Optimizing proliferation and assessment of valak morpho-phenological traits; an endangered nutritious Allium endemic to Iran

Optimizing proliferation and assessment of valak morpho-phenological traits; an endangered nutritious Allium endemic to Iran | 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 Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Optimizing proliferation and assessment of valak morpho-phenological traits; an endangered nutritious Allium endemic to Iran Sajad Jafari, Mohammadreza Hassandokht, Madi Taheri This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-250915/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Allium elburzense W. (subg. Melanocrommyum ) is an endangered and endemic species to Iran, which is called “valak” and known as a valuable and pricy vegetable with high nutritional and medicinal properties. This study was carried out in order to domestication, removing barriers of sexual reproduction, micropropagation and investigation of phenological stages and growth cycle of the species. The results indicated that fresh seeds had better germination than one year old seeds and appropriate germination temperature in A. elburzense W. seed was 12 °C. According to ANOVA results, the triple interaction of scarification (Sc), stratification period (StP) and GA 3 had a significant effect on seedling length, seedling fresh and dry weight, bulb height and diameter (p<0.01). Comparison of means showed that the treatments in which the StP (45 days) and scarification was applied had better effects on seed germination (66.5% germination) than other treatments. Murashige and Skoog with 300 mg L -1 KH2PO4 (MSP) medium caused a significant increase in the diameter of the basal plate compared to MS medium. Efficient system for in vitro propagation and conservation of valak genetic resources were the use of MSP culture medium supplemented with 0.6 mg l -1 NAA, 4 mg l -1 BA and 120 g l -1 of sucrose, which resulted in a direct propagation coefficient of 26.83 seedlings and bulblets at 18 weeks. As regards its life cycle, the juvenile phase in a low proportion of individuals (13.33%) lasted 2 years from seeding, while 87.66% of individuals were capable of producing inflorescence in the third year. Results showed that this plant is a metamorphosis and cross pollinator species. Results of present study can be used in domestication, conservation of genetic resources and proper exploitation of valak species. Plant Physiology and Morphology Allium elburzense W. Efficient rapid micropropagation Morpho-phenology Organogenesis Phyto-hormones Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Key Message In this study, a very valuable and widely used plant in Iran that was in danger of extinction due to excessive harvesting from wild and lack of cultivation was studied, and a rapid and low-cost micropropagation system was introduced. In addition, by reviewing life cycle and behavior of its various organs, the barriers to sexual reproduction were removed. According to the results obtained and high price of this species in Iran, not only will its extinction be prevented, but it will be a very good choice for cultivation and investment. Introduction The subgenus Melanocrommyum (Webb & Berthel.) Rouy is the second largest subgenus of the genus Allium , which consists of 10 section and has over 170 species. Allium elburzense W. belongs to the Asteroprason section and called “valak” in Iran and used as fresh vegetable and medical agent (Jafari et al. 2017). This species is found only in mountainous areas of northern Iran in sandy slopes of the Alborz mountain range (Frisch and Abbasi, 2013). Valak is a very expensive vegetable in Iran and its leaves uses for the preparation of a variety of special soups and “valak pilaw”. Its inflorescence uses to prepare pickles, spices and food decoration, and its bulb is consumed in the pharmaceutical industry (Jafari, 2017). This species is not cultivated, but collected from the nature (Akhavan et al. 2015). Indiscriminate harvesting of valak from natural habitats, harvesting pre-flower and before seed formation with bulb (permanent organ) and low percentage of seed germination has caused the endemic species is exposure to extinction. Reducing the number of individuals in a habitat has led to a decrease in genetic diversity, which in turn will increase the vulnerability of the species to environmental changes, genetic drift and natural selection. Domestication of these valuable species is a suitable way to preserve desirable species and prevent their extinction (Ebrahimi, 2014). One of the main steps in the process is to study the methods of reproduction and preservation of genetic resources of these species. Therefore, awareness and promotion of knowledge about the life cycle of a species, the effect of the environment on its reproduction and its phenological and genetic traits are of great importance. As such knowledge will provide a better understanding of their phenology in order to predict the most favorable periods for seedling establishment in the field and facilitate the provision of nursery plants from seed and organ in vitro culture (Lentz and Johnson, 1998). Low seed germination speed and rate, slow seedling growth and low energy storage of seeds are some of the germination problems of some Allium species. Seed dormancy has been reported in a large number of wild Alliums, so it seems necessary to study methods to eliminate it in this species (Dashti et al. 2012; Ebrahimi et al. 2014; Phillips 2010). Seed and bulb dormancy in wild allium species is an adaptation to environmental conditions to avoid germination and growth in inappropriate seasons, so, in order to eliminate the dormancy of these organs, their natural environment should be modeled. In addition to sexual reproduction, which has problems such as segregation of traits, time consuming and more costly, asexual methods such as tissue and organ culture can be used to produce these species. Tissue culture is the most effective virus-free and rapid propagation technology and provides an efficient tool for producing virus-free Allium plants (Gimenez et al. 2016; Taşkın et al. 2013). There is no reliable information on the dormancy of seeds and bulbs and how to facilitate its propagation and conservation of A. elburzense W.. Therefore, due to the high nutritional, pharmaceutical and economic value of valak on the one hand, and its lack of cultivation on the other hand, this study was design and performed in order to domestication (evaluation of phenological behaviors of growth and reproduction stages by both bulb and seed pathway), elimination barriers to sexual and asexual reproduction and achievement a rapid and low-cost proliferation per unit of time. Material And Methods Plant material Bulb and seed of species A. elburzense W. (Iranian Valak) were collected from the Kalha habitat (in Alborz province at an altitude 2454 m, 36°44ʹ N and 51°17ʹ E with average precipitation 411.2 mm and average temperature of 11.8°C (herbarium no. 006467). Sexual propagation Stratification period (StP), Germination temperature (GT) and seed age (SAg) Seeds were disinfected into sodium hypochlorite 1% before use for 5 minutes and then were washed 3 times with distilled water. A factorial experiment was conducted in a completely randomized design (CRD) with 3 factors and 3 replications (each replicate containing 100 seeds). The factors were included stratification period (StP) (a1= 15, a2= 30, a3= 45 and a4= 60 days of moist chilling at 5 °C), seed age (SAg) (b1= fresh seed and b2= one year old seed) and germination temperature (GT) (c1= 5, c2= 12 and c3= 20 °C). Percent and rate of germination were measured in the test. Scarification (sc) , stratification, GA 3 and KNO 3 A factorial experiment was conducted as a CRD design with 4 factors in 3 replications. The factors were included scarification (Sc) (a1= without scarification, a2= scarification with 95% sulfuric acid for 10 minutes and a3= scarification with 95% sulfuric acid for 20 minutes, and a4= sandpaper for 30 seconds), moist stratification (St) (b1= 0 and b2= 45 days at 5°C), GA 3 (immersed seeds in c1= 0 and c2= 500 ppm for 12 h) and KNO 3 (immersed seeds in d1= 0 and d2= 0.2% for 24 h). Percent and rate of germination, seedling length, seedling fresh and dry weight and bulb height and width were measured. The seeds and filter papers were kept wet with distilled water. The petri dishes were placed in a germinator at 20°C, 16 h light and 8 h dark conditions for 20 days. Seeds in which the tip of the root appeared were considered germinated (radicle protrusion) (Albert et al. 2002). Treated seeds were planted in pots (25 seeds/plot) containing coco-peat and perlite (with a ratio 3:1). Asexual propagation ( In vitro ) Plant material The collected bulbs were placed in a cool temperature (15-18 °C) for a week in order to drying the surface moisture and avoidance of rotting, and then placed in a refrigerator of 5 °C (June 2015). Bulbs were reviewed in November and explants prepared after observing the sprouts grown. The bulb central bud with basal plate were used as explant. Disinfection test In order to find the best disinfectant concentration and duration of treatment, a factorial experiment based on CRD was conducted with 3 factors in five replications included sodium hypochlorite (NaOCl), immersion time and type of explants (cut out before or after treatment (Fig. 1)). Culture conditions and establishment medium MS medium (Murashige and Skoog, 1962) containing 3 % (w v −1 ) sucrose and 0.7 % agar was used. According to past researches, MS is the most suitable medium for in vitro culture of Alliums (yan et al. 2009; Ebrahimi et al. 2014; Farhadi et al. 2017). The medium pH was adjusted to 6 before autoclaving at 121 ° C for 20 minutes. The cultivated explant were placed in a growth chamber at a light intensity of 40 μmol m -2 s -1 (a combination of cool fluorescent lamps with white and yellow light with a ratio of 50:50) at 25 ± 1 °C. The explants were cultured in MSP and MS media in six replicates to determination of establishment medium. The amount of phosphorus in MSP (with 300 mg L -1 KH2PO4) medium was twice the MS (170 mg L-1 KH2PO4) medium, approximately (Kahane et al. 1992). Diameter of basal plate was measured after 45 days with digital caliper and its increase was calculated as a percentage. Effect of growth regulators on micro propagation indices The explants were divided into two parts from the base plate after 45 days and then transferred to MS containing growth regulators. A factorial experiment based on CRD was conducted with two factors in six replications, including NAA (a1= 0, a2= 0.2, a3= 0.4, a4= 0.6 and a5= 0.8 mg l -1 ) and cytokinin contains benzyl adenine and Kinetin (b1= 0, b2= 2BA, b3= 4BA, b4= 6BA, b5= 1Kin, b6= 2Kin and b7= 3Kin mg l -1 ). Explants were placed in 14 hours light and 10 hours dark (based on pre-treatment) and characteristics of callus growth index (equation 1) (Fan et al. 2017), number of days to callus emergence, number of bulblet and shoot per explant were calculated. Callus growth index= (final callus fresh weight - initial callus fresh weight)]/final callus fresh weight× 100 Effect of sucrose concentration on micro propagation indices Effect of sucrose different concentrations (30, 60, 90, 120 and 150 g l -1 ) on propagation indices of A. elburzense W. was evaluated in a completely randomized design with six replications. Explants were placed in 14 hours light and 10 hours dark. After 8 weeks, callus growth index, number of days to callus emergence, number of bulblet and shoot per explant were calculated. Calculation of in vitro propagation coefficient (direct organogenesis) Explant of A. elburzense W. was cultured in the best condition (establishment medium, BA, cytokinin, sucrose) based on the results of previous experiments and propagation coefficient was calculated for an explant after 18 weeks. Propagation coefficient was total number of shoot and bulblet composed of a complete explant, in a given period of time. Acclimation and transfer to non-controlling condition The formed bulblet were separated and cultured in pots containing perlite and cocopeat with a 3:1 ratio and were transferred to ambient temperature (non-controlling condition) in late February 2016. Morpho-phenological evaluation of A. elburzense W. This experiment was carried out in order to evaluation of growth phenology and stages of reproductive organs production of A. elburzense W., duration 3 years, at the Horticultural Sciences Research Station of the University of Tehran (long. 51° 10′ 35′′ N, lat. 35° 42′ 18′′ E, height 1297 m), in the both form of sowing seeds and bulbs. The dried and mature inflorescences (previously marked) and bulbs were collected from the Kalha habitat in May 2014 and kept in the shade at 25 °C. After separating the hollow and immature seeds, 30 seeds were planted in trays of 30 cells with a mixture combination of peat moss, coco peat and perlite bed in a proportion of 6:1:1 in 3 replications. Observations were recorded until the formation of inflorescences and seeds. The bulbs from the seedlings harvested and were kept at 17 °C in a place with low humidity and then planted in November 2015 10 cm pots (10 bulbs per pot) in mixture of soil and sand in proportion of 1:1. Simultaneously with sowing seeds, bulbs collected from this habitat were also planted in the field to study bulb behavior and asexual reproduction. Developmental phases from flower formation to seed maturity were examined using a stereomicroscope (S21, Berlin, Germany). After the end of the growing season and drying of the aerial part, the bulbs were harvested and weighed before planting at the beginning of the cold season in both planting methods (seed and bulb). In June 2016 percent of inflorescence formation, bulb diameter and weight, number of leaf and plant height were evaluated. In early autumn 2016 the harvested bulbs were planted. In June 2017, bulbs were harvested and morphological traits were measured. Statistical analysis Data on indices of seed germination and micropropagation were analyzed with the statistical analysis software SPSS v.21 and MSTATC (version 2.1 USA). Comparisons between establishment media were performed using independent t-test. All data were subjected to analysis of variance and means were compared using Duncan’s multiple range test at P < 0.01. Results And Discussion Facilitate germination and remove obstacles Effect of germination temperature, seed age and stratification period The results showed that the highest germination rate was obtained in 45 and 60 day stratification (Fig. 2). Based on the comparison of means, the highest germination percentage was observed in treatments of 45 days StP × GT 12 °C (62.42%) and 60 days StP × GT 20 °C (61.1%) (Table 1). Therefore, it can be said that the seeds of A. elburzense W. need at least 45 days StP at 5 °C for germination. In the treatment of SAg×GT, the highest germination percentage and rate were observed in fresh seed and GT 12 °C with 43.24 % and 4.31 s d -1 , respectively (Table 2). Effect of scarification, stratification, GA 3 and KNO 3 According to the results of the previous experiment, the GT 12 °C and Stp 45 days were considered for this experiment. The use of GA 3 improved the studied traits compared to the control. KNO 3 had no significant effect on the studied traits. No seeds germinated in 10 minutes using of sulfuric acid without StP. The results showed that the interaction effect of GA 3 and KNO 3 led to a significant increase in germination percentage (19.98 %) and germination rate (2.17 s d -1 ) (Table 3). The results of ANOVA showed that the triple interaction of Sc, StP and GA 3 had a significant effect on seedling length, seedling fresh and dry weight, bulb height and diameter at the statistical level of 1%. Comparison of means showed that the treatments in which the StP (45 days) was applied had better effects on the studied traits than other treatments (Table 4). Although all Allium species follow the same germination pattern, the mechanisms of seed dormancy differ in taxonomic groups and in different habitats (Kamenetsky and Rabinoswitch 2006). Specht and Keller (1997) examined the germination temperatures of 94 Allium species and the appropriate seed germination temperature for Allium subspecies was 16 °C, for Rhiziridium subspecies between 16-26 °C, and 5-15 °C was also reported for species belonging to the Melanocrommyum section. The use of sulfuric acid removed the black seed coat, but had no significant effect on seed germination indices. However, sandpaper had a better effect than sulfuric acid and caused significant seed germination and improved seedling quality indices (Table 4). In the present study, it was observed that without using the appropriate StP, the seeds did not germinate significantly. These results were also reported by Ebrahimi et al. (2014) in shallot ( A. hirtifolium Boiss). According to the obtained results, dormancy of A. elburzense W. seed can be considered as physiological and physical type. GA 3 increases the growth potential and strength of the embryo, overcoming resistant shells and mechanical barriers (pericarp, seed coat and endosperm) for germination and seedling growth (Finch-Savage and Leubner-Metzger 2006). Endosperm attenuation is regulated by phyto-hormones and environmental factors. Radicle emerging can be accelerated by GA 3 and inhibited by abscisic acid (Silva et al. 2004; Finch-Savage and Leubner-Metzger 2006; Linkies et al. 2010). The hard and thick endosperm in species of Melanocrommyum section is a type of plant acclimation to low temperatures in the main habitats, which protects the embryo during the winter cold. Micropropagation Disinfection of explants The results showed that the lowest contamination (0%) was observed in the treatment of 3% active chlorine and 15 minutes immersion time, and the highest contamination in the explant (78%) belonged to the treatment of 1% active chlorine for 5 minutes in cut explants (Table 5). As a result, 3% active chlorine treatment and 15 minutes of immersion were used for uncut explants. Establishment medium The results of T-test showed that there was a significant difference between the two used establishment culture medium, basal plate in MSP medium indicated an increase with an average of 7.36% than explant cultured in MS medium (Table 6). The amount of phosphorus in MSP medium (300 mg l -1 KH 2 PO 4 ) is nearly twice that of MS medium (170 mg l -1 KH 2 PO 4 ). High phosphorus in medium causes the growth of the basal plate and with increasing the surface of the explant, its potential for organogenesis increases (Kahane et al. 1992). Effect of growth regulators on micropropagation indices Comparison of means showed that the best treatment in terms of callus production was 0.2 mg l -1 NAA with 1 mg l -1 Kin (47.14% callus growth) (p< 0.01) (Table 7). Farhadi et al. (2017) reported that the highest amount of callus in shallot explants ( A. hirtifolium ) was obtained in MS medium with 1.5 mg l -1 2.4, D and 0.5 mg l -1 BAP. In the present study, it was observed that with increasing cytokinin content, callus growth decreased, which was reported by Toaima et al. (2003) in A. ampeloprasum , Tiwari et al. (2004) in A. cepa and Yan et al. (2009) in A. sativum ,. Based on results, it was observed that much time is required for callus emergence. Callus induction in monocots requires more time than in dicotyledons (Zheng et al. 1998; Luciani et al. 2006). Therefore, in this study, direct organogenesis was targeted. The results of analysis of variance indicated that the effect of auxin and cytokinin on the number of seedlings per explant was significant (p<0.01). BA was better than Kin in direct seedling production. The highest number of seedlings per explant (7.17 per explant) was obtained in the treatment of 0.6 mg l -1 NAA in combination with 4 mg l -1 BA. Cytokinin plays a key role in shoot production in in vitro culture medium (Xu et al. 2008; Hailekidan et al. 2013; Fan et al. 2017). Kahane et al. (1992) reported that in the absence of cytokinin no shoots were formed in onion explants and the highest number of shoots obtained at 100 μM BA. However, in this study, some albino seedlings emerged from some cytokinin-free treatments (Fig. 3). Plant tissues contain varying amounts of different types of plant growth regulators that affect their response to external growth regulators in the culture medium. As a result, different species will react differently to regulators and even at different times (Winson et al. 2020). Pelkonen and Kauppi (1999) were investigated the effect of different auxins on direct bulblet emerging of Lillium Regale Wil and was reported that among the various auxins (NAA, 2,4, D and IAA) used, NAA had the best results in terms of organogenesis and especially direct bulblet formation. The superiority of NAA over other types of auxins in organ regeneration in onion plants has been proven in many studies (Ghosh and Sen 1991; Wang et al. 1993; Buiteveld et al. 1993; Mizuguchi and Ohkawa 1994; Cid et al. 1994; Yan et al. 2009; Farhadi et al. 2017). Therefore, in this study only NAA was used and it was observed that alone (without the use of cytokinin) it is able to directly from bulblets and seedlings and caused high callus growth. The highest number of bulblets per explant was observed in the treatment of 0.6 mg l -1 NAA and 4 mg l -1 BA (6.17 bulblets). Based on the results, the best combination of growth regulators for direct organogenesis (seedling and bulblet) in A.elburzense W. explants was 0.6 mg l -1 NAA and 4 mg l -1 BA (Table 7). Naik and Nayak (2005) investigated the direct organogenesis of Ornithogalum virens and reported that the highest number of bulblets (12-15 bulblets) was obtained in the treatment of 1 mg l -1 NAA, 2 mg l -1 BA and 60 g.l -1 sucrose during 8 weeks. Direct production of bulblets from bulb explants (basal plate and scales) in other Allium species including A. hirtifolium Boiss. (Ebrahimi et al. 2014), A. sativum (Yan et al., 2009) and other bulbous plant (Bach, 1992; Slabbert and Niederwieser, 1999) have been reported. Carbohydrates effect The results showed that no seedlings were formed at any of the concentrations of sucrose used. Concentrations of 120 and 150 g.l -1 sucrose caused the highest increase in the diameter of the basal plate and concentrations of 60 and 30 g.l -1 showed the lowest (Fig. 4 (left)). The highest number of bulblets per explant was obtained in 120 and 150 g.l -1 sucrose treatments (Fig. 4 (right)). Sucrose is stored as starch in the storage organs (such as bulb scales) of most bulbous plants and has been reported that sucrose to increase the formation of various storage organs (bulbs, corms, tubers and rhizomes) in plants with these organs (Arora et al. 1996; Nayak 2000; Naik and Nayak 2005). Proliferation index The explant multiplication coefficient was considered as the sum of seedlings and bulblets consisting of one explant at a certain period. Explants were cultured in the best treatments based on previous experiments (MSP medium completed with 0.6 m.l -1 NAA, 4 m.l -1 BA, 120 g.l -1 sucrose) and observed that an average of 26.83 seedlings and bulblets were obtained from an A. elburzense W. bulb during 18 weeks. The bulblets that formed the seedlings were transferred to the pot and were placed for 10 days at 20 °C, 10 hours of light and 14 hours of darkness, at a light intensity of 40 µmol m -2 s -1 (a combination of cool fluorescent lamps with white and yellow light with a ratio of 50:50), then were transferred to the open space in March. Eighty two percent of the seedlings continued to grow and formed a leaf, after 19 days were dried. The leaves are the main organ used by humans, reaching full growth 3 weeks after emergence. It was observed that after transferring the seedlings to the pot, they acted like mother plants and reached their maximum growth after 2 to 3 weeks. In results, by using obtained protocol, the production of high-performance and low cost is possible. Fig. 5 shows images of direct organogenesis in valak explants. Morpho-Phenology traits and life cycle After the emergence of the germination hook in early march 2015 (seed method), it took between 10-14 days for the length of the seedling (root and aerial part) to reach a constant value (11-20 cm). After 3 weeks of germination and emergence of the radicle, a small bulb with a diameter of 3-4 mm and a length of 3-5 mm was formed at a depth of 6-8 cm (Fig. 6). After another 2 weeks, the bulb size increased (5-7 mm in diameter and 6-8 mm in length) and finally the aerial part dried (35-40 days from germination to aerial drying). Based on results in the first year, seedlings from seeds formed only one leaf and seedlings from bulb yielded 1.33 ± 0.02 leaves (Table 8). Plants grown from seed did not form inflorescence in the first year and the average weight of the formed bulbs was 1.05 ± 0.04 gr. 73.33 ± 1.05 % of the plants from bulb cultivation formed inflorescences with an average diameter of 5.07 ± 0.11 cm. The weight of bulbs at the first year was 11.65 ± 0.42 gr. In early March 2016, plant grown from seed sowing formed 1.48 ± 0.05 leaves and 13.33 ± 0.11% of them formed inflorescences with a diameter of 4.4 ± 0.07cm. The average weight of bulbs obtained from seed cultivation in the second year (2015/2016) was 6.76 ± 0.13 gr. In the third year of growth, the plants obtained from seed cultivation reached full maturity and formed 1.71 ± 0.08 leaves, 87.66 ± 1.35 % formed inflorescences with a diameter of 5.85 ± 0.16 and the bulbs weighed 12.92 ± 0.21 gr. The mean of studied traits in plants from bulb cultivation in the second and third years were not significantly different (Table 8). Therefore, it can be concluded that with bulb cultivation, in the first and second year, the desired result (leaf and bulb harvest or seed production) was achieved, but from seed cultivation to seed production in A. elburzense W. taken 3 years. After the third year, was observed that the bulbs did not reproduce. While Kamenetsky and Rabinoswitch (2006) reported that the bulb of most Allium species is a permanent organ, which will be completely replaced by a new bulb after two years, the study found that one bulb per seed it was formed and the bulb remained stable and was not replaced by another bulb for 3 years. Flower and seed phenology The time of flowering in Karaj farm varied from April 6 (the earliest specimen) to April 17 (the latest specimen). The growth stages of inflorescences and flowers in A. elburzense W. species can be seen in Figure 11. Like other alliums, A. elburzense W. seeds are initially flat and round, and as they matured, their surface becomes shrink and black due to the presence of phytomelanin (Rahn 1998) (Fig. 7). Study of reproductive stages from flower bud formation to opening and seed formation revealed that A. elburzense W. is a protander plant (Fig. 8). According to protander nature of this species, it can be acknowledged that this plant is a metamorphosis and is pollinated by flowers from other plants. Conclusion The results showed that stratification period alone caused the germination of A. elburzense W. seeds. However, due to the hardness of the endosperm and the inhibition of radicle emerging, a combination of stratification (45 days at 5˚C) and scarification (sandpaper) led to better results. Based on results, it was determined that A. elburzense W. seeds had physiological and mechanical dormancy. Indiscriminate harvesting of valak from wild in the vegetative stage and before seed formation, lack of cultivation and low percentage of seed germination, it has made this valuable and endemic plant endangered. The results of the present study and the appropriate proliferation coefficient obtained, can be considered in 1) preserving the genetic resources of these species, 2) low cost production of virus-free plants, 3) reproduction facilitating, 4) Initiation and development of its cultivation and 5) preventing the extinction of these species. According to phenological evaluations, it takes 3 years from seed sowing to seed formation in A. elburzense W., while with bulb sowing, usable seeds and leaves can be obtained in the same year. Also, it was determined that A. elburzense W. is a protander and cross pollination species. Declarations Ethical statement The research meets all applicable standards with regard to the ethics of experimentation and research integrity, and the following is being certified true. As an expert scientist and along with co-authors of concerned field, the paper has been submitted with full responsibility, following due ethical procedure, and there is no duplicate publication, fraud, plagiarism, or concerns about animals or human experimentation. A disclosure/ conflict of interest statement None of the authors of this paper has a financial or personal relationship with other people or organization that could inappropriately influence or bias the content of the paper. It is to specifically state that “No Competing interests are at stake and there is No Conflict of Interest” with other people or organization that could inappropriately influence or bias the content of the paper. 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Indian J Vegetable Sci 31: 107-111. https://doi.org/10.1007/s11627-013-9510-3 Toaima N, Novak E, Schumann G (2003) Callus induction from different explants of commercial cultivars of leek, Allium ampeloprasum var. porrum L.. Acta Hortic (ISHS) 597: 303-309. https://doi.org/10.17660/ActaHortic.2003.597.43 Wang L, Ding H, Lin J (1993) The influence of actinomycin D and cycloheximide on the formation of the masses of embryogenic cells and the globular embryos and on metabolic kinesis of macromolecule in embryogenic calli of F. pallidiflora Schrenk. Acta Gen Sci 20:159–166. https://doi.org/10.1038/2251022a0 Winson KWS, Chew BL, Kathiresan Sathasivam K, Subramaniam S (2020) The establishment of callus and cell suspension cultures of Hylocereus costaricensis for the production of betalain pigments with antioxidant potential. 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Plant Cell Tissue Organ Cult PCTOC 53:99–105. https://doi.org/10.1023/A:1006034623942 Tables Table 1 Effect of stratification period and germination temperature on germination (%) of A. elburzense W. seeds StP (day) GT (°C) Germination (%) 15 5 3.33 g 15 12 1.11 h 15 20 0 i 30 5 18.88 ef 30 12 31.1 d 30 20 14.44 f 45 5 43.99 c 45 12 62.42 a 45 20 47.44 bc 60 5 45.55 bc 60 12 48.3 bc 60 20 61.1 a Means followed by the same letter in column are not significantly different at 5 % (based on Duncan multiple range test) Table 2 Effect of seed age and germination temperature on germination traits of A. elburzense W. seeds SAg GT (°C) Germination (%) Germination rate (s d -1 ) fresh 5 27.03 de 2.58 e fresh 12 43.24 a 4.31 a fresh 20 37.31 b 3.99 abc One year old 5 25.51 e 2.91 de One year old 12 35.73 bc 3.62 bc One year old 20 29.99 cde 3.2 cd Means followed by the same letter in column are not significantly different at 5 % (based on Duncan multiple range test) Table 3 Effect of GA 3 and KNO 3 on germination traits of A. elburzense W. seeds Germination rate (s d -1 ) Germination (%) KNO 3 (%) GA 3 (ppm) 1.4 b 12.91 b 0 0 1.28 b 11.63 b 0.2 0 1.45 b 12.33 b 0 500 2.17 a 19.98 a 0.2 500 Means followed by the same letter in column are not significantly different at 5 % (based on Duncan multiple range test) Table 4 Effect of stratification period and germination temperature on germination traits of A. elburzense W. seeds Sc StP (day) GA 3 (ppm) Germination (%) Germination rate (s/d) Seedling length (cm) Seedling FW (mg) Seedling DW (mg) Bulb height (mm) Bulb width (mm) Without Sc 0 0 2.6 k 0.23 h 4.95 hi 29.83 ef 2.31 f 1.33 d 1.14 e Without Sc 0 500 19.42 gh 1.88 ef 11.85 cd 85.83 cd 6.64 cd 3.79 b 3.18 cd Without Sc 45 0 55.14 c 5.69 b 16.76 b 107.66 b 8.52 a 5.43 a 4.65 a Without Sc 45 500 60.08 b 6.34 a 17.11 b 110.81 ab 8.75 a 5.68 a 4.67 a 10 min H 2 So 4 0 0 13.91 i 1.25 fg 6.01 g 30 e 2.42 f 2.01 cd 1.8 de 10 min H 2 So 4 0 500 28.2 f 2.73 d 10.66 e 26.5 fg 2.03 f 2.18 c 1.99 de 10 min H 2 So 4 45 0 59.7 b 5.6 b 17.01 b 105.74 b 7.61 bc 5.44 a 4.06 ab 10 min H 2 So 4 45 500 64.35 a 5.93 ab 17.44 b 113.5 a 8.26 ab 5.82 a 4.57 a 20 min H 2 So 4 0 0 3.46 k 0.29 h 4.35 i 21.13 g 1.62 g 1.42 d 1.35 e 20 min H 2 So 4 0 500 16.33 hi 1.46 fg 6.93 g 33.55 e 2.12 f 2.71 bc 2.23 d 20 min H 2 So 4 45 0 36.81 de 3.17 c 9.13 f 61.33 e 4.75 e 2.15 c 1.87 de 20 min H 2 So 4 45 500 33.5 e 3.11 c 12.6 c 91.15 c 6.74 cd 2.98 b 2.12 d Sandpaper 0 0 7.8 j 0.6 h 5.5 h 23.5 g 1.95 fg 1.6 d 1.33 e Sandpaper 0 500 24.5 fg 2.11 de 10.36 e 82.66 d 5.28 de 3.2 b 2.64 d Sandpaper 45 0 63.17 ab 5.82 ab 17.33 b 111 ab 8.32 ab 5.28 a 4.24 a Sandpaper 45 500 66.5 a 6.17 a 18.75 a 114 a 8.89 a 5.79 a 4.81 a Means followed by the same letter in column are not significantly different at 5 % (based on Duncan multiple range test) Table 5 Percentage of explant contamination under the influence of 3 factors NaOCl, immersion time and explant type NaOCl (% Cl active) Immersion Time (Min) Explant Type Contamination (%) 1 5 Non cut 68.22 bc 1 5 Cut out 81.39 a 1 10 Non cut 49.16 e 1 10 Cut out 72.5 b 1 15 Non cut 37.8 fg 1 15 Cut out 59.17 d 2 5 Non cut 43.04 ef 2 5 Cut out 66.25 cd 2 10 Non cut 22.93 h 2 10 Cut out 41.31 f 2 15 Non cut 13.76 i 2 15 Cut out 34.67 g 3 5 Non cut 18.33 hi 3 5 Cut out 42.4 f 3 10 Non cut 2.67 j 3 10 Cut out 19.5 hi 3 15 Non cut 0 j 3 15 Cut out 3.14 j Means followed by the same letter in column are not significantly different at 5 % (based on Duncan multiple range test) Table 6 Comparing the mean of two established culture media on the percentage of increase in basal plate width of valak bulb based on the independent T-test Trait MS MSP T Pr>t Diameter of basal plate 4.58 ± 0.32 7.36 ± 0.89 2.93 0.013 Table 7 Mean comparison of the interaction of NAA, BA and Kin on micropropagation indices of A. elburzense W. NAA (mg l -1 ) Cytokinin (mg l -1 ) Callus (%) Number of seedlings in explant Number of bulblets in explant 0 0 0 l 0.33 k 0.33 i 2 BA 0 l 0.66jk 0.67 hi 4 BA 5.7 kl 1.17 ij 0.33 i 6 BA 3.95 kl 0.33 k 0.17 i 1 Kin 10.11 i-k 0.46 k 0.67 hi 2 Kin 1.91 l 0.5 jk 0.33 i 3 Kin 2.11 l 0.17k 0.17 i 0.2 0 33.98 b 0.66jk 0.83 hi 2 BA 37.44 b 1.67 hi 1.33 gh 4 BA 23.24 d-e 4.83 c 1.67 fg 6 BA 8.79 i-k 2.17 gh 1.83 fg 1 Kin 47.14 a 0.33 k 2 ef 2 Kin 11.91 h-j 1.67 hi 0.67 hi 3 Kin 8.98 i-k 0.83 ij 0.33 i 0.4 0 15.3 f-i 1.67 hj 1.67 fg 2 BA 18.48 f-h 5.83 b 2.33 ef 4 BA 9.6 i-k 5.66 b 1.83 fg 6 BA 6.46 jk 2.17 gh 1.67 fg 1 Kin 29.27 b-d 1.5 hi 1.33 gh 2 Kin 17.19 f-h 1.83 gh 1.5 fg 3 Kin 11.93 h-j 1 ij 0.5 i 0.6 0 30.29 b-d 0.17 k 1.83 fg 2 BA 19.27 f-g 4.17 cd 3.67 c 4 BA 3.56 kl 7.17 a 6.17 a 6 BA 19.88 e-g 2.67 fg 3.17 cd 1 Kin 37.89 b 0.83 ij 1.17 hi 2 Kin 13.29 g-j 2.67 fg 2.67 de 3 Kin 0 l 2.17 gh 1.67 fg 0.8 0 27.86 c-e 0.5 jk 1.5 fg 2 BA 32.21 bc 1.33 hi 2.17 ef 4 BA 27.05 c-e 2.5 fg 3.33 cd 6 BA 0 l 3.67 de 4.67 b 1 Kin 12.37 g-j 2 gh 1.83 fg 2 Kin 7.3jk 0.83 ij 2.17 ef 3 Kin 0 l 0.5 jk 1.17 hi Means followed by the same letter in column are not significantly different at 5 % (based on Duncan multiple range test) Table 8 Morphological properties of A. elburzense W. seedlings grown from seed and bulb over 3 years. Propagation method Traits 2014/2015 2015/2016 2016/2017 Seed Leaf number 1 ± 0 c 1.48 ± 0.05 b 1.71 ± 0.08 a Inflorescence formation (%) - 13.33 ± 0.11 b 87.66 ± 1.35 a Inflorescence diameter (cm) - 4.4 ± 0.07 b 5.58 ± 0.16 a Bulb weight (gr) 1.05 ± 0.04 c 6.76 ± 0.13 b 12.92 ± 0.21 a Bulb Leaf number 1.33 ± 0.02 b 1.76 ± 0.03 a 1.67 ± 0.12 a Inflorescence formation (%) 73.33 ± 1.17 b 89.35 ± 1.3 a 84.2 ± 1.22a Inflorescence diameter (cm) 5.07 ± 0.11 b 5.63 ± 0.06 a 5.49 ± 0.18 a Bulb weight (gr) 11.65 ± 0.42 b 13.59 ± 0.24 a 12.69 ± 0.23 ab Means followed by the same letter in row are not significantly different at P < 0.05 level Cite Share Download PDF Status: Posted Version 1 posted 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. 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Also discoverable on Platform About In Review Editorial Policies 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-250915","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":14100832,"identity":"d4081660-2456-4245-8ba0-7d7db232b129","order_by":0,"name":"Sajad Jafari","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA7UlEQVRIie3PsarCMBTG8S8Idgl2rQjtK0ScHMRXiRTqouvlDg6ddAm4OvgupgR0qc4ddXFyqJvgRTyK4JY6XjD/odCQHzkHcLn+ZxIcdXgpoMvXkf6IcLqWLSA+IiCCJ6nxF7EWzfShPKERcW97ML3pH/yZZubXQkQu4+YS9bbiQ2HGU4Egl8hyG4GULdqFrZDgSVDQUqltsPk+vhLpK/8I0yUSVREUMnm8MlABvcKIiCoiin3SpV1iFRyRqV2Ht/NBWjHYKC5OWPeUn9TKy08YhhtjztbBwCXYbf3+BZgdAJ6mz6Tiksvlcn11d67cS+9qIf41AAAAAElFTkSuQmCC","orcid":"","institution":"University of Tehran","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Sajad","middleName":"","lastName":"Jafari","suffix":""},{"id":14100833,"identity":"937d3a8a-6f10-428f-b4e4-eeeb57554cb8","order_by":1,"name":"Mohammadreza Hassandokht","email":"","orcid":"","institution":"Univarsity of Tehran","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Mohammadreza","middleName":"","lastName":"Hassandokht","suffix":""},{"id":14100834,"identity":"48531d49-85b6-42d8-8ba2-02afeb2e3d12","order_by":2,"name":"Madi Taheri","email":"","orcid":"","institution":"Agricultural Research Education and Extension Organization (Zanjan)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Madi","middleName":"","lastName":"Taheri","suffix":""}],"badges":[],"createdAt":"2021-02-17 09:57:18","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-250915/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-250915/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":7568863,"identity":"0ab60421-8cdf-4dff-a555-3b7b9b7b4f0c","added_by":"auto","created_at":"2021-04-01 11:33:28","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":633620,"visible":true,"origin":"","legend":"Explant of A. elburzense W. bulb, A without cutting the ends of bud and B cutting of the bud.","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-250915/v1/68978227ce15acbeca93577e.png"},{"id":7569060,"identity":"f5a0b0a6-9904-4e3c-b763-8080176b395a","added_by":"auto","created_at":"2021-04-01 11:36:28","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":30079,"visible":true,"origin":"","legend":"Effect of stratification period on germination rate of A. elburzense W. seeds (Bars followed by the same letter are not significantly different at 1 % (based on Duncan multiple range test))\n\n","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-250915/v1/0d8b17504e27bf26f3b70375.png"},{"id":7569063,"identity":"29aa7ac8-1aa8-4a11-942f-d2dd85d515b7","added_by":"auto","created_at":"2021-04-01 11:36:28","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":720888,"visible":true,"origin":"","legend":"Albino seedlings of species A. elburzense W. A medium without growth regulators B medium containing 0.2 mg l-1 NAA.","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-250915/v1/e1d7cc083b3d704444576103.png"},{"id":7569061,"identity":"92474246-7c2c-4cc3-9ffd-6cbb3db7c7d0","added_by":"auto","created_at":"2021-04-01 11:36:28","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":43977,"visible":true,"origin":"","legend":"The effect of sucrose different concentrations on the percentage increase in diameter of basal plate (left) and the number of regenerated bulblet per explant (right) in A. elburzense W. (Bars followed by the same letter are not significantly different at 5 % (based on Duncan multiple range test))","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-250915/v1/23190090bc6e048f9353a6c9.png"},{"id":7568867,"identity":"2efbfcda-5084-4f87-a210-f743abd50c0a","added_by":"auto","created_at":"2021-04-01 11:33:28","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":1497964,"visible":true,"origin":"","legend":"A callus proliferation from basal plate, B production of bulblets and emergence plantlets, C plantlet direct production from scales and basal plate, D bulblets and plantlets grown from an explant, E plantlets formation from a bulblet, F Plantlet production from sub cultured bulblet, G Regenerated plantlets and bulbs from a complete explant (one basal plate), H In vitro acclimatization of A. elburzense W. (pretreatment of bulblet on ½ MS + 0.6 m.l-1 NAA for 10 days) and I 3 week-old acclimatized plantlet in peatmoss, cocopeat and perlite (3:1:1)","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-250915/v1/444dbb35e491c788e5e4dcdd.png"},{"id":7569186,"identity":"38778a7a-a4cf-4b02-8ce5-cbddc5f664f5","added_by":"auto","created_at":"2021-04-01 11:39:28","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":1754927,"visible":true,"origin":"","legend":"A Germination hook, after 11 days B leaf, after 18 days C bulb formation, after 20 days from seed sowing in A. elburzense W.","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-250915/v1/6d5b67783186c5021f1cf63b.png"},{"id":7569064,"identity":"a42a5b36-43ca-4134-a1d5-bde1e25972db","added_by":"auto","created_at":"2021-04-01 11:36:28","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":1958194,"visible":true,"origin":"","legend":"A Inflorescence development stages from the time of emergence B flower development stages from the time of inflorescence opening C, D and E seed (C, D and E using stereomicroscope) of A. elburzense W.","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-250915/v1/346ab025c23c552a0c0d8f8b.png"},{"id":7569187,"identity":"8f765707-ad96-48b9-b37d-41884ebddf99","added_by":"auto","created_at":"2021-04-01 11:39:28","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":1046138,"visible":true,"origin":"","legend":"Protandry in A. elburzense W. A mature anthers and immature pistil B flowers with mature pistil and anthers dried and dropped.","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-250915/v1/271dfea2e4d978c95f3411e0.png"},{"id":15672147,"identity":"6ac8e7cc-3ece-47ce-a7e4-8acb6c13efb0","added_by":"auto","created_at":"2021-11-18 14:10:12","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":7037940,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-250915/v1/0f33e531-e00b-4771-abad-79b380dae4e4.pdf"}],"financialInterests":"","formattedTitle":"\u003cp\u003eOptimizing proliferation and assessment of valak morpho-phenological traits; an endangered nutritious \u003cem\u003eAllium \u003c/em\u003eendemic to Iran\u003c/p\u003e","fulltext":[{"header":"Key Message","content":"\u003cp\u003eIn this study, a very valuable and widely used plant in Iran that was in danger of extinction due to excessive harvesting from wild and lack of cultivation was studied, and a rapid and low-cost micropropagation system was introduced. In addition, by reviewing life cycle and behavior of its various organs, the barriers to sexual reproduction were removed. According to the results obtained and high price of this species in Iran, not only will its extinction be prevented, but it will be a very good choice for cultivation and investment.\u003c/p\u003e"},{"header":"Introduction","content":"\u003cp\u003eThe subgenus \u003cem\u003eMelanocrommyum\u003c/em\u003e (Webb \u0026amp; Berthel.) Rouy is the second largest subgenus of the genus \u003cem\u003eAllium\u003c/em\u003e, which consists of 10 section and has over 170 species. \u003cem\u003eAllium elburzense\u003c/em\u003e W. belongs to the Asteroprason section and called \u0026ldquo;valak\u0026rdquo; in Iran and used as fresh vegetable and medical agent (Jafari et al. 2017). This species is found only in mountainous areas of northern Iran in sandy slopes of the Alborz mountain range (Frisch and Abbasi, 2013). Valak is a very expensive vegetable in Iran and its leaves uses for the preparation of a variety of special soups and \u0026ldquo;valak pilaw\u0026rdquo;. Its inflorescence uses to prepare pickles, spices and food decoration, and its bulb is consumed in the pharmaceutical industry (Jafari, 2017). This species is not cultivated, but collected from the nature (Akhavan et al. 2015). Indiscriminate harvesting of valak from natural habitats, harvesting pre-flower and before seed formation with bulb (permanent organ) and low percentage of seed germination has caused the endemic species is exposure to extinction. Reducing the number of individuals in a habitat has led to a decrease in genetic diversity, which in turn will increase the vulnerability of the species to environmental changes, genetic drift and natural selection. Domestication of these valuable species is a suitable way to preserve desirable species and prevent their extinction (Ebrahimi, 2014). One of the main steps in the process is to study the methods of reproduction and preservation of genetic resources of these species. Therefore, awareness and promotion of knowledge about the life cycle of a species, the effect of the environment on its reproduction and its phenological and genetic traits are of great importance. As such knowledge will provide a better understanding of their phenology in order to predict the most favorable periods for seedling establishment in the field and facilitate the provision of nursery plants from seed and organ \u003cem\u003ein vitro\u003c/em\u003e culture (Lentz and Johnson, 1998).\u003c/p\u003e\n\u003cp\u003eLow seed germination speed and rate, slow seedling growth and low energy storage of seeds are some of the germination problems of some \u003cem\u003eAllium\u003c/em\u003e species. Seed dormancy has been reported in a large number of wild Alliums, so it seems necessary to study methods to eliminate it in this species (Dashti et al. 2012; Ebrahimi et al. 2014; Phillips 2010). Seed and bulb dormancy in wild allium species is an adaptation to environmental conditions to avoid germination and growth in inappropriate seasons, so, in order to eliminate the dormancy of these organs, their natural environment should be modeled. In addition to sexual reproduction, which has problems such as segregation of traits, time consuming and more costly, asexual methods such as tissue and organ culture can be used to produce these species. Tissue culture is the most effective virus-free and rapid propagation technology and provides an efficient tool for producing virus-free \u003cem\u003eAllium\u003c/em\u003e plants (Gimenez et al. 2016; Taşkın et al. 2013).\u003c/p\u003e\n\u003cp\u003eThere is no reliable information on the dormancy of seeds and bulbs and how to facilitate its propagation and conservation of \u003cem\u003eA. elburzense\u003c/em\u003e W.. Therefore, due to the high nutritional, pharmaceutical and economic value of valak on the one hand, and its lack of cultivation on the other hand, this study was design and performed in order to domestication (evaluation of phenological behaviors of growth and reproduction stages by both bulb and seed pathway), elimination barriers to sexual and asexual reproduction and achievement a rapid and low-cost proliferation per unit of time.\u003c/p\u003e"},{"header":"Material And Methods","content":"\u003cp\u003e\u003cstrong\u003ePlant material\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBulb and seed of species \u003cem\u003eA. elburzense\u003c/em\u003e W. (Iranian Valak) were collected from the Kalha habitat (in Alborz province at an altitude 2454 m, 36\u0026deg;44ʹ N and 51\u0026deg;17ʹ E with average precipitation 411.2 mm and average temperature of 11.8\u0026deg;C (herbarium no. 006467).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSexual propagation\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStratification period (StP), Germination temperature (GT) and seed age (SAg)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSeeds were disinfected into sodium hypochlorite 1% before use for 5 minutes and then were washed 3 times with distilled water. A factorial experiment was conducted in a completely randomized design (CRD) with 3 factors and 3 replications (each replicate containing 100 seeds). The factors were included stratification period (StP) (a1= 15, a2= 30, a3= 45 and a4= 60 days of moist chilling at 5 \u0026deg;C), seed age (SAg) (b1= fresh seed and b2= one year old seed) and germination temperature (GT) (c1= 5, c2= 12 and c3= 20 \u0026deg;C). Percent and rate of germination were measured in the test.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eScarification (sc) , stratification, GA\u003csub\u003e3\u003c/sub\u003e and KNO\u003csub\u003e3\u003c/sub\u003e \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA factorial experiment was conducted as a CRD design with 4 factors in 3 replications. The factors were included scarification (Sc) (a1= without scarification, a2= scarification with 95% sulfuric acid for 10 minutes and a3= scarification with 95% sulfuric acid for 20 minutes, and a4= sandpaper for 30 seconds), moist stratification (St) (b1= 0 and b2= 45 days at 5\u0026deg;C), GA\u003csub\u003e3 \u003c/sub\u003e(immersed seeds in c1= 0 and c2= 500 ppm for 12 h) and KNO\u003csub\u003e3\u003c/sub\u003e (immersed seeds in d1= 0 and d2= 0.2% for 24 h). Percent and rate of germination, seedling length, seedling fresh and dry weight and bulb height and width were measured. The seeds and filter papers were kept wet with distilled water. The petri dishes were placed in a germinator at 20\u0026deg;C, 16 h light and 8 h dark conditions for 20 days. Seeds in which the tip of the root appeared were considered germinated (radicle protrusion) (Albert et al. 2002). Treated seeds were planted in pots (25 seeds/plot) containing coco-peat and perlite (with a ratio 3:1).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAsexual propagation (\u003cem\u003eIn vitro\u003c/em\u003e)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePlant material\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe collected bulbs were placed in a cool temperature (15-18 \u0026deg;C) for a week in order to drying the surface moisture and avoidance of rotting, and then placed in a refrigerator of 5 \u0026deg;C (June 2015). Bulbs were reviewed in November and explants prepared after observing the sprouts grown. The bulb central bud with basal plate were used as explant.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDisinfection test\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn order to find the best disinfectant concentration and duration of treatment, a factorial experiment based on CRD was conducted with 3 factors in five replications included sodium hypochlorite (NaOCl), immersion time and type of explants (cut out before or after treatment (Fig. 1)).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCulture conditions and establishment medium\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMS medium (Murashige and Skoog, 1962) containing 3 % (w v\u003csup\u003e\u0026minus;1\u003c/sup\u003e) sucrose and 0.7 % agar was used. According to past researches, MS is the most suitable medium for \u003cem\u003ein vitro\u003c/em\u003e culture of \u003cem\u003eAlliums\u003c/em\u003e (yan et al. 2009; Ebrahimi et al. 2014; Farhadi et al. 2017). The medium pH was adjusted to 6 before autoclaving at 121 \u0026deg; C for 20 minutes. The cultivated explant were placed in a growth chamber at a light intensity of 40 \u0026mu;mol m\u003csup\u003e-2\u003c/sup\u003e s\u003csup\u003e-1\u003c/sup\u003e (a combination of cool fluorescent lamps with white and yellow light with a ratio of 50:50) at 25 \u0026plusmn; 1 \u0026deg;C. The explants were cultured in MSP and MS media in six replicates to determination of establishment medium. The amount of phosphorus in MSP (with 300 mg L\u003csup\u003e-1\u003c/sup\u003e KH2PO4) medium was twice the MS (170 mg L-1 KH2PO4) medium, approximately (Kahane et al. 1992). Diameter of basal plate was measured after 45 days with digital caliper and its increase was calculated as a percentage.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEffect of growth regulators on micro propagation indices \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe explants were divided into two parts from the base plate after 45 days and then transferred to MS containing growth regulators. A factorial experiment based on CRD was conducted with two factors in six replications, including NAA (a1= 0, a2= 0.2, a3= 0.4, a4= 0.6 and a5= 0.8 mg l\u003csup\u003e-1\u003c/sup\u003e) and cytokinin contains benzyl adenine and Kinetin (b1= 0, b2= 2BA, b3= 4BA, b4= 6BA, b5= 1Kin, b6= 2Kin and b7= 3Kin mg l\u003csup\u003e-1\u003c/sup\u003e). Explants were placed in 14 hours light and 10 hours dark (based on pre-treatment) and characteristics of callus growth index (equation 1) (Fan et al. 2017), number of days to callus emergence, number of bulblet and shoot per explant were calculated.\u003c/p\u003e\n\u003cp\u003eCallus growth index= (final callus fresh weight - initial callus fresh weight)]/final callus fresh weight\u0026times; 100 \u0026nbsp;\u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEffect of sucrose concentration on micro propagation indices\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eEffect of sucrose different concentrations (30, 60, 90, 120 and 150 g l\u003csup\u003e-1\u003c/sup\u003e) on propagation indices of \u003cem\u003eA. elburzense\u003c/em\u003e W. was evaluated in a completely randomized design with six replications. Explants were placed in 14 hours light and 10 hours dark. After 8 weeks, callus growth index, number of days to callus emergence, number of bulblet and shoot per explant were calculated.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCalculation of \u003cem\u003ein vitro\u003c/em\u003e propagation coefficient (direct organogenesis)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eExplant of \u003cem\u003eA. elburzense W.\u003c/em\u003e was cultured in the best condition (establishment medium, BA, cytokinin, sucrose) based on the results of previous experiments and propagation coefficient was calculated for an explant after 18 weeks. Propagation coefficient was total number of shoot and bulblet composed of a complete explant, in a given period of time.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcclimation and transfer to non-controlling condition\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe formed bulblet were separated and cultured in pots containing perlite and cocopeat with a 3:1 ratio and were transferred to ambient temperature (non-controlling condition) in late February 2016.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMorpho-phenological evaluation of \u003cem\u003eA. elburzense\u003c/em\u003e W.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis experiment was carried out in order to evaluation of growth phenology and stages of reproductive organs production of \u003cem\u003eA. elburzense\u003c/em\u003e W., duration 3 years, at the Horticultural Sciences Research Station of the University of Tehran (long. 51\u0026deg; 10\u0026prime; 35\u0026prime;\u0026prime; N, lat. 35\u0026deg; 42\u0026prime; 18\u0026prime;\u0026prime; E, height 1297 m), in the both form of sowing seeds and bulbs. The dried and mature inflorescences (previously marked) and bulbs were collected from the Kalha habitat in May 2014 and kept in the shade at 25 \u0026deg;C. After separating the hollow and immature seeds, 30 seeds were planted in trays of 30 cells with a mixture combination of peat moss, coco peat and perlite bed in a proportion of 6:1:1 in 3 replications. Observations were recorded until the formation of inflorescences and seeds. The bulbs from the seedlings harvested and were kept at 17 \u0026deg;C in a place with low humidity and then planted in November 2015 10 cm pots (10 bulbs per pot) in mixture of soil and sand in proportion of 1:1. Simultaneously with sowing seeds, bulbs collected from this habitat were also planted in the field to study bulb behavior and asexual reproduction. Developmental phases from flower formation to seed maturity were examined using a stereomicroscope (S21, Berlin, Germany). After the end of the growing season and drying of the aerial part, the bulbs were harvested and weighed before planting at the beginning of the cold season in both planting methods (seed and bulb). In June 2016 percent of inflorescence formation, bulb diameter and weight, number of leaf and plant height were evaluated. In early autumn 2016 the harvested bulbs were planted. In June 2017, bulbs were harvested and morphological traits were measured.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData on indices of seed germination and micropropagation were analyzed with the statistical analysis software SPSS v.21 and MSTATC (version 2.1 USA). Comparisons between establishment media were performed using independent t-test. All data were subjected to analysis of variance and means were compared using Duncan\u0026rsquo;s multiple range test at \u003cem\u003eP \u003c/em\u003e\u0026lt; 0.01.\u003c/p\u003e"},{"header":"Results And Discussion","content":"\u003cp\u003e\u003cstrong\u003eFacilitate germination and remove obstacles\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEffect of germination temperature, seed age and stratification period\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe results showed that the highest germination rate was obtained in 45 and 60 day stratification (Fig. 2). Based on the comparison of means, the highest germination percentage was observed in treatments of 45 days StP \u0026times; GT 12 \u0026deg;C (62.42%) and 60 days StP \u0026times; GT 20 \u0026deg;C (61.1%) (Table 1).\u003c/p\u003e\n\u003cp\u003eTherefore, it can be said that the seeds of \u003cem\u003eA. elburzense\u003c/em\u003e W. need at least 45 days StP at 5 \u0026deg;C for germination. In the treatment of SAg\u0026times;GT, the highest germination percentage and rate were observed in fresh seed and GT 12 \u0026deg;C with 43.24 % and 4.31 s d\u003csup\u003e-1\u003c/sup\u003e, respectively (Table 2).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEffect of scarification, stratification, GA\u003csub\u003e3\u003c/sub\u003e and KNO\u003csub\u003e3\u003c/sub\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAccording to the results of the previous experiment, the GT 12 \u0026deg;C and Stp 45 days were considered for this experiment. The use of GA\u003csub\u003e3\u003c/sub\u003e improved the studied traits compared to the control. KNO\u003csub\u003e3 \u003c/sub\u003ehad no significant effect on the studied traits. No seeds germinated in 10 minutes using of sulfuric acid without StP. The results showed that the interaction effect of GA\u003csub\u003e3\u003c/sub\u003e and KNO\u003csub\u003e3\u003c/sub\u003e led to a significant increase in germination percentage (19.98 %) and germination rate (2.17 s d\u003csup\u003e-1\u003c/sup\u003e) (Table 3).\u003c/p\u003e\n\u003cp\u003eThe results of ANOVA showed that the triple interaction of Sc, StP and GA\u003csub\u003e3\u003c/sub\u003e had a significant effect on seedling length, seedling fresh and dry weight, bulb height and diameter at the statistical level of 1%. Comparison of means showed that the treatments in which the StP (45 days) was applied had better effects on the studied traits than other treatments (Table 4). Although all \u003cem\u003eAllium\u003c/em\u003e species follow the same germination pattern, the mechanisms of seed dormancy differ in taxonomic groups and in different habitats (Kamenetsky and Rabinoswitch 2006). Specht and Keller (1997) examined the germination temperatures of 94 \u003cem\u003eAllium\u003c/em\u003e species and the appropriate seed germination temperature for \u003cem\u003eAllium\u003c/em\u003e subspecies was 16 \u0026deg;C, for \u003cem\u003eRhiziridium\u003c/em\u003e subspecies between 16-26 \u0026deg;C, and 5-15 \u0026deg;C was also reported for species belonging to the \u003cem\u003eMelanocrommyum\u003c/em\u003e section. The use of sulfuric acid removed the black seed coat, but had no significant effect on seed germination indices. However, sandpaper had a better effect than sulfuric acid and caused significant seed germination and improved seedling quality indices (Table 4). In the present study, it was observed that without using the appropriate StP, the seeds did not germinate significantly. These results were also reported by Ebrahimi et al. (2014) in shallot (\u003cem\u003eA. hirtifolium\u003c/em\u003e Boiss). According to the obtained results, dormancy of \u003cem\u003eA. elburzense\u003c/em\u003e W. seed can be considered as physiological and physical type. GA\u003csub\u003e3\u003c/sub\u003e increases the growth potential and strength of the embryo, overcoming resistant shells and mechanical barriers (pericarp, seed coat and endosperm) for germination and seedling growth (Finch-Savage and Leubner-Metzger 2006). Endosperm attenuation is regulated by phyto-hormones and environmental factors. Radicle emerging can be accelerated by GA\u003csub\u003e3\u003c/sub\u003e and inhibited by abscisic acid (Silva et al. 2004; Finch-Savage and Leubner-Metzger 2006; Linkies et al. 2010). The hard and thick endosperm in species of \u003cem\u003eMelanocrommyum\u003c/em\u003e section is a type of plant acclimation to low temperatures in the main habitats, which protects the embryo during the winter cold.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMicropropagation\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDisinfection of explants\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe results showed that the lowest contamination (0%) was observed in the treatment of 3% active chlorine and 15 minutes immersion time, and the highest contamination in the explant (78%) belonged to the treatment of 1% active chlorine for 5 minutes in cut explants (Table 5). As a result, 3% active chlorine treatment and 15 minutes of immersion were used for uncut explants.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEstablishment medium\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe results of T-test showed that there was a significant difference between the two used establishment culture medium, basal plate in MSP medium indicated an increase with an average of 7.36% than explant cultured in MS medium (Table 6). The amount of phosphorus in MSP medium (300 mg l\u003csup\u003e-1\u003c/sup\u003e KH\u003csub\u003e2\u003c/sub\u003ePO\u003csub\u003e4\u003c/sub\u003e) is nearly twice that of MS medium (170 mg l\u003csup\u003e-1 \u003c/sup\u003eKH\u003csub\u003e2\u003c/sub\u003ePO\u003csub\u003e4\u003c/sub\u003e). High phosphorus in medium causes the growth of the basal plate and with increasing the surface of the explant, its potential for organogenesis increases (Kahane et al. 1992).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEffect of growth regulators on micropropagation indices\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eComparison of means showed that the best treatment in terms of callus production was 0.2 mg l\u003csup\u003e-1\u003c/sup\u003e NAA with 1 mg l\u003csup\u003e-1\u003c/sup\u003e Kin (47.14% callus growth) (p\u0026lt; 0.01) (Table 7). Farhadi et al. (2017) reported that the highest amount of callus in shallot explants (\u003cem\u003eA. hirtifolium\u003c/em\u003e) was obtained in MS medium with 1.5 mg l\u003csup\u003e-1\u0026nbsp; \u003c/sup\u003e2.4, D and 0.5 mg l\u003csup\u003e-1 \u003c/sup\u003eBAP. In the present study, it was observed that with increasing cytokinin content, callus growth decreased, which was reported by Toaima et al. (2003) in \u003cem\u003eA. ampeloprasum\u003c/em\u003e, Tiwari et al. (2004) in \u003cem\u003eA. cepa\u003c/em\u003e and Yan et al. (2009) in \u003cem\u003eA. sativum\u003c/em\u003e,. Based on results, it was observed that much time is required for callus emergence. Callus induction in monocots requires more time than in dicotyledons (Zheng et al. 1998; Luciani et al. 2006). Therefore, in this study, direct organogenesis was targeted. The results of analysis of variance indicated that the effect of auxin and cytokinin on the number of seedlings per explant was significant (p\u0026lt;0.01). \u003ca name=\"_Toc486331988\"\u003e\u003c/a\u003eBA was better than Kin in direct seedling production. The highest number of seedlings per explant (7.17 per explant) was obtained in the treatment of 0.6 mg l\u003csup\u003e-1\u003c/sup\u003e NAA in combination with 4 mg l\u003csup\u003e-1 \u003c/sup\u003eBA. \u0026nbsp;Cytokinin plays a key role in shoot production in \u003cem\u003ein vitro\u003c/em\u003e culture medium (Xu et al. 2008; Hailekidan et al. 2013; Fan et al. 2017). Kahane et al. (1992) reported that in the absence of cytokinin no shoots were formed in onion explants and the highest number of shoots obtained at 100 \u0026mu;M BA. However, in this study, some albino seedlings emerged from some cytokinin-free treatments (Fig. 3). Plant tissues contain varying amounts of different types of plant growth regulators that affect their response to external growth regulators in the culture medium. As a result, different species will react differently to regulators and even at different times (Winson et al. 2020).\u003c/p\u003e\n\u003cp\u003e\u003ca name=\"_Toc486331989\"\u003e\u003c/a\u003ePelkonen and Kauppi (1999) were investigated the effect of different auxins on direct bulblet emerging of \u003cem\u003eLillium Regale\u003c/em\u003e Wil and was reported that among the various auxins (NAA, 2,4, D and IAA) used, NAA had the best results in terms of organogenesis and especially direct bulblet formation. The superiority of NAA over other types of auxins in organ regeneration in onion plants has been proven in many studies (Ghosh and Sen 1991; Wang et al. 1993; Buiteveld et al. 1993; Mizuguchi and Ohkawa 1994; Cid et al. 1994; Yan et al. 2009; Farhadi et al. 2017). Therefore, in this study only NAA was used and it was observed that alone (without the use of cytokinin) it is able to directly from bulblets and seedlings and caused high callus growth.\u003c/p\u003e\n\u003cp\u003eThe highest number of bulblets per explant was observed in the treatment of 0.6 mg l\u003csup\u003e-1\u003c/sup\u003e NAA and 4 mg l\u003csup\u003e-1\u003c/sup\u003e BA (6.17 bulblets). Based on the results, the best combination of growth regulators for direct organogenesis (seedling and bulblet) in \u003cem\u003eA.elburzense\u003c/em\u003e W. explants was 0.6 mg l\u003csup\u003e-1\u003c/sup\u003e NAA and 4 mg l\u003csup\u003e-1\u003c/sup\u003e BA (Table 7). Naik and Nayak (2005) investigated the direct organogenesis of \u003cem\u003eOrnithogalum virens\u003c/em\u003e and reported that the highest number of bulblets (12-15 bulblets) was obtained in the treatment of 1 mg l\u003csup\u003e-1\u003c/sup\u003e NAA, 2 mg l\u003csup\u003e-1\u003c/sup\u003e BA and 60 g.l\u003csup\u003e-1\u003c/sup\u003e sucrose during 8 weeks. Direct production of bulblets from bulb explants (basal plate and scales) in other \u003cem\u003eAllium\u003c/em\u003e species including \u003cem\u003eA. hirtifolium\u003c/em\u003e Boiss. (Ebrahimi et al. 2014), \u003cem\u003eA. sativum\u003c/em\u003e (Yan et al., 2009) and other bulbous plant (Bach, 1992; Slabbert and Niederwieser, 1999) have been reported.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCarbohydrates effect\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe results showed that no seedlings were formed at any of the concentrations of sucrose used. Concentrations of 120 and 150 g.l\u003csup\u003e-1\u003c/sup\u003e sucrose caused the highest increase in the diameter of the basal plate and concentrations of 60 and 30 g.l\u003csup\u003e-1\u003c/sup\u003e showed the lowest (Fig. 4 (left)). The highest number of bulblets per explant was obtained in 120 and 150 g.l\u003csup\u003e-1 \u003c/sup\u003esucrose treatments (Fig. 4 (right)). Sucrose is stored as starch in the storage organs (such as bulb scales) of most bulbous plants and has been reported that sucrose to increase the formation of various storage organs (bulbs, corms, tubers and rhizomes) in plants with these organs (Arora et al. 1996; Nayak 2000; Naik and Nayak 2005).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eProliferation index \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe explant multiplication coefficient was considered as the sum of seedlings and bulblets consisting of one explant at a certain period. Explants were cultured in the best treatments based on previous experiments (MSP medium completed with 0.6 m.l\u003csup\u003e-1\u003c/sup\u003e NAA, 4 m.l\u003csup\u003e-1\u003c/sup\u003e BA, 120 g.l\u003csup\u003e-1\u003c/sup\u003e sucrose) and observed that an average of 26.83 seedlings and bulblets were obtained from an \u003cem\u003eA. elburzense\u003c/em\u003e W. bulb during 18 weeks. The bulblets that formed the seedlings were transferred to the pot and were placed for 10 days at 20 \u0026deg;C, 10 hours of light and 14 hours of darkness, at a light intensity of 40 \u0026micro;mol m\u003csup\u003e-2\u003c/sup\u003e s\u003csup\u003e-1\u003c/sup\u003e (a combination of cool fluorescent lamps with white and yellow light with a ratio of 50:50), then were transferred to the open space in March. Eighty two percent of the seedlings continued to grow and formed a leaf, after 19 days were dried. The leaves are the main organ used by humans, reaching full growth 3 weeks after emergence. It was observed that after transferring the seedlings to the pot, they acted like mother plants and reached their maximum growth after 2 to 3 weeks. In results, by using obtained protocol, the production of high-performance and low cost is possible. Fig. 5 shows images of direct organogenesis in valak explants.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMorpho-Phenology traits and life cycle\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAfter the emergence of the germination hook in early march 2015 (seed method), it took between 10-14 days for the length of the seedling (root and aerial part) to reach a constant value (11-20 cm). After 3 weeks of germination and emergence of the radicle, a small bulb with a diameter of 3-4 mm and a length of 3-5 mm was formed at a depth of 6-8 cm (Fig. 6). After another 2 weeks, the bulb size increased (5-7 mm in diameter and 6-8 mm in length) and finally the aerial part dried (35-40 days from germination to aerial drying). Based on results in the first year, seedlings from seeds formed only one leaf and seedlings from bulb yielded 1.33 \u0026plusmn; 0.02 leaves (Table 8).\u003c/p\u003e\n\u003cp\u003ePlants grown from seed did not form inflorescence in the first year and the average weight of the formed bulbs was 1.05 \u0026plusmn; 0.04 gr. 73.33 \u0026plusmn; 1.05 % of the plants from bulb cultivation formed inflorescences with an average diameter of 5.07 \u0026plusmn; 0.11 cm. The weight of bulbs at the first year was 11.65 \u0026plusmn; 0.42 gr. In early March 2016, plant grown from seed sowing formed 1.48 \u0026plusmn; 0.05 leaves and 13.33 \u0026plusmn; 0.11% of them formed inflorescences with a diameter of 4.4 \u0026plusmn; 0.07cm. The average weight of bulbs obtained from seed cultivation in the second year (2015/2016) was 6.76 \u0026plusmn; 0.13 gr. In the third year of growth, the plants obtained from seed cultivation reached full maturity and formed 1.71 \u0026plusmn; 0.08 leaves, 87.66 \u0026plusmn; 1.35 % formed inflorescences with a diameter of 5.85 \u0026plusmn; 0.16 and the bulbs weighed 12.92 \u0026plusmn; 0.21 gr. The mean of studied traits in plants from bulb cultivation in the second and third years were not significantly different (Table 8). Therefore, it can be concluded that with bulb cultivation, in the first and second year, the desired result (leaf and bulb harvest or seed production) was achieved, but from seed cultivation to seed production in \u003cem\u003eA. elburzense\u003c/em\u003e W. taken 3 years. After the third year, was observed that the bulbs did not reproduce. While Kamenetsky and Rabinoswitch (2006) reported that the bulb of most \u003cem\u003eAllium\u003c/em\u003e species is a permanent organ, which will be completely replaced by a new bulb after two years, the study found that one bulb per seed it was formed and the bulb remained stable and was not replaced by another bulb for 3 years.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFlower and seed phenology\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe time of flowering in Karaj farm varied from April 6 (the earliest specimen) to April 17 (the latest specimen). The growth stages of inflorescences and flowers in \u003cem\u003eA. elburzense\u003c/em\u003e W. species can be seen in Figure 11. Like other alliums, \u003cem\u003eA. elburzense\u003c/em\u003e W. seeds are initially flat and round, and as they matured, their surface becomes shrink and black due to the presence of phytomelanin (Rahn 1998) (Fig. 7).\u003ca name=\"_Toc486336140\"\u003e\u003c/a\u003e\u003c/p\u003e\n\u003cp\u003eStudy of reproductive stages from flower bud formation to opening and seed formation revealed that \u003cem\u003eA. elburzense\u003c/em\u003e W. is a protander plant (Fig. 8). According to protander nature of this species, it can be acknowledged that this plant is a metamorphosis and is pollinated by flowers from other plants.\u003cstrong\u003e\u0026nbsp; \u003c/strong\u003e\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe results showed that stratification period alone caused the germination of \u003cem\u003eA. elburzense\u003c/em\u003e W. seeds. However, due to the hardness of the endosperm and the inhibition of radicle emerging, a combination of stratification (45 days at 5˚C) and scarification (sandpaper) led to better results. Based on results, it was determined that \u003cem\u003eA. elburzense\u003c/em\u003e W. seeds had physiological and mechanical dormancy. Indiscriminate harvesting of valak from wild in the vegetative stage and before seed formation, lack of cultivation and low percentage of seed germination, it has made this valuable and endemic plant endangered. The results of the present study and the appropriate proliferation coefficient obtained, can be considered in 1) preserving the genetic resources of these species, 2) low cost production of virus-free plants, 3) reproduction facilitating, 4) Initiation and development of its cultivation and 5) preventing the extinction of these species. According to phenological evaluations, it takes 3 years from seed sowing to seed formation in \u003cem\u003eA. elburzense\u003c/em\u003e W., while with bulb sowing, usable seeds and leaves can be obtained in the same year. Also, it was determined that \u003cem\u003eA. elburzense\u003c/em\u003e W. is a protander and cross pollination species.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthical statement\u003c/strong\u003e\u003c/p\u003e\n\u003cul\u003e\n\u003cli\u003eThe research meets all applicable standards with regard to the ethics of experimentation and research integrity, and the following is being certified true.\u003c/li\u003e\n\u003cli\u003eAs an expert scientist and along with co-authors of concerned field, the paper has been submitted with full responsibility, following due ethical procedure, and there is no duplicate publication, fraud, plagiarism, or concerns about animals or human experimentation.\u003c/li\u003e\n\u003c/ul\u003e\n\u003cp\u003e\u003cstrong\u003eA disclosure/ conflict of interest statement\u003c/strong\u003e\u003c/p\u003e\n\u003cul\u003e\n\u003cli\u003eNone of the authors of this paper has a financial or personal relationship with other people or organization that could inappropriately influence or bias the content of the paper.\u003c/li\u003e\n\u003cli\u003eIt is to specifically state that \u0026ldquo;No Competing interests are at stake and there is No Conflict of Interest\u0026rdquo; with other people or organization that could inappropriately influence or bias the content of the paper.\u003c/li\u003e\n\u003c/ul\u003e"},{"header":"References","content":"\u003cp\u003eAkhavan A, Saeidi H, Rahiminejad MR, Zarre Sh, Blattner FR (2015) Interspecific Relationships in Allium Subgenus Melanocrommyum Sections Acanthoprason and Asteroprason (Amaryllidaceae) Revealed Using ISSR Markers. 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Acta Gen Sci 20:159\u0026ndash;166. \u003ca href=\"https://doi.org/10.1038/2251022a0\"\u003ehttps://doi.org/10.1038/2251022a0\u003c/a\u003e\u003c/p\u003e\n\u003cp\u003eWinson KWS, Chew BL, Kathiresan Sathasivam K, Subramaniam S (2020) The establishment of callus and cell suspension cultures of \u003cem\u003eHylocereus costaricensis\u003c/em\u003e for the production of betalain pigments with antioxidant potential. Indus Crops Prod 155: 112750. \u003ca href=\"https://doi.org/10.1016/j.indcrop.2020.112750\"\u003ehttps://doi.org/10.1016/j.indcrop.2020.112750\u003c/a\u003e\u003c/p\u003e\n\u003cp\u003eXu Z, Yeong-Cheol YC, Kim CH (2008) Effect of plant growth regulators,temperature and sucrose on shoot proliferation from the stem disc of Chinese jiaotou (\u003cem\u003eAllium chinense\u003c/em\u003e) and in vitro bulblet formation. Acta Physiol Planta 30:521\u0026ndash;528. \u003ca href=\"https://doi.org/10.1007/s11738-008-0150-x\"\u003ehttps://doi.org/10.1007/s11738-008-0150-x\u003c/a\u003e\u003c/p\u003e\n\u003cp\u003eYan M, Xu Ch, Kim Ch, Um YC, Bah AA, Guo D (2009) Effects of explant type, culture media and growth regulators on callus induction and plant regeneration of Chinese jiaotou (\u003cem\u003eAllium chinense\u003c/em\u003e). Scientia Horti 123: 124\u0026ndash;128. \u003ca href=\"https://doi.org/10.1016/j.scienta.2009.07.021\"\u003ehttps://doi.org/10.1016/j.scienta.2009.07.021\u003c/a\u003e\u003c/p\u003e\n\u003cp\u003eZheng SJ, Henken B, Sofiari E, Jacobsen E, Krens FA, Kik C (1998) Factors influencing induction, propagation and regeneration of mature zygoticembryo-derived callus from \u003cem\u003eAllium cepa\u003c/em\u003e. Plant Cell Tissue Organ Cult PCTOC 53:99\u0026ndash;105. \u003ca href=\"https://doi.org/10.1023/A:1006034623942\"\u003ehttps://doi.org/10.1023/A:1006034623942\u003c/a\u003e\u003c/p\u003e"},{"header":"Tables","content":"\u003cp\u003e\u003cstrong\u003eTable 1\u003c/strong\u003e Effect of stratification period and germination temperature on germination (%) of \u003cem\u003eA. elburzense\u003c/em\u003e W. seeds\u003c/p\u003e\n\u003ctable border=\"1\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003eStP (day)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"60\"\u003e\n\u003cp\u003eGT (\u0026deg;C)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003eGermination (%)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"60\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e3.33 g\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"60\"\u003e\n\u003cp\u003e12\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e1.11 h\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"60\"\u003e\n\u003cp\u003e20\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e0 i\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e30\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"60\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e18.88 ef\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e30\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"60\"\u003e\n\u003cp\u003e12\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e31.1 d\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e30\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"60\"\u003e\n\u003cp\u003e20\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e14.44 f\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e45\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"60\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e43.99 c\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e45\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"60\"\u003e\n\u003cp\u003e12\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e62.42 a\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e45\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"60\"\u003e\n\u003cp\u003e20\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e47.44 bc\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e60\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"60\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e45.55 bc\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e60\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"60\"\u003e\n\u003cp\u003e12\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e48.3 bc\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e60\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"60\"\u003e\n\u003cp\u003e20\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e61.1 a\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eMeans followed by the same letter in column are not significantly different at 5 % (based on Duncan multiple range test)\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2\u003c/strong\u003e Effect of seed age and germination temperature on germination traits of \u003cem\u003eA. elburzense\u003c/em\u003e W. seeds\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"1\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003eSAg\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"36\"\u003e\n\u003cp\u003eGT (\u0026deg;C)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003eGermination (%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003eGermination rate (s d\u003csup\u003e-1\u003c/sup\u003e)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003efresh\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"36\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e27.03 de\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e2.58 e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003efresh\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"36\"\u003e\n\u003cp\u003e12\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e43.24 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e4.31 a\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003efresh\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"36\"\u003e\n\u003cp\u003e20\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e37.31 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e3.99 abc\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003eOne year old\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"36\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e25.51 e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e2.91 de\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003eOne year old\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"36\"\u003e\n\u003cp\u003e12\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e35.73 bc\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e3.62 bc\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003eOne year old\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"36\"\u003e\n\u003cp\u003e20\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e29.99 cde\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e3.2 cd\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eMeans followed by the same letter in column are not significantly different at 5 % (based on Duncan multiple range test)\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3\u003c/strong\u003e Effect of GA\u003csub\u003e3 \u003c/sub\u003eand KNO\u003csub\u003e3 \u003c/sub\u003eon germination traits of \u003cem\u003eA. elburzense\u003c/em\u003e W. seeds\u003c/p\u003e\n\u003ctable border=\"1\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"82\"\u003e\n\u003cp\u003eGermination rate (s d\u003csup\u003e-1\u003c/sup\u003e)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"82\"\u003e\n\u003cp\u003eGermination (%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"64\"\u003e\n\u003cp\u003eKNO\u003csub\u003e3 \u003c/sub\u003e(%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003eGA\u003csub\u003e3 \u003c/sub\u003e(ppm)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"82\"\u003e\n\u003cp\u003e1.4 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"82\"\u003e\n\u003cp\u003e12.91 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"64\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"82\"\u003e\n\u003cp\u003e1.28 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"82\"\u003e\n\u003cp\u003e11.63 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"64\"\u003e\n\u003cp\u003e0.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"82\"\u003e\n\u003cp\u003e1.45 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"82\"\u003e\n\u003cp\u003e12.33 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"64\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003e500\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"82\"\u003e\n\u003cp\u003e2.17 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"82\"\u003e\n\u003cp\u003e19.98 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"64\"\u003e\n\u003cp\u003e0.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003e500\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eMeans followed by the same letter in column are not significantly different at 5 % (based on Duncan multiple range test)\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 4\u003c/strong\u003e Effect of stratification period and germination temperature on germination traits of \u003cem\u003eA. elburzense\u003c/em\u003e W. seeds\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"1\" width=\"641\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"75\"\u003e\n\u003cp\u003eSc\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003eStP (day)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"62\"\u003e\n\u003cp\u003eGA\u003csub\u003e3 \u003c/sub\u003e(ppm)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003eGermination (%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003eGermination rate (s/d)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003eSeedling length (cm)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003eSeedling FW (mg)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003eSeedling DW (mg)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003eBulb height (mm)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003eBulb width (mm)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"75\"\u003e\n\u003cp\u003eWithout Sc\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"62\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e2.6 k\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e0.23 h\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e4.95 hi\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e29.83 ef\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e2.31 f\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e1.33 d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e1.14 e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"75\"\u003e\n\u003cp\u003eWithout Sc\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"62\"\u003e\n\u003cp\u003e500\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e19.42 gh\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e1.88 ef\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e11.85 cd\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e85.83 cd\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e6.64 cd\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e3.79 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e3.18 cd\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"75\"\u003e\n\u003cp\u003eWithout Sc\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003e45\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"62\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e55.14 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e5.69 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e16.76 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e107.66 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e8.52 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e5.43 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e4.65 a\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"75\"\u003e\n\u003cp\u003eWithout Sc\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003e45\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"62\"\u003e\n\u003cp\u003e500\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e60.08 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e6.34 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e17.11 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e110.81 ab\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e8.75 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e5.68 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e4.67 a\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"75\"\u003e\n\u003cp\u003e10 min H\u003csub\u003e2\u003c/sub\u003eSo\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"62\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e13.91 i\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e1.25 fg\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e6.01 g\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e30 e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e2.42 f\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e2.01 cd\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e1.8 de\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"75\"\u003e\n\u003cp\u003e10 min H\u003csub\u003e2\u003c/sub\u003eSo\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"62\"\u003e\n\u003cp\u003e500\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e28.2 f\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e2.73 d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e10.66 e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e26.5 fg\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e2.03 f\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e2.18 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e1.99 de\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"75\"\u003e\n\u003cp\u003e10 min H\u003csub\u003e2\u003c/sub\u003eSo\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003e45\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"62\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e59.7 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e5.6 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e17.01 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e105.74 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e7.61 bc\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e5.44 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e4.06 ab\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"75\"\u003e\n\u003cp\u003e10 min H\u003csub\u003e2\u003c/sub\u003eSo\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003e45\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"62\"\u003e\n\u003cp\u003e500\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e64.35 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e5.93 ab\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e17.44 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e113.5 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e8.26 ab\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e5.82 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e4.57 a\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"75\"\u003e\n\u003cp\u003e20 min H\u003csub\u003e2\u003c/sub\u003eSo\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"62\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e3.46 k\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e0.29 h\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e4.35 i\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e21.13 g\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e1.62 g\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e1.42 d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e1.35 e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"75\"\u003e\n\u003cp\u003e20 min H\u003csub\u003e2\u003c/sub\u003eSo\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"62\"\u003e\n\u003cp\u003e500\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e16.33 hi\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e1.46 fg\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e6.93 g\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e33.55 e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e2.12 f\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e2.71 bc\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e2.23 d\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"75\"\u003e\n\u003cp\u003e20 min H\u003csub\u003e2\u003c/sub\u003eSo\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003e45\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"62\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e36.81 de\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e3.17 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e9.13 f\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e61.33 e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e4.75 e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e2.15 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e1.87 de\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"75\"\u003e\n\u003cp\u003e20 min H\u003csub\u003e2\u003c/sub\u003eSo\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003e45\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"62\"\u003e\n\u003cp\u003e500\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e33.5 e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e3.11 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e12.6 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e91.15 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e6.74 cd\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e2.98 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e2.12 d\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"75\"\u003e\n\u003cp\u003eSandpaper\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"62\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e7.8 j\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e0.6 h\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e5.5 h\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e23.5 g\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e1.95 fg\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e1.6 d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e1.33 e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"75\"\u003e\n\u003cp\u003eSandpaper\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"62\"\u003e\n\u003cp\u003e500\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e24.5 fg\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e2.11 de\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e10.36 e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e82.66 d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e5.28 de\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e3.2 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e2.64 d\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"75\"\u003e\n\u003cp\u003eSandpaper\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003e45\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"62\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e63.17 ab\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e5.82 ab\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e17.33 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e111 ab\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e8.32 ab\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e5.28 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e4.24 a\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"75\"\u003e\n\u003cp\u003eSandpaper\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"51\"\u003e\n\u003cp\u003e45\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"62\"\u003e\n\u003cp\u003e500\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e66.5 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e6.17 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e18.75 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e114 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"59\"\u003e\n\u003cp\u003e8.89 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e5.79 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"65\"\u003e\n\u003cp\u003e4.81 a\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eMeans followed by the same letter in column are not significantly different at 5 % (based on Duncan multiple range test)\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 5\u003c/strong\u003e Percentage of explant contamination under the influence of 3 factors NaOCl, immersion time and explant type\u003c/p\u003e\n\u003ctable border=\"1\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e\u003cstrong\u003eNaOCl \u0026nbsp;(% Cl active)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e\u003cstrong\u003eImmersion Time (Min)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e\u003cstrong\u003eExplant Type\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e\u003cstrong\u003eContamination (%)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003eNon cut\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e68.22 bc\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003eCut out\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e81.39 a\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003eNon cut\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e49.16 e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003eCut out\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e72.5 b\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003eNon cut\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e37.8 fg\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003eCut out\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e59.17 d\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003eNon cut\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e43.04 ef\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003eCut out\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e66.25 cd\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003eNon cut\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e22.93 h\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003eCut out\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e41.31 f\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003eNon cut\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e13.76 i\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003eCut out\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e34.67 g\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003eNon cut\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e18.33 hi\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003eCut out\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e42.4 f\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003eNon cut\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e2.67 j\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003eCut out\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e19.5 hi\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003eNon cut\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e0 j\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003eCut out\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e3.14 j\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eMeans followed by the same letter in column are not significantly different at 5 % (based on Duncan multiple range test)\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 6\u003c/strong\u003e Comparing the mean of two established culture media on the percentage of increase in basal plate width of valak bulb based on the independent T-test\u003c/p\u003e\n\u003ctable border=\"1\" width=\"360\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"129\"\u003e\n\u003cp\u003e\u003cstrong\u003eTrait\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e\u003cstrong\u003eMS\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"75\"\u003e\n\u003cp\u003e\u003cstrong\u003eMSP\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"37\"\u003e\n\u003cp\u003e\u003cstrong\u003eT\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"48\"\u003e\n\u003cp\u003e\u003cstrong\u003ePr\u0026gt;t\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"129\"\u003e\n\u003cp\u003eDiameter of basal plate\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"70\"\u003e\n\u003cp\u003e4.58 \u0026plusmn; 0.32\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"75\"\u003e\n\u003cp\u003e7.36 \u0026plusmn; 0.89\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"37\"\u003e\n\u003cp\u003e2.93\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"48\"\u003e\n\u003cp\u003e0.013\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 7\u003c/strong\u003e Mean comparison of the interaction of NAA, BA and Kin on micropropagation indices of \u003cem\u003eA. elburzense\u003c/em\u003e W.\u003c/p\u003e\n\u003ctable border=\"1\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"84\"\u003e\n\u003cp\u003e\u003cstrong\u003eNAA (mg l\u003csup\u003e-1\u003c/sup\u003e)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e\u003cstrong\u003eCytokinin (mg l\u003csup\u003e-1\u003c/sup\u003e)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e\u003cstrong\u003eCallus (%)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e\u003cstrong\u003eNumber of seedlings in explant\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e\u003cstrong\u003eNumber of bulblets in explant\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"7\" width=\"84\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e0 l\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e0.33 k\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e0.33 i\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e2 BA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e0 l\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e0.66jk\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e0.67 hi\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e4 BA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e5.7 kl\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e1.17 ij\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e0.33 i\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e6 BA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e3.95 kl\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e0.33 k\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e0.17 i\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e1 Kin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e10.11 i-k\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e0.46 k\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e0.67 hi\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e2 Kin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e1.91 l\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e0.5 jk\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e0.33 i\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e3 Kin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e2.11 l\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e0.17k\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e0.17 i\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"7\" width=\"84\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e0.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e33.98 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e0.66jk\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e0.83 hi\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e2 BA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e37.44 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e1.67 hi\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e1.33 gh\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e4 BA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e23.24 d-e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e4.83 c\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e1.67 fg\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e6 BA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e8.79 i-k\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e2.17 gh\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e1.83 fg\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e1 Kin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e47.14 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e0.33 k\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e2 ef\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e2 Kin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e11.91 h-j\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e1.67 hi\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e0.67 hi\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e3 Kin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e8.98 i-k\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e0.83 ij\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e0.33 i\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"7\" width=\"84\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e0.4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e15.3 f-i\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e1.67 hj\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e1.67 fg\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e2 BA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e18.48 f-h\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e5.83 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e2.33 ef\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e4 BA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e9.6 i-k\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e5.66 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e1.83 fg\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e6 BA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e6.46 jk\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e2.17 gh\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e1.67 fg\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e1 Kin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e29.27 b-d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e1.5 hi\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e1.33 gh\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e2 Kin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e17.19 f-h\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e1.83 gh\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e1.5 fg\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e3 Kin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e11.93 h-j\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e1 ij\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e0.5 i\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"7\" width=\"84\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e0.6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e30.29 b-d\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e0.17 k\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e1.83 fg\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e2 BA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e19.27 f-g\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e4.17 cd\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e3.67 c\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e4 BA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e3.56 kl\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e7.17 a\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e6.17 a\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e6 BA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e19.88 e-g\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e2.67 fg\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e3.17 cd\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e1 Kin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e37.89 b\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e0.83 ij\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e1.17 hi\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e2 Kin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e13.29 g-j\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e2.67 fg\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e2.67 de\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e3 Kin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e0 l\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e2.17 gh\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e1.67 fg\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"7\" width=\"84\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e0.8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e27.86 c-e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e0.5 jk\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e1.5 fg\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e2 BA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e32.21 bc\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e1.33 hi\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e2.17 ef\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e4 BA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e27.05 c-e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e2.5 fg\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e3.33 cd\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e6 BA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e0 l\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e3.67 de\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e4.67 b\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e1 Kin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e12.37 g-j\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e2 gh\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e1.83 fg\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e2 Kin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e7.3jk\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e0.83 ij\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e2.17 ef\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"108\"\u003e\n\u003cp\u003e3 Kin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"78\"\u003e\n\u003cp\u003e0 l\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"120\"\u003e\n\u003cp\u003e0.5 jk\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"114\"\u003e\n\u003cp\u003e1.17 hi\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eMeans followed by the same letter in column are not significantly different at 5 % (based on Duncan multiple range test)\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 8 \u003c/strong\u003eMorphological properties of \u003cem\u003eA. elburzense\u003c/em\u003e W. seedlings grown from seed and bulb over 3 years.\u003c/p\u003e\n\u003ctable border=\"1\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"123\"\u003e\n\u003cp\u003e\u003cstrong\u003ePropagation method\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"153\"\u003e\n\u003cp\u003e\u003cstrong\u003eTraits\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"96\"\u003e\n\u003cp\u003e\u003cstrong\u003e2014/2015\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e\u003cstrong\u003e2015/2016\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e\u003cstrong\u003e2016/2017\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"4\" width=\"123\"\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSeed\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"153\"\u003e\n\u003cp\u003eLeaf number\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"96\"\u003e\n\u003cp\u003e1 \u0026plusmn; 0\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e1.48 \u0026plusmn; 0.05\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e1.71 \u0026plusmn; 0.08\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"153\"\u003e\n\u003cp\u003eInflorescence formation (%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"96\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e13.33 \u0026plusmn; 0.11\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e87.66 \u0026plusmn; 1.35\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"153\"\u003e\n\u003cp\u003eInflorescence diameter (cm)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"96\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e4.4 \u0026plusmn; 0.07\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e5.58 \u0026plusmn; 0.16\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"153\"\u003e\n\u003cp\u003eBulb weight (gr)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"96\"\u003e\n\u003cp\u003e1.05 \u0026plusmn; 0.04\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e6.76 \u0026plusmn; 0.13\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e12.92 \u0026plusmn; 0.21\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"4\" width=\"123\"\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eBulb\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"153\"\u003e\n\u003cp\u003eLeaf number\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"96\"\u003e\n\u003cp\u003e1.33 \u0026plusmn; 0.02\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e1.76 \u0026plusmn; 0.03\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e1.67 \u0026plusmn; 0.12\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"153\"\u003e\n\u003cp\u003eInflorescence formation (%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"96\"\u003e\n\u003cp\u003e73.33 \u0026plusmn; 1.17\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e89.35 \u0026plusmn; 1.3\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e84.2 \u0026plusmn; 1.22a\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"153\"\u003e\n\u003cp\u003eInflorescence diameter (cm)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"96\"\u003e\n\u003cp\u003e5.07 \u0026plusmn; 0.11\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e5.63 \u0026plusmn; 0.06\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e5.49 \u0026plusmn; 0.18\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"153\"\u003e\n\u003cp\u003eBulb weight (gr)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"96\"\u003e\n\u003cp\u003e11.65 \u0026plusmn; 0.42\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e13.59 \u0026plusmn; 0.24\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"102\"\u003e\n\u003cp\u003e12.69 \u0026plusmn; 0.23\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eMeans followed by the same letter in row are not significantly different at P \u0026lt; 0.05 level\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Allium elburzense W., Efficient rapid micropropagation, Morpho-phenology, Organogenesis, Phyto-hormones","lastPublishedDoi":"10.21203/rs.3.rs-250915/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-250915/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Allium elburzense W. (subg. Melanocrommyum ) is an endangered and endemic species to Iran, which is called “valak” and known as a valuable and pricy vegetable with high nutritional and medicinal properties. This study was carried out in order to domestication, removing barriers of sexual reproduction, micropropagation and investigation of phenological stages and growth cycle of the species. The results indicated that fresh seeds had better germination than one year old seeds and appropriate germination temperature in A. elburzense W. seed was 12 °C. According to ANOVA results, the triple interaction of scarification (Sc), stratification period (StP) and GA 3 had a significant effect on seedling length, seedling fresh and dry weight, bulb height and diameter (p\u0026lt;0.01). Comparison of means showed that the treatments in which the StP (45 days) and scarification was applied had better effects on seed germination (66.5% germination) than other treatments. Murashige and Skoog with 300 mg L -1 KH2PO4 (MSP) medium caused a significant increase in the diameter of the basal plate compared to MS medium. Efficient system for in vitro propagation and conservation of valak genetic resources were the use of MSP culture medium supplemented with 0.6 mg l -1 NAA, 4 mg l -1 BA and 120 g l -1 of sucrose, which resulted in a direct propagation coefficient of 26.83 seedlings and bulblets at 18 weeks. As regards its life cycle, the juvenile phase in a low proportion of individuals (13.33%) lasted 2 years from seeding, while 87.66% of individuals were capable of producing inflorescence in the third year. Results showed that this plant is a metamorphosis and cross pollinator species. Results of present study can be used in domestication, conservation of genetic resources and proper exploitation of valak species.","manuscriptTitle":"Optimizing proliferation and assessment of valak morpho-phenological traits; an endangered nutritious Allium endemic to Iran","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2021-04-01 11:33:26","doi":"10.21203/rs.3.rs-250915/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"81c3acb3-ba8a-4982-835e-dc2281dbf3bd","owner":[],"postedDate":"April 1st, 2021","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":3349892,"name":"Plant Physiology and Morphology"}],"tags":[],"updatedAt":"2021-08-01T09:11:27+00:00","versionOfRecord":[],"versionCreatedAt":"2021-04-01 11:33:26","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-250915","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-250915","identity":"rs-250915","version":["v1"]},"buildId":"qQ7_6M8ijIrYJ9CiyUnPg","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}