cultivated species ( L. usitatissimum L., 2n = 30) of Linum for this investigation. The wild species used under the current study involved its wild progenitor, L. bienne M. (2n = 30) (accession EC993391) and the other distant CWR L. grandiflorum L. (2n = 16). Since, L. grandiflorum is distylic but unlike most distylic plants the level of the anthers is not greatly different in the two types 37 , 48 , 11 , we used a mixed population of originally procured L. grandiflorum (IC633096) to ensure the legitimate PG and subsequent PTG in the pistils of manually pollinated flower buds. Details of the plant materials (Fig. 1 ) used and its characteristic features are listed in Table 1 . Field experiment and hand emasculation The field experiment was conducted at Indian Council of Agricultural Research - National Bureau of Plant Genetic Resources, experimental farm, IARI, New Delhi (28° 38′ 53.7′′ N, 77° 09′ 05.4′′ E and 218 m above mean sea level) in paired row of 2m length and row spacing of 90cm during cropping season from November to April, 2021-22. Self and cross pollination were attempted manually during the season using cultivated species IC268345, wild species EC993391 ( L. bienne M.) and IC633096 ( L. grandiflorum L.) with reciprocal cross combinations as represented in Table 2 . To assure the continuous availability of flowers, all the three species were staggered planted on three sowing dates, after 3–5 days of intervals. Artificial pollination was carried out following the method suggested by Bolling et al. (1961) 4 with slight modifications. A day before pollination, the un-opened flower buds with exposed petals were hand emasculated during the evening hours, between 5–7 PM. For emasculation, all the five anthers were exposed for removal using forceps. The petals were left intact in the buds to allow anthesis next morning. This helped exposure of stigmas naturally avoiding any damage to the stigmas while making efforts to separate the sepals covering stigmas next morning. This slight modification also helped transferring the pollens grains on the surface of naturally exposed stigma while performing pollination manually, that otherwise needed extra efforts to keep the sepals stay apart from each other while pollen transfer. After all the five anthers were removed, the pistils were closed back with petals and bagged with waxy pollination bags to avoid drying up of stigmas. Bagging of donor flower buds was also done the previous evening to obtain fresh non-contaminated pollen grains to hybridize the emasculated flower buds the next morning to avoid cross-contamination. The next morning between 7–9 AM, the pollen was transferred using freshly opened bagged flower by directly placing the pollen grains onto the exposed stigma of emasculated flower by contact method. Transfer of a sufficient number of pollen grains was ensured. The pollinated flowers were tagged with all the relevant information and bagged immediately to avoid cross contamination. Self-pollination was done in the same manner for all the three species. Pollen vitality test The vitality of pollen grains in L. usitatissimum , L. bienne , and L. grandiflorum was assessed using a Leica light microscope. To begin, mature flower buds were carefully collected and kept in an ice container. Subsequently, the anthers were delicately excised, placed on slides, and gently crushed in a 1% (w/v) acetocarmine solution with the aid of forceps and a needle. After removing any debris, a coverslip was carefully positioned on the slide before observation under the microscope. The captured images were then utilized to quantify and evaluate the stained and unstained pollen grains. During assessment, nonviable pollen, showing signs of shriveling or remaining unstained, was distinguished from viable, round, plump, and stained pollen (Fig. S1 ). Each of the species under scrutiny was thoroughly examined using four to six slides, each divided into 4 to 8 nonoverlapping sections (Table 3). Standardization of protocol for the study of pollen-pistil interaction in wide crosses of Linum Sample collection and storage The pistils of the artificially selfed and cross-pollinated flower buds were collected without separating them from petals and sepals after 2, 4, 6, 8, 10, 24 and 48 Hours after pollination (HAP) followed by fixation in acetic acid: alcohol (1:3 v/v) fixative in amber colored 5ml glass vials and stored at 4°C until further analysis. For every collection time, self and cross-pollinated flower buds were used for microscopic observations. For each self and cross combination 15 artificially pollinated flower buds were collected. Thus, for all the seven cross combinations of each duration, a total of 735 flower buds were collected and studied further. Sample processing To study PPI, aniline blue fluorescence method 51 was modified slightly for Linum for softening period by altering sodium hydroxide (NaOH) concentration. This helped to reduce the time duration of softening treatment for dissected pistils thereby, increasing the efficiency of sample handling with reduced time duration and minimal damage to the fixed pistils. Instead of direct fixation of pistils over the fixed flower buds, was proved advantageous in terms of keeping the fibrous Linum pistils retaining their integrity while sample processing without breaking the fragile tissues. The fixed pistils along with the intact flower buds were washed 2 to 3 times with distilled water for 5 minutes each followed by dissection. The pistils were dissected out from the flower buds with the help of pointed forceps and needles available in biological experiments dissection box to separate out the attached stigmatic regions. Unlike, Parani, 1998 51 , where higher concentration of softening agent (8N NaOH) was used for longer duration (8h), we reduced the concentration to 6N NaOH with 1h time duration to clear and soften the tissue at room temperature. The softened pistils were kept undisturbed in the distilled water for 30 min followed by mild washing to avoid any damage to the softened tissue and to remove all traces of softening agent. This enhanced the absorption of fluorescent dye in the target tissue. The softened tissue was then subjected to staining buffer overnight under dark conditions for further analysis through fluorescent microscopy. Fluorescence microscopy For preparation of staining buffer, 0.1M dibasic potassium phosphate (K 2 HPO 4 ) mixed with 0.1% water-soluble aniline blue dye (WS Color Index 42780) was used to stain the processed pistils. Staining with 0.1% water-soluble aniline blue prepared in 0.1M dibasic potassium phosphate (K 2 HPO 4 ) overnight, gave good results in terms of dye absorption by the target tissue. The stained pistil was then placed on glass slide without glycerol treatment to avoid intertwining of fibrous pistils. Pistils were covered with cover slip and pressed gently into a thin layer. The slides were placed in dark till further observation. The image was captured under dark room condition and freely definable black balance available with the Leica DM6 B Upright Fluorescent Microscope using 390–420 nm barrier filter coupled with a 450 nm excitation filter. Pollen germination and tube growth image thus captured with a Leica DFC7000 T camera installed with the microscope was used to process the image with Lieca-X Software for taking various parameters such as pollen germination and determination of PTG in different time intervals. Conclusion The present study revealed that the interaction between the pollen and pistil is inhibited in the distant crosses among Lu X Lg due to both temporal and spatial pre-fertilization barriers. Callose deposition at the stigmatic surface of foreign pistils within 2HAP leading to inability of pollen grains to penetrate the stylar tissue appeared as the major barrier. Various kinds of aberrations started appearing during the 2-4HAP, in the developing PTs imposing barriers in further growth. Delayed PTG was observed mainly due to the discordant conditions appearing in the various stylar regions. The complexity of interspecific hybridization was observed in terms of arrest of pollen tube (PT) growth in the ovary, ruptured, twisted and swollen pollen tube tip, tube growth in reverse direction, convoluted and terminated growth patterns. Although interspecies crossover is often less efficient, it can be made more effective by using embryo rescue and cultivating isolated, immature embryos in vitro . We propose that the disruption of genes responsible for producing callose on the stigmatic surface when encountering non-self-pollen grains could offer a promising opportunity to overcome pre-fertilization barriers associated with callose deposition. Further studies are recommended to investigate the suppression of these genes and the potential impact on surmounting callose-related barriers. When pollen-stigma incompatibility prevents natural interspecific hybridization in Linum from occurring, the application of cutting-edge methods like protoplast fusion and embryo rescue presents great potential for producing interspecific hybrids that succeed. The advanced studies on isolated immature flax embryos from distant hybridization and protoplast culture will surely provide important information on choosing the best culture medium and circumstances, opening the door to ground-breaking discoveries in this area. Declarations Author Contribution V.K.R: Experimentation and writing—original draft, Data collection; A.Y. & M.R.: Supervision; V.K. & D.W.: Investigation, review, and editing; P.P.: Experimentation; M.S.: Conceptualization, supervision, writing—original draft; funding acquisition; G.P.S.: funding acquisition; project administration. Acknowledgement Authors acknowledge the funding support for the project (No. BT/Ag/Network/Linseed/2019-20) from the Department of Biotechnology (DBT), Government of India. The authors also acknowledge the funding and research facilities available at the Indian Council of Agricultural Research-National Bureau of Plant Genetic Resources (ICAR-NBPGR), New Delhi. The authors thank the Head, Division of Germplasm Evaluation, NBPGR for the research facilities and guidance to carry out field experiments. The authors are thankful to Dr. B.L. Meena (ACTO & Farm manager, NBPGR farm) for assistance in field experiments. The authors are also thankful to Mr. Raghvendra Pratap Singh, Mr. Santosh, and other project staff for assistance in field and laboratory experiments. Data Availability All the data have been provided in the main manuscript and supplementary files. References Allaby, R. G., Peterson, G. W., Merriwether, D. A. & Fu, Y. B. Evidence of the domestication history of flax ( Linum usitatissimum L.) from genetic diversity of the sad2 locus. Theoretical and applied genetics. 112 , 58-65. doi: 10.1007/s00122-005-0103-3 (2005). Badiger, M., Yadav, R. K. & Sharma, B. B . Pollen germination, pollen–pistil interaction and crossability studies in interspecific and induced colchiploid population of Abelmoschus species. Genet Resour Crop Evol . 71 , 107-127. https://doi.org/10.1007/s10722-023-01610-y (2024). Bedinger, P. A., Broz, A. K., Tovar-Mendez, A. & McClure, B. Pollen-pistil interactions and their role in mate selection. Plant Physiol . 173 , 79-90. doi: 10.1104/pp.16.01286 (2017). Bolling, M., Sander, D. A. & Matlock, R. S. Mungbean hybridization technique. Agron. J . 53 , 54-55 (1961). Chen, X. Y. & Kim, J. Y. Callose synthesis in higher plants: Mini-review. Plant Signaling and Behavior . 4 , 489-492 (2009). Cheng, L. L., Guan, F. Z., Wu, G. W., Yu, Y., Huang, W. G. & Zhao, D. S. Interspecific hybridization and immature embryo rescure between Linum usitatissimum and Linum grandiflorum . Plant Fiber Sci . 37 , 1-4. doi: 10.3969/j.issn.1671-3532.2015.01.001 (2015). Cheung, A. Y., Duan, Q., Li, C., James Liu, M. C. & Wu, H. M. Pollen-pistil interactions: It takes two to tangle but a molecular cast of many to deliver. Curr. Opin. Plant Biol . 69 , 102279. https://doi.org/10.1016/j.pbi.2022.102279 (2022). Chung, T. D. & Plonka, F. Differential germination in flax pollination. Agronomie, EDP Sciences . 6(4) , 379-386 (1986). Cisneros, L. M. E. et al . Pollen–pistil interaction, pistil histology and seed production in A×B grain sorghum crosses under chilling field temperatures. The Journal of Agricultural Science . 148 , 73-82. 10.1017/S0021859609990396 (2010). Darlington, C. D. & Wylie, A. P. Chromosome atlas of flowering plants. George Allen and Unwin, London (1955). Dickinson, H. G. & Lewis, D. Changes in the Pollen Grain Wall of Linum grandiflorum following Compatible and Incompatible Intraspecific Pollinations. Ann. Bot . 38 , 23-9 (1974). Diederichsen, A. & Hammer, K.. Variation of cultivated flax ( Linum usitatissimum L. subsp. usitatissimum) and its wild progenitor pale flax (subsp. angustifolium (Huds.) Thell.). Genet. Resour. Crop Evolut . 42 , 263-272. https://doi.org/10.1007/BF02431261 (1995). Diederichsen, A. & Richards, K. Cultivated flax and the genus Linum L. Taxonomy and germplasm conservation. In: Muir, A. D. and Westcott, N. D. (Eds). Flax : The genus Linum. CRC Press, London, 22-54 (2003). Doebley, J. F., Brandon, S. G. & Bruce, D. S. The molecular genetics of crop domestication. Cell . 127(7) , 1309-1321. https://doi.org/10.1016/j.cell.2006.12.006 (2006). Duigou, A. & Castro, M. Evaluation of force generation mechanisms in natural, passive hydraulic actuators. Scientific Reports . 6 , 18105. 10.1038/srep18105 (2016). Duk, M. et al . The Genetic Landscape of Fiber Flax. Frontiers in Plant Science . 12 , 764612. 10.3389/fpls.2021.764612 (2021). Dumas, C. & Knox, R. B. Callose and determination of pistil viability and incompatibility. Theoretical and Applied Genetics . 67 , 1-10 (1983). FAO, Statistics Division, Food and Agriculture Organization of the United Nations, Viale delle Terme di Caracalla, 00153 Rome, Italy. http://www.fao.org/faostat/en/#home (2018). Fayaz, Z. et al . Parthenocarpy: ‘‘A Potential Trait to Exploit in Vegetable Crops’’. Environment and Ecology . 39(4A) , 1332-1346 (2021). Feller, A. F., Burgin, G., Lewis, N., Prabhu, R. & Hopkins, R. Mismatch between pollen and pistil size causes asymmetric mechanical reproductive isolation across Phlox species. bioRxiv . 593106. doi: 10.1101/2024.05.08.593106 (2024). Flint-Garcia, S. A. Genetics and consequences of crop domestication. J. Agric. Food Chem . 61(35) , 8267-76. doi: 10.1021/jf305511d (2013). Fu, Y. B. Pale Flax ( Linum Bienne ): an Underexplored Flax Wild Relative. In: You, F.M., Fofana, B. (eds) The Flax Genome. Compendium of Plant Genomes. Springer, Cham . https://doi.org/10.1007/978-3-031-16061-5_3 (2023). Ganesh Ram, S., Sundaravelpandian, K., Kumar, M., Vinod, K. K., KannanBapu, J. R. & Raveendran, T. S. Pollen pistil interaction in the inter-specific crosses of Sesamum sp . Euphytica . 152 , 379-385 (2006). Ghosh, S. & Shivanna, K. R. Pollen-pistil interaction in Linum grandiflorum : Scanning electron microscopic observations and proteins of the stigma surface. Planta . 149(3) , 257-61. doi: 10.1007/BF00384562 (1980). Gill, K. S. Objectives, breeding approaches and achievements in linseed ( Linum usitatissimum ). In: Gill, K.S. (Ed.), Breeding Oilseed Crops. Springer, Dordrecht , pp. 212e225 (1980). Gill, K. S. Linseed. Indian Council of Agricultural Research, New Delhi, India (1987). Gong, H., Chang, Y., Xu, J., Yu, X. & Gong, W. Unilateral cross-incompatibility between Camellia oleifera and C. yuhsienensis provides new insights for hybridization in Camellia spp. Front Plant Sci . 14 , 1182745. doi: 10.3389/fpls.2023.1182745 (2023). Heer, O. Prehistoric culture of flax. Nature . 7 , 453. https://doi.org/10.1038/007453a0 (1873). Helback, H. Domestication of Food Plants in the Old World: Joint efforts by botanists and archeologists illuminate the obscure history of plant domestication. Science . 14 , 130 : 365-372. doi: 10.1126/science.130.3372.365 (1959). Herbig, C. & Maier, U. Flax for oil or fibre? Morphometric analysis of flax seeds and new aspects of flax cultivation in Late Neolithic wetland settlements in southwest Germany. Vegetation History and Archaeobotany . 20 , 527-533. 10.1007/s00334-011-0289-z (2011). Hodnett, G. L., Burson, B. L., Rooney, W. L., Dillon, S. L. & Price, H. J. Pollen-pistil interactions result in reproductive isolation between Sorghum bico lor and divergent Sorghum species. Crop. Sci . 45 , 1403-1409 (2005). Jhala, A. J., Hall, L. M. & Hall, J. C. Potential hybridization of flax with wild and weedy species: An avenue for movement of engineered genes? Crop Science . 48(2) , 825-840 (2008). Kaur, V. et al . Multi-environment phenotyping of linseed ( Linum usitatissimum L.) germplasm for morphological and seed quality traits to assemble a core collection. Industrial Crops and Products . 206 , 117657. https://doi.org/10.1016/j.indcrop.2023.117657 (2023). Krishnasamy, T., Palaniappan, J. & Nagasamy, N. Pollen pistil interaction in interspecific crosses of Vigna spp . Cytologia . 73 , 251-257 (2008). Kvavadze, E. et al . 30,000-Year-Old Wild Flax Fibers. Science (New York, N.Y.) . 325 , 1359. 10.1126/science.1175404 (2009). Lee, H. K., Macgregor, S. & Goring, D. R. A Toolkit for Teasing Apart the Early Stages of Pollen–Stigma Interactions in Arabidopsis thaliana . In: Geitmann, A. (eds) Pollen and Pollen Tube Biology. Methods in Molecular Biology . 2160. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0672-8_2 (2020). Lewis, D. The physiology of incompatibility in plants. 2. Linum grandiflorum . Annals of Botany . 7 , 115-122 (1943). Lewis, D. Genetic control of specificity and activity of the S antigen implants. Proe. R. Soc., Ser. B . 151 , 468-477 (1960). Li, N., Lin, Z., Yu, P., Zeng, Y., Du, S. & Huang, L. J. The multifarious role of callose and callose synthase in plant development and environment interactions. Front. Plant Sci . 14 , 1183402. doi: 10.3389/fpls.2023.1183402 (2023). Li, Y. et al . Pollen-Specific Protein PSP231 Activates Callose Synthesis to Govern Male Gametogenesis and Pollen Germination. Plant Physiology . 184(2) , 1024-1041. https://doi.org/10.1104/pp.20.00297 (2020). Mahoney, J. D. & Brand, M. H. Pre and post zygotic barriers associated with intergeneric hybridization between Aronia melanocarpa (Michx.) Elliott × Pyrus communis L. and × Sorbaronia dippelii (Zabel) CK Schneid. × Pyrus communis . HortScience . 56 , 177-184 (2021). Mei, J. et al . MAP3Kε1/2 Interact with MOB1A/1B and Play Important Roles in Control of Pollen Germination through Crosstalk with JA Signaling in Arabidopsis . Int. J. Mol. Sci . 23 , 2683. doi: 10.3390/ijms23052683 (2022). Melelli, A. et al . Lessons on textile history and fibre durability from a 4,000-year-old Egyptian flax yarn. Nature Plants . 7 , 1-7. 10.1038/s41477-021-00998-8 (2021). Mohanty, A. K., Vivekanandhan, S., Pin, J. M. & Misra, M. Composites from renewable and sustainable resources: challenges and innovations. Science . 36 , 536-542. 10.1126/science.aat9072 (2018). Moreira, D., Kaur, D., Pereira, A. M., Held, M. A., Showalter, A. M. & Coimbra, S. Type II arabinogalactans initiated by hydroxyproline‐O‐galactosyltransferases play important roles in pollen–pistil interactions. The Plant Journal . 114(2) , 371-389. doi: 10.1111/tpj.16141 (2023). Morimoto, T., Matsuda, Y., Sekiguchi, R. & Itai, A. Comprehensive Assessment of Intergeneric Cross-compatibility of Six Fruit Tree Species in the Tribe Maleae ( Rosaceae ) Based on In Vivo Pollen Tube Growth and Field Pollination. The Horticulture Journal . 92(1) , 13-21. https://doi.org/10.2503/hortj.UTD-383 (2023). Mosquera, D. J. C., Salinas, D. G. C. & Moreno, G. A. L. Pollen viability and germination in Elaeis oleifera , Elaeis guineensis and their interspecific hybrid. Pesqui. Agropecuária Trop . 51 , e68076. 10.1590/1983-40632021v5168076 (2021). Murray, B. G. Floral Biology and Self-Incompatibility in Linum . Botanical Gazette . 147(3) , 327-333. http://www.jstor.org/stable/2474401 (1986). Nair, B. et al . Multi-environment screening of Linum germplasm collection for dissecting the potential of bud fly (Dasyneura lini Barnes) resistance and assembling a reference set for efficient utilization in genetic improvement. Industrial Crops and Products . 207(1) , 117743. https://doi.org/10.1016/j.indcrop.2023.117743 (2024). Pandiyan, M. et al . Interspecific Hybridization in Direct and Reciprocal Crosses of Vigna radiata and Vigna aconitifolia . Ann Agric Crop Sci . 5(2) , 1062 (2020). Parani, M. Incompatibility in direct and reciprocal cross between S. indicum L. and S. alatum (Thonn). J. Indian Bot. Soc . 77 , 9-12 (1998). Pascual, M., Ruiz, J., Posas, J. & Niño, M. Pollen viability and incompatibility in indigenous rice bean ( Vigna umbellata (Thunb.) Ohwi & Ohashi). Plant Science Today . 9(1) , 157-166. 10.14719/pst.1375 (2022). Patil, P. et al . Pollen germination characteristics, pollen-pistil interaction and reproductive behaviour in interspecific crosses among Abelmoschus esculentus Moench and its wild relatives. Grana . 52 , 1-14. DOI: 10.1080/00173134.2013.768699 (2013). Pavelek, M., Vrbová-Prokopová, M., Ondráčková, E., Ludvíková, M. & Griga, M. 17- Developments in fibrous flax and linseed breeding and cultivation, Editor(s): Ryszard M. Kozłowski, Maria Mackiewicz-Talarczyk, In: Woodhead Publishing Series in Textiles, Handbook of Natural Fibres (Second Edition), Woodhead Publishing, 605-692. https://doi.org/10.1016/B978-0-12-818398-4.00019-0 (2020). Picarella, M. E. & Mazzucato, A. The Occurrence of Seedlessness in Higher Plants; Insights on Roles and Mechanisms of Parthenocarpy. Front Plant Sci . 9 , 1997. doi: 10.3389/fpls.2018.01997 (2019). Qi, Y et al . Phenotypic analysis of Longya-10 × pale flax hybrid progeny and identification of candidate genes regulating prostrate/erect growth in flax plants. Front. Plant Sci . 13 , 1044415. doi: 10.3389/fpls.2022.1044415 (2022). Radice, S. & Arena, M. Characterization and evaluation of Berberis microphylla G. Forst pollen grains. Advances in Horticultural Science . 30(1) , 31-37. 10.13128/ahs-18699 (2016). Ravali, P., Kalaiyarasi, R., Hemavathy A. T. & Sudha M. Cytological investigation on pre and post fertilization barriers in interspecific cross of pigeonpea. Electronic Journal of Plant Breeding . 13(2) , 724-730. https://doi.org/10.37992/2022.1302.095 (2022). Ren, L. et al . OsSPL regulates meiotic fate acquisition in rice. New Phytol . 218(2) , 789-803. doi: 10.1111/nph.15017 (2018). Robichaux, K. J. & Wallace, I. S. Signaling at Physical Barriers during Pollen-Pistil Interactions. Int. J. Mol. Sci. 22 , 11230 (2021). Rosa, M. P. et al . Tomato POLLEN DEFICIENT 2 encodes a G-type lectin receptor kinase required for viable pollen grain formation. Journal of Experimental Botany . 74(1) , 178-193, https://doi.org/10.1093/jxb/erac419 (2023). Seale, M. Callose Deposition during Pollen Development. Plant Physiol . 184(2) , 564-565. doi: 10.1104/pp.20.01143 (2020). Seetharam, A. Interspecific Hybridization in Linum. Euphytica . 21 , 489-495 (1972). Smith, D. K.. et al. A putative protein O-fucosyltransferase facilitates pollen tube penetration through the stigma-style interface. Plant Physiol . 176 , 2804-2818. doi: 10.1104/pp.17.01577 (2018). Somashekar, H., Mimura, M., Tsuda, K. & Nonomura, K. I. Rice GLUCAN SYNTHASE-LIKE5 promotes anther callose deposition to maintain meiosis initiation and progression. Plant Physiol . 191 , 400-413. doi: 10.1093/plphys/kiac488 (2023). Soto-Cerda, B. J., Urbina, S. H., Navarro, C. & Mora O. P. Characterization of novel genetic SSR markers in Linum usitatissimum L. and their transferability across eleven Linum species. Electronic Journal of Biotechnology . 14(2) (2011). Sruthi, S. R., Kalaiyarasi, R. R., Sasikala, R. & Sudha, M. Pollen pistil interaction in the interspecific cross of Sesamum indicum and S. radiatum . Electronic Journal of Plant Breeding . 13(4) , 1288-1296. 10.37992/2022.1304.162 (2023). Suryawanshi, S. N., Wasnik, S. B., Makesar, A. D. & Walke, R. D. Economics of linseed marketing in Bhandara district. Journal of Pharmacognosy and Phytochemistry . 9(5) , 803-806 (2020). Unal, M., Vardar, F. & Ayturk, Z. Callose in Plant Sexual Reproduction. Current Progress in Biological Research . InTech. doi: 10.5772/53001 (2013). Ushijima, K. Et al . Isolation of the floral morph-related genes in heterostylous flax ( Linum grandiflorum ): the genetic polymorphism and the transcriptional and post-transcriptional regulations of the S locus. Plant J . 69(2) , 317-31. doi: 10.1111/j.1365-313X.2011.04792.x (2012). Van Zeist, W. & Bakker-Heeres, J. A. H. Evidence for linseed cultivation before 6000 BC. J. Archaeol. Sci . 2(3) , 215-219. https://doi.org/10.1016/0305-4403(75)90059-X (1975). Vavilov, N. I. Studies on the Origin of Cultivated Plants. Leningrad: des Plantes, State Press (1926). Wang, Y. F. The present situation and suggestions about the study of flax germplasm resource in China. J. Life Sci . 2(1) , 67e71 (2008). Wang, Y., Li, X., Fan, B., Zhu, C. & Chen, Z. Regulation and Function of Defense-Related Callose Deposition in Plants. International Journal of Molecular Sciences . 22(5) , 2393. doi: 10.3390/ijms22052393 (2021). Ye, Y., Zhou, Y., Tan, J., Zhu, G., Liu, J. & Xu, Y. Cross-compatibility in interspecific hybridization of different Curcuma accessions. Plants (Basel) . 12(10) , 1961. doi: 10.3390/plants12101961 (2023). Zheng, Y. Y., Lin, X. J., Liang, H. M., Wang, F. F. & Chen, L. Y. The Long Journey of Pollen Tube in the Pistil. Int J Mol Sci . 19(11) , 3529. doi: 10.3390/ijms19113529 (2018). Zhukovsky, P. M. Cultivated plants and their wild relatives. Systematics, geography, cytogenetics, resistance, ecology, origin and use. Moscow: Kolos. Leningrad, 751 p (1971). Tables Table 1 . Details of plant material used in the study Scientific Name L. usitatissimum L. L. bienne Mill. L. grandiflorum L. Common name Flax/linseed Pale flax Red flax/ Scarlet flax Biological Status Germplasm (Registered genetic stock) Wild progenitor Distant CWR National ID IC268345 EC993391 IC633096 Chromosome number (2n) 30 30 16 Potential use High oil content (42.5%) Fibre quality and more primary branches Bud fly resistance Growth habit Bushy to erect Prostrate growth habit Semi-spreading Cross compatibility with cultivated species Yes Yes No (Shriveled seeds) Geographic distribution/origin Southern Europe, the Near East, or Central Asia Mediterranean Sea, Iran, and the Canary Islands Native to Algeria but occurrence is found in India, Northern Africa, Southern Europe and North America Procured from National Genebank, ICAR-NBPGR, New Delhi, India CGN Gene Bank, Plant Research International Wageningen, Netherlands National Genebank, ICAR-NBPGR, New Delhi, India Table 2. Details of cross combinations attempted for wide hybridization including self-pollination S. No. Nature of Cross Species involved Accessions 1. Self-pollination L. usitatissimum X L. usitatissimum ( Lu X Lu ) IC268345 X IC268345 2. Self-pollination L. bienne X L. bienne ( Lb X Lb ) EC993391X EC993391 3. Self-pollination L. grandiflorum X L. grandiflorum ( Lg X Lg ) IC633096 X IC633096 4. Cross pollination L. usitatissimum X L. bienne ( Lu X Lb ) IC268345 X EC993391 5. Cross pollination L. bienne X L. usitatissimum ( Lb X Lu ) EC993391 X IC268345 6. Cross pollination L. usitatissimum X L. grandiflorum ( Lu X Lg ) IC268345 X IC633096 7. Cross pollination L. grandiflorum X L. usitatissimum ( Lg X Lu ) IC633096 X IC268345 Table 3. Pollen vitality in studied Linum species ( L. usitatissimum, L. bienne and L. grandiflorum ) Section 1 Section 2 Section 3 Section 4 Section 5 Section 6 Section 7 Section 8 F S F S F S F S F S F S F S F S Total F Total S Total PMC PV% L. usitatissimum Slide 1 48 41 47 40 41 31 51 39 - - - - - - - - 187 151 338 55.33 Slide 2 25 12 34 13 24 10 19 13 27 16 - - - - - - 129 64 193 66.84 Slide 3 28 15 28 17 24 6 21 14 28 10 36 21 43 23 30 13 238 119 357 66.67 Slide 4 78 10 91 19 67 16 43 9 43 8 97 18 120 22 62 13 601 115 716 83.94 Slide 5 92 39 25 9 73 9 43 19 73 30 79 33 57 21 57 34 499 194 693 72.01 Slide 6 99 29 70 29 74 28 65 15 48 16 62 31 71 28 117 29 606 205 811 74.72 Total PMCs 3108 - Avg. PV% - 69.92 L. bienne Slide 1 47 13 31 12 40 15 30 10 50 9 - - - - - - 198 59 257 77.04 Slide 2 63 5 48 4 103 5 67 9 59 6 - - - - - - 340 29 369 92.14 Slide 3 48 7 37 7 35 8 28 5 60 10 34 6 39 7 39 10 320 60 380 84.21 Slide 4 46 7 61 3 54 3 25 3 46 3 25 3 34 3 42 4 333 29 362 91.99 Total PMCs 1368 - Avg. PV% - 86.35 L. grandiflorum Slide 1 31 6 33 3 42 5 31 3 - - - - - - - - 137 17 154 88.96 Slide 2 88 6 110 4 95 7 48 4 - - - - - - - - 341 21 362 94.20 Slide 3 39 1 79 7 46 5 58 11 - - - - - - - - 222 24 246 90.24 Slide 4 40 1 34 1 31 1 79 5 - - - - - - - - 184 8 192 95.83 Slide 5 85 5 44 3 45 3 63 1 74 0 55 1 38 1 36 2 440 16 456 96.49 Slide 6 158 31 162 34 78 19 82 17 66 17 78 18 42 9 38 9 704 154 858 82.05 Total PMCs 2268 - Avg. PV% - 91.30 Abbreviations: F: Fertile; S: Sterile; PMC: Pollen Mother Cell; PV%: Pollen vitality percent; Table 4. Pollen germination and pollen tube growth for studied cross combinations in Linum Lu X Lu Lb X Lb Lg X Lg Lu X Lb Lb X Lu Lu X Lg Lg X Lu Pollen germination (%) Max. 95.99 (10HAP) 95.58 (8HAP) 94.82 (2HAP) 97.00 (4HAP) 96.19 (4HAP) 10.92 (4HAP) 10.33 (6HAP) Min. 92.83 (8HAP) 92.17 (24HAP) 93.54 (10HAP) 91.91 (8HAP) 93.03 (48HAP) 6.66 (48HAP) 7.71 (8HAP) Average 94.51 94.27 94.12 94.00 94.87 8.15 8.82 Variance 1.68 1.54 0.23 3.24 0.98 2.13 0.72 Tubes to Style (No.) Max. 75 (4HAP) 62 (4HAP) 49 (8HAP) 44 (4HAP) 40 (8HAP) 39 (4HAP) 23 (4HAP) Min. 13 (48HAP) 29 (48HAP) 10 (2HAP) 13 (24HAP) 19 (24HAP) 12 (8HAP) 14 (6HAP) Average 13.36 4.44 1.36 8.09 2.80 43.53 14.92 Variance 582.29 122.48 147.29 149.62 46.67 104.29 9.24 Tubes to Ovary (No.) Max. 35 (2HAP) 36 (4HAP) 43 (8HAP) 18 (2HAP) 30 (8HAP) 22 (48HAP) 21 (4HAP) Min. 8 (48HAP) 11 (48HAP) 7 (2HAP) 5 (48HAP) 10 (2HAP) 8 (8HAP) 12 (10HAP) Average 6.34 2.46 1.20 3.33 1.73 22.82 12.95 Variance 106.81 64.67 123.57 25.62 43.00 23.14 11.00 Tubes to Ovule (No.) Max. 12 (8HAP) 14 (4/8HAP) 40 (8HAP) 8 (8HAP) 16 (8HAP) 15 (48HAP) 14 (4HAP) Min. 3 (24HAP) 5 (24HAP) 6 (2HAP) 2 (48HAP) 2 (6HAP) 0 (2HAP) 0 (2HAP) Average 2.10 1.16 1.12 1.29 0.82 12.00 8.01 Variance 10.67 11.48 107.24 4.14 20.95 21.24 20.57 Abbreviations: ( Lu X Lu ): L. usitatissimum X L. usitatissimum , ( Lb X Lb ): L. bienne X L. bienne , ( Lg X Lg ): L. grandiflorum X L. grandiflorum , ( Lu X Lb ): L. usitatissimum X L. bienne , ( Lb X Lu ): L. bienne X L. usitatissimum , ( Lu X Lg ): L. usitatissimum X L. grandiflorum , ( Lg X Lu ): L. grandiflorum X L. usitatissimum , HAP: Hours after pollination, No.: Numbers; %: Percentage, Max.: Maximum value, Min.: Minimum value Additional Declarations No competing interests reported. Supplementary Files FigS1.jpg Fig. S1. Pollen vitality status in L. usitatissimum, L. bienne and L. grandiflorum . Dark stained circles represent viable pollen grains. Light stained or unstained circles represent non-viable pollen grains. FigS2.jpg Fig. S2. Representation of capsule and seed set among crosses in L. usitatissimum with its distant relative i.e., L. grandiflorum under field conditions. (a & b): Normal capsule development; (c & d): Threshed capsules with inviable shriveled seeds; (e & f): Non-viable seed formation TableS1.docx TableS2.docx Cite Share Download PDF Status: Published Journal Publication published 25 Feb, 2025 Read the published version in Scientific Reports → Version 1 posted Editorial decision: Revision requested 20 Dec, 2024 Reviews received at journal 20 Dec, 2024 Reviewers agreed at journal 12 Dec, 2024 Reviews received at journal 06 Nov, 2024 Reviewers agreed at journal 05 Nov, 2024 Reviewers agreed at journal 05 Nov, 2024 Reviewers invited by journal 04 Nov, 2024 Editor assigned by journal 04 Nov, 2024 Editor invited by journal 04 Nov, 2024 Submission checks completed at journal 01 Nov, 2024 First submitted to journal 17 Oct, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5280537","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":375073885,"identity":"8f8e2a5e-2d0b-4a34-90a3-2250014040fc","order_by":0,"name":"Vijaykumar Kailasrao Raut","email":"","orcid":"","institution":"Rama University","correspondingAuthor":false,"prefix":"","firstName":"Vijaykumar","middleName":"Kailasrao","lastName":"Raut","suffix":""},{"id":375073886,"identity":"7e121625-f92e-494c-a2ef-1d47ae59c157","order_by":1,"name":"Aneeta Yadav","email":"","orcid":"","institution":"Rama University","correspondingAuthor":false,"prefix":"","firstName":"Aneeta","middleName":"","lastName":"Yadav","suffix":""},{"id":375073887,"identity":"43ac41ff-c442-4490-8b61-d8d470aead33","order_by":2,"name":"Vikender Kaur","email":"","orcid":"","institution":"Indian Council of Agricultural Research-National Bureau of Plant Genetic Resources (ICAR-NBPGR)","correspondingAuthor":false,"prefix":"","firstName":"Vikender","middleName":"","lastName":"Kaur","suffix":""},{"id":375073888,"identity":"b3560470-b680-4f3f-97d0-16f91b3e84c8","order_by":3,"name":"Mahesh Rao","email":"","orcid":"","institution":"Indian Council of Agricultural Research-National Institute for Plant Biotechnology, (ICAR-NIPB)","correspondingAuthor":false,"prefix":"","firstName":"Mahesh","middleName":"","lastName":"Rao","suffix":""},{"id":375073889,"identity":"f8df7430-700c-42b0-84bb-6f1515494e14","order_by":4,"name":"Pooja Pathania","email":"","orcid":"","institution":"Indian Council of Agricultural Research-National Bureau of Plant Genetic Resources (ICAR-NBPGR)","correspondingAuthor":false,"prefix":"","firstName":"Pooja","middleName":"","lastName":"Pathania","suffix":""},{"id":375073890,"identity":"d3dfea7d-15b7-4f22-86a2-c1bebb9fc6d2","order_by":5,"name":"Dhammaprakash Wankhede","email":"","orcid":"","institution":"Indian Council of Agricultural Research-National Bureau of Plant Genetic Resources (ICAR-NBPGR)","correspondingAuthor":false,"prefix":"","firstName":"Dhammaprakash","middleName":"","lastName":"Wankhede","suffix":""},{"id":375073891,"identity":"01960ad5-bc9b-4b0d-8dee-f0c906d53d91","order_by":6,"name":"Mamta Singh","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA+UlEQVRIiWNgGAWjYBADHiBifJAA5xvgVc3YANXCbECSFpAuNgmiHKTbfjr9wQ+GOhlz/rXHKh7U2OUz8C8+JsFQcAenFrMzuRsbexgO81jOeJd2I+FYsmWDxLM0CQaDZ7i1HMjd2MDDcIDH4MYZsxuJDcwGDBJnjA0YDA7j1nL+7cbGPwx1YC0FiQ31RGi5kbuxmYeBmcfgfI8ZQ2LDYQMG/h7DB/i1vN04W8bgMNAWHmOJhGPHDdgk2BIfJOB1WO6Gj28q6uwNzp8x/PijptqAn//wgQMf/uDWAgGgiJNIgLDZQIwEAhoggP8AOmMUjIJRMApGAQQAANflVV7PWGj2AAAAAElFTkSuQmCC","orcid":"","institution":"Indian Council of Agricultural Research-National Bureau of Plant Genetic Resources (ICAR-NBPGR)","correspondingAuthor":true,"prefix":"","firstName":"Mamta","middleName":"","lastName":"Singh","suffix":""},{"id":375073892,"identity":"ebc2baa4-2b8f-4f0f-9d14-f3ea120b87d3","order_by":7,"name":"Gyanendra Pratap Singh","email":"","orcid":"","institution":"Indian Council of Agricultural Research-National Bureau of Plant Genetic Resources (ICAR-NBPGR)","correspondingAuthor":false,"prefix":"","firstName":"Gyanendra","middleName":"Pratap","lastName":"Singh","suffix":""}],"badges":[],"createdAt":"2024-10-17 07:23:30","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5280537/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5280537/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41598-025-90046-8","type":"published","date":"2025-02-25T15:57:05+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":68936702,"identity":"9f01d20c-59f1-4995-885a-4ca8cfc840f7","added_by":"auto","created_at":"2024-11-13 16:51:13","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":1036066,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eLinum\u003c/em\u003e species used in the study. (a): \u003cem\u003eL. usitatissimum\u003c/em\u003eL.; (b)\u003cem\u003e: L. bienne\u003c/em\u003e Mill.; (c): \u003cem\u003eL. grandiflorum\u003c/em\u003e L.\u003c/p\u003e","description":"","filename":"Fig1.png","url":"https://assets-eu.researchsquare.com/files/rs-5280537/v1/0582cc3c50ee8c21427403c4.png"},{"id":68937532,"identity":"a908ae2c-f549-48a9-8783-3d72107002c5","added_by":"auto","created_at":"2024-11-13 16:59:13","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":19893142,"visible":true,"origin":"","legend":"\u003cp\u003ePollen germination and pollen tube growth in various cross combinations in \u003cem\u003eLinum \u003c/em\u003espp.\u003c/p\u003e","description":"","filename":"Fig2.png","url":"https://assets-eu.researchsquare.com/files/rs-5280537/v1/b92c3d6cc55319a934635976.png"},{"id":68936695,"identity":"869499a4-1795-4f33-a192-117022fecfd4","added_by":"auto","created_at":"2024-11-13 16:51:13","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":510940,"visible":true,"origin":"","legend":"\u003cp\u003eVisualization of pollen germination and pollen tube growth in different hours after pollination (HAP) in various cross combinations in \u003cem\u003eLinum\u003c/em\u003e spp.\u003c/p\u003e","description":"","filename":"Fig3.png","url":"https://assets-eu.researchsquare.com/files/rs-5280537/v1/c39022d06d9869b6acced2ae.png"},{"id":68938718,"identity":"4c9e32e3-98fb-41d3-8607-0ffff86478e4","added_by":"auto","created_at":"2024-11-13 17:23:13","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":3482894,"visible":true,"origin":"","legend":"\u003cp\u003ePollen-pistil interaction in self and cross combination. (a, b, c, d): Pollen germination, pollen tube growth in style, PTG in attachment region and, pollen tube navigation to ovule in self-pollinated pistils of \u003cem\u003eL. usitatissimum\u003c/em\u003e (2HAP); (e, f, g, h): Pollen germination, pollen tube growth in style, PTG in attachment region and, pollen tube navigation to ovule in self-pollinated pistils of \u003cem\u003eL. bienne\u003c/em\u003e (2HAP); (i, j, k, l): Pollen germination, pollen tube growth in style, PTG in attachment region and, pollen tube navigation to ovule in self-pollinated pistils of \u003cem\u003eL. grandiflorum\u003c/em\u003e (2HAP); (m, n, o, p): Pollen germination, pollen tube growth in style, PTG in attachment region and, pollen tube navigation to ovule in pistils of \u003cem\u003eL. usitatissimum\u003c/em\u003e pollinated with \u003cem\u003eL. bienne \u003c/em\u003e(2HAP); (q, r, s, t): Pollen germination, pollen tube growth in style, PTG in attachment region and, pollen tube navigation to ovule in pistils of \u003cem\u003eL. bienne\u003c/em\u003e pollinated with \u003cem\u003eL. usitatissimum\u003c/em\u003e (2HAP).\u003c/p\u003e","description":"","filename":"Fig4.png","url":"https://assets-eu.researchsquare.com/files/rs-5280537/v1/22c056d4c1bc908dbbc6f5ae.png"},{"id":68936692,"identity":"613b9673-1588-40d9-ac1d-75b46c0c8093","added_by":"auto","created_at":"2024-11-13 16:51:13","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":315299,"visible":true,"origin":"","legend":"\u003cp\u003ePollen-pistil interaction in crosses with \u003cem\u003eL. grandiflorum\u003c/em\u003e (a): Callose formation (2HAP) \u003cem\u003eLu \u003c/em\u003eX\u003cem\u003e Lg\u003c/em\u003e; (b): Callose formation (2HAP) \u003cem\u003eLg\u003c/em\u003e X \u003cem\u003eLu\u003c/em\u003e; (c): Ungerminated pollen grains seen during longer hours after pollination in \u003cem\u003eLu\u003c/em\u003e X \u003cem\u003eLg\u003c/em\u003e(10HAP); (d, e): Ungerminated pollen grains seen during longer hours after pollination in \u003cem\u003eLg \u003c/em\u003eX \u003cem\u003eLu\u003c/em\u003e (24HAP) that start degradation/deformation; (f, g): Limited/restricted pollen grain germination on stigmatic surfaces in \u003cem\u003eLg\u003c/em\u003e X \u003cem\u003eLu\u003c/em\u003e (4HAP) and \u003cem\u003eLu\u003c/em\u003e X \u003cem\u003eLg\u003c/em\u003e (6HAP) wide crosses respectively.\u003c/p\u003e","description":"","filename":"Fig5.png","url":"https://assets-eu.researchsquare.com/files/rs-5280537/v1/7705cb04429b2a9fe3ed82b4.png"},{"id":68936696,"identity":"dc16a9fb-c8c6-4c75-b856-e7a4df9f3655","added_by":"auto","created_at":"2024-11-13 16:51:13","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":1063468,"visible":true,"origin":"","legend":"\u003cp\u003eRepresentation of activities taking place in \u003cem\u003eL. usitatissimum\u003c/em\u003e during pollen-pistil interaction in wide crosses of \u003cem\u003eL. usitatissimum\u003c/em\u003e X \u003cem\u003eL. grandiflorum\u003c/em\u003e. (a): Partially elongated/restricted pollen tube growth due to callose deposition on the stigma in \u003cem\u003eLu\u003c/em\u003e X \u003cem\u003eLg\u003c/em\u003e (4HAP); (b): Swollen pollen tube tip in mid stylar region (4HAP in \u003cem\u003eLu \u003c/em\u003eX \u003cem\u003eLg\u003c/em\u003e); (c): Swollen pollen tube tip at the attachment region (4HAP in \u003cem\u003eLu\u003c/em\u003e X \u003cem\u003eLg\u003c/em\u003e); (d, e): Twisted pollen tube growth in the upper and mid stylar region of pistil (4HAP); (f): Swollen pollen tube tip at attachment region of ovary (4HAP in \u003cem\u003eLu \u003c/em\u003eX \u003cem\u003eLg\u003c/em\u003e); (g): Inhibited pollen tubes at the attachment region of ovary (4HAP in \u003cem\u003eLu\u003c/em\u003e X \u003cem\u003eLg\u003c/em\u003e); (h): Appearance of pollen tubes growing in reverse direction while reaching towards ovule (6HAP); (i): Complex and convoluted pollen tube growth at middle stylar region (6HAP); (j): Ungerminated pollen grains and terminated pollen tube growth (10HAP).\u003c/p\u003e","description":"","filename":"Fig6.png","url":"https://assets-eu.researchsquare.com/files/rs-5280537/v1/f43bac09693f4e2c312efcaa.png"},{"id":68936694,"identity":"35690a38-76ff-4e5f-abe4-74955d617128","added_by":"auto","created_at":"2024-11-13 16:51:13","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":1264634,"visible":true,"origin":"","legend":"\u003cp\u003eRepresentation of activities taking place in \u003cem\u003eL. grandiflorum\u003c/em\u003e during pollen-pistil interaction in wide crosses of \u003cem\u003eL. grandiflorum\u003c/em\u003e X \u003cem\u003eL. usitatissimum\u003c/em\u003e. (a): Restricted and twisted pollen tube growth (2HAP); (b): Twisted pollen tube growth at attachment region of ovary (2HAP); (c, d): Under-germinated pollen grains with terminated, twisted and swollen pollen tube (4HAP); (e): Ungerminated pollen grains are observed even after 4HAP; (f): Reduced numbers of pollen tubes growing in the stylar region along with non-germinated pollen grains (4HAP); (g): Ruptured/bifurcated pollen tube tip in the stylar region (4HAP); (h): Convoluted pollen tube growth in the mid-stylar region (10HAP); (i): Pollen grains are seen ungerminated on foreign stigma of \u003cem\u003eL. grandiflorum\u003c/em\u003e (10HAP); (j): Considerable pollen tube growth is visible only in 6HAP with elongated pollen tube growth in the stylar region. Some pollen tubes can still be observed with partial growth at this stage; (k): Delayed pollen tube growth with swollen tip (6HAP). Pollen tube growth termination at mid stylar region with swollen tip 6HAP \u003cem\u003eLg\u003c/em\u003e X \u003cem\u003eLu\u003c/em\u003e delayed pollen germination can be observed even after 6HAP; (l): Considerably non germinated pollen grains observed on stigmatic surface (8HAP); (m): Many pollen grains germinated with swollen pollen tube tips and twisted growth with random growth patterns can be seen in the stylar region even 10HAP; (n): Complex, random pollen tube growth pattern (10HAP); (o): Non-germinated pollen grains start degradation (24HAP).\u003c/p\u003e","description":"","filename":"Fig7.png","url":"https://assets-eu.researchsquare.com/files/rs-5280537/v1/ac6fe2eafcaa96681a681e76.png"},{"id":77622528,"identity":"831e4762-a8b8-441a-b7e1-6b4716b69c3f","added_by":"auto","created_at":"2025-03-03 16:07:59","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":50326387,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5280537/v1/8b87c7bd-038d-4e21-a41f-bbacffd23c52.pdf"},{"id":68938716,"identity":"89ddff84-2240-432e-a4cd-bc64626a1e5c","added_by":"auto","created_at":"2024-11-13 17:23:13","extension":"jpg","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":156183,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFig. S1\u003c/strong\u003e. Pollen vitality status in \u003cem\u003eL. usitatissimum, L. bienne and L. grandiflorum\u003c/em\u003e. Dark stained circles represent viable pollen grains. Light stained or unstained circles represent non-viable pollen grains.\u003c/p\u003e","description":"","filename":"FigS1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-5280537/v1/e0962215c23c3a6d0ff32afd.jpg"},{"id":68938515,"identity":"476568fb-2dd4-427c-ad6c-8c126f5e490f","added_by":"auto","created_at":"2024-11-13 17:15:13","extension":"jpg","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":9401741,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFig. S2. \u003c/strong\u003eRepresentation of capsule and seed set among crosses in \u003cem\u003eL. usitatissimum\u003c/em\u003e with its distant relative i.e., \u003cem\u003eL. grandiflorum\u003c/em\u003e under field conditions. (a \u0026amp; b): Normal capsule development; (c \u0026amp; d): Threshed capsules with inviable shriveled seeds; (e \u0026amp; f): Non-viable seed formation\u003c/p\u003e","description":"","filename":"FigS2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-5280537/v1/21799bed729d075577914bef.jpg"},{"id":68937887,"identity":"8e85df2b-9f8a-42bd-a9ee-a92b75be08b4","added_by":"auto","created_at":"2024-11-13 17:07:13","extension":"docx","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":36826,"visible":true,"origin":"","legend":"","description":"","filename":"TableS1.docx","url":"https://assets-eu.researchsquare.com/files/rs-5280537/v1/ab0d35a910c6cec695659a98.docx"},{"id":68936693,"identity":"89110555-1ac3-4000-91af-4b2e380ea8dc","added_by":"auto","created_at":"2024-11-13 16:51:13","extension":"docx","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":17415,"visible":true,"origin":"","legend":"","description":"","filename":"TableS2.docx","url":"https://assets-eu.researchsquare.com/files/rs-5280537/v1/5abf24a793e3165e25892867.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Pollen–pistil interactions in divergent wide crosses lead to spatial and temporal pre-fertilization reproductive barrier in flax (Linum usitatissimum L.)","fulltext":[{"header":"Introduction","content":"\u003cp\u003eFlaxseed, also known as linseed has evolved as one of the predominant industrial crops during the last few decades\u003csup\u003e\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e\u003c/sup\u003e and originally domesticated for its oilseed and natural fiber usage\u003csup\u003e\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e\u003c/sup\u003e. Grown for dual purpose, the two separate main directions have been developed for utilizing the plant's products, leading to the cultivation of fiber flax and oil flax (linseed)\u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e. A rich and interesting history surrounds flax. The Fertile Crescent region, saw the domestication of this plant approximately 8,000 BCE\u003csup\u003e\u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e71\u003c/span\u003e\u003c/sup\u003e first for its seeds and later for its fibres\u003csup\u003e\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u003c/sup\u003e. It\u0026rsquo;s fascinating that its usage existed even before domestication. Caucasus area was investigated to report flax yarn residues dated 30,000 years ago\u003csup\u003e\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e\u003c/sup\u003e. Nonetheless, the cultivation of flax as a crop for textile fiber was made possible by Ancient Egypt\u003csup\u003e\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e\u003c/sup\u003e. Vavilov, (1926)\u003csup\u003e\u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e72\u003c/span\u003e\u003c/sup\u003e highlighted the Indian subcontinent, Abyssinia, and the Mediterranean area as major regions of flax diversity. In these regions, the wild ancestor \u003cem\u003eLinum bienne\u003c/em\u003e was cultivated, likely in isolation, leading to the domestication of \u003cem\u003eL. usitatissimum\u003c/em\u003e. Thus, these areas could be associated with multiple domestication events of flax. These days, high-end fabrics, natural actuators\u003csup\u003e\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/sup\u003e, and composite material reinforcements\u003csup\u003e\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e\u003c/sup\u003e all use flax fibres. Therefore, flax is a link between cultures and periods\u003csup\u003e\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e\u003c/sup\u003e. As a vital industrial resource, linseed oil is employed for multiple purposes, primarily as a fundamental component in paints, resins, printing inks, varnishes, and linoleum\u003csup\u003e\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u003c/sup\u003e. Advances in material science have brought fiber flax back into the spotlight, as its fibers are now being harnessed for a multitude of environmentally sustainable industrial uses, such as composites, geotextiles, insulation, and specialty papers. In India during the winter season, linseed is the second most important oilseed crop after rapeseed mustard in terms of both area and production. It holds the top position in the country for industrial oil production and holds the top spot globally in terms of acreage, representing 23.8 percent of the world total, and ranks third in production, accounting for 10.2 percent of the world total\u003csup\u003e\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e\u003c/sup\u003e. Every part of the linseed plant is commercially utilized, either directly or after processing. The seed contains 33 to 47% of oil, containing around 58% omega-3 fatty acid. Flax is cultivated on an area of about 2.31\u0026nbsp;million hectares globally. Asian countries contribute about 49.2% area in the world\u003csup\u003e\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003e. India, Canada, China, the USA, and Ethiopia are the main countries for linseed production.\u003c/p\u003e \u003cp\u003eThe existence of modern-day flax varieties mainly includes domestication and selection as important historical breeding phenomenon. The initial bred flax varieties to surface in Southern Europe, especially in the Danube valley, were winter oil crops\u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e. Evidence from history shows that bast fiber textiles extended to the Nile Valley and even to present-day Britain by 4,000 BC. In Eastern Europe, the creation of spring-planted fiber varieties is thought to have originated from either a unique domestication event in Central Asia (Indo-Afghan region) or from the existing European varieties found in the south\u003csup\u003e\u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e77\u003c/span\u003e,\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e. At the dawn of the 20th century, Russia established itself as the foremost supplier of top-quality fiber flax to Europe. The tireless efforts of local peasants in cultivation gave rise to \u0026ldquo;kryazh,\u0026rdquo; a set of heritage landraces that developed over centuries through both natural and artificial selection, focusing on the creation of tall fiber flax varieties. These kryazh varieties were considered premium commodity crops, leading to their active trade and incorporation into breeding programs worldwide. It is widely recognized that the origins of all modern fiber flax varieties can be traced back to Eastern Europe\u003csup\u003e\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eFlaxseed, being a self-pollinated crop, has faced the narrowing down of its genetic base due to selection during domestication and plant breeding just like all other cultivated crops\u003csup\u003e\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/sup\u003e. The wild progenitor species gave rise to all the major crops around the world, mainly about 4000 to 10000 years ago through domestication\u003csup\u003e\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u003c/sup\u003e. New variants can be derived from wild relatives or intermediary landraces, and a portion of these can be beneficial for enhancing crops through traditional breeding techniques or biotechnology. \u003cem\u003eLinum\u003c/em\u003e is one of the largest genus of \u003cem\u003eLinaceae\u003c/em\u003e family, containing more than 200 species\u003csup\u003e\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e\u003c/sup\u003e growing in diverse geographical regions of world. In India, apart from cultivated flax viz., \u003cem\u003eL. usitatissimum\u003c/em\u003e, the occurrence of other five Crop Wild Relatives (CWR\u0026rsquo;s) namely, \u003cem\u003eL. strictrum\u003c/em\u003e, \u003cem\u003eL. bienne\u003c/em\u003e, \u003cem\u003eL. grandiflorum, L. mysorense\u003c/em\u003e and \u003cem\u003eL. perenne\u003c/em\u003e\u003csup\u003e\u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e66\u003c/span\u003e\u003c/sup\u003e is reported. However, utilization of wild linseed species in crop improvement programs has been limited partly because of lack of collection and conservation of diverse species in gene banks and partly because of the degree of genetic divergence that exists among the interspecific ecotypes causing crossability issues.\u003c/p\u003e \u003cp\u003eLarge number of CWR\u0026rsquo;s of flax are available worldwide in genus \u003cem\u003eLinum\u003c/em\u003e that could be of immense potential for their inclusion as trait donors under various flax improvement programmes. However, the genus \u003cem\u003eLinum\u003c/em\u003e has many diploid species that exhibit a remarkable diversity in chromosome number including n\u0026thinsp;=\u0026thinsp;8, 9, 10, 12, 14, 15, 16, 18, 30, and \u0026gt;\u0026thinsp;30\u003csup\u003e10,26\u003c/sup\u003e. No absolute studies have yet emerged as a reflective of phylogenetic relationships among \u003cem\u003eLinum\u003c/em\u003e species. There are uncertainties regarding the chromosome numbers of some wild species. The differences in chromosome size and their number pose immense hurdles in hybridizing CWRs with cultivated flax. These genetic differences between wild and cultivated species have their unfavorable impact on feasibility of hybridization among flax wild species in both or at least one direction. Successful production of fertile hybrids to an extent, is assumed to occur only in species with equal chromosome numbers. A comprehensive detail on this aspect has been represented by Jhala et al. (2008)\u003csup\u003e\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003ePotential usage of some wild relatives of \u003cem\u003eLinum\u003c/em\u003e includes fiber quality and core hardness (\u003cem\u003eL. bienne\u003c/em\u003e), bud fly resistance, lignan and neolignans synthesis (\u003cem\u003eL. grandiflorum\u003c/em\u003e), nutritious and oily quality (\u003cem\u003eL. lewisii\u003c/em\u003e), drought and rust tolerance (\u003cem\u003eL. marginale\u003c/em\u003e). Despite their huge potential, the use of these CWR\u0026rsquo;s under flax improvement programme is very scarce. Studies focused on the crossability potential\u003csup\u003e\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e,\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e\u003c/sup\u003e or elaborating the reasons behind the reproductive isolation in flax are limited. Interspecific hybridization between wild and cultivated species can play a major part to add economically important characters in \u003cem\u003eLinum\u003c/em\u003e.\u003c/p\u003e \u003cp\u003ePollen-pistil interaction (PPI) offers enormous potential for the manipulation of pollen screening which is obviously for the compatibility of pollen. PPI can be considered as an important contributory factor in introducing genetic diversity to the flowering plants from their wild and weedy counterparts. Assessing the compatibility of foreign pollen on cultivated species during pollination and fertilization offers enormous potential to study the PPI and understand incompatibility barriers\u003csup\u003e\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e,\u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e67\u003c/span\u003e,\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e. This may broaden the scope for bringing wild species under crop breeding programs for the improvement of traits of interests or introducing genetic diversity.\u003c/p\u003e \u003cp\u003eSelf-incompatibility due to the occurrence of heterostyly is very well reported in different species related to cultivated \u003cem\u003eLinum\u003c/em\u003e \u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e,\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e,\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e. Also, the mechanism of self-incompatibility between different floral morphs has been investigated\u003csup\u003e\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e,\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e,\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e\u003c/sup\u003e. However, to the best of our knowledge, there is rarely any reference literature available pertaining to germination and tube growth of foreign pollen grain onto the stigmatic surface and stylar regions of the parent belonging to another species in \u003cem\u003eLinum\u003c/em\u003e. Therefore, the present investigation was carried out to (i.) assess the major pre-zygotic barriers and their effect on the germination of pollen grains on foreign stigma using fluorescent microscopy of aniline blue stain aided technology, (ii.) understand how the species barriers operate on pollen germination (PG) and pollen tube growth (PTG) when different wild species are used as the female parents and (iii.) account for the impact of maternal cytoplasm on pollen rejection, PTG, and fertilization in different cross combinations.\u003c/p\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n \u003ch2\u003ePollen vitality\u003c/h2\u003e\n \u003cp\u003ePollen vitality test revealed an average 69.92% vital pollen grains in \u003cem\u003eL. usitatissimum\u003c/em\u003e in 3108 studied pollen mother cells (PMCs) ranging from 55.33\u0026ndash;83.94%. In \u003cem\u003eL. bienne\u003c/em\u003e, pollen vitality percentage (PV%) ranged from 77.04\u0026ndash;92.14% with an average of 86.35% in 1368 studied PMCs. In case of \u003cem\u003eL. grandiflorum\u003c/em\u003e, pollen vitality percentage (PV%) ranged from 82.05\u0026ndash;96.49% in 2268 studied pollen grains with an average of 91.30% (Table\u0026nbsp;3).\u003c/p\u003e\n\u003c/div\u003e\n\u003ch3\u003ePollen germination on stigmatic surface of self-pollinated buds\u003c/h3\u003e\n\u003cp\u003eAnalysis of PG in self- and cross-pollinated stigmatic surfaces of processed pistils was done as the percent PG (%) for the respective duration. The analysis of the captured images showed more than 90% PG on the stigmas in all the three species in case of self-pollination in each time duration (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003ea, b and c). Overall PG in self-pollination for \u003cem\u003eL. usitatissimum\u003c/em\u003e X \u003cem\u003eL. usitatissimum\u003c/em\u003e (\u003cem\u003eLu\u003c/em\u003e X \u003cem\u003eLu\u003c/em\u003e) was 94.51% for all HAP, maximum 95.99% for 10HAP and minimum 92.83% for 8HAP (Table \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e, Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003ea, Table \u003cspan class=\"InternalRef\"\u003eS1\u003c/span\u003e). Overall PG in self-pollination for \u003cem\u003eL. bienne\u003c/em\u003e X \u003cem\u003eL. bienne\u003c/em\u003e (\u003cem\u003eLb\u003c/em\u003e X \u003cem\u003eLb\u003c/em\u003e) was 94.27% for all HAP, maximum 95.58% for 8HAP and minimum 92.17% for 24HAP. Overall PG in legitimate mating for \u003cem\u003eL. grandiflorum\u003c/em\u003e X \u003cem\u003eL. grandiflorum\u003c/em\u003e (\u003cem\u003eLg\u003c/em\u003e X \u003cem\u003eLg\u003c/em\u003e) was 94.12% for all HAP, maximum 94.82% for 2HAP and minimum 93.54% for 10HAP.\u003c/p\u003e\n\u003ch3\u003ePollen germination on stigmatic surface of cross-pollinated buds\u003c/h3\u003e\n\u003cp\u003eFor the cross combinations involving \u003cem\u003eL. bienne\u003c/em\u003e, the PG percentage on the foreign stigmas was more than 90% in both direct and reciprocal combinations. Overall PG in cross pollination for \u003cem\u003eL. usitatissimum\u003c/em\u003e X \u003cem\u003eL. bienne\u003c/em\u003e (\u003cem\u003eLu\u003c/em\u003e X \u003cem\u003eLb\u003c/em\u003e) was 94.00% for all HAP (Table \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e), maximum 97.00% for 4HAP and minimum 91.91% for 8HAP. For the first time duration of observation i.e., 2HAP, the PG percent was 93.71% (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003ed). Overall PG in reciprocal cross combination for \u003cem\u003eL. bienne\u003c/em\u003e X \u003cem\u003eL. usitatissimum\u003c/em\u003e (\u003cem\u003eLb\u003c/em\u003e X \u003cem\u003eLu\u003c/em\u003e) was 94.87% for all HAP, maximum 96.19% for 4HAP and minimum 93.03% for 48HAP. For the first time duration of observation i.e., 2HAP, the PG percent was 95.20% (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003ee). For the cross combinations involving \u003cem\u003eL. grandiflorum\u003c/em\u003e, the PG percentage on the foreign stigmas was restricted to about 10% in both direct and reciprocal combinations. Overall PG in cross pollination for \u003cem\u003eL. usitatissimum\u003c/em\u003e X \u003cem\u003eL. grandiflorum\u003c/em\u003e (\u003cem\u003eLu\u003c/em\u003e X \u003cem\u003eLg\u003c/em\u003e) was 8.15% for all HAP (Table \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e), maximum 10.92% for 4HAP and minimum 6.66% for 48HAP. While or the first-time duration of observation i.e., 2HAP, the PG percent was 7.77% (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003ef). For its reciprocal combination \u003cem\u003eL. grandiflorum\u003c/em\u003e X \u003cem\u003eL. usitatissimum\u003c/em\u003e (\u003cem\u003eLg\u003c/em\u003e X \u003cem\u003eLu\u003c/em\u003e), the overall PG was 8.82% for all HAP, ranging from a maximum of 10.33% for 6HAP to a minimum of 7.71% for 8HAP. While for the first-time duration of observation i.e., 2HAP, the PG percent was 8.65% (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003eg). Overall, in self-pollination, PG ranged from 92.17% (\u003cem\u003eLb\u003c/em\u003e X \u003cem\u003eLb\u003c/em\u003e for 24HAP) \u0026minus;\u0026thinsp;95.99% (\u003cem\u003eLu\u003c/em\u003e X \u003cem\u003eLu\u003c/em\u003e for 10HAP) and in cross combination it ranged from 6.66% (\u003cem\u003eLu\u003c/em\u003e X \u003cem\u003eLg\u003c/em\u003e for 48HAP) \u0026minus;\u0026thinsp;97.00% (\u003cem\u003eLu\u003c/em\u003e X \u003cem\u003eLb\u003c/em\u003e for 4HAP) (Table \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e\n\u003ch3\u003ePollen tube growth in self-pollinated pistils\u003c/h3\u003e\n\u003cp\u003eIn case of self-pollination in all the three species, it took only 2HAP to grow through the stylar region and reach to up-to the ovule (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eb, c and d, Table \u003cspan class=\"InternalRef\"\u003eS1\u003c/span\u003e). The maximum number of pollen tubes (PTs) observed growing in the stylar region, ovary and tubes reaching to ovules were 75, 35 and 12 in 4HAP, 2HAP and 8HAP respectively for \u003cem\u003eL. usitatissimum\u003c/em\u003e. The germinated PTs were observed growing with different rates in pistil ranging from 4.74% (24HAP) to 23.17% (8HAP) in stylar region, 2.37% (24HAP) to 11.38% (8HAP) in attachment region of ovary and, 0.79% (24HAP) to 4.88% (8HAP) seen penetrating ovular region inside the ovary (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003ea). For \u003cem\u003eL. bienne\u003c/em\u003e, maximum number of PTs observed growing in the stylar region, ovary and tubes reaching to ovules were 62 (4HAP), 36 (4HAP) and 14 (4 and 8HAP) respectively. The germinated PTs were observed growing with different rates in pistil ranging from 3.35% (6HAP) to 6.15% (24HAP) in stylar region, 1.32% (48HAP) to 3.28% (4HAP) in attachment region of ovary and, 0.79% (6HAP) to 1.54% (8 and 10HAP) seen penetrating ovular region inside the ovary (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003eb). In case of \u003cem\u003eL. grandiflorum\u003c/em\u003e, the PTG was comparatively lower in style (10), ovary (7) and ovule (6) during the initial two hours of observation (2HAP). The maximum numbers could be observed only during 8HAP in the stylar region (49), ovary (43) and tubes reaching to ovules (40). The germinated PTs were observed growing with different rates in pistil ranging from 0.93% (48HAP) to 1.95% (2HAP) in stylar region, 0.87% (48HAP) to 1.67% (4 HAP) in attachment region of ovary and, 0.87% (48HAP) to 1.48% (4HAP) seen penetrating ovular region inside the ovary (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003ec). The overall extent of PTs observed growing in the stylar region, ovary and ovule were 13.36%, 6.34% and 2.10% respectively for \u003cem\u003eL. usitatissimum\u003c/em\u003e over the germinated pollen grains. For \u003cem\u003eL. bienne\u003c/em\u003e. overall extent of PTs observed growing in the stylar region, ovary and ovule were 4.44%, 2.46% and 1.16% respectively. For \u003cem\u003eL. grandiflorum\u003c/em\u003e these parameters were 1.36%, 1.20% and 1.12% for the respective pistil regions (Table \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e\n\u003ch3\u003ePollen tube growth in cross-pollinated pistils\u003c/h3\u003e\n\u003cp\u003eIn case of wide cross combination, with \u003cem\u003eLu\u003c/em\u003e X \u003cem\u003eLb\u003c/em\u003e and its reciprocal \u003cem\u003eLb\u003c/em\u003e X \u003cem\u003eLu\u003c/em\u003e it took only 2HAP to reach to the stylar region (40 and 21 respectively) and ovary (18 and 10 respectively). However, the tube could be seen reaching up to the ovular region only in about 2HAP (5 and 3 respectively) (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eb, c and d, Table \u003cspan class=\"InternalRef\"\u003eS1\u003c/span\u003e). The maximum number of PTs observed growing in the stylar region, ovary and tubes reaching to ovules were 44, 18 and 8 in 4HAP, 2HAP and 8HAP respectively for \u003cem\u003eLu\u003c/em\u003e X \u003cem\u003eLb.\u003c/em\u003e The average number of PTs observed growing in the stylar region, ovary and ovule were 8.10%, 3.33% and 1.29% respectively over the germinate pollen grains for all the studied hours (Table \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). The germinated PTs were observed growing with different rates in pistil ranging from 4.39% (24HAP) to 12.78% (2HAP) in stylar region, 2.10% (48HAP) to 5.75% (2HAP) in attachment region of ovary and, 0.84% (48HAP) to 2.82% (8HAP) seen penetrating ovular region inside the ovary (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003ed). For its reciprocal cross combination \u003cem\u003eLb\u003c/em\u003e X \u003cem\u003eLu\u003c/em\u003e, the maximum number of pollen tubes observed growing in the stylar region (40), ovary (30) and tubes reaching to ovules (16) could be observed only in 8HAP. The average number of PTs observed growing in the stylar region, ovary and ovule were 2.80%, 1.73% and 0.82% respectively for overall time duration considered under the current study, over the germinated pollen grains \u003cstrong\u003e(\u003c/strong\u003eTable \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). The germinated PTs were observed growing with different rates in pistil ranging from 2.11% (10HAP) to 3.47% (4 and 48HAP) in stylar region, 1.07 (2HAP) to 2.11% (4HAP) in attachment region of ovary and, 0.27% (6HAP) to 1.25% (48HAP) seen penetrating ovular region inside the ovary (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003ee). With other distant cross combination i.e., \u003cem\u003eLu\u003c/em\u003e X \u003cem\u003eLg\u003c/em\u003e and its reciprocal \u003cem\u003eLg\u003c/em\u003e X \u003cem\u003eLu\u003c/em\u003e, again it took only 2HAP to reach to the stylar region (32 and 19 respectively) and ovary (16 and 15 respectively) (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eb, c). However, the tube could be seen reaching up to the ovular region only in about 4HAP in \u003cem\u003eLu\u003c/em\u003e X \u003cem\u003eLg\u003c/em\u003e (10) and in \u003cem\u003eLg\u003c/em\u003e X \u003cem\u003eLu\u003c/em\u003e (14) (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003ed). The maximum number of PTs observed growing in the stylar region, ovary and tubes reaching to ovules were 39 (4HAP), 22 (48HAP) and 15(48HAP) for \u003cem\u003eLu\u003c/em\u003e X \u003cem\u003eLg.\u003c/em\u003e The average number of PTs observed growing in the stylar region, ovary and ovule were 43.53%, 22.82% and 12.00% respectively over the germinate pollen grains for all the studied hours (Table \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). The germinated PTs were observed growing with different rates in pistil ranging from 27.91% (8HAP) to 68.09% (2HAP) in stylar region, 13.54% (6HAP) to 40.00% (24HAP) in attachment region of ovary and, none in 2HAP to 28% (24HAP) seen penetrating ovular region inside the ovary (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003ef). For its reciprocal cross combination \u003cem\u003eLg\u003c/em\u003e X \u003cem\u003eLu\u003c/em\u003e, the maximum number of PTs observed growing in the stylar region (23), ovary (21) and tubes reaching to ovules (14) was observed in 4HAP. The average number of PTs observed growing in the stylar region, ovary and ovule were 14.92%, 12.95% and 8.01%, respectively over the germinated pollen grains for all the studied hours (Table \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). The germinated PTs growing with different rates in pistil ranging from 11.59% (24HAP) to 22.22% (8HAP) in stylar region, 9.42% (24HAP) to 22.22% (8HAP) in attachment region of ovary and none in 2HAP to 16.05% (8HAP) were observed penetrating ovular region inside the ovary (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003eg).\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eFertilization between diverse species within the same genus is expected to be successful only if there is some means of compatibility between both the parents. Once there happens a successful completion of a series on sequential events following pollination, as allowed by the coordinated gene and gene complexes of both pollen and pollen parents, successful fertilization takes place\u003csup\u003e\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e,\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e,\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e. As per our past experiences, in case of interspecific hybridization in \u003cem\u003eL. usitatissimum\u003c/em\u003e and its wild progenitor i.e., \u003cem\u003eL. bienne\u003c/em\u003e, the seed set was successful up to 81.5 to 83.3% including reciprocals (Table \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003e). However, despite many attempts for making wide crosses in \u003cem\u003eL. usitatissimum\u003c/em\u003e with its distant relative i.e., \u003cem\u003eL. grandiflorum\u003c/em\u003e, we had been unsuccessful getting any viable seeds under field conditions (Fig. \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003e). Earlier studies also report unsuccessful attempts for obtaining interspecific hybrids between cultivated flax and \u003cem\u003eL. grandiflorum\u003c/em\u003e\u003csup\u003e\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e\u003c/sup\u003e and attempts to get hybrids with embryo rescue have been also done in the past\u003csup\u003e\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u003c/sup\u003e. It is worth mentioning here that, capsules formation was observed during most of the attempts made under field conditions to an extent of about 70% in direct crosses and 22% in reciprocal cross combinations with \u003cem\u003eL. grandiflorum\u003c/em\u003e (Table \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003e). Since, capsules were formed without any viable seed development; we assume the occurrence of parthenocarpic development of fruits without affecting successful fertilization resulting into capsules with non-viable seeds. Wide hybridization as a driving force for the development of seedless or rudimentary fruits in higher plants have been discussed thoroughly\u003csup\u003e\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e,\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/sup\u003e. Recently, Badiger et al. (2024)\u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e reported that formation of aborted embryo with partially filled seeds suffered from severe pre-zygotic barriers, because of interspecific hybridization in the crosses in \u003cem\u003eAbelmoschus\u003c/em\u003e species.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eOn the other hand, pollen vitality is another important factor affecting the pollen germination percentage. Pollen represents a critical stage in the life cycle of plants, as viable pollen is crucial for efficient sexual plant reproduction. Reproductive fitness in wild plants is often determined by the quantity and quality of pollen grains produced. Both parameters can be considered as survival strategy in deciding reproductive fitness of natural populations growing spontaneously under resource limited marginal environments\u003csup\u003e\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e\u003c/sup\u003e. High pollen vitality in \u003cem\u003eL. bienne\u003c/em\u003e (86.35%) and \u003cem\u003eL. grandiflorum\u003c/em\u003e (91.30%) than the cultivated \u003cem\u003eL. usitatissimum\u003c/em\u003e (69.92%) indicates towards the competitive ability of these wild species to capture more opportunities to affect efficient fertilization during sexual reproduction. Though, pollen vitality is an important factor for pollen germination and tube growth, these parameters were seemed to be drastically reduced (Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e) in case of wide crosses with \u003cem\u003eL. grandiflorum\u003c/em\u003e, despite appreciable pollen vitality percentage in \u003cem\u003eL. usitatissimum\u003c/em\u003e (69.92%) and \u003cem\u003eL. grandiflorum\u003c/em\u003e (91.30%) (Table\u0026nbsp;3). This emphasizes that high pollen vitality does not guarantee successful pollen germination and subsequent pollen tube growth inside the foreign style's transmitting tract when dealing with genetically distant parents. Pollen germination is a vital indicator of viability and provides valuable estimation for pollen tube growth. It is crucial to note that germination loss does not necessarily mean the pollen has died, but rather underscores the need for optimal germination conditions\u003csup\u003e\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e\u003c/sup\u003e. The genetic differences between cultivated and wild \u003cem\u003eL. grandiflorum\u003c/em\u003e seems violating the congenial conditions required for optimal germination of pollen grains and subsequent pollen tube growth.\u003c/p\u003e \u003cp\u003eSelf-incompatibility arising from heterostyly is widely documented in various species linked to cultivated \u003cem\u003eLinum\u003c/em\u003e, as mentioned previously. Mechanism of self-incompatibility (SI) between different floral morphs are also studied and the mechanism of SI among various floral forms in \u003cem\u003eLinum grandiflorum\u003c/em\u003e\u003csup\u003e\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e,\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e\u003c/sup\u003e, \u003cem\u003eLinum usitatissimum\u003c/em\u003e\u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u003c/sup\u003e have been investigated in the past. Based on what we know, there is a lack of available literature on the germination and tube growth of foreign pollen grain on the stigma and style of a different species in flaxseed.\u003c/p\u003e\n\u003ch3\u003ePollen germination on stigmatic surface\u003c/h3\u003e\n\u003cp\u003eIn case of self-pollination for the cultivated species \u003cem\u003eL. usitatissimum\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003ea), its wild progenitor \u003cem\u003eL. bienne\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eb) and, legitimate mating with its distant relative \u003cem\u003eL. grandiflorum\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003ec), the PG percentage were as high as 95% (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003ea, Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ea, b and c, Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003ea-l). The occurrence of high germination percentage may be explained by the hypothesis proposed by Lewis (1960)\u003csup\u003e\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e\u003c/sup\u003e that resulted into adaptation from allogamy to autogamy in flax following two phenomena. These may include either a mutation of activation of pollen specificities, the styles maintaining their incompatibility reaction or, the development of secondary fertility genes thwarting the incompatibility reaction or partial allogamy and the lack of complete auto-sterility. PG percentages and pollen tube (PT) lengths varied in flax styles for different crosses, including reciprocals and selfings, using cultivated \u003cem\u003eL. usitatissimum\u003c/em\u003e species\u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u003c/sup\u003e. Similarly, the PG percentage on the foreign stigma, when \u003cem\u003eL. bienne\u003c/em\u003e was used either as pollen or pistil parent, were as good as in case of self-pollination of individual species (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003em, q). This can be accounted to the genetic similarity between the cultivated \u003cem\u003eLinum\u003c/em\u003e species and its wild progenitor which is the immediate ancestor of modern cultivated flax\u003csup\u003e\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e. However, when \u003cem\u003eL. grandiflorum\u003c/em\u003e was used as pollen or pistil parent, the PG percentage were drastically reduced during different HAP (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003ea-g, Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003ea). The maximum PG on the stigmatic surface of cultivated \u003cem\u003eLinum\u003c/em\u003e, when \u003cem\u003eL. grandiflorum\u003c/em\u003e served as pollen parent, could reach to a maximum of 10.92% only in 4HAP. When used as pistil parent, the PG reached to a maximum of 10.33% only in 6HAP (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ef, g). Varied response and reciprocal differences of pollen and pistil parents, on PG, PTG and compatibility parameters have been studied in wide hybridization of \u003cem\u003eVigna\u003c/em\u003e species\u003csup\u003e\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e\u003c/sup\u003e, pigeon pea\u003csup\u003e\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e\u003c/sup\u003e, \u003cem\u003eCurcuma\u003c/em\u003e\u003csup\u003e\u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e75\u003c/span\u003e\u003c/sup\u003e. It is evident that there is a reproductive barrier imposed on the landed pollen grain during the pre-fertilization stage. Analysis of captured images under fluorescent microscope, indicated the formation of a thick callose layer on the stigma surface (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003ea, b). The callose deposition could be seen within the 2HAP. In the incompatible cross-combinations of \u003cem\u003eLu\u003c/em\u003e X \u003cem\u003eLg\u003c/em\u003e, and its reciprocal, larger proportions of pollen grains lying ungerminated on the foreign stigmatic surface could be observed even longer hours after (10 to 24HAP) pollen landing (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003ec, d and e) that started showing up some form of degradation. The main obstacles for PG included the callose layer forming within 2HAP, which limited the germination of foreign pollen grains on stigmatic surfaces, taking 4 to 6 hours (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003ef, g). All flowering plants rely on the crucial process of sexual reproduction. This essential process is initiated when a pollen grain reaches the stigma of a pistil and gives rise to a vital pollen tube\u003csup\u003e\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e\u003c/sup\u003e. The journey to hybridization between different species within the same genus may encounter challenges, but groundbreaking researchers such as Dumas and Knox (1983)\u003csup\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e and Chen and Kim (2009)\u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e have championed the use of aniline blue to unravel the callose response, shedding light on the phenomena of rejection. Researchers have stressed the importance of utilizing aniline blue to track callose response, associated with pollen rejection. The Aniline Blue Fluorescent (ABF) method and fluorescence microscopy have been employed to extensively investigate PPI in a variety of crops such as sorghum\u003csup\u003e\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e, sesame\u003csup\u003e\u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e67\u003c/span\u003e\u003c/sup\u003e. However, studies for analyzing unsuccessful seed set because of pre-fertilization barrier due to incompatible PPI in \u003cem\u003eLinum\u003c/em\u003e interspecific crosses is scarce.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eCallose is a crucial component of the cell wall, being dynamically deposited and degraded throughout pollen development\u003csup\u003e\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e\u003c/sup\u003e. In many angiosperms, a brief callose-rich cell wall forms around each microsporocyte in the anthers, while callose is also present on the outer pollen wall. Callose effectively separates microspores and acts as an impermeable barrier\u003csup\u003e\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e\u003c/sup\u003e. In species with callose-rich walls, mutants lacking callose production consistently produce inviable pollen\u003csup\u003e\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e\u003c/sup\u003e. Visible ungerminated pollen grains on the stigmatic surfaces in case of incompatible interspecific hybridization after longer hours of pollen landing may be explained due to deposition of the callose layer as an attempt to retain longer pollen viability (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003ea-e).\u003c/p\u003e \u003cp\u003eCallose provides a useful phenotypic bioassay in plant breeding to determine incompatibility system, pollen competition, stigma, and ovule viability and to understand the pathogen induced defense mechanism triggered in the plant systems\u003csup\u003e\u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e74\u003c/span\u003e,\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e. The role of callose in pollen-stigma interactions has many analogies with host-parasite interactions as callose deposition is viewed as an indicator of biocommunication between pollen and stigma during PPI\u003csup\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e. This pathogen-induced callose deposition functions as a chemical and physical defense mechanism for reinforcing plant cell wall and plays an essential role in the defense response to invading pathogens\u003csup\u003e\u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e74\u003c/span\u003e\u003c/sup\u003e. In \u003cem\u003eLinum\u003c/em\u003e, it is assumed that callose deposition in response to non-self-pollen grains serves as a formidable barrier to pollen germination and pollen tube growth in incompatible distant crosses under the current study. The response of stigma callose is distinctly influenced by informational molecules from self or interspecific pollen grains. Dumas and Knox's (1983)\u003csup\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e proposed model establishes a compelling connection between callose, boron, and inhibitor synthesis akin to phytoalexins.\u003c/p\u003e \u003cp\u003eIn the model plant \u003cem\u003eArabidopsis thaliana\u003c/em\u003e two independent research groups identified twelve genes encoding putative callose synthase namely CalS (callose synthase) and GSL (glucan synthase-like) genes\u003csup\u003e\u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e69\u003c/span\u003e\u003c/sup\u003e. Callose wall plays an important role in normal pollen mother cell development. Recently, through genetic approach it is revealed that \u003cem\u003eOsGSL5\u003c/em\u003e gene is responsible for callose deposition in anther locules and \u003cem\u003eOsGSL5\u003c/em\u003e gene mutation resulted in anthers with less callose deposition, aberrant pollen mother cells and abnormal microspores\u003csup\u003e\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e\u003c/sup\u003e. Global transcriptome analysis showed that expression of \u003cem\u003eOsGSL5\u003c/em\u003e was downregulated in the \u003cem\u003eOsspl\u003c/em\u003e (\u003cem\u003eOsSPOROCYTELESS\u003c/em\u003e) mutant which was defective in meiosis-specific callose deposition\u003csup\u003e\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e,\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e\u003c/sup\u003e. Similarly, we posit that a mutated or malfunctioning expression of genes responsible for synthesizing callose on the stigmatic surface when encountering non-self-pollen grains may present an opportunity to overcome pre-fertilization barriers arising from callose deposition. It is recommended to conduct further studies to explore the suppression of such genes and its potential impact on enabling the overcoming of callose-related barriers.\u003c/p\u003e\n\u003ch3\u003ePollen tube growth within the pistil\u003c/h3\u003e\n\u003cp\u003eIrrespective of self-or cross-pollination, in all the studied three \u003cem\u003eLinum\u003c/em\u003e species, and their wide cross-combinations, the germinated pollen grains were able to grow and become visible within the various stylar regions of the pistil parent in different time intervals. The number of growing PTs kept reducing in all the cases of self and cross pollination while growing towards the ovular region from stylar region (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003eb, c and d). Just like self-pollinated pistils, wide crosses with \u003cem\u003eL. bienne\u003c/em\u003e, were able to be seen growing PTs within various stylar regions in foreign pistils and seemed navigating ovular region within 2HAP (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003em-p). However, the pollen grains that could successfully cross the germination barriers due to callose deposition on the stigmatic surface in \u003cem\u003eLu\u003c/em\u003e X \u003cem\u003eLg\u003c/em\u003e and its reciprocal, which were seen growing their PTs in the stylar regions of pistil parent within 2HAP were unable to reach up to the ovular region of foreign pistil parent. Appearance of PTs growing in the ovular region for distant crosses with \u003cem\u003eL. grandiflorum\u003c/em\u003e could be noticed only during 4HAP irrespective of pistil parent (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003ed). The delayed growth of PT in the foreign style was explained due to predominant resistance force due to a high intensity of PTs in the stigmatic surface\u003csup\u003e\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e\u003c/sup\u003e. Unlike self-pollinated pistils and, crosses with its wild progenitor \u003cem\u003ei.e\u003c/em\u003e., \u003cem\u003eL. bienne\u003c/em\u003e, various kinds of aberrations were observed in the developing PTs, imposing barriers in the path of further growth in wide crosses with \u003cem\u003eL. grandiflorum\u003c/em\u003e (Figs.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e and \u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe major PTG inhibition occurred in the stylar region was, due to appearance of emerging deformed growth patterns in the developing tubes. These discordant growth patterns may be attributed to cause delayed arrival of PTs to ovular region of foreign pistil in wide crosses with \u003cem\u003eL. grandiflorum\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003e). According to Hodnett et al. (2005)\u003csup\u003e\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e\u003c/sup\u003e, unfavorable conditions hindered the normal metabolism of the PT, leading to its malformation and subsequent reduction in growth towards the micropyle. Comprehensive analysis of PTG in the interspecific and intergeneric crosses as an indicator of pre-zygotic barrier is not just limited to field crops but have been extensively implied in horticultural crops too\u003csup\u003e\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e,\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eMost of the nonconductive circumstances started appearing during the 2-4HAP (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003ea-g). Partially elongated PTs along with the non-germinated pollen grains could still be seen in the upper stylar regions of \u003cem\u003eL. usitatissimum\u003c/em\u003e during this time duration (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003ea). Molecular aspects better explain the reasons behind the rejection of incompatible PTG. Some studies mention role of putative protein O-fucosyltransferase facilitating PT penetration through the stigma-style interface\u003csup\u003e\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e\u003c/sup\u003e. Stylar region comprises various chemical signals that reside in the extracellular matrix of transmitting tract play crucial roles in the recognition of self or non-self-PT. There are various substances existing in the extracellular matrix of the transmitting tract that reject non-self-PTs and the loss-of-function of the pistil-side barrier will result in self-incompatibility transition to self-compatibility\u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e. Various biomolecules involved in guiding the PT through the transmitting tract have been extensively elaborated and reviewed by Zheng et al. (2018)\u003csup\u003e\u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e76\u003c/span\u003e\u003c/sup\u003e and recently by Cheung et al. (2022)\u003csup\u003e\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eSwelling of PTs at the tip was traced through the florescent image analysis in the mid-stylar (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eb) and attachment regions of the ovary (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003ec) in the pistil. Non-penetration of the ovular region even at 96HAP was reported in the studies of Gong et al. (2023)\u003csup\u003e\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e, where they reported that PT barely entered the ovule, but remained at the base of the style and became swollen in another oilseed plant species \u003cem\u003eCamellia oleifera.\u003c/em\u003e The control of PTG by protein factors was addressed by Mei et al. (2022)\u003csup\u003e\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e\u003c/sup\u003e in model plant \u003cem\u003eArabidopsis thaliana.\u003c/em\u003e Twisted PTG was witnessed in the upper (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003ed) and mid-stylar (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003ee) regions along with severe inhibition of the growing PT in the ovary (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003ef, g). The appearance of PTs growing in reverse direction (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eh) while reaching towards the ovule, complex and convoluted PTG at the mid-stylar region (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003ei) were noticed during 6HAP. Partially germinated pollen grains with terminated PTG (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003ej) were still visible even after longer hours of pollination (10HAP). Thus, the initial 4 hours were proved critical and crucial from the PG and PTG point of view. In the interspecific crosses, there were frequent observations of delayed growth of PTs, as well as other structural abnormalities such as twisting, swelling, high branching, a bi-furcated tip, PTG before reaching the ovary and inability to navigate to micropyle and variations in callose form. These variations included reverse orientation and irregularity in callose plugs along the PT. Such abnormalities attributed for delayed pollination and delayed pollen tube growth resulting into incompatible interaction\u003csup\u003e\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eIn case of distant crosses made with \u003cem\u003eL. grandiflorum\u003c/em\u003e as pistil parent, almost similar abnormality patterns were observed during PTG of \u003cem\u003eL. usitatissimum\u003c/em\u003e. Within 2HAP, the pre-fertilization barriers started appearing in the form of restricted and twisted PTG in the upper stylar region (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003ea). As soon as the PT arrives at the attachment region in of ovary at the lower stylar region, some PT faced twisted growth patterns (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eb). Under-germinated pollen grains with terminated, twisted and, swollen PT was most frequent during the 4HAP (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003ec, d). Non-germinated pollen grains were quite obviously visible during different HAP from 2 to 24HAP (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003ee). Large proportions of ungerminated pollen grains along with consistently growing PTs were noticed in the stylar region (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003ef). Reduced number of PTs passing through ovular regions affecting fertilization after longer hours of pollination taking up to 18 to 24 hours have been reported in wide crosses of \u003cem\u003eVigna\u003c/em\u003e spp.\u003csup\u003e\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e\u003c/sup\u003e. Ruptured and bifurcated PT tip in the stylar region could also be witnessed as soon as 4 hours after pollen landed and germinated on foreign pistil (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eg). Convoluted PTG (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eh), partial or delayed PTG (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003ei) with swollen tip (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003ej) were visible even after longer durations following pollination. Many pollen grains germinated with swollen PT tips and twisted growth patterns showing random growth patterns could be seen in the stylar region even 10-12HAP (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003em, n).\u003c/p\u003e \u003cp\u003eThe pre-fertilization barriers were predominant in wide crosses with \u003cem\u003eL. grandiflorum\u003c/em\u003e, that operated in all the stages resulting into delayed PTG. These barriers not only acted on the stigmatic surfaces but also had operated gradually and mildly at various growth stages. No PTs could be noticed penetrating ovular region in wide crosses with \u003cem\u003eL. grandiflorum\u003c/em\u003e till 4HAP as opposed in crosses attempted with \u003cem\u003eL. bienne\u003c/em\u003e, where the ovular penetration occurred within 2HAP (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003em-t). Low levels of PT penetration and PTG in the wide crosses due to operative pre-fertilization barriers have been advocated by Krishnasamy et al. (2008)\u003csup\u003e\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e\u003c/sup\u003e in \u003cem\u003eVigna\u003c/em\u003e spp., Ganesh Ram et al. (2006)\u003csup\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e in Sesame spp., Badiger et al. (2024)\u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e in \u003cem\u003eAbelmoschus\u003c/em\u003e species. Based on image analysis, the non-arrival of the PT within the first four HAP could be attributed to the various kinds of pre-fertilization barriers and deformities appearing during PTG. The first two to four hours seemed critical for the PT growth in wide crosses with \u003cem\u003eL. grandiflorum\u003c/em\u003e. Though, the PT was able to reach up to the attachment region of ovary at the lower stylar region of pistil within the first 2HAP, they remained unable to penetrate the ovular region until 4HAP.\u003c/p\u003e \u003cp\u003eInterspecific hybridization in \u003cem\u003eLinum\u003c/em\u003e has been advocated in the past and has been successfully used in both West European and East European cultivars. For example, \u003cem\u003eLinum crepitans\u003c/em\u003e L. was employed to introduce early maturity traits into the \u003cem\u003eL. usitatissimum\u003c/em\u003e L. However, the potential of these early maturing types was not fully realized in subsequent breeding efforts. While interspecies crossing generally exhibits low effectiveness, this can be improved through the application of embryo tissue culture and the cultivation of immature isolated embryos under \u003cem\u003ein vitro\u003c/em\u003e conditions\u003csup\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e,\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e\u003c/sup\u003e. In situations where natural interspecific hybridization in \u003cem\u003eLinum\u003c/em\u003e fails due to pollen-stigma incompatibility, the utilization of advanced techniques such as protoplast fusion and embryo rescue holds immense promise for obtaining successful interspecific hybrids. The ongoing research on protoplast culture and isolated immature embryos of distant hybridization of flax will undoubtedly yield valuable insights into the selection of optimal medium and culture conditions, paving the way for groundbreaking advancements in this field\u003csup\u003e\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e\u003c/sup\u003e. In Wang's (2008)\u003csup\u003e\u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e73\u003c/span\u003e\u003c/sup\u003e study, interspecific hybridization between \u003cem\u003eL. usitatissimum\u003c/em\u003e and \u003cem\u003eL. perenne\u003c/em\u003e was conducted. The results demonstrated that repeat-pollination alongside the use of plant growth regulators (such as GA\u003csub\u003e3\u003c/sub\u003e, NAA, and 2,4-D) proved to be an effective method for overcoming interspecific cross-incompatibility in flax. Production of haploids followed by genotype stabilization in the case of interspecific hybridization for extension of genetic variability in \u003cem\u003eLinum\u003c/em\u003e is one of the unconventional methods for producing interspecific hybrids. For this purpose, the application of biochemical methods has been studied and suggested.\u003c/p\u003e"},{"header":"Material and Methods","content":"\u003cp\u003e \u003cb\u003eDetails of\u003c/b\u003e \u003cb\u003eLinum\u003c/b\u003e \u003cb\u003especies used in the study\u003c/b\u003e\u003c/p\u003e \u003cp\u003eLinseed accession IC268345 represented cultivated species (\u003cem\u003eL. usitatissimum\u003c/em\u003e L., 2n\u0026thinsp;=\u0026thinsp;30) of \u003cem\u003eLinum\u003c/em\u003e for this investigation. The wild species used under the current study involved its wild progenitor, \u003cem\u003eL. bienne\u003c/em\u003e M. (2n\u0026thinsp;=\u0026thinsp;30) (accession EC993391) and the other distant CWR \u003cem\u003eL. grandiflorum\u003c/em\u003e L. (2n\u0026thinsp;=\u0026thinsp;16). Since, \u003cem\u003eL. grandiflorum\u003c/em\u003e is distylic but unlike most distylic plants the level of the anthers is not greatly different in the two types\u003csup\u003e\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e,\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e,\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e, we used a mixed population of originally procured \u003cem\u003eL. grandiflorum\u003c/em\u003e (IC633096) to ensure the legitimate PG and subsequent PTG in the pistils of manually pollinated flower buds. Details of the plant materials (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e1\u003c/span\u003e) used and its characteristic features are listed in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eField experiment and hand emasculation\u003c/h2\u003e \u003cp\u003eThe field experiment was conducted at Indian Council of Agricultural Research - National Bureau of Plant Genetic Resources, experimental farm, IARI, New Delhi (28\u0026deg; 38\u0026prime; 53.7\u0026prime;\u0026prime; N, 77\u0026deg; 09\u0026prime; 05.4\u0026prime;\u0026prime; E and 218 m above mean sea level) in paired row of 2m length and row spacing of 90cm during cropping season from November to April, 2021-22. Self and cross pollination were attempted manually during the season using cultivated species IC268345, wild species EC993391 (\u003cem\u003eL. bienne\u003c/em\u003e M.) and IC633096 (\u003cem\u003eL. grandiflorum\u003c/em\u003e L.) with reciprocal cross combinations as represented in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e2\u003c/span\u003e. To assure the continuous availability of flowers, all the three species were staggered planted on three sowing dates, after 3\u0026ndash;5 days of intervals. Artificial pollination was carried out following the method suggested by Bolling et al. (1961)\u003csup\u003e\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e with slight modifications. A day before pollination, the un-opened flower buds with exposed petals were hand emasculated during the evening hours, between 5\u0026ndash;7 PM. For emasculation, all the five anthers were exposed for removal using forceps. The petals were left intact in the buds to allow anthesis next morning. This helped exposure of stigmas naturally avoiding any damage to the stigmas while making efforts to separate the sepals covering stigmas next morning. This slight modification also helped transferring the pollens grains on the surface of naturally exposed stigma while performing pollination manually, that otherwise needed extra efforts to keep the sepals stay apart from each other while pollen transfer. After all the five anthers were removed, the pistils were closed back with petals and bagged with waxy pollination bags to avoid drying up of stigmas. Bagging of donor flower buds was also done the previous evening to obtain fresh non-contaminated pollen grains to hybridize the emasculated flower buds the next morning to avoid cross-contamination. The next morning between 7\u0026ndash;9 AM, the pollen was transferred using freshly opened bagged flower by directly placing the pollen grains onto the exposed stigma of emasculated flower by contact method. Transfer of a sufficient number of pollen grains was ensured. The pollinated flowers were tagged with all the relevant information and bagged immediately to avoid cross contamination. Self-pollination was done in the same manner for all the three species.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003ePollen vitality test\u003c/h2\u003e \u003cp\u003eThe vitality of pollen grains in \u003cem\u003eL. usitatissimum\u003c/em\u003e, \u003cem\u003eL. bienne\u003c/em\u003e, and \u003cem\u003eL. grandiflorum\u003c/em\u003e was assessed using a Leica light microscope. To begin, mature flower buds were carefully collected and kept in an ice container. Subsequently, the anthers were delicately excised, placed on slides, and gently crushed in a 1% (w/v) acetocarmine solution with the aid of forceps and a needle. After removing any debris, a coverslip was carefully positioned on the slide before observation under the microscope. The captured images were then utilized to quantify and evaluate the stained and unstained pollen grains. During assessment, nonviable pollen, showing signs of shriveling or remaining unstained, was distinguished from viable, round, plump, and stained pollen (Fig. \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). Each of the species under scrutiny was thoroughly examined using four to six slides, each divided into 4 to 8 nonoverlapping sections (Table\u0026nbsp;3).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eStandardization of protocol for the study of pollen-pistil interaction in wide crosses of\u003c/b\u003e \u003cb\u003eLinum\u003c/b\u003e\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eSample collection and storage\u003c/h2\u003e \u003cp\u003eThe pistils of the artificially selfed and cross-pollinated flower buds were collected without separating them from petals and sepals after 2, 4, 6, 8, 10, 24 and 48 Hours after pollination (HAP) followed by fixation in acetic acid: alcohol (1:3 v/v) fixative in amber colored 5ml glass vials and stored at 4\u0026deg;C until further analysis. For every collection time, self and cross-pollinated flower buds were used for microscopic observations. For each self and cross combination 15 artificially pollinated flower buds were collected. Thus, for all the seven cross combinations of each duration, a total of 735 flower buds were collected and studied further.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eSample processing\u003c/h2\u003e \u003cp\u003eTo study PPI, aniline blue fluorescence method\u003csup\u003e\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e\u003c/sup\u003e was modified slightly for \u003cem\u003eLinum\u003c/em\u003e for softening period by altering sodium hydroxide (NaOH) concentration. This helped to reduce the time duration of softening treatment for dissected pistils thereby, increasing the efficiency of sample handling with reduced time duration and minimal damage to the fixed pistils. Instead of direct fixation of pistils over the fixed flower buds, was proved advantageous in terms of keeping the fibrous \u003cem\u003eLinum\u003c/em\u003e pistils retaining their integrity while sample processing without breaking the fragile tissues. The fixed pistils along with the intact flower buds were washed 2 to 3 times with distilled water for 5 minutes each followed by dissection. The pistils were dissected out from the flower buds with the help of pointed forceps and needles available in biological experiments dissection box to separate out the attached stigmatic regions. Unlike, Parani, 1998\u003csup\u003e51\u003c/sup\u003e, where higher concentration of softening agent (8N NaOH) was used for longer duration (8h), we reduced the concentration to 6N NaOH with 1h time duration to clear and soften the tissue at room temperature. The softened pistils were kept undisturbed in the distilled water for 30 min followed by mild washing to avoid any damage to the softened tissue and to remove all traces of softening agent. This enhanced the absorption of fluorescent dye in the target tissue. The softened tissue was then subjected to staining buffer overnight under dark conditions for further analysis through fluorescent microscopy.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eFluorescence microscopy\u003c/h2\u003e \u003cp\u003eFor preparation of staining buffer, 0.1M dibasic potassium phosphate (K\u003csub\u003e2\u003c/sub\u003eHPO\u003csub\u003e4\u003c/sub\u003e) mixed with 0.1% water-soluble aniline blue dye (WS Color Index 42780) was used to stain the processed pistils. Staining with 0.1% water-soluble aniline blue prepared in 0.1M dibasic potassium phosphate (K\u003csub\u003e2\u003c/sub\u003eHPO\u003csub\u003e4\u003c/sub\u003e) overnight, gave good results in terms of dye absorption by the target tissue. The stained pistil was then placed on glass slide without glycerol treatment to avoid intertwining of fibrous pistils. Pistils were covered with cover slip and pressed gently into a thin layer. The slides were placed in dark till further observation. The image was captured under dark room condition and freely definable black balance available with the Leica DM6 B Upright Fluorescent Microscope using 390\u0026ndash;420 nm barrier filter coupled with a 450 nm excitation filter. Pollen germination and tube growth image thus captured with a Leica DFC7000 T camera installed with the microscope was used to process the image with Lieca-X Software for taking various parameters such as pollen germination and determination of PTG in different time intervals.\u003c/p\u003e \u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe present study revealed that the interaction between the pollen and pistil is inhibited in the distant crosses among \u003cem\u003eLu\u003c/em\u003e X \u003cem\u003eLg\u003c/em\u003e due to both temporal and spatial pre-fertilization barriers. Callose deposition at the stigmatic surface of foreign pistils within 2HAP leading to inability of pollen grains to penetrate the stylar tissue appeared as the major barrier. Various kinds of aberrations started appearing during the 2-4HAP, in the developing PTs imposing barriers in further growth. Delayed PTG was observed mainly due to the discordant conditions appearing in the various stylar regions. The complexity of interspecific hybridization was observed in terms of arrest of pollen tube (PT) growth in the ovary, ruptured, twisted and swollen pollen tube tip, tube growth in reverse direction, convoluted and terminated growth patterns. Although interspecies crossover is often less efficient, it can be made more effective by using embryo rescue and cultivating isolated, immature embryos \u003cem\u003ein vitro\u003c/em\u003e. We propose that the disruption of genes responsible for producing callose on the stigmatic surface when encountering non-self-pollen grains could offer a promising opportunity to overcome pre-fertilization barriers associated with callose deposition. Further studies are recommended to investigate the suppression of these genes and the potential impact on surmounting callose-related barriers. When pollen-stigma incompatibility prevents natural interspecific hybridization in \u003cem\u003eLinum\u003c/em\u003e from occurring, the application of cutting-edge methods like protoplast fusion and embryo rescue presents great potential for producing interspecific hybrids that succeed. The advanced studies on isolated immature flax embryos from distant hybridization and protoplast culture will surely provide important information on choosing the best culture medium and circumstances, opening the door to ground-breaking discoveries in this area.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eV.K.R: Experimentation and writing\u0026mdash;original draft, Data collection; A.Y. \u0026amp; M.R.: Supervision; V.K. \u0026amp; D.W.: Investigation, review, and editing; P.P.: Experimentation; M.S.: Conceptualization, supervision, writing\u0026mdash;original draft; funding acquisition; G.P.S.: funding acquisition; project administration.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eAuthors acknowledge the funding support for the project (No. BT/Ag/Network/Linseed/2019-20) from the Department of Biotechnology (DBT), Government of India. The authors also acknowledge the funding and research facilities available at the Indian Council of Agricultural Research-National Bureau of Plant Genetic Resources (ICAR-NBPGR), New Delhi. The authors thank the Head, Division of Germplasm Evaluation, NBPGR for the research facilities and guidance to carry out field experiments. The authors are thankful to Dr. B.L. Meena (ACTO \u0026amp; Farm manager, NBPGR farm) for assistance in field experiments. The authors are also thankful to Mr. Raghvendra Pratap Singh, Mr. Santosh, and other project staff for assistance in field and laboratory experiments.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eAll the data have been provided in the main manuscript and supplementary files.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAllaby, R. G., Peterson, G. W., Merriwether, D. A. \u0026amp; Fu, Y. B. Evidence of the domestication history of flax (\u003cem\u003eLinum usitatissimum\u003c/em\u003e L.) from genetic diversity of the sad2 locus. \u003cem\u003eTheoretical and applied genetics.\u003c/em\u003e\u003cstrong\u003e112\u003c/strong\u003e, 58-65. doi: 10.1007/s00122-005-0103-3 (2005).\u003c/li\u003e\n\u003cli\u003eBadiger, M., Yadav, R. K. \u0026amp; Sharma, B. B\u003cem\u003e.\u003c/em\u003e Pollen germination, pollen\u0026ndash;pistil interaction and crossability studies in interspecific and induced colchiploid population of \u003cem\u003eAbelmoschus\u003c/em\u003e species. \u003cem\u003eGenet Resour Crop Evol\u003c/em\u003e. \u003cstrong\u003e71\u003c/strong\u003e, 107-127. https://doi.org/10.1007/s10722-023-01610-y (2024).\u003c/li\u003e\n\u003cli\u003eBedinger, P. A., Broz, A. K., Tovar-Mendez, A. \u0026amp; McClure, B. Pollen-pistil interactions and their role in mate selection. \u003cem\u003ePlant Physiol\u003c/em\u003e. \u003cstrong\u003e173\u003c/strong\u003e, 79-90. doi: 10.1104/pp.16.01286 (2017).\u003c/li\u003e\n\u003cli\u003eBolling, M., Sander, D. A. \u0026amp; Matlock, R. S. Mungbean hybridization technique. \u003cem\u003eAgron. J\u003c/em\u003e. \u003cstrong\u003e53\u003c/strong\u003e, 54-55 (1961).\u003c/li\u003e\n\u003cli\u003eChen, X. Y. \u0026amp; Kim, J. Y. Callose synthesis in higher plants: Mini-review. \u003cem\u003ePlant Signaling and Behavior\u003c/em\u003e. \u003cstrong\u003e4\u003c/strong\u003e, 489-492 (2009).\u003c/li\u003e\n\u003cli\u003eCheng, L. L., Guan, F. Z., Wu, G. W., Yu, Y., Huang, W. G. \u0026amp; Zhao, D. S. Interspecific hybridization and immature embryo rescure between \u003cem\u003eLinum usitatissimum\u003c/em\u003e and \u003cem\u003eLinum grandiflorum\u003c/em\u003e. \u003cem\u003ePlant Fiber Sci\u003c/em\u003e. \u003cstrong\u003e37\u003c/strong\u003e, 1-4. doi: 10.3969/j.issn.1671-3532.2015.01.001 (2015).\u003c/li\u003e\n\u003cli\u003eCheung, A. Y., Duan, Q., Li, C., James Liu, M. C. \u0026amp; Wu, H. M. Pollen-pistil interactions: It takes two to tangle but a molecular cast of many to deliver. \u003cem\u003eCurr. Opin. Plant Biol\u003c/em\u003e. \u003cstrong\u003e69\u003c/strong\u003e, 102279. https://doi.org/10.1016/j.pbi.2022.102279 (2022).\u003c/li\u003e\n\u003cli\u003eChung, T. D. \u0026amp; Plonka, F. Differential germination in flax pollination. \u003cem\u003eAgronomie, EDP Sciences\u003c/em\u003e. \u003cstrong\u003e6(4)\u003c/strong\u003e, 379-386 (1986).\u003c/li\u003e\n\u003cli\u003eCisneros, L. M. E. \u003cem\u003eet al\u003c/em\u003e. Pollen\u0026ndash;pistil interaction, pistil histology and seed production in A\u0026times;B grain sorghum crosses under chilling field temperatures. \u003cem\u003eThe Journal of Agricultural Science\u003c/em\u003e. \u003cstrong\u003e148\u003c/strong\u003e, 73-82. 10.1017/S0021859609990396 (2010).\u003c/li\u003e\n\u003cli\u003eDarlington, C. D. \u0026amp; Wylie, A. P. Chromosome atlas of flowering plants. George Allen and Unwin, London (1955).\u003c/li\u003e\n\u003cli\u003eDickinson, H. G. \u0026amp; Lewis, D. Changes in the Pollen Grain Wall of \u003cem\u003eLinum grandiflorum\u003c/em\u003e following Compatible and Incompatible Intraspecific Pollinations. \u003cem\u003eAnn. Bot\u003c/em\u003e. \u003cstrong\u003e38\u003c/strong\u003e, 23-9 (1974).\u003c/li\u003e\n\u003cli\u003eDiederichsen, A. \u0026amp; Hammer, K.. Variation of cultivated flax (\u003cem\u003eLinum usitatissimum\u003c/em\u003e L. subsp. usitatissimum) and its wild progenitor pale flax (subsp. angustifolium (Huds.) Thell.). \u003cem\u003eGenet. Resour. Crop Evolut\u003c/em\u003e. \u003cstrong\u003e42\u003c/strong\u003e, 263-272. https://doi.org/10.1007/BF02431261 (1995).\u003c/li\u003e\n\u003cli\u003eDiederichsen, A. \u0026amp; Richards, K. Cultivated flax and the genus \u003cem\u003eLinum \u003c/em\u003eL. Taxonomy and germplasm conservation. In: Muir, A. D. and Westcott, N. D. (Eds). Flax\u003cem\u003e: \u003c/em\u003eThe genus \u003cem\u003eLinum. \u003c/em\u003eCRC Press, London, 22-54 (2003).\u003c/li\u003e\n\u003cli\u003eDoebley, J. F., Brandon, S. G. \u0026amp; Bruce, D. S. The molecular genetics of crop domestication. \u003cem\u003eCell\u003c/em\u003e. \u003cstrong\u003e127(7)\u003c/strong\u003e, 1309-1321. https://doi.org/10.1016/j.cell.2006.12.006 (2006).\u003c/li\u003e\n\u003cli\u003eDuigou, A. \u0026amp; Castro, M. Evaluation of force generation mechanisms in natural, passive hydraulic actuators. \u003cem\u003eScientific Reports\u003c/em\u003e. \u003cstrong\u003e6\u003c/strong\u003e, 18105. 10.1038/srep18105 (2016).\u003c/li\u003e\n\u003cli\u003eDuk, M. \u003cem\u003eet al\u003c/em\u003e. The Genetic Landscape of Fiber Flax. \u003cem\u003eFrontiers in Plant Science\u003c/em\u003e. \u003cstrong\u003e12\u003c/strong\u003e, 764612. 10.3389/fpls.2021.764612 (2021).\u003c/li\u003e\n\u003cli\u003eDumas, C. \u0026amp; Knox, R. B. Callose and determination of pistil viability and incompatibility. \u003cem\u003eTheoretical and Applied Genetics\u003c/em\u003e. \u003cstrong\u003e67\u003c/strong\u003e, 1-10 (1983).\u003c/li\u003e\n\u003cli\u003eFAO, Statistics Division, Food and Agriculture Organization of the United Nations, Viale delle Terme di Caracalla, 00153 Rome, Italy. http://www.fao.org/faostat/en/#home (2018).\u003c/li\u003e\n\u003cli\u003eFayaz, Z. \u003cem\u003eet al\u003c/em\u003e. Parthenocarpy: \u0026lsquo;\u0026lsquo;A Potential Trait to Exploit in Vegetable Crops\u0026rsquo;\u0026rsquo;. \u003cem\u003eEnvironment and Ecology\u003c/em\u003e. \u003cstrong\u003e39(4A)\u003c/strong\u003e, 1332-1346 (2021).\u003c/li\u003e\n\u003cli\u003eFeller, A. F., Burgin, G., Lewis, N., Prabhu, R. \u0026amp; Hopkins, R. Mismatch between pollen and pistil size causes asymmetric mechanical reproductive isolation across Phlox species. \u003cem\u003ebioRxiv\u003c/em\u003e. 593106. doi: 10.1101/2024.05.08.593106 (2024).\u003c/li\u003e\n\u003cli\u003eFlint-Garcia, S. A. Genetics and consequences of crop domestication. \u003cem\u003eJ. Agric. Food Chem\u003c/em\u003e. \u003cstrong\u003e61(35)\u003c/strong\u003e, 8267-76. doi: 10.1021/jf305511d (2013).\u003c/li\u003e\n\u003cli\u003eFu, Y. B. Pale Flax (\u003cem\u003eLinum Bienne\u003c/em\u003e): an Underexplored Flax Wild Relative. In: You, F.M., Fofana, B. (eds) The Flax Genome. Compendium of Plant Genomes. \u003cem\u003eSpringer, Cham\u003c/em\u003e. https://doi.org/10.1007/978-3-031-16061-5_3 (2023).\u003c/li\u003e\n\u003cli\u003eGanesh Ram, S., Sundaravelpandian, K., Kumar, M., Vinod, K. K., KannanBapu, J. R. \u0026amp; Raveendran, T. S. Pollen pistil interaction in the inter-specific crosses of \u003cem\u003eSesamum sp\u003c/em\u003e. \u003cem\u003eEuphytica\u003c/em\u003e. \u003cstrong\u003e152\u003c/strong\u003e, 379-385 (2006).\u003c/li\u003e\n\u003cli\u003eGhosh, S. \u0026amp; Shivanna, K. R. Pollen-pistil interaction in \u003cem\u003eLinum grandiflorum\u003c/em\u003e: Scanning electron microscopic observations and proteins of the stigma surface. \u003cem\u003ePlanta\u003c/em\u003e. \u003cstrong\u003e149(3)\u003c/strong\u003e, 257-61. doi: 10.1007/BF00384562 (1980).\u003c/li\u003e\n\u003cli\u003eGill, K. S. Objectives, breeding approaches and achievements in linseed (\u003cem\u003eLinum usitatissimum\u003c/em\u003e). In: Gill, K.S. (Ed.), Breeding Oilseed Crops. \u003cem\u003eSpringer, Dordrecht\u003c/em\u003e, pp. 212e225 (1980).\u003c/li\u003e\n\u003cli\u003eGill, K. S. Linseed. Indian Council of Agricultural Research, New Delhi, India (1987).\u003c/li\u003e\n\u003cli\u003eGong, H., Chang, Y., Xu, J., Yu, X. \u0026amp; Gong, W. Unilateral cross-incompatibility between \u003cem\u003eCamellia oleifera\u003c/em\u003e and \u003cem\u003eC. yuhsienensis\u003c/em\u003e provides new insights for hybridization in \u003cem\u003eCamellia\u003c/em\u003e spp. \u003cem\u003eFront Plant Sci\u003c/em\u003e. \u003cstrong\u003e14\u003c/strong\u003e, 1182745. doi: 10.3389/fpls.2023.1182745 (2023).\u003c/li\u003e\n\u003cli\u003eHeer, O. Prehistoric culture of flax. \u003cem\u003eNature\u003c/em\u003e. \u003cstrong\u003e7\u003c/strong\u003e, 453. https://doi.org/10.1038/007453a0 (1873).\u003c/li\u003e\n\u003cli\u003eHelback, H. Domestication of Food Plants in the Old World: Joint efforts by botanists and archeologists illuminate the obscure history of plant domestication. \u003cem\u003eScience\u003c/em\u003e. \u003cstrong\u003e14\u003c/strong\u003e, \u003cstrong\u003e130\u003c/strong\u003e: 365-372. doi: 10.1126/science.130.3372.365 (1959).\u003c/li\u003e\n\u003cli\u003eHerbig, C. \u0026amp; Maier, U. Flax for oil or fibre? Morphometric analysis of flax seeds and new aspects of flax cultivation in Late Neolithic wetland settlements in southwest Germany. \u003cem\u003eVegetation History and Archaeobotany\u003c/em\u003e. \u003cstrong\u003e20\u003c/strong\u003e, 527-533. 10.1007/s00334-011-0289-z (2011).\u003c/li\u003e\n\u003cli\u003eHodnett, G. L., Burson, B. L., Rooney, W. L., Dillon, S. L. \u0026amp; Price, H. J. Pollen-pistil interactions result in reproductive isolation between \u003cem\u003eSorghum bico\u003c/em\u003elor and divergent Sorghum species. \u003cem\u003eCrop. Sci\u003c/em\u003e.\u003cem\u003e \u003c/em\u003e\u003cstrong\u003e45\u003c/strong\u003e, 1403-1409 (2005).\u003c/li\u003e\n\u003cli\u003eJhala, A. J., Hall, L. M. \u0026amp; Hall, J. C. Potential hybridization of flax with wild and weedy species: An avenue for movement of engineered genes? \u003cem\u003eCrop Science\u003c/em\u003e. \u003cstrong\u003e48(2)\u003c/strong\u003e, 825-840 (2008).\u003c/li\u003e\n\u003cli\u003eKaur, V. \u003cem\u003eet al\u003c/em\u003e. Multi-environment phenotyping of linseed (\u003cem\u003eLinum usitatissimum\u003c/em\u003e L.) germplasm for morphological and seed quality traits to assemble a core collection. \u003cem\u003eIndustrial Crops and Products\u003c/em\u003e. \u003cstrong\u003e206\u003c/strong\u003e, 117657. https://doi.org/10.1016/j.indcrop.2023.117657 (2023).\u003c/li\u003e\n\u003cli\u003eKrishnasamy, T., Palaniappan, J. \u0026amp; Nagasamy, N. Pollen pistil interaction in interspecific crosses of \u003cem\u003eVigna spp\u003c/em\u003e. \u003cem\u003eCytologia\u003c/em\u003e. \u003cstrong\u003e73\u003c/strong\u003e, 251-257 (2008).\u003c/li\u003e\n\u003cli\u003eKvavadze, E. \u003cem\u003eet al\u003c/em\u003e. 30,000-Year-Old Wild Flax Fibers. \u003cem\u003eScience (New York, N.Y.)\u003c/em\u003e. \u003cstrong\u003e325\u003c/strong\u003e, 1359. 10.1126/science.1175404 (2009).\u003c/li\u003e\n\u003cli\u003eLee, H. K., Macgregor, S. \u0026amp; Goring, D. R. A Toolkit for Teasing Apart the Early Stages of Pollen\u0026ndash;Stigma Interactions in \u003cem\u003eArabidopsis thaliana\u003c/em\u003e. In: Geitmann, A. (eds) Pollen and Pollen Tube Biology. \u003cem\u003eMethods in Molecular Biology\u003c/em\u003e. 2160. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0672-8_2 (2020).\u003c/li\u003e\n\u003cli\u003eLewis, D. The physiology of incompatibility in plants. 2. \u003cem\u003eLinum grandiflorum\u003c/em\u003e. \u003cem\u003eAnnals of Botany\u003c/em\u003e. \u003cstrong\u003e7\u003c/strong\u003e, 115-122 (1943).\u003c/li\u003e\n\u003cli\u003eLewis, D. Genetic control of specificity and activity of the S antigen implants. \u003cem\u003eProe. R. Soc., Ser. B\u003c/em\u003e. \u003cstrong\u003e151\u003c/strong\u003e, 468-477 (1960).\u003c/li\u003e\n\u003cli\u003eLi, N., Lin, Z., Yu, P., Zeng, Y., Du, S. \u0026amp; Huang, L. J. The multifarious role of callose and callose synthase in plant development and environment interactions. \u003cem\u003eFront. Plant Sci\u003c/em\u003e. \u003cstrong\u003e14\u003c/strong\u003e, 1183402. doi: 10.3389/fpls.2023.1183402 (2023).\u003c/li\u003e\n\u003cli\u003eLi, Y. \u003cem\u003eet al\u003c/em\u003e. Pollen-Specific Protein PSP231 Activates Callose Synthesis to Govern Male Gametogenesis and Pollen Germination. \u003cem\u003ePlant Physiology\u003c/em\u003e. \u003cstrong\u003e184(2)\u003c/strong\u003e, 1024-1041. https://doi.org/10.1104/pp.20.00297 (2020).\u003c/li\u003e\n\u003cli\u003eMahoney, J. D. \u0026amp; Brand, M. H. Pre and post zygotic barriers associated with intergeneric hybridization between \u003cem\u003eAronia melanocarpa\u003c/em\u003e (Michx.) Elliott \u0026times; \u003cem\u003ePyrus communis\u003c/em\u003e L. and \u0026times; \u003cem\u003eSorbaronia dippelii\u003c/em\u003e (Zabel) CK Schneid. \u0026times; \u003cem\u003ePyrus communis\u003c/em\u003e. \u003cem\u003eHortScience\u003c/em\u003e. \u003cstrong\u003e56\u003c/strong\u003e, 177-184 (2021).\u003c/li\u003e\n\u003cli\u003eMei, J. \u003cem\u003eet al\u003c/em\u003e. MAP3K\u0026epsilon;1/2 Interact with MOB1A/1B and Play Important Roles in Control of Pollen Germination through Crosstalk with JA Signaling in \u003cem\u003eArabidopsis\u003c/em\u003e. \u003cem\u003eInt. J. Mol. Sci\u003c/em\u003e.\u003cem\u003e \u003c/em\u003e\u003cstrong\u003e23\u003c/strong\u003e, 2683. doi: 10.3390/ijms23052683 (2022).\u003c/li\u003e\n\u003cli\u003eMelelli, A. \u003cem\u003eet al\u003c/em\u003e. Lessons on textile history and fibre durability from a 4,000-year-old Egyptian flax yarn. \u003cem\u003eNature Plants\u003c/em\u003e. \u003cstrong\u003e7\u003c/strong\u003e, 1-7. 10.1038/s41477-021-00998-8 (2021).\u003c/li\u003e\n\u003cli\u003eMohanty, A. K., Vivekanandhan, S., Pin, J. M. \u0026amp; Misra, M. Composites from renewable and sustainable resources: challenges and innovations. \u003cem\u003eScience\u003c/em\u003e. \u003cstrong\u003e36\u003c/strong\u003e, 536-542. 10.1126/science.aat9072 (2018).\u003c/li\u003e\n\u003cli\u003eMoreira, D., Kaur, D., Pereira, A. M., Held, M. A., Showalter, A. M. \u0026amp; Coimbra, S. Type II arabinogalactans initiated by hydroxyproline‐O‐galactosyltransferases play important roles in pollen\u0026ndash;pistil interactions. \u003cem\u003eThe Plant Journal\u003c/em\u003e. \u003cstrong\u003e114(2)\u003c/strong\u003e, 371-389. doi: 10.1111/tpj.16141 (2023).\u003c/li\u003e\n\u003cli\u003eMorimoto, T., Matsuda, Y., Sekiguchi, R. \u0026amp; Itai, A. Comprehensive Assessment of Intergeneric Cross-compatibility of Six Fruit Tree Species in the Tribe Maleae (\u003cem\u003eRosaceae\u003c/em\u003e) Based on In Vivo Pollen Tube Growth and Field Pollination. \u003cem\u003eThe Horticulture Journal\u003c/em\u003e. \u003cstrong\u003e92(1)\u003c/strong\u003e, 13-21. https://doi.org/10.2503/hortj.UTD-383 (2023).\u003c/li\u003e\n\u003cli\u003eMosquera, D. J. C., Salinas, D. G. C. \u0026amp; Moreno, G. A. L. Pollen viability and germination in \u003cem\u003eElaeis oleifera\u003c/em\u003e, \u003cem\u003eElaeis guineensis\u003c/em\u003e and their interspecific hybrid. \u003cem\u003ePesqui. Agropecu\u0026aacute;ria Trop\u003c/em\u003e. \u003cstrong\u003e51\u003c/strong\u003e, e68076. 10.1590/1983-40632021v5168076 (2021).\u003c/li\u003e\n\u003cli\u003eMurray, B. G. Floral Biology and Self-Incompatibility in \u003cem\u003eLinum\u003c/em\u003e. \u003cem\u003eBotanical Gazette\u003c/em\u003e. \u003cstrong\u003e147(3)\u003c/strong\u003e, 327-333. http://www.jstor.org/stable/2474401 (1986).\u003c/li\u003e\n\u003cli\u003eNair, B. \u003cem\u003eet al\u003c/em\u003e. Multi-environment screening of \u003cem\u003eLinum\u003c/em\u003e germplasm collection for dissecting the potential of bud fly (Dasyneura lini Barnes) resistance and assembling a reference set for efficient utilization in genetic improvement. \u003cem\u003eIndustrial Crops and Products\u003c/em\u003e. \u003cstrong\u003e207(1)\u003c/strong\u003e, 117743. https://doi.org/10.1016/j.indcrop.2023.117743 (2024).\u003c/li\u003e\n\u003cli\u003ePandiyan, M. \u003cem\u003eet al\u003c/em\u003e. Interspecific Hybridization in Direct and Reciprocal Crosses of \u003cem\u003eVigna radiata\u003c/em\u003e and \u003cem\u003eVigna aconitifolia\u003c/em\u003e. \u003cem\u003eAnn Agric Crop Sci\u003c/em\u003e. \u003cstrong\u003e5(2)\u003c/strong\u003e, 1062 (2020).\u003c/li\u003e\n\u003cli\u003eParani, M. Incompatibility in direct and reciprocal cross between \u003cem\u003eS. indicum\u003c/em\u003e L. and \u003cem\u003eS. alatum \u003c/em\u003e(Thonn). \u003cem\u003eJ. Indian Bot. Soc\u003c/em\u003e.\u003cem\u003e \u003c/em\u003e\u003cstrong\u003e77\u003c/strong\u003e, 9-12 (1998).\u003c/li\u003e\n\u003cli\u003ePascual, M., Ruiz, J., Posas, J. \u0026amp; Ni\u0026ntilde;o, M. Pollen viability and incompatibility in indigenous rice bean (\u003cem\u003eVigna umbellata\u003c/em\u003e (Thunb.) Ohwi \u0026amp; Ohashi). \u003cem\u003ePlant Science Today\u003c/em\u003e. \u003cstrong\u003e9(1)\u003c/strong\u003e, 157-166. 10.14719/pst.1375 (2022).\u003c/li\u003e\n\u003cli\u003ePatil, P. \u003cem\u003eet al\u003c/em\u003e. Pollen germination characteristics, pollen-pistil interaction and reproductive behaviour in interspecific crosses among \u003cem\u003eAbelmoschus esculentus\u003c/em\u003e Moench and its wild relatives. \u003cem\u003eGrana\u003c/em\u003e. \u003cstrong\u003e52\u003c/strong\u003e, 1-14. DOI: 10.1080/00173134.2013.768699 (2013).\u003c/li\u003e\n\u003cli\u003ePavelek, M., Vrbov\u0026aacute;-Prokopov\u0026aacute;, M., Ondr\u0026aacute;čkov\u0026aacute;, E., Ludv\u0026iacute;kov\u0026aacute;, M. \u0026amp; Griga, M. 17- Developments in fibrous flax and linseed breeding and cultivation, Editor(s): Ryszard M. Kozłowski, Maria Mackiewicz-Talarczyk, In: Woodhead Publishing Series in Textiles, Handbook of Natural Fibres (Second Edition), Woodhead Publishing, 605-692. https://doi.org/10.1016/B978-0-12-818398-4.00019-0 (2020).\u003c/li\u003e\n\u003cli\u003ePicarella, M. E. \u0026amp; Mazzucato, A. The Occurrence of Seedlessness in Higher Plants; Insights on Roles and Mechanisms of Parthenocarpy. \u003cem\u003eFront Plant Sci\u003c/em\u003e. \u003cstrong\u003e9\u003c/strong\u003e, 1997. doi: 10.3389/fpls.2018.01997 (2019).\u003c/li\u003e\n\u003cli\u003eQi, Y \u003cem\u003eet al\u003c/em\u003e. Phenotypic analysis of Longya-10 \u0026times; pale flax hybrid progeny and identification of candidate genes regulating prostrate/erect growth in flax plants. \u003cem\u003eFront. Plant Sci\u003c/em\u003e. \u003cstrong\u003e13\u003c/strong\u003e, 1044415. doi: 10.3389/fpls.2022.1044415 (2022).\u003c/li\u003e\n\u003cli\u003eRadice, S. \u0026amp; Arena, M. Characterization and evaluation of \u003cem\u003eBerberis microphylla\u003c/em\u003e G. Forst pollen grains. \u003cem\u003eAdvances in Horticultural Science\u003c/em\u003e. \u003cstrong\u003e30(1)\u003c/strong\u003e, 31-37. 10.13128/ahs-18699 (2016).\u003c/li\u003e\n\u003cli\u003eRavali, P., Kalaiyarasi, R., Hemavathy A. T. \u0026amp; Sudha M. Cytological investigation on pre and post fertilization barriers in interspecific cross of pigeonpea. \u003cem\u003eElectronic Journal of Plant Breeding\u003c/em\u003e. \u003cstrong\u003e13(2)\u003c/strong\u003e, 724-730. https://doi.org/10.37992/2022.1302.095 (2022).\u003c/li\u003e\n\u003cli\u003eRen, L. \u003cem\u003eet al\u003c/em\u003e. OsSPL regulates meiotic fate acquisition in rice. \u003cem\u003eNew Phytol\u003c/em\u003e. \u003cstrong\u003e218(2)\u003c/strong\u003e, 789-803. doi: 10.1111/nph.15017 (2018).\u003c/li\u003e\n\u003cli\u003eRobichaux, K. J. \u0026amp; Wallace, I. S. Signaling at Physical Barriers during Pollen-Pistil Interactions. \u003cem\u003eInt. J. Mol. Sci.\u003c/em\u003e\u003cstrong\u003e22\u003c/strong\u003e, 11230 (2021).\u003c/li\u003e\n\u003cli\u003eRosa, M. P. \u003cem\u003eet al\u003c/em\u003e. Tomato \u003cem\u003ePOLLEN DEFICIENT 2\u003c/em\u003e encodes a G-type lectin receptor kinase required for viable pollen grain formation. \u003cem\u003eJournal of Experimental Botany\u003c/em\u003e. \u003cstrong\u003e74(1)\u003c/strong\u003e, 178-193, https://doi.org/10.1093/jxb/erac419 (2023).\u003c/li\u003e\n\u003cli\u003eSeale, M. Callose Deposition during Pollen Development. \u003cem\u003ePlant Physiol\u003c/em\u003e. \u003cstrong\u003e184(2)\u003c/strong\u003e, 564-565. doi: 10.1104/pp.20.01143 (2020).\u003c/li\u003e\n\u003cli\u003eSeetharam, A. Interspecific Hybridization in \u003cem\u003eLinum. Euphytica\u003c/em\u003e. \u003cstrong\u003e21\u003c/strong\u003e, 489-495 (1972).\u003c/li\u003e\n\u003cli\u003eSmith, D. K.. \u003cem\u003eet al. \u003c/em\u003eA putative protein O-fucosyltransferase facilitates pollen tube penetration through the stigma-style interface. \u003cem\u003ePlant Physiol\u003c/em\u003e. \u003cstrong\u003e176\u003c/strong\u003e, 2804-2818. doi: 10.1104/pp.17.01577 (2018).\u003c/li\u003e\n\u003cli\u003eSomashekar, H., Mimura, M., Tsuda, K. \u0026amp; Nonomura, K. I. Rice GLUCAN SYNTHASE-LIKE5 promotes anther callose deposition to maintain meiosis initiation and progression. \u003cem\u003ePlant Physiol\u003c/em\u003e. \u003cstrong\u003e191\u003c/strong\u003e, 400-413. doi: 10.1093/plphys/kiac488 (2023).\u003c/li\u003e\n\u003cli\u003eSoto-Cerda, B. J., Urbina, S. H., Navarro, C. \u0026amp; Mora O. P. Characterization of novel genetic SSR markers in \u003cem\u003eLinum usitatissimum\u003c/em\u003e L. and their transferability across eleven \u003cem\u003eLinum\u003c/em\u003e species. \u003cem\u003eElectronic Journal of Biotechnology\u003c/em\u003e.\u003cstrong\u003e 14(2)\u003c/strong\u003e (2011).\u003c/li\u003e\n\u003cli\u003eSruthi, S. R., Kalaiyarasi, R. R., Sasikala, R. \u0026amp; Sudha, M. Pollen pistil interaction in the interspecific cross of \u003cem\u003eSesamum indicum\u003c/em\u003e and \u003cem\u003eS. radiatum\u003c/em\u003e. \u003cem\u003eElectronic Journal of Plant Breeding\u003c/em\u003e. \u003cstrong\u003e13(4)\u003c/strong\u003e, 1288-1296. 10.37992/2022.1304.162 (2023).\u003c/li\u003e\n\u003cli\u003eSuryawanshi, S. N., Wasnik, S. B., Makesar, A. D. \u0026amp; Walke, R. D. Economics of linseed marketing in Bhandara district. \u003cem\u003eJournal of Pharmacognosy and Phytochemistry\u003c/em\u003e. \u003cstrong\u003e9(5)\u003c/strong\u003e, 803-806 (2020).\u003c/li\u003e\n\u003cli\u003eUnal, M., Vardar, F. \u0026amp; Ayturk, Z. Callose in Plant Sexual Reproduction. \u003cem\u003eCurrent Progress in Biological Research\u003c/em\u003e. InTech. doi: 10.5772/53001 (2013).\u003c/li\u003e\n\u003cli\u003eUshijima, K. \u003cem\u003eEt al\u003c/em\u003e. Isolation of the floral morph-related genes in heterostylous flax (\u003cem\u003eLinum grandiflorum\u003c/em\u003e): the genetic polymorphism and the transcriptional and post-transcriptional regulations of the S locus. \u003cem\u003ePlant J\u003c/em\u003e. \u003cstrong\u003e69(2)\u003c/strong\u003e, 317-31. doi: 10.1111/j.1365-313X.2011.04792.x (2012).\u003c/li\u003e\n\u003cli\u003eVan Zeist, W. \u0026amp; Bakker-Heeres, J. A. H. Evidence for linseed cultivation before 6000 BC. \u003cem\u003eJ. Archaeol. Sci\u003c/em\u003e. \u003cstrong\u003e2(3)\u003c/strong\u003e, 215-219. https://doi.org/10.1016/0305-4403(75)90059-X (1975).\u003c/li\u003e\n\u003cli\u003eVavilov, N. I. Studies on the Origin of Cultivated Plants. Leningrad: des Plantes, State Press (1926).\u003c/li\u003e\n\u003cli\u003eWang, Y. F. The present situation and suggestions about the study of flax germplasm resource in China. \u003cem\u003eJ. Life Sci\u003c/em\u003e. \u003cstrong\u003e2(1)\u003c/strong\u003e, 67e71 (2008).\u003c/li\u003e\n\u003cli\u003eWang, Y., Li, X., Fan, B., Zhu, C. \u0026amp; Chen, Z. Regulation and Function of Defense-Related Callose Deposition in Plants. \u003cem\u003eInternational Journal of Molecular Sciences\u003c/em\u003e. \u003cstrong\u003e22(5)\u003c/strong\u003e, 2393. doi: 10.3390/ijms22052393 (2021).\u003c/li\u003e\n\u003cli\u003eYe, Y., Zhou, Y., Tan, J., Zhu, G., Liu, J. \u0026amp; Xu, Y. Cross-compatibility in interspecific hybridization of different \u003cem\u003eCurcuma\u003c/em\u003e accessions. \u003cem\u003ePlants (Basel)\u003c/em\u003e. \u003cstrong\u003e12(10)\u003c/strong\u003e, 1961. doi: 10.3390/plants12101961 (2023).\u003c/li\u003e\n\u003cli\u003eZheng, Y. Y., Lin, X. J., Liang, H. M., Wang, F. F. \u0026amp; Chen, L. Y. The Long Journey of Pollen Tube in the Pistil. \u003cem\u003eInt J Mol Sci\u003c/em\u003e. \u003cstrong\u003e19(11)\u003c/strong\u003e, 3529. doi: 10.3390/ijms19113529 (2018).\u003c/li\u003e\n\u003cli\u003eZhukovsky, P. M. Cultivated plants and their wild relatives. Systematics, geography, cytogenetics, resistance, ecology, origin and use. Moscow: Kolos. Leningrad, 751 p (1971).\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003e\u003cstrong\u003eTable 1\u003c/strong\u003e\u003cstrong\u003e. Details of plant material used in the study\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"664\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 27.259%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eScientific Name\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.7831%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eL. usitatissimum\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003eL.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6988%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eL. bienne\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003eMill.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27.259%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eL. grandiflorum\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003eL.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 27.259%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCommon name\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.7831%;\"\u003e\n \u003cp\u003eFlax/linseed\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6988%;\"\u003e\n \u003cp\u003ePale flax\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27.259%;\"\u003e\n \u003cp\u003eRed flax/ Scarlet flax\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 27.259%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eBiological Status\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.7831%;\"\u003e\n \u003cp\u003eGermplasm (Registered genetic stock)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6988%;\"\u003e\n \u003cp\u003eWild progenitor\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27.259%;\"\u003e\n \u003cp\u003eDistant CWR\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 27.259%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNational ID\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.7831%;\"\u003e\n \u003cp\u003eIC268345\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6988%;\"\u003e\n \u003cp\u003eEC993391\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27.259%;\"\u003e\n \u003cp\u003eIC633096\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 27.259%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eChromosome number (2n)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.7831%;\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6988%;\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27.259%;\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 27.259%;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePotential use\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.7831%;\"\u003e\n \u003cp\u003eHigh oil content (42.5%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6988%;\"\u003e\n \u003cp\u003eFibre quality and more primary branches\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27.259%;\"\u003e\n \u003cp\u003eBud fly resistance\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 27.259%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eGrowth habit\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.7831%;\"\u003e\n \u003cp\u003eBushy to erect\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6988%;\"\u003e\n \u003cp\u003eProstrate growth habit\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27.259%;\"\u003e\n \u003cp\u003eSemi-spreading\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 27.259%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCross compatibility with cultivated species\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.7831%;\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6988%;\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27.259%;\"\u003e\n \u003cp\u003eNo (Shriveled seeds)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 27.259%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eGeographic distribution/origin\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.7831%;\"\u003e\n \u003cp\u003eSouthern Europe, the Near East, or Central Asia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6988%;\"\u003e\n \u003cp\u003eMediterranean Sea, Iran, and the Canary Islands\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27.259%;\"\u003e\n \u003cp\u003eNative to Algeria but occurrence is found in India, Northern Africa, Southern Europe and North America\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 27.259%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eProcured from\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20.7831%;\"\u003e\n \u003cp\u003eNational Genebank, ICAR-NBPGR, New Delhi, India\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 24.6988%;\"\u003e\n \u003cp\u003eCGN Gene Bank, Plant Research International Wageningen, Netherlands\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 27.259%;\"\u003e\n \u003cp\u003eNational Genebank, ICAR-NBPGR, New Delhi, India\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2. Details of cross combinations attempted for wide hybridization including self-pollination\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"622\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 8.5209%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eS. No.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.9711%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNature of Cross\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47.2669%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSpecies involved\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 25.2412%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAccessions\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 8.5209%;\"\u003e\n \u003cp\u003e1.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.9711%;\"\u003e\n \u003cp\u003eSelf-pollination\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47.2669%;\"\u003e\n \u003cp\u003e\u003cem\u003eL. usitatissimum\u0026nbsp;\u003c/em\u003eX \u003cem\u003eL. usitatissimum\u0026nbsp;\u003c/em\u003e(\u003cem\u003eLu\u0026nbsp;\u003c/em\u003eX \u003cem\u003eLu\u003c/em\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 25.2412%;\"\u003e\n \u003cp\u003eIC268345 X IC268345\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 8.5209%;\"\u003e\n \u003cp\u003e2.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.9711%;\"\u003e\n \u003cp\u003eSelf-pollination\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47.2669%;\"\u003e\n \u003cp\u003e\u003cem\u003eL. bienne\u0026nbsp;\u003c/em\u003eX \u003cem\u003eL. bienne\u0026nbsp;\u003c/em\u003e(\u003cem\u003eLb\u0026nbsp;\u003c/em\u003eX \u003cem\u003eLb\u003c/em\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 25.2412%;\"\u003e\n \u003cp\u003eEC993391X EC993391\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 8.5209%;\"\u003e\n \u003cp\u003e3.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.9711%;\"\u003e\n \u003cp\u003eSelf-pollination\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47.2669%;\"\u003e\n \u003cp\u003e\u003cem\u003eL. grandiflorum\u0026nbsp;\u003c/em\u003eX\u003cem\u003e\u0026nbsp;L. grandiflorum\u0026nbsp;\u003c/em\u003e(\u003cem\u003eLg\u0026nbsp;\u003c/em\u003eX \u003cem\u003eLg\u003c/em\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 25.2412%;\"\u003e\n \u003cp\u003eIC633096 X IC633096\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 8.5209%;\"\u003e\n \u003cp\u003e4.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.9711%;\"\u003e\n \u003cp\u003eCross pollination\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47.2669%;\"\u003e\n \u003cp\u003e\u003cem\u003eL. usitatissimum\u0026nbsp;\u003c/em\u003eX\u003cem\u003e\u0026nbsp;L. bienne\u0026nbsp;\u003c/em\u003e(\u003cem\u003eLu\u0026nbsp;\u003c/em\u003eX \u003cem\u003eLb\u003c/em\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 25.2412%;\"\u003e\n \u003cp\u003eIC268345 X EC993391\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 8.5209%;\"\u003e\n \u003cp\u003e5.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.9711%;\"\u003e\n \u003cp\u003eCross pollination\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47.2669%;\"\u003e\n \u003cp\u003e\u003cem\u003eL. bienne\u0026nbsp;\u003c/em\u003eX \u003cem\u003eL. usitatissimum\u0026nbsp;\u003c/em\u003e(\u003cem\u003eLb\u0026nbsp;\u003c/em\u003eX \u003cem\u003eLu\u003c/em\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 25.2412%;\"\u003e\n \u003cp\u003eEC993391 X\u003cem\u003e\u0026nbsp;\u003c/em\u003eIC268345\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 8.5209%;\"\u003e\n \u003cp\u003e6.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.9711%;\"\u003e\n \u003cp\u003eCross pollination\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47.2669%;\"\u003e\n \u003cp\u003e\u003cem\u003eL. usitatissimum\u0026nbsp;\u003c/em\u003eX \u003cem\u003eL. grandiflorum\u0026nbsp;\u003c/em\u003e(\u003cem\u003eLu\u0026nbsp;\u003c/em\u003eX \u003cem\u003eLg\u003c/em\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 25.2412%;\"\u003e\n \u003cp\u003eIC268345 X IC633096\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 8.5209%;\"\u003e\n \u003cp\u003e7.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18.9711%;\"\u003e\n \u003cp\u003eCross pollination\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 47.2669%;\"\u003e\n \u003cp\u003e\u003cem\u003eL. grandiflorum\u0026nbsp;\u003c/em\u003eX\u003cem\u003e\u0026nbsp;L. usitatissimum\u0026nbsp;\u003c/em\u003e(\u003cem\u003eLg\u0026nbsp;\u003c/em\u003eX \u003cem\u003eLu\u003c/em\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 25.2412%;\"\u003e\n \u003cp\u003eIC633096 X\u003cem\u003e\u0026nbsp;\u003c/em\u003eIC268345\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3. Pollen vitality in studied\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eLinum\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;species (\u003c/strong\u003e\u003cstrong\u003eL. usitatissimum, L. bienne and L. grandiflorum\u003c/strong\u003e\u003cstrong\u003e)\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"905\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 68px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSection 1\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 71px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSection 2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 75px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSection 3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 73px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSection 4\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 69px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSection 5\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 75px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSection 6\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 71px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSection 7\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 73px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSection 8\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eF\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eS\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eF\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eS\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eF\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eS\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eF\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eS\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eF\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eS\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eF\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eS\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eF\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eS\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 43px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eF\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 30px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eS\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal F\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal S\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal PMC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePV%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"21\" style=\"width: 905px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eL. usitatissimum\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eSlide 1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 35px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 35px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 42px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 30px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\n \u003cp\u003e187\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e151\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e338\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e55.33\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eSlide 2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 42px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 30px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\n \u003cp\u003e129\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e193\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e66.84\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eSlide 3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 43px;\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 30px;\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\n \u003cp\u003e238\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e119\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e357\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e66.67\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eSlide 4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e120\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 43px;\"\u003e\n \u003cp\u003e62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 30px;\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\n \u003cp\u003e601\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e115\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e716\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e83.94\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eSlide 5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 43px;\"\u003e\n \u003cp\u003e57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 30px;\"\u003e\n \u003cp\u003e34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\n \u003cp\u003e499\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e194\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e693\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e72.01\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eSlide 6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e99\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e71\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 43px;\"\u003e\n \u003cp\u003e117\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 30px;\"\u003e\n \u003cp\u003e29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\n \u003cp\u003e606\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e205\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e811\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e74.72\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 43px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 30px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal PMCs\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3108\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 43px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 30px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAvg. PV%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e69.92\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"21\" style=\"width: 905px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eL. bienne\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eSlide 1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 42px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 30px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\n \u003cp\u003e198\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e59\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e257\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e77.04\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eSlide 2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e103\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e59\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 42px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 30px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\n \u003cp\u003e340\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e369\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e92.14\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eSlide 3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 43px;\"\u003e\n \u003cp\u003e39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 30px;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\n \u003cp\u003e320\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e380\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e84.21\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eSlide 4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e46\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e61\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e46\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 43px;\"\u003e\n \u003cp\u003e42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 30px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\n \u003cp\u003e333\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e362\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e91.99\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 43px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 30px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal PMCs\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1368\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 43px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 30px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAvg. PV%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e86.35\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"21\" style=\"width: 905px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eL. grandiflorum\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eSlide 1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 35px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 35px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 42px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 30px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\n \u003cp\u003e137\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e154\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e88.96\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eSlide 2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e110\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e95\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 35px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 35px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 42px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 30px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\n \u003cp\u003e341\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e362\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e94.20\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eSlide 3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e46\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 35px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 35px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 42px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 30px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\n \u003cp\u003e222\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e246\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e90.24\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eSlide 4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 35px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 35px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 37px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 42px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 43px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 30px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\n \u003cp\u003e184\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e192\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e95.83\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eSlide 5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 43px;\"\u003e\n \u003cp\u003e36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 30px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\n \u003cp\u003e440\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e456\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e96.49\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003eSlide 6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e158\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e162\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 43px;\"\u003e\n \u003cp\u003e38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 30px;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\n \u003cp\u003e704\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e154\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e858\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e82.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 43px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 30px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal PMCs\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2268\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 52px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 28px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 44px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 31px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 35px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 37px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 42px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 29px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 43px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 30px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 63px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAvg. PV%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 95px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 46px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e91.30\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eAbbreviations: F: Fertile; S: Sterile; PMC: Pollen Mother Cell; PV%: Pollen vitality percent;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 4. Pollen germination and pollen tube growth for studied cross combinations in \u003cem\u003eLinum\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"607\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 67px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eLu X Lu\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eLb X Lb\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eLg X Lg\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eLu X Lb\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eLb X Lu\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eLu X Lg\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eLg X Lu\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003ePollen germination (%)\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003eMax.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 67px;\"\u003e\n \u003cp\u003e95.99\u003c/p\u003e\n \u003cp\u003e(10HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e95.58\u003c/p\u003e\n \u003cp\u003e(8HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e94.82\u003c/p\u003e\n \u003cp\u003e(2HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e97.00\u003c/p\u003e\n \u003cp\u003e(4HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e96.19\u003c/p\u003e\n \u003cp\u003e(4HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e10.92\u003c/p\u003e\n \u003cp\u003e(4HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e10.33\u003c/p\u003e\n \u003cp\u003e(6HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003eMin.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 67px;\"\u003e\n \u003cp\u003e92.83\u003c/p\u003e\n \u003cp\u003e(8HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e92.17\u003c/p\u003e\n \u003cp\u003e(24HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e93.54\u003c/p\u003e\n \u003cp\u003e(10HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e91.91\u003c/p\u003e\n \u003cp\u003e(8HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e93.03\u003c/p\u003e\n \u003cp\u003e(48HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e6.66\u003c/p\u003e\n \u003cp\u003e(48HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e7.71\u003c/p\u003e\n \u003cp\u003e(8HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003eAverage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 67px;\"\u003e\n \u003cp\u003e94.51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e94.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e94.12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e94.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e94.87\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e8.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e8.82\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003eVariance\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 67px;\"\u003e\n \u003cp\u003e1.68\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e1.54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e0.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e3.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e0.98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e2.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e0.72\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003eTubes to Style (No.)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003eMax.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 67px;\"\u003e\n \u003cp\u003e75\u003c/p\u003e\n \u003cp\u003e(4HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e62\u003c/p\u003e\n \u003cp\u003e(4HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e49\u003c/p\u003e\n \u003cp\u003e(8HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e44\u003c/p\u003e\n \u003cp\u003e(4HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e40\u003c/p\u003e\n \u003cp\u003e(8HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e39\u003c/p\u003e\n \u003cp\u003e(4HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e23\u003c/p\u003e\n \u003cp\u003e(4HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003eMin.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 67px;\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003cp\u003e(48HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e29\u003c/p\u003e\n \u003cp\u003e(48HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003cp\u003e(2HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003cp\u003e(24HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e19\u003c/p\u003e\n \u003cp\u003e(24HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003cp\u003e(8HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003cp\u003e(6HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003eAverage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 67px;\"\u003e\n \u003cp\u003e13.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e4.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e1.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e8.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e2.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e43.53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e14.92\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003eVariance\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 67px;\"\u003e\n \u003cp\u003e582.29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e122.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e147.29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e149.62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e46.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e104.29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e9.24\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003eTubes to Ovary (No.)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003eMax.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 67px;\"\u003e\n \u003cp\u003e35\u003c/p\u003e\n \u003cp\u003e(2HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e36\u003c/p\u003e\n \u003cp\u003e(4HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e43\u003c/p\u003e\n \u003cp\u003e(8HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003cp\u003e(2HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003cp\u003e(8HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003cp\u003e(48HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003cp\u003e(4HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003eMin.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 67px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003cp\u003e(48HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003cp\u003e(48HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003cp\u003e(2HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003cp\u003e(48HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003cp\u003e(2HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003cp\u003e(8HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003cp\u003e(10HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003eAverage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 67px;\"\u003e\n \u003cp\u003e6.34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e2.46\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e1.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e3.33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e1.73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e22.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e12.95\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003eVariance\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 67px;\"\u003e\n \u003cp\u003e106.81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e64.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e123.57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e25.62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e43.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e23.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e11.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003eTubes to Ovule (No.)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003eMax.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 67px;\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003cp\u003e(8HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003cp\u003e(4/8HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e40\u003c/p\u003e\n \u003cp\u003e(8HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003cp\u003e(8HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003cp\u003e(8HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e15\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e(48HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003cp\u003e(4HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003eMin.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 67px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003cp\u003e(24HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003cp\u003e(24HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003cp\u003e(2HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003cp\u003e(48HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003cp\u003e(6HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e(2HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003cp\u003e(2HAP)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003eAverage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 67px;\"\u003e\n \u003cp\u003e2.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e1.16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e1.12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e1.29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e0.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e12.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e8.01\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 96px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003eVariance\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 67px;\"\u003e\n \u003cp\u003e10.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e11.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e107.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e4.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e20.95\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 66px;\"\u003e\n \u003cp\u003e21.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 60px;\"\u003e\n \u003cp\u003e20.57\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eAbbreviations:\u0026nbsp;(\u003cem\u003eLu\u0026nbsp;\u003c/em\u003eX \u003cem\u003eLu\u003c/em\u003e): \u003cem\u003eL. usitatissimum\u0026nbsp;\u003c/em\u003eX \u003cem\u003eL. usitatissimum\u003c/em\u003e, (\u003cem\u003eLb\u0026nbsp;\u003c/em\u003eX \u003cem\u003eLb\u003c/em\u003e): \u003cem\u003eL. bienne\u0026nbsp;\u003c/em\u003eX \u003cem\u003eL. bienne\u003c/em\u003e,\u0026nbsp;(\u003cem\u003eLg\u0026nbsp;\u003c/em\u003eX\u003cem\u003e\u0026nbsp;Lg\u003c/em\u003e): \u003cem\u003eL. grandiflorum\u0026nbsp;\u003c/em\u003eX\u003cem\u003e\u0026nbsp;L. grandiflorum\u003c/em\u003e, (\u003cem\u003eLu\u0026nbsp;\u003c/em\u003eX\u003cem\u003e\u0026nbsp;Lb\u003c/em\u003e): \u003cem\u003eL. usitatissimum\u0026nbsp;\u003c/em\u003eX\u003cem\u003e\u0026nbsp;L. bienne\u003c/em\u003e, (\u003cem\u003eLb\u0026nbsp;\u003c/em\u003eX \u003cem\u003eLu\u003c/em\u003e): \u003cem\u003eL. bienne\u0026nbsp;\u003c/em\u003eX \u003cem\u003eL. usitatissimum\u003c/em\u003e, (\u003cem\u003eLu\u0026nbsp;\u003c/em\u003eX \u003cem\u003eLg\u003c/em\u003e): \u003cem\u003eL. usitatissimum\u0026nbsp;\u003c/em\u003eX \u003cem\u003eL. grandiflorum\u003c/em\u003e, (\u003cem\u003eLg\u0026nbsp;\u003c/em\u003eX\u003cem\u003e\u0026nbsp;Lu\u003c/em\u003e): \u003cem\u003eL. grandiflorum\u0026nbsp;\u003c/em\u003eX\u003cem\u003e\u0026nbsp;L. usitatissimum\u003c/em\u003e, HAP: Hours after pollination, No.: Numbers; %: Percentage, Max.: Maximum value, Min.: Minimum value\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Pollen-pistil interaction, Linum, interspecific, Pre-fertilization barriers, fluorescent microscopy","lastPublishedDoi":"10.21203/rs.3.rs-5280537/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5280537/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eLinseed, has been a source of natural fiber for terxtile industries since its domestication. However, despite being the potential source of trait reservoir, the use of \u003cem\u003eLinum\u003c/em\u003e wild genetic resources for the improvement of economic traits are not exploited widely. This is mainly due to the degree of genetic divergence that exists among the interspecific ecotypes causing crossability issues. Self-incompatibility due to the occurrence of heterostyly is very well reported in distantly related crop wild relatives of \u003cem\u003eLinum\u003c/em\u003e and, the mechanism of self-incompatibility between different floral morphs is also studied. However, pollen germination and tube growth responses in the interspecific crosses are rarely studied. Thus, the present study was exclusively carried out to assess the major pre-zygotic barriers and their effect on pollen germination on foreign stigma using fluorescent microscopy of aniline blue stain-aided technology, to understand how the species barriers operate on pollen germination and pollen tube growth. The study revealed that the pollen-pistil interaction in the wide crosses among \u003cem\u003eL. usitatissimum\u003c/em\u003e X \u003cem\u003eL. grandiflorum\u003c/em\u003e was regulated by both temporal and spatial pre-fertilization barriers. Callose deposition within 2 hours after pollination (HAP) at the stigma surface, was the major cause inhibiting pollen germination. Various kinds of aberrations started appearing during the 2-4HAP. The complexity of interspecific hybridization was observed in terms of arrest of pollen tube (PT) growth in the ovary, ruptured, twisted and swollen pollen tube tip, tube growth in reverse direction, convoluted and terminated growth patterns. The results suggest that although, distant hybridization is usually less efficient, hybridization success can be improved by advanced techniques such as embryo rescue and \u003cem\u003ein vitro\u003c/em\u003e culture of isolated immature embryos that will undoubtedly yield crucial information on selecting the ideal culture conditions and medium, paving the way for groundbreaking discoveries in this field.\u003c/p\u003e","manuscriptTitle":"Pollen–pistil interactions in divergent wide crosses lead to spatial and temporal pre-fertilization reproductive barrier in flax (Linum usitatissimum L.)","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-11-13 16:51:08","doi":"10.21203/rs.3.rs-5280537/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-12-20T11:50:16+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-12-20T10:00:46+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"249265595327666484331756996711930683349","date":"2024-12-13T00:58:42+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-11-07T02:01:24+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"203302544870448078680068814384631294557","date":"2024-11-05T07:49:03+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"43986810978896184277583927175824568251","date":"2024-11-05T06:22:23+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-11-04T11:20:13+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-11-04T11:14:22+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2024-11-04T06:20:12+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-11-01T04:36:31+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2024-10-17T07:14:53+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"777d6d7a-00e3-41ed-a7c5-ff2b3820a521","owner":[],"postedDate":"November 13th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":39925134,"name":"Biological sciences/Plant sciences/Plant breeding"},{"id":39925135,"name":"Biological sciences/Plant sciences/Plant reproduction"},{"id":39925136,"name":"Biological sciences/Plant sciences"}],"tags":[],"updatedAt":"2025-03-03T16:01:30+00:00","versionOfRecord":{"articleIdentity":"rs-5280537","link":"https://doi.org/10.1038/s41598-025-90046-8","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2025-02-25 15:57:05","publishedOnDateReadable":"February 25th, 2025"},"versionCreatedAt":"2024-11-13 16:51:08","video":"","vorDoi":"10.1038/s41598-025-90046-8","vorDoiUrl":"https://doi.org/10.1038/s41598-025-90046-8","workflowStages":[]},"version":"v1","identity":"rs-5280537","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5280537","identity":"rs-5280537","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}