Introgression of Desirable Alien Chromatin from Wild Avena Species into Cultivated Oat (Avena sativa L.) and Its Identification through Agro-Morphological and Molecular Techniques

Introgression of Desirable Alien Chromatin from Wild Avena Species into Cultivated Oat (Avena sativa L.) and Its Identification through Agro-Morphological and Molecular Techniques | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Introgression of Desirable Alien Chromatin from Wild Avena Species into Cultivated Oat (Avena sativa L.) and Its Identification through Agro-Morphological and Molecular Techniques Sawan Kumar, VK Sood, Sanjay Kumar Sanadya, Gaurav Sharma, Vinaykumar Rachappanavar, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5057298/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 The genetic enhancement of polyploid oats poses a significant challenge for crop production improvement. This study aimed to introgress desirable agronomic and quality traits from wild ‘ Avena ’ species into cultivated A. sativa L. We utilized three diploid species ( A. strigosa L., A. longiglumis L., and A. orientalis L.), one tetraploid species ( A. barbata L.), and three hexaploid species ( A. sterilis L., A. byzantina L., and A. sativa L.) for interspecific and interploidy crosses. Interspecific crosses were advanced to BC 1 F 1 and BC 2 F 1 generations over three Rabi seasons at CSK Himachal Pradesh Agricultural University, India. Phytohormone application, embryo culture, and colchicine treatment facilitated interploidy crosses, while intraploidy crosses were performed without manipulation. We developed 41 embryos from A. strigosa × A. sativa , 18 from A. longiglumis × A. sativa , and 14 from A. orientalis × A. sativa , with 15 plants surviving colchicine treatment (34.88% survival rate). SSR primers confirmed hybridity, and molecular screening of BC 1 F 1 and BC 2 F 1 generations showed donor alleles in 16 of 26 BC 2 F 1 plants. The BC 2 F 1 derivatives exhibited significant improvements over the recurrent parent, including up to 75% higher beta-glucan content, 41% higher protein content, 11.28% reduced maturity days, and 70.96% higher seed yield. These advancements underscore the potential of BC 2 F 1 derivatives in oat breeding programs for trait introgression and crop improvement. Avena species Interspecific crosses Embryo culture Oat Figures Figure 1 1. Introduction Cereal production is strategically crucial to the global economy. The primary aim of breeding programs has traditionally been to develop cultivars with enhanced agronomic performance, such as high grain yield, superior grain quality, and resistance to diseases and lodging (Tadesse et al., 2019). However, in today's rapidly changing environment, adaptability to varying environmental conditions has become an increasingly important trait (Nelson et al., 2007 ). While genetic improvement of diploid cereal species like rice, barley, and maize is relatively straightforward(Hisano et al., 2021 ), the enhancement of polyploid crops such as hexaploidy wheat and oats presents significant challenges. Oat ( A. sativa L.), an important annual forage crop of the Rabi season, demonstrates wide adaptability to diverse growing conditions. It offers excellent nutritional value for both animal and human consumption, making it a valuable dual-purpose crop (Ahmad et al., 2014). Oats are well-suited to a variety of soil types and outperform many other small-seeded cereals, particularly on acidic and alkaline soils (Wibbererley, 2006). The genus ‘Avena’ is extensive, encompassing both wild and cultivated polyploid species with a basic chromosome number of n = 7 (Liu et al., 2017 ). The regions of the Himalayas, in particular, provide an ideal environment for oat cultivation, thanks to the crop's adaptability, rapid regrowth, and superior nutritional value.In many parts of the Himalayan region, oats serve as a vital forage crop in summer and are also grown as a multipurpose crop for fodder and grain (Sood et al., 2016). Over time, elite germplasm tends to lose valuable genetic variation, prompting breeders to explore the rich allele diversity found in crop wild relatives (CWR) and landraces stored in seed banks (Mohler et al., 2023 ). Despite the challenges involved, these ancestral species hold the potential to introduce new alleles that improve disease resistance, abiotic stress tolerance, and overall yield and quality in cultivated crops (Zhang et al., 2017). Wild species related to cultivated oats often contain desirable genetic variation that can be transferred into cultivated oats through interspecific crossing techniques (Imam and Allard, 1965 ). Progenitor species and wild relatives, especially those at lower ploidy levels, have preserved a high degree of useful genetic variation. The success of plant breeders in utilizing this alien variation for crop improvement depends largely on the degree of relatedness between the cultivated species and the wild donor species (Kashyap et al., 2022 ). However, transferring desirable genes from diploid and tetraploid ‘ Avena ’ species into hexaploid oat cultivars is a complex and challenging task(Mohler et al., 2023 ). The integration of beneficial genes from wild relatives into cultivated oat species is a key objective for plant breeders aiming to broaden the genetic base and enhance desirable traits (Mohler et al., 2023 ). Despite strong sterility barriers and the difficulty of chromosome pairing between species with different ploidy levels, several studies have reported significant improvements in agronomic and quality traits through crosses with these wild species. Techniques such as embryo rescue (Rogo et al., 2023 ) and colchicine treatment (Eng and Ho, 2019 ) are often required to overcome the challenges of interploidy transfer, with greater success typically observed when the lower ploidy species is used as the female parent (Huang et al., 2022 ). In recent years, considerable progress has been made in this area, with several successful gene transfers reported. Pollination followed by auxin treatment and embryo rescue has become a well-established approach for producing interspecific hybrids in oats. Given these advancements and the potential benefits, the present investigation aims to introgress desirable alien chromatin from wild ‘Avena’ species into cultivated oat, utilizing both agro-morphological and molecular techniques to evaluate and identify the successful introgressions. This research holds promise for enhancing the genetic diversity and agronomic traits of cultivated oats, contributing to the development of more resilient and productive oat cultivars. 2. Materials and Methods The present investigation was carried out during the Rabi seasons from 2017-18 to 2020-21 at the Experimental Farm and Molecular Cytogenetics and Tissue Culture Lab, CSK HP Agricultural University, Palampur (India). The experimental material consisting of three accessions of wild diploid species viz ., A. strigosa L., A. longiglumis L. and A. orientalis L., one tetraploid species i.e. A. Barbata L. and two hexaploid wild species viz ., A. sterilis L., A. byzantina L. and one cultivated A. sativa cv. HJ-8 were used to develop the interspecific crosses (Table 1 ). Table 1 Detail of the material used in the present study Sr. No. Species Accession No. Chromosome number (2n) Source 1. Avena barbata HFO 58 28 CCS HAU, Hisar 2. Avena byzantina HFO 60 42 CCS HAU, Hisar 3. Avena orientalis HFO 103 14 CCS HAU, Hisar 4. Avena longiglumis HFO 498 14 CCS HAU, Hisar 5. Avena strigosa HFO 505 14 CCS HAU, Hisar 6. Avena sterilis HFO 508 42 CCS HAU, Hisar 7. Avena sterilis HFO 878 42 CCS HAU, Hisar 8. Avena sativa HJ-8 42 CSKHPKV, Palampur 2.1 Crossing Plan During Rabi 2017-18, interspecific crosses between wild ‘ Avena’ species and cultivated A. sativa cv. HJ-8 were attempted (Fig. 1 ). Hybrid embryos developed through crossing diploid and tetraploid species with A. sativa were rescued and cultured on suitable MS medium followed by colchicine treatment under controlled conditions. The procedure to develop interploidy crosses, hormone treatment, embryo culture and colchicines treatment was used as per Rines et. al. ( 2007 ). Plants developed through embryo culture were maintained in the lab under artificial conditions till maturity and F 1 seeds were harvested after ripening. The crosses between wild hexaploid and cultivated A. sativa were developed and harvested at maturity as they are fully compatible. F 1 plants were grown and back crossed twice with cultivated A. sativa cv. HJ-8 to generate BC 1 F 1 and BC 2 F 1 generation during Rabi , 2018-19 and 2019-20, respectively in the field conditions (Fig. 1 ). BC 2 F 1 generation along with parents were evaluated in the pots during Rabi , 2020-21 for various agro-morphological and quality traits under field conditions. 2.2 Embryo culture of interploidy hybrids The caryopses from crossed panicles were harvested and washed thoroughly using Tween-20 under tap water to avoid any sort of contamination/infection. At the time of embryo rescue in Laminar Air Flow Chamber, surface sterilization of caryopses was done using 0.1 per cent HgCl 2 for 1–2 minutes followed by three washing with autoclaved distilled water. The embryos were then excised from sterilized seeds and transferred to the test tubes containing working MS medium. Half strength Murashige and Skoog (MS) medium(Murashige and Skoog 1962 ) supplemented with essential amino acids (Table 3.2 and 3.3) was used for the rescue of interploidy hybrid embryos as per suggested by Rines et al., 2003.Cultured embryos were placed in the dark section of Plant Growth Chamber (PGC) at 20 ± 2°C with 75 per cent humidity for regeneration for about a week till the roots and shoots initiated. The regenerated plantlets were then shifted to the other section of the PGC at 20 ± 2°C with 10/14 hrs light/dark profile with 75 per cent relative humidity for proper development of plantlets.The plantlets regenerated from embryo culture were treated with IBA (Indole butyric acid) and NAA (Naphthalene acetic acid) solution of 100 ppm concentration each for 2 minutes for profuse rooting.Potting mixture was prepared by mixing of coco peat, vermiculite and perlite in the proportion of 3:1:1 v/v . The plantlets were then transferred to the soil less media in small pots (5 cm diameter) for proper rooting and shooting. Appropriate conditions were maintained for proper growth.At 3–4 leaf stage, some plantlets were treated with 0.05% and remaining were treated with 0.10% colchicine with 2% DMSO for 4 hours. DMSO acts as osmoregulator. After attaining 10–12 leaf stage, the treated plants were transferred to potting mixture. 2.3 Data recorded Observations were recorded on five randomly selected competitive plants of parents for all the traits, and each plant data recorded in case of BC 2 F 1 generation. The agro-morphological characters were included days to 50% flowering, plant height (cm), number of leaves per plant, tillers per plant, leaf area (cm 2 ), spikelets per panicle, green fodder yield/ plant (g), leaf: stem ratio, dry matter yield/ plant (g), days to 75% maturity, biological yield/ plant (g), seed yield/ plant (g), harvest index, 1000 grain weight (g), seed length (mm), protein content (%) and beta- glucan content (%). The protein content for each genotype was analysed by the quantitative method given by Lowry et. al. 1951 .Beta-glucan content (%) was determined by the extraction method given by Wood et al. (1977) and estimation method given by Semedo et al. ( 2015 ). 2.4 Molecular marker analysis To detect the hybridity of F 1 plants from different interspecific crosses, 25 SSR primer pairs were screened for polymorphism among the parental ‘Avena’ species. Details of these primers, including their sequences, are provided in Supplementary Table 1.Polymorphic SSR markers were used for hybridity test in F 1 and tracing the introgressions in BC 1 F 1 and BC 2 F 1 . Extraction of Plant genomic DNA, PCR amplification and Agarose gel electrophoresis were done in Molecular Cytogenetics and Tissue Culture Lab, Department of Genetics and Plant Breeding, CSK HPKV, Palampur. The PCR amplification was performed in a mixture containing 1.5 ul of DNA template, 0.5 ul of each forward and reverse primers, 0.1 ul of Taq polymerase (Takara Pvt. Ltd.), 2.0 ul of dNTP mix, 2 ul of PCR Buffer + mgcl 2 and 6.4 ul of sterile water with final volume of 12 ul per reaction. 2.5 Statistical Analysis Data recorded for various traits was pertained to test whether the mean difference between mean values of plants is significant or not over individual cross mean as well as overall mean of BC 2 F 1 generation using simple t-test performed as: Where, = mean-difference between two sets of related samples SE (X d ) = Standard error of mean difference n = Number of related samples The significance of backcross derivatives and parents was tested at tabulated value of (n-1) degree of freedom at 5% level. 3. Results 3.1 Interspecific hybridization in Avena species A total of 73 F 1 hybrid plants were successfully developed from six interspecific crosses (Table 2 ). All crosses were successful except for the A. barbata × A. sativa cross, which did not result in embryo formation. The successful hybrids were backcrossed with the recipient parent twice, resulting in 24 plants in the BC 1 F 1 generation and 26 plants in the BC 2 F 1 generation. For the cross between A. orientalis and A. sativa , 5 seeds were produced in the F 1 generation, followed by 2 seeds in BC 1 F 1 and 3 seeds in BC 2 F 1 . A greater number of seeds were obtained from the A. longiglumis × A. sativa cross, with 28 F 1 seeds, 9 BC 1 F 1 seeds, and 5 BC 2 F 1 seeds. Similarly successful was the A. strigosa × A. sativa cross, which yielded 27 F 1 seeds, 6 BC 1 F 1 seeds, and 6 BC 2 F 1 seeds. When crossing A. sativa with A. byzantina , the resulting generations produced 5 F 1 seeds, 2 BC 1 F 1 seeds, and 4 BC 2 F 1 seeds. Two different cultivars of A. sterilis were crossed with A. sativa : cv. HFO 508 yielded 4, 3, and 4 seeds in F 1 , BC 1 F 1 , and BC 2 F 1 generations respectively, while cv. HFO 878 produced 4, 2, and 4 seeds in the corresponding generations. Table 2 Plants developed in F 1 , BC 1 F 1 and BC 2 F 1 generations of different interspecific crosses Crosses Number of Plants F 1 BC 1 F 1 BC 2 F 1 A. barbata × A. sativa 0 0 0 A. orientalis × A. sativa 5 2 3 A. longiglumis × A. sativa 28 9 5 A. strigosa × A. sativa 27 6 6 A. sativa × A. byzantine 5 2 4 A. sativa × A. sterilis cv. HFO 508 4 3 4 A. sativa × A. sterilis cv. HFO 878 4 2 4 Total 73 24 26 3.2 Embryo culture in Interploidy hybridization In the interploidy crossing program, species with lower ploidy levels were used as female parents, while species with higher ploidy levels were used as male parents, as this approach has a higher success rate according to previous studies. From 839 caryopses, a total of 73 hybrid embryos were developed. Specifically, 41 embryos were derived from the A. strigosa × A. sativa cross, 18 from A. longiglumis × A. sativa , and 14 from A. orientalis × A. sativa . These embryos were cultured on suitable MS medium and incubated in the dark for germination. The hybrid plant regeneration frequency was 61.64%, with 45 of the 73 embryos germinating within 7–10 days. Of these, 43 embryos developed into seedlings with 3–4 leaves. 3.3 Evaluation of BC 2 F 1 derivatives The BC 2 F 1 derivatives were assessed for a range of agro-morphological and quality traits during the Rabi season of 2020-21, alongside parent lines. The evaluation revealed significant variations in key traits among the derivatives (Tables 3 and 4 ). Days to 50% flowering ranged from 117 to 142 days, with several plants, including P 1 of A. orientalis × A. sativa (124 days) and P 2 and P 3 of A. longiglumis × A. Sativa (122 and 124 days, respectively), flowering earlier than the overall BC 2 F 1 mean (130.30 days). Whereas, A. strigosa (123 days) was significantly superior for this trait among the parents. Table 5 Promising BC 2 F 1 plants selected for important agro-morphological and quality traits. Sr. No. Cross Plant number Seed yield per plant (g) Biological yield per plant (g) Days to 75% maturity Beta-glucan content (%) 1. A. orientalis × A. sativa P 1 P 1 , P 3 P 1 P 3 2. A. longiglumis × A. sativa P 3 , P 4, P 5 P 3 P 1 , P 2 , P 3 P 1 , P 2 , P 3 3. A. strigosa × A. sativa P 1 , P 2 , P 5 P 1 , P 4 P 1 , P 2 , P 3 , P 5 , P 6 P 1 , P 3 , P 4 4. A. sativa × A. byzantina P 1 P 1 , P 3 P 1 P 3 5. A. sativa × A. sterilis cv. HFO-508 P 1 , P 4 P 1 , P 4 - P 1 6. A. sativa × A. sterilis cv. HFO-878 P 3 P 2 , P 3 - - Plant height among the BC 2 F 1 derivatives varied from 58 cm to 95 cm. Plants P 1 of A. orientalis × A. sativa (80.00 cm) and P 3 of A. longiglumis × A. Sativa (95.00 cm) were notably taller than the overall mean (74.0 cm). A. strigosa (82.00 cm) and A. sterilis cv. HFO-508 (83.00 cm) showed significant superiority for plant height. Similarly, leaf number ranged from 22 to 60, with A. strigosa (48.00), A. byzantina (52.00), and A. sterilis cv. HFO-878 (52.00) exhibiting superior performance. The number of tillers per plant varied from 5 to 14, with plants P 3 of A. longiglumis × A. sativa (14.00) and P 1 and P 5 of A. strigosa × A. sativa (12.00 and 10.00, respectively) showing significant superiority. Leaf area ranged from 13.87 to 56.00 cm², with plants P 3 of A. orientalis × A. sativa (26.50 cm 2 ), and P 4 and P 5 of A. longiglumis × A. sativa (56.00 and 37.33 cm 2 , respectively) demonstrating superior values. Number of spikelets per plant varied from 28 to 76, with several plants across different crosses showing significant superiority. Green fodder yield per plant ranged from 72 to 118 g. A. strigosa (110.00 g) and A. longiglumis (106.00 g) were exhibited superiority among the parents. The leaf:stem ratio ranged from 0.22 to 0.65, with significant superiority in several plants including P 3 of A. orientalis × A. sativa (0.39), P 2 of A. strigsa × A. sativa (0.40). Dry matter yield per plant varied from 15.54 to 32.96 g, with A. byzantina (24.09 g), A. strigosa (29.77 g) and A. longiglumis (31.10 g) showing significant superiority. Days to 75% maturity ranged from 155 to 178 days, with several BC 2 F 1 derivatives showing earlier maturity than the overall mean. Biological yield per plant varied from 27.06 to 42.90 g, with A. sterilis cv. HFO-508 (36.87 g) and A. sterilis cv. HFO-878 (36.34 g) showing significant superiority. Seed yield per plant ranged from 3.11 to 7.30 g, with several plants, including those from A. orientalis × A. sativa , showing superior values. Harvest index ranged from 8.64 to 17.97%, with significant superiority in several plants from cross A. orientalis × A. sativa and A. longiglumis × A. sativa . Thousand-grain weight varied from 28.30 to 35.40 g, with A. orientalis (32.70 g), A. byzantina (34.40 g), and A. longiglumis (35.70 g) showing significant superiority. Seed length ranged from 11.95 to 14.15 mm, with A. orientalis (15.12 mm), A. byzantina (14.60 mm), A. longiglumis (10.40 mm), and A. strigosa (10.15 mm) showing superior values. Protein content ranged from 7.3–11.00%, and all wild accessions except A. sterilis cv. HFO-878 demonstrating superior values than overall mean and cultivated species A. sativa . Beta-glucan content varied from 2.89–5.42%, with A. longiglumis (4.43%) and A. strigosa (4.62%) showing significant superiority. Plant P 3 (3.97%) of cross A. orientalis × A. sativa ; P 1 (5.42%), P 2 (4.31%) and P 5 (4.70%) of cross A. longiglumis × A. sativa ; P 1 (4.74%), P 3 (4.00%) and P 4 (4.95%) of A. strigosa × A. sativa ; P 3 (3.94%) of A. byzantina × A. sativa ; P 1 (4.10%) of A. sativa × A. sterilis cv. HFO-508 showed significant superiority for this trait over overall mean. Overall, the BC 2 F 1 derivatives exhibited promising traits for introgression into cultivated oat genotypes, potentially enhancing their performance. These findings align with previous research highlighting the benefits of interspecific crosses in improving plant traits (Murphy, 1981). 3.4 Molecular Screening Molecular characterization of all ‘ Avena ’ species accessions utilized 25 SSR primers. Among these, five SSR primers displayed polymorphism between wild species and the cultivated oat variety, A. sativa cv. HJ-8. Each wild species exhibited unique polymorphic primers, except for MAMA_4, which was polymorphic in both A. orientalis and A. sterilis cv. HFO-878 (Table 6 ). The polymorphic primers confirmed the hybridity of F 1 plants from interploidy crosses, demonstrating the presence of alleles from both parent species (Plate 2). For intraploidy crosses, hybridity was verified in five plants of A. sativa × A. byzantina , four plants of A. sativa × A. sterilis HFO-508, and four plants of A. sativa × A. sterilis HFO-878, all showing alleles from both parent species. Table 6 List of SSR primers polymorphic for different Avena species S. No. Primer Cross Annealing Temp. (ºC) No. of Alleles 1. AM87 A. sativa × A. byzantina 52.59 2 2. MAMA_4 A. orientalis × A. sativa 52.30 2 3. MAMA_5 A. longiglumis × A. sativa 53.09 3 4. AM115 A. strigosa × A. sativa 51.17 2 5. Astavea A. sativa × A. sterilis cv. HFO 508 54.35 2 6. MAMA_4 A. sativa × A. sterilis cv. HFO 878 52.30 2 In the BC 1 F 1 generation, all plants from intraploidy crosses presented amplicon segments from both parent species, whereas in interploidy crosses, all plants of A. orientalis × A. sativa possessed alleles from both parents. However, in the cross A. longiglumis × A. sativa , plants 4, 7, and 8 out of 9 lacked the amplicon region from the donor parent P 1 . In the BC 2 F 1 generation, the cross A. sativa × A. byzantina yielded four plants, three of which exhibited the amplified region of primer AM87 from both parents (Plate 3). Similarly, two plants each in the crosses A. sativa × A. sterilis cv. HFO-508 and A. sativa × A. sterilis cv. HFO-878 demonstrated amplicon regions from the donor parent. These plants were notably superior in seed yield per plant, biological yield per plant, and plant height. In the interploidy cross A. orientalis × A. sativa , only one plant showed the allelic region for SSR marker MAMA_4 from both parents, and this plant excelled in days to 50% flowering, seed yield per plant, and tillers per plant compared to the recurrent parent A. sativa cv. HJ-8. Similarly, in the cross A. longiglumis × A. sativa , only plant P 1 exhibited the allelic region from the donor parent, and it was significantly superior in beta-glucan content, protein content, and days to 75% maturity over the recurrent parent. In the cross A. strigosa × A. sativa , three plants showed the allelic region from the donor parent and were also significantly superior in beta-glucan, protein content, and days to 75% maturity. 4. Discussion The successful introgression of desirable chromatin from wild ‘ Avena ’ species into cultivated oat underscores the potential of interspecific and interploidy hybridization in enhancing the genetic diversity and agronomic traits of cultivated oats (Tinker et al., 2022 ). CWRs can significantly expand the genetic diversity of crops and introduce beneficial traits (Brozynska et al., 2016; Zhang et al., 2017). However, their direct use in breeding programs is often challenging due to the presence of undesirable traits like, linkage drag (peng et al., 2014) and common breeding barriers (Kashyap et al., 2022 ) with cultivated crops.Cross compatibility is notably low in hexaploid × diploid ‘Avena’ hybrids (Latta et al., 2019 ). A significant challenge in transferring genes from wild diploid and tetraploid oat species into hexaploid cultivars is successfully producing healthy, vigorous inter-ploidy F 1 hybrids (Aung et al., 2010). These hybrids must maintain all chromosomes from both the cultivated and wild parents, ensuring that the gene of interest is effectively expressed (Aung et al., 2010).Few studies have focused on transferring traits from wild relatives to cultivated oats, particularly for agronomic characteristics (Hermann et al., 2014), nutritional traits (Ahokas and Manninen, 2000 ), crown rust resistance (Rines et al., 2007 ; Aung et al., 2010), and powdery mildew resistance (Yu and Hermann, 2006) and they got positive result by successful introgression. 4.1 Molecular Screening This study generated a significant number of F 1 hybrids from various interspecific crosses, although the cross between A. barbata and A. sativa was unsuccessful due to the absence of embryo formation. The subsequent backcrossing efforts yielded promising BC 1 F 1 and BC 2 F 1 generations, indicating the feasibility of incorporating wild ‘Avena’ chromatin into cultivated backgrounds. Embryo culture proved crucial in rescuing hybrid embryos from interploidy crosses, with a notable regeneration frequency of 61.64%. The development of 43 viable seedlings from 73 germinated embryos highlights the effectiveness of this technique in overcoming barriers associated with interploidy hybridization. These results align with previous studies, which suggest that using species with lower ploidy levels as females in crosses increases the likelihood of success. The hybrids demonstrated a broad range of agro-morphological variations, with several BC 2 F 1 derivatives exhibiting significant improvements in traits such as plant height, number of tillers, leaf area, green fodder yield, seed yield, and other agronomic characteristics. 4.2 Evaluation of BC 2 F 1 derivatives The evaluation of BC 2 F 1 derivatives in our study revealed significant variations in key agro-morphological and quality traits, highlighting the potential of introgressing desirable traits from wild ‘Avena’ species into cultivated oat ( A. sativa L.). The observed variability in days to 50% flowering, ranging from 117 to 142 days, demonstrates the potential for modifying flowering times through interspecific crosses. Notably, plants derived from A.orientalis × A. sativa and A.longiglumis × A. sativa exhibited earlier flowering compared to the overall BC 2 F 1 mean, suggesting that these crosses can be used to accelerate flowering times in cultivated oats, an important trait for adapting to varying climatic conditions (Smith et al., 2018 ). The significant variation in plant height among the derivatives, with values ranging from 58 cm to 95 cm, underscores the potential for introducing height traits from wild species such as A. strigosa and A. sterilis cv. HFO-508. Taller plants may offer advantages in forage yield and mechanical harvesting efficiency, consistent with findings from previous studies (Semchenko and Zobel, 2005 ; Rosser, 2014; Coblentz et al., 2014 ). Similarly, the variation in leaf number and leaf area suggests that wild species can contribute to improved canopy architecture and productivity, aligning with the observations of leaf trait improvement in other interspecific crosses (Carlson et al., 2021 ). The variation in number of tillers per plant and green fodder yield per plant further emphasizes the potential for improving these traits through wild species introgression. The observed superiority of certain plants for these traits, such as those from A.longiglumis × A. sativa and A. strigosa × A. sativa , indicates that these wild species can enhance forage yield and quality, which is critical for sustainable oat production (Humphreys, 2005 ; Kumar et al., 2018 ; Chand et al., 2022 ; Sood et al., 2022 ). In terms of yield components, the significant differences in biological yield, seed yield, harvest index, thousand-grain weight, and seed length suggest that wild species introgression can positively impact yield and grain quality. The superior values observed in plants from crosses involving A. orientalis , A. byzantina , and A. longiglumis highlight the potential for developing high-yielding oat cultivars with desirable agronomic characteristics (Beavis and Frey, 1987 ; Sharma et al., 2018 ). Additionally, the higher protein and beta-glucan content in certain wild accessions, particularly A. longiglumis and A. strigosa , aligns with findings from previous research that emphasize the nutritional benefits of integrating wild species into cultivated crops (Frison et al., 2011 ; Kahane et al., 2013 ; Ebert, 2014 ). The promising traits observed in the BC 2 F 1 derivatives underscore the value of utilizing wild ‘ Avena ’ species for improving cultivated oat performance. This study's findings support previous research highlighting the benefits of interspecific crosses in enhancing plant traits and emphasize the potential for developing improved oat cultivars through continued exploration of wild species (Murphy, 1981; Johnson et al., 2004). 4.3 Molecular Screening The molecular characterization of ‘ Avena ’ species accessions using 25 SSR primers provided valuable insights into the genetic introgression from wild species into cultivated oat ( A. sativa L.). Notably, five SSR primers exhibited polymorphism between the wild species and the cultivated variety, A. sativa cv. HJ-8. This finding underscores the potential of SSR markers in distinguishing between wild and cultivated accessions, offering a robust tool for genetic analysis in oat breeding programs (Gunnaiah et al., 2021 ; Thapa et al., 2021 ; Liu et al., 2022). The distinct polymorphic patterns observed in wild species, except for the MAMA_4 primer which was shared between A. orientalis and A. sterilis cv. HFO-878, highlight the genetic diversity present within the wild ‘ Avena ’ species. Such variability is crucial for the introgression of beneficial traits from wild relatives into cultivated varieties. Previous studies have similarly demonstrated the utility of SSR markers in revealing genetic differences between species and in assessing genetic diversity (Yarvaan et al., 2020; Arora et al., 2021 ; Koroluk et al., 2022; Kaur et al., 2024 ). The confirmation of hybridity in F 1 plants from interploidy crosses, as indicated by the presence of alleles from both parent species, reflects successful genetic transfer. This aligns with findings from other research where SSR markers were used to validate hybridization and the introgression of alien chromatin (Boutin et al., 2006; Zhang et al., 2009). Specifically, the detection of polymorphic alleles in intraploidy crosses, such as A. sativa × A. byzantina and A. sativa × A. sterilis , further supports the effectiveness of these markers in tracking genetic introgression and verifying the presence of alien genes. However, the cross between A. longiglumis and A. sativa revealed a notable exception. In this cross, plants 4, 7, and 8 out of 9 lacked the amplicon region from the donor parent (P 1 ). This observation highlights the challenges associated with introgressing genetic material from certain wild relatives. It suggests that the introgression from A. longiglumis may be less efficient or that specific genetic barriers might be present (Jones et al., 2022). Such variability in successful introgression has been reported in other studies, where the presence or absence of donor chromatin in progeny varied significantly (Lee et al., 2020; Kim & Park, 2021). The differential success in introgression observed in this study underscores the need for a tailored approach when selecting wild relatives for breeding programs and highlights the importance of comprehensive molecular screening to ensure the retention of desirable chromatin segments. The successful integration of traits from wild ‘ Avena ’ species into cultivated oat demonstrates the value of molecular techniques in enhancing the genetic base of crop varieties. This approach aligns with other studies that emphasize the role of wild relatives in broadening genetic diversity and improving traits such as disease resistance and stress tolerance (Perrino et al., 2020; Bohra et al., 2022 ; Kashyap et al., 2022 ). Future research should focus on understanding the underlying genetic mechanisms influencing successful introgression and developing strategies to overcome barriers associated with specific wild species. The overall study's findings emphasize the importance of utilizing wild relatives and landraces to broaden the genetic base of cultivated oat crop. The introgressed lines developed in this study not only possess enhanced agronomic traits but also offer potential for further genetic improvement through conventional breeding and molecular techniques. The observed transgressive segregation and superior performance of certain BC 2 F 1 plants underscore the potential for creating new oat cultivars with improved yield, quality, and resilience to biotic and abiotic stresses. The introgression of desirable alien chromatin from wild ‘Avena’ species into cultivated oat has yielded promising results, demonstrating the value of wild species in oat improvement programs (Boczowska et al., 2015; 2016). Several past research studies indicates that genetic diversity within modern oat gene pools is limited. Over time, valuable alleles have been lost through selection, introduction, and domestication making it essential to explore obsolete cultivars, wild ancestors, and landraces for new germplasm to advance oat breeding efforts (Okoń et al., 2014 ; Koroluk et al., 2022). The combination of agro-morphological evaluation and molecular characterization has provided a robust framework for selecting superior lines (Ghazy et al., 2024 ; Bayahi and Rezgui, 2015; Guizani et al., 2024 ; khan et al., 2024 ), paving the way for the development of new oat varieties that meet the challenges of modern agriculture (Kumar et al., 2023 ; Kebede et al., 2023 ). Further research should focus on the continued evaluation and utilization of these introgressed lines in breeding programs, with particular attention to their performance under diverse environmental conditions. 5. Conclusion This study successfully demonstrated the potential of interspecific and interploidy hybridization in ‘ Avena ’ species for enhancing cultivated oat ( A. sativa L.) through the introgression of desirable alien chromatin. A total of 73 F 1 hybrids were generated, and subsequent backcrossing produced BC 1 F 1 and BC 2 F 1 generations, revealing significant agro-morphological and quality trait variations. Key traits such as plant height, leaf number, green fodder yield, and protein and β-glucan content showed notable improvements in several BC 2 F 1 derivatives compared to the recurrent parent. These newely developed interspecific derivatives will be screened for ploidy level and alien introgressions using cytological techniques such as Genomic In situ Hybridization, after two more successive bakcrossings to achieve the stable useful introgressions. Molecular screening using SSR markers confirmed the introgression of chromatin from wild species into cultivated oat, validating the hybridity and the presence of alleles from both parent species. This integration of wild genetic material into cultivated oat shows promise for improving yield, stress tolerance, and nutritional quality, aligning with the benefits observed in previous research. The findings highlight the potential for utilizing wild ‘Avena’ species to enhance the genetic base and performance of cultivated oats. Declarations Authors contribution statement: Conceptualization, Methodology, Software, Validation, Formal analysis, Resources, Data curation, Writing-original draft- SK, VKS, SKS, Writing-review and editing, Visualization, Supervision- GS, VR, RK. References Ahokas H, Manninen ML (2000) Introgressive hexaploid oats from the Avena abyssinia (♀) × A. sativa hybrid: performance, grain lipids and proteins. Euphytica 113:153–160 Arora A, Sood VK, Chaudhary HK et al (2021) Genetic diversity analysis of oat ( Avena sativa L.) germplasm revealed by agro-morphological and SSR markers. Range Manage Agrofor 42:38–48 Aung T, Zwer P, Park R et al (2010a) Hybrids of Avena sativa with two diploid wild oats (CIav6956) and (CIav7233) resistant to crown rust. Euphytica 174:189–198. https://doi.org/10.1007/s10681-009-0111-5 Aung T, Zwer P, Park R et al (2010b) Hybrids of Avena sativa with two diploid wild oats (CIav6956) and (CIav7233) resistant to crown rust. Euphytica 174:189–198. https://doi.org/10.1007/s10681-009-0111-5 Bayahi K, Rezgui S (2017) Genetic Study of Nutritional and Physicochemical Characters of Chickpea Lines and Cultivars ( Cicer arietinum L). J Agric Sci 9:151. https://doi.org/10.5539/jas.v9n3p151 Beavis WD, Frey KJ (1987) Expression of nuclear-cytoplasmic interactions and heterosis in quantitative traits of oats ( Avena spp). Euphytica 36:877–886. https://doi.org/10.1007/bf00051873 Boczkowska M, Harasimiuk M, Onyśk A (2015) Studies on genetic variation within old Polish cultivars of common oat. Cereal Res Commun 43:12–21. https://doi.org/10.1556/crc.2014.0025 Boczkowska M, Łapiński B, Kordulasińska I et al (2016) Promoting the Use of Common Oat Genetic Resources through Diversity Analysis and Core Collection Construction. PLoS ONE 11:e0167855. https://doi.org/10.1371/journal.pone.0167855 Bohra A, Kilian B, Sivasankar S et al (2022) Reap the crop wild relatives for breeding future crops. Trends Biotechnol 40:412–431. https://doi.org/10.1016/j.tibtech.2021.08.009 Brozynska M, Furtado A, Henry RJ (2015) Genomics of crop wild relatives: expanding the gene pool for crop improvement. Plant Biotechnol J 14:1070–1085. https://doi.org/10.1111/pbi.12454 Carlson CH, Stack GM, Jiang Y et al (2021) Morphometric relationships and their contribution to biomass and cannabinoid yield in hybrids of hemp ( Cannabis sativa ). J Exp Bot 72:7694–7709. https://doi.org/10.1093/jxb/erab346 Chand S, Indu, Singhal RK, Govindasamy P (2022) Agronomical and breeding approaches to improve the nutritional status of forage crops for better livestock productivity. Grass Forage Sci 77:11–32. https://doi.org/10.1111/gfs.12557 Chhuneja P, Kaur S, Dhaliwal HS (2016) Introgression and Exploitation of Biotic Stress Tolerance from Related Wild Species in Wheat Cultivars. Sustainable Development and Biodiversity. Springer International Publishing, Cham, pp 269–324 Coblentz WK, Jokela WE, Bertram MG (2014) Cultivar, Harvest Date, and Nitrogen Fertilization Affect Production and Quality of Fall Oat. Agron J 106:2075–2086. https://doi.org/10.2134/agronj13.0436 Curto M, Winter S, Seiter A et al (2019) Application of a SSR-GBS marker system on investigation of European Hedgehog species and their hybrid zone dynamics. Ecol Evol 9:2814–2832. https://doi.org/10.1002/ece3.4960 Ebert A (2014) Potential of Underutilized Traditional Vegetables and Legume Crops to Contribute to Food and Nutritional Security, Income and More Sustainable Production Systems. Sustainability 6:319–335. https://doi.org/10.3390/su6010319 Eng W-H, Ho W-S (2019) Polyploidization using colchicine in horticultural plants: A review. Sci Hort 246:604–617. https://doi.org/10.1016/j.scienta.2018.11.010 Frison EA, Cherfas J, Hodgkin T (2011) Agricultural Biodiversity Is Essential for a Sustainable Improvement in Food and Nutrition Security. Sustainability 3:238–253. https://doi.org/10.3390/su3010238 Ghazy AI, Ali MA, Ibrahim EI et al (2024) Characterization of Improved Barley Germplasm under Desert Environments Using Agro-Morphological and SSR Markers. Agronomy 14:1716. https://doi.org/10.3390/agronomy14081716 Guizani A, Babay E, Askri H et al (2024) Screening for drought tolerance and genetic diversity of wheat varieties using agronomic and molecular markers. Mol Biol Rep 51. https://doi.org/10.1007/s11033-024-09340-9 Gunnaiah R, Jagadeesha RC, Cholin S et al (2021) Genetic diversity assessment and population structure analysis of pomegranate cultivars from different countries and Himalayan wild accessions. J Hortic Sci Biotechnol 96:614–623. https://doi.org/10.1080/14620316.2021.1899854 Herrmann MH, Yu J, Beuch S, Weber WE (2014) Quantitative trait loci for quality and agronomic traits in two advanced backcross populations in oat ( Avena sativa L). Plant Breeding 133:588–601. https://doi.org/10.1111/pbr.12188 Hisano H, Abe F, Hoffie RE, Kumlehn J (2021) Targeted genome modifications in cereal crops. Breed Sci 71:405–416. https://doi.org/10.1270/jsbbs.21019 Huang X-Y, Shang J, Zhong Y-H et al (2022) Disaggregation of Ploidy, Gender, and Genotype Effects on Wood and Fiber Traits in a Diploid and Triploid Hybrid Poplar Family. Front Plant Sci 13. https://doi.org/10.3389/fpls.2022.866296 Humphreys MO (2005) Genetic improvement of forage crops – past, present and future. J Agricultural Sci 143:441–448. https://doi.org/10.1017/s0021859605005599 Imam AG, Allard RW (1965) Population studies in predominantly self-pollinated species. Vi. Genetic variability between and within natural populations of wild oats from differing habitats in california. Genetics 51:49–62. https://doi.org/10.1093/genetics/51.1.49 Kahane R, Hodgkin T, Jaenicke H et al (2013) Agrobiodiversity for food security, health and income. Agron Sustain Dev 33:671–693. https://doi.org/10.1007/s13593-013-0147-8 Kashyap A, Garg P, Tanwar K et al (2022) Strategies for utilization of crop wild relatives in plant breeding programs. Theor Appl Genet 135:4151–4167. https://doi.org/10.1007/s00122-022-04220-x Kaur R, Kapoor R, Vikal Y, Amandeep A (2024) Molecular characterization of oats (Avena sativa L.) diversity: Implications for dual-purpose breeding. Genetika 56:103–120. https://doi.org/10.2298/gensr2401103k Kebede G, Worku W, Feyissa F, Jifar H (2023) Agro-morphological traits-based genetic diversity assessment on oat ( Avena sativa L.) genotypes in the central highlands of Ethiopia. All Life 16. https://doi.org/10.1080/26895293.2023.2236313 Khan T, Rashid R, Shah L et al (2024) Genetic and phenotypic diversity in Solanum lycopersicum genotypes: insights from morpho-molecular and biochemical analyses. Plant Biotechnol Rep 18:207–221. https://doi.org/10.1007/s11816-024-00894-5 Koroluk A, Paczos-Grzęda E, Sowa S et al (2022a) Diversity of Polish Oat Cultivars with a Glance at Breeding History and Perspectives. Agronomy 12:2423. https://doi.org/10.3390/agronomy12102423 Koroluk A, Paczos-Grzęda E, Sowa S et al (2022b) Diversity of Polish Oat Cultivars with a Glance at Breeding History and Perspectives. Agronomy 12:2423. https://doi.org/10.3390/agronomy12102423 Kumar R, Kumar D, Datt C et al (2018) Forage Yield and Nutritional Characteristics of Cultivated Fodders as Affected by Agronomic Interventions: A Review. Indian J Anim Nutr 35:373. https://doi.org/10.5958/2231-6744.2018.00057.9 Kumar R, Varghese S, Jayaswal D et al (2023) Agro-morphological and genetic variability analysis in oat germplasms with special emphasis on food and feed. PLoS ONE 18:e0280450. https://doi.org/10.1371/journal.pone.0280450 Latta RG, Bekele WA, Wight CP, Tinker NA (2019) Comparative linkage mapping of diploid, tetraploid, and hexaploid Avena species suggests extensive chromosome rearrangement in ancestral diploids. Sci Rep 9. https://doi.org/10.1038/s41598-019-48639-7 Liu Q, Lin L, Zhou X et al (2017) Unraveling the evolutionary dynamics of ancient and recent polyploidization events in Avena (Poaceae). Sci Rep 7. https://doi.org/10.1038/srep41944 Liu R, Huang Y, Yang T et al (2021) Population genetic structure and classification of cultivated and wild pea ( Pisum sp.) based on morphological traits and SSR markers. J Syst Evol 60:85–100. https://doi.org/10.1111/jse.12710 Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275. https://doi.org/10.1016/s0021-9258(19)52451-6 Maluszynski M, Kasha K, Forster BP, Szarejko I (2013) Doubled Haploid Production in Crop Plants: A Manual. Springer Science & Business Media Mohler V, Paczos-Grzęda E, Sowa S (2023) Loving the Alien: The Contribution of the Wild in Securing the Breeding of Cultivated Hexaploid Wheat and Oats. Agriculture 13:2060. https://doi.org/10.3390/agriculture13112060 Murashige T, Skoog F (1962) A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures. Physiol Plant 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x Nelson DR, Adger WN, Brown K (2007) Adaptation to Environmental Change: Contributions of a Resilience Framework. Annu Rev Environ Resour 32:395–419. https://doi.org/10.1146/annurev.energy.32.051807.090348 Okoń SM, Chrząstek M, Kowalczyk K, Koroluk A (2014) Identification of new sources of resistance to powdery mildew in oat. Eur J Plant Pathol 139:9–12. https://doi.org/10.1007/s10658-013-0367-4 Peng T, Sun X, Mumm RH (2013) Optimized breeding strategies for multiple trait integration: I. Minimizing linkage drag in single event introgression. Mol Breeding 33:89–104. https://doi.org/10.1007/s11032-013-9936-7 Perrino EV, Perrino P (2020) Crop wild relatives: know how past and present to improve future research, conservation and utilization strategies, especially in Italy: a review. Genet Resour Crop Evol 67:1067–1105. https://doi.org/10.1007/s10722-020-00930-7 Rey E, Molnár I, Doležel J (2015) Genomics of Wild Relatives and Alien Introgressions. Alien Introgression in Wheat. Springer International Publishing, Cham, pp 347–381 Rines HW (2003) Oat haploids from wide hybridization. Doubled Haploid Production in Crop Plants. Springer Netherlands, Dordrecht, pp 155–159 Rines HW, Porter HL, Carson ML, Ochocki GE (2007) Introgression of crown rust resistance from diploid oat Avena strigosa into hexaploid cultivated oat A. sativa by two methods: direct crosses and through an initial 2x·4x synthetic hexaploid. Euphytica 158:67–79. https://doi.org/10.1007/s10681-007-9426-2 Rogo U, Fambrini M, Pugliesi C (2023) Embryo Rescue in Plant Breeding. Plants 12:3106. https://doi.org/10.3390/plants12173106 Rosser CL, Beattie AD, Block HC et al (2016) Effect of maturity at harvest for whole-crop barley and oat on dry matter intake, sorting, and digestibility when fed to beef cattle1. J Anim Sci 94:697–708. https://doi.org/10.2527/jas.2015-0063 Semchenko M, Zobel K (2005) The effect of breeding on allometry and phenotypic plasticity in four varieties of oat ( Avena sativa L). Field Crops Res 93:151–168. https://doi.org/10.1016/j.fcr.2004.09.019 Semedo MC, Karmali A, Fonseca L (2015) A high throughput colorimetric assay of β-1,3-d-glucans by Congo red dye. J Microbiol Methods 109:140–148. https://doi.org/10.1016/j.mimet.2014.12.020 Sharma D, Tiwari A, Sood S et al (2018) Genome wide association mapping of agro-morphological traits among a diverse collection of finger millet ( Eleusine coracana L.) genotypes using SNP markers. PLoS ONE 13:e0199444. https://doi.org/10.1371/journal.pone.0199444 Smith EG, Neil Harker K, O’Donovan JT et al (2018) The profitability of diverse crop rotations and other cultural methods that reduce wild oat ( Avena fatua ). Can J Plant Sci 98:1094–1101. https://doi.org/10.1139/cjps-2018-0019 Sood VK, Sanadya SK, Kumar S et al (2022) Health benefits of oat ( Avena sativa ) and nutritional improvement through plant breeding interventions. Crop Pasture Sci 74:993–1013. https://doi.org/10.1071/cp22268 Tadsse W, Sanchez-Garcia M (2019) Genetic Gains in Wheat Breeding and Its Role in Feeding the World. Crop Breeding, Genetics and Genomics. https://doi.org/10.20900/cbgg20190005 Thapa R, Edwards M, Blair MW (2021) Relationship of Cultivated Grain Amaranth Species and Wild Relative Accessions. Genes 12:1849. https://doi.org/10.3390/genes12121849 Tinker NA, Wight CP, Bekele WA et al (2022) Genome analysis in Avena sativa reveals hidden breeding barriers and opportunities for oat improvement. Commun Biology 5. https://doi.org/10.1038/s42003-022-03256-5 wood PJ, Paton D, Siddiqui R (2006) Determination of beta-glucan in oats and barley. Cereal Chem 54:524–533 Yarvaan M, Zongwen Z (2020) Study of genetic diversity in oat accessions identified using of SSR markers. Mongolian J Agricultural Sci 29:120–124. https://doi.org/10.5564/mjas.v29i1.1380 Zhang H, Mittal N, Leamy LJ et al (2016) Back into the wild—Apply untapped genetic diversity of wild relatives for crop improvement. Evol Appl 10:5–24. https://doi.org/10.1111/eva.12434 Tables Tables 3 and 4 are available in the Supplementary Files section. Plates Plates 1 and 3 are available in the Supplementary Files section Supplementary Files Table34.docx Plates.docx SupplementaryTable1.docx 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. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5057298","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":380072225,"identity":"e064cad9-f1a8-423c-8ee7-552d5fff558d","order_by":0,"name":"Sawan Kumar","email":"","orcid":"","institution":"CSK HPKV: CSK Himachal Pradesh Krishi Vishvavidyalaya","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Sawan","middleName":"","lastName":"Kumar","suffix":""},{"id":380072226,"identity":"977cdd96-55f9-4a45-9cef-f710d85dc2c4","order_by":1,"name":"VK Sood","email":"","orcid":"","institution":"CSK HPKV: CSK Himachal 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Introduction","content":"\u003cp\u003eCereal production is strategically crucial to the global economy. The primary aim of breeding programs has traditionally been to develop cultivars with enhanced agronomic performance, such as high grain yield, superior grain quality, and resistance to diseases and lodging (Tadesse et al., 2019). However, in today's rapidly changing environment, adaptability to varying environmental conditions has become an increasingly important trait (Nelson et al., \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). While genetic improvement of diploid cereal species like rice, barley, and maize is relatively straightforward(Hisano et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), the enhancement of polyploid crops such as hexaploidy wheat and oats presents significant challenges.\u003c/p\u003e \u003cp\u003eOat (\u003cem\u003eA. sativa\u003c/em\u003e L.), an important annual forage crop of the \u003cem\u003eRabi\u003c/em\u003e season, demonstrates wide adaptability to diverse growing conditions. It offers excellent nutritional value for both animal and human consumption, making it a valuable dual-purpose crop (Ahmad et al., 2014). Oats are well-suited to a variety of soil types and outperform many other small-seeded cereals, particularly on acidic and alkaline soils (Wibbererley, 2006). The genus \u0026lsquo;Avena\u0026rsquo; is extensive, encompassing both wild and cultivated polyploid species with a basic chromosome number of n\u0026thinsp;=\u0026thinsp;7 (Liu et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The regions of the Himalayas, in particular, provide an ideal environment for oat cultivation, thanks to the crop's adaptability, rapid regrowth, and superior nutritional value.In many parts of the Himalayan region, oats serve as a vital forage crop in summer and are also grown as a multipurpose crop for fodder and grain (Sood et al., 2016).\u003c/p\u003e \u003cp\u003eOver time, elite germplasm tends to lose valuable genetic variation, prompting breeders to explore the rich allele diversity found in crop wild relatives (CWR) and landraces stored in seed banks (Mohler et al., \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Despite the challenges involved, these ancestral species hold the potential to introduce new alleles that improve disease resistance, abiotic stress tolerance, and overall yield and quality in cultivated crops (Zhang et al., 2017). Wild species related to cultivated oats often contain desirable genetic variation that can be transferred into cultivated oats through interspecific crossing techniques (Imam and Allard, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e1965\u003c/span\u003e). Progenitor species and wild relatives, especially those at lower ploidy levels, have preserved a high degree of useful genetic variation. The success of plant breeders in utilizing this alien variation for crop improvement depends largely on the degree of relatedness between the cultivated species and the wild donor species (Kashyap et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). However, transferring desirable genes from diploid and tetraploid \u0026lsquo;\u003cem\u003eAvena\u003c/em\u003e\u0026rsquo; species into hexaploid oat cultivars is a complex and challenging task(Mohler et al., \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe integration of beneficial genes from wild relatives into cultivated oat species is a key objective for plant breeders aiming to broaden the genetic base and enhance desirable traits (Mohler et al., \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Despite strong sterility barriers and the difficulty of chromosome pairing between species with different ploidy levels, several studies have reported significant improvements in agronomic and quality traits through crosses with these wild species. Techniques such as embryo rescue (Rogo et al., \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) and colchicine treatment (Eng and Ho, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) are often required to overcome the challenges of interploidy transfer, with greater success typically observed when the lower ploidy species is used as the female parent (Huang et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). In recent years, considerable progress has been made in this area, with several successful gene transfers reported. Pollination followed by auxin treatment and embryo rescue has become a well-established approach for producing interspecific hybrids in oats.\u003c/p\u003e \u003cp\u003eGiven these advancements and the potential benefits, the present investigation aims to introgress desirable alien chromatin from wild \u0026lsquo;Avena\u0026rsquo; species into cultivated oat, utilizing both agro-morphological and molecular techniques to evaluate and identify the successful introgressions. This research holds promise for enhancing the genetic diversity and agronomic traits of cultivated oats, contributing to the development of more resilient and productive oat cultivars.\u003c/p\u003e"},{"header":"2. Materials and Methods","content":"\u003cp\u003eThe present investigation was carried out during the \u003cem\u003eRabi\u003c/em\u003e seasons from 2017-18 to 2020-21 at the Experimental Farm and Molecular Cytogenetics and Tissue Culture Lab, CSK HP Agricultural University, Palampur (India). The experimental material consisting of three accessions of wild diploid species \u003cem\u003eviz\u003c/em\u003e., \u003cem\u003eA. strigosa\u003c/em\u003e L., \u003cem\u003eA. longiglumis\u003c/em\u003e L. and \u003cem\u003eA. orientalis\u003c/em\u003e L., one tetraploid species i.e. \u003cem\u003eA. Barbata\u003c/em\u003eL. and two hexaploid wild species \u003cem\u003eviz\u003c/em\u003e., \u003cem\u003eA. sterilis\u003c/em\u003e L., \u003cem\u003eA. byzantina\u003c/em\u003e L. and one cultivated \u003cem\u003eA. sativa\u003c/em\u003ecv. HJ-8 were used to develop the interspecific crosses (Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n \u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n \u003ctable id=\"Tab1\" border=\"1\"\u003e\n \u003ccaption\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eDetail of the material used in the present study\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSr. No.\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSpecies\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eAccession No.\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eChromosome number (2n)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSource\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eAvena barbata\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHFO 58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCCS HAU, Hisar\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eAvena byzantina\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHFO 60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCCS HAU, Hisar\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eAvena orientalis\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHFO 103\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCCS HAU, Hisar\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eAvena longiglumis\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHFO 498\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCCS HAU, Hisar\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eAvena strigosa\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHFO 505\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCCS HAU, Hisar\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eAvena sterilis\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHFO 508\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCCS HAU, Hisar\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eAvena sterilis\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHFO 878\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCCS HAU, Hisar\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eAvena sativa\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHJ-8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCSKHPKV, Palampur\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n \u003ch2\u003e2.1 Crossing Plan\u003c/h2\u003e\n \u003cp\u003eDuring \u003cem\u003eRabi\u003c/em\u003e 2017-18, interspecific crosses between wild \u0026lsquo;\u003cem\u003eAvena\u0026rsquo;\u003c/em\u003e species and cultivated \u003cem\u003eA. sativa\u003c/em\u003e cv. HJ-8 were attempted (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). Hybrid embryos developed through crossing diploid and tetraploid species with \u003cem\u003eA. sativa\u003c/em\u003e were rescued and cultured on suitable MS medium followed by colchicine treatment under controlled conditions. The procedure to develop interploidy crosses, hormone treatment, embryo culture and colchicines treatment was used as per Rines et. al. (\u003cspan class=\"CitationRef\"\u003e2007\u003c/span\u003e). Plants developed through embryo culture were maintained in the lab under artificial conditions till maturity and F\u003csub\u003e1\u003c/sub\u003e seeds were harvested after ripening. The crosses between wild hexaploid and cultivated \u003cem\u003eA. sativa\u003c/em\u003e were developed and harvested at maturity as they are fully compatible.\u003c/p\u003e\n \u003cp\u003eF\u003csub\u003e1\u003c/sub\u003e plants were grown and back crossed twice with cultivated \u003cem\u003eA. sativa\u003c/em\u003e cv. HJ-8 to generate BC\u003csub\u003e1\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003eand BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e generation during \u003cem\u003eRabi\u003c/em\u003e, 2018-19 and 2019-20, respectively in the field conditions (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e generation along with parents were evaluated in the pots during \u003cem\u003eRabi\u003c/em\u003e, 2020-21 for various agro-morphological and quality traits under field conditions.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\n \u003ch2\u003e2.2 Embryo culture of interploidy hybrids\u003c/h2\u003e\n \u003cp\u003eThe caryopses from crossed panicles were harvested and washed thoroughly using \u003cem\u003eTween-20\u003c/em\u003e under tap water to avoid any sort of contamination/infection. At the time of embryo rescue in Laminar Air Flow Chamber, surface sterilization of caryopses was done using 0.1 per cent HgCl\u003csub\u003e2\u003c/sub\u003e for 1\u0026ndash;2 minutes followed by three washing with autoclaved distilled water. The embryos were then excised from sterilized seeds and transferred to the test tubes containing working MS medium. Half strength Murashige and Skoog (MS) medium(Murashige and Skoog \u003cspan class=\"CitationRef\"\u003e1962\u003c/span\u003e) supplemented with essential amino acids (Table\u0026nbsp;3.2 and 3.3) was used for the rescue of interploidy hybrid embryos as per suggested by Rines et al., 2003.Cultured embryos were placed in the dark section of Plant Growth Chamber (PGC) at 20\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C with 75 per cent humidity for regeneration for about a week till the roots and shoots initiated. The regenerated plantlets were then shifted to the other section of the PGC at 20\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C with 10/14 hrs light/dark profile with 75 per cent relative humidity for proper development of plantlets.The plantlets regenerated from embryo culture were treated with IBA (Indole butyric acid) and NAA (Naphthalene acetic acid) solution of 100 ppm concentration each for 2 minutes for profuse rooting.Potting mixture was prepared by mixing of coco peat, vermiculite and perlite in the proportion of 3:1:1\u003cem\u003ev/v\u003c/em\u003e. The plantlets were then transferred to the soil less media in small pots (5 cm diameter) for proper rooting and shooting. Appropriate conditions were maintained for proper growth.At 3\u0026ndash;4 leaf stage, some plantlets were treated with 0.05% and remaining were treated with 0.10% colchicine with 2% DMSO for 4 hours. DMSO acts as osmoregulator. After attaining 10\u0026ndash;12 leaf stage, the treated plants were transferred to potting mixture.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\n \u003ch2\u003e2.3 Data recorded\u003c/h2\u003e\n \u003cp\u003eObservations were recorded on five randomly selected competitive plants of parents for all the traits, and each plant data recorded in case of BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e generation. The agro-morphological characters were included days to 50% flowering, plant height (cm), number of leaves per plant, tillers per plant, leaf area (cm\u003csup\u003e2\u003c/sup\u003e), spikelets per panicle, green fodder yield/ plant (g), leaf: stem ratio, dry matter yield/ plant (g), days to 75% maturity, biological yield/ plant (g), seed yield/ plant (g), harvest index, 1000 grain weight (g), seed length (mm), protein content (%) and beta- glucan content (%). The protein content for each genotype was analysed by the quantitative method given by Lowry et. al. \u003cspan class=\"CitationRef\"\u003e1951\u003c/span\u003e.Beta-glucan content (%) was determined by the extraction method given by Wood et al. (1977) and estimation method given by Semedo et al. (\u003cspan class=\"CitationRef\"\u003e2015\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\n \u003ch2\u003e2.4 Molecular marker analysis\u003c/h2\u003e\n \u003cp\u003eTo detect the hybridity of F\u003csub\u003e1\u003c/sub\u003e plants from different interspecific crosses, 25 SSR primer pairs were screened for polymorphism among the parental \u0026lsquo;Avena\u0026rsquo; species. Details of these primers, including their sequences, are provided in Supplementary Table 1.Polymorphic SSR markers were used for hybridity test in F\u003csub\u003e1\u003c/sub\u003e and tracing the introgressions in BC\u003csub\u003e1\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e and BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e. Extraction of Plant genomic DNA, PCR amplification and Agarose gel electrophoresis were done in Molecular Cytogenetics and Tissue Culture Lab, Department of Genetics and Plant Breeding, CSK HPKV, Palampur. The PCR amplification was performed in a mixture containing 1.5 ul of DNA template, 0.5 ul of each forward and reverse primers, 0.1 ul of Taq polymerase (Takara Pvt. Ltd.), 2.0 ul of dNTP mix, 2 ul of PCR Buffer\u0026thinsp;+\u0026thinsp;mgcl\u003csub\u003e2\u003c/sub\u003e and 6.4 ul of sterile water with final volume of 12 ul per reaction.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\n \u003ch2\u003e2.5 Statistical Analysis\u003c/h2\u003e\n \u003cp\u003eData recorded for various traits was pertained to test whether the mean difference between mean values of plants is significant or not over individual cross mean as well as overall mean of BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e generation using simple t-test performed as:\u003c/p\u003e\n \u003cp\u003e\u003cimg src=\"data:image/png;base64,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\" style=\"width: 279px;\"\u003e\u003c/p\u003e\n \u003cdiv class=\"BlockQuote\"\u003e\n \u003cp\u003eWhere,\u003c/p\u003e\n \u003c/div\u003e\n \u003cp\u003e\u003cimg src=\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAEUAAAAuCAYAAAB+vtSwAAAAAXNSR0IArs4c6QAAAARnQU1BAACxjwv8YQUAAAAJcEhZcwAAFiUAABYlAUlSJPAAAAIRSURBVGhD7ZlBssFAEIYnb23HCahyAyfgCKqspNzBDWwsUOUMFi7AAZyAG1iysmKfl3/ezKtGi+dVehKqv6pUxmRmkU93p0OUpBjlii93VggqhUGlMKgUBpXCoFIYVAqDSmFQKQwqhSH3Nj+KIjf6H2V469B3HwZNHwaVwqBSGESkXC4X0+l0bNGt1Wpmt9u5KzzH49E0Gg27Hvuwv1BQaCU4HA5JvV5HEU/a7XZyPp/dlXviOLbrsB77ikZMClitVvZmcUynUzd7Deb9GqwvA6JSgI+CarWabLdbN/sDPmM+S1oRiEuhaUTT45X0Co24FEAjApEDylZHKEGkAFo7er3e77gsdYQSTArSA2niZeAoUx2hBJMCaBqVMW08QTva+XxuTqeTHe/3e7NcLu34FdbrtW300PBJEUzKbDYzi8XCjpvNpj2Px+On3W4RBJGCG4cAkD51zGazMWn62KjpdruZ3zp9BfjLK0MuuDQSgxZY2o/Qbtc/pm/xe31B9r2NdD0Sl5LV0T5r8TF3K4CbyxtRKTQauMcvjSLuRrGHRheA2FarJSpFrKYg9weDgR2jjgyHQzumVCoVM5lMTBpF9mnU7/eL/9kAODm58iwCbnkUUR+VPll15BHcHi8X1wBEQMhbSskTGnUQNhqNxKXoXxwMQdv8d0GlMKgUBpXCoFIYVAqDSmFQKQwqhUGl3GHMNy4pY33uMMuKAAAAAElFTkSuQmCC\" style=\"width: 36px;\"\u003e= mean-difference between two sets of related samples\u003c/p\u003e\n \u003cdiv class=\"BlockQuote\"\u003e\n \u003cp\u003eSE (X\u003csub\u003ed\u003c/sub\u003e)\u0026thinsp;=\u0026thinsp;Standard error of mean difference\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;Number of related samples\u003c/p\u003e\n \u003cp\u003eThe significance of backcross derivatives and parents was tested at tabulated value of (n-1) degree of freedom at 5% level.\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\n\u003ch2\u003e3.1 Interspecific hybridization in \u003cem\u003eAvena\u003c/em\u003e species\u003c/h2\u003e\n\u003cp\u003eA total of 73 F\u003csub\u003e1\u003c/sub\u003e hybrid plants were successfully developed from six interspecific crosses (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). All crosses were successful except for the \u003cem\u003eA. barbata\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e cross, which did not result in embryo formation. The successful hybrids were backcrossed with the recipient parent twice, resulting in 24 plants in the BC\u003csub\u003e1\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e generation and 26 plants in the BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e generation. For the cross between \u003cem\u003eA. orientalis\u003c/em\u003e and \u003cem\u003eA. sativa\u003c/em\u003e, 5 seeds were produced in the F\u003csub\u003e1\u003c/sub\u003e generation, followed by 2 seeds in BC\u003csub\u003e1\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e and 3 seeds in BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e. A greater number of seeds were obtained from the \u003cem\u003eA. longiglumis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e cross, with 28 F\u003csub\u003e1\u003c/sub\u003e seeds, 9 BC\u003csub\u003e1\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e seeds, and 5 BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e seeds. Similarly successful was the \u003cem\u003eA. strigosa\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e cross, which yielded 27 F\u003csub\u003e1\u003c/sub\u003e seeds, 6 BC\u003csub\u003e1\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e seeds, and 6 BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e seeds. When crossing \u003cem\u003eA. sativa\u003c/em\u003e with \u003cem\u003eA. byzantina\u003c/em\u003e, the resulting generations produced 5 F\u003csub\u003e1\u003c/sub\u003e seeds, 2 BC\u003csub\u003e1\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e seeds, and 4 BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e seeds. Two different cultivars of \u003cem\u003eA. sterilis\u003c/em\u003e were crossed with \u003cem\u003eA. sativa\u003c/em\u003e: cv. HFO 508 yielded 4, 3, and 4 seeds in F\u003csub\u003e1\u003c/sub\u003e, BC\u003csub\u003e1\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e, and BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e generations respectively, while cv. HFO 878 produced 4, 2, and 4 seeds in the corresponding generations.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003ctable id=\"Tab2\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003ePlants developed in F\u003csub\u003e1\u003c/sub\u003e, BC\u003csub\u003e1\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e and BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e generations of different interspecific crosses\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eCrosses\u003c/p\u003e\n\u003c/th\u003e\n\u003cth colspan=\"3\" align=\"left\"\u003e\n\u003cp\u003eNumber of Plants\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eF\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eBC\u003csub\u003e1\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eBC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eA. barbata \u0026times; A. sativa\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eA. orientalis \u0026times; A. sativa\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eA. longiglumis \u0026times; A. sativa\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e28\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eA. strigosa \u0026times; A. sativa\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e27\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e6\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eA. sativa \u0026times; A. byzantine\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e4\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eA. sativa \u0026times; A. sterilis\u003c/strong\u003e \u003cstrong\u003ecv. HFO 508\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e4\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eA. sativa \u0026times; A. sterilis\u003c/strong\u003e \u003cstrong\u003ecv. HFO 878\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e4\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eTotal\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e\u003cstrong\u003e73\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e\u003cstrong\u003e24\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e\u003cstrong\u003e26\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e\n\u003ch2\u003e3.2 Embryo culture in Interploidy hybridization\u003c/h2\u003e\n\u003cp\u003eIn the interploidy crossing program, species with lower ploidy levels were used as female parents, while species with higher ploidy levels were used as male parents, as this approach has a higher success rate according to previous studies. From 839 caryopses, a total of 73 hybrid embryos were developed. Specifically, 41 embryos were derived from the \u003cem\u003eA. strigosa\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e cross, 18 from \u003cem\u003eA. longiglumis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e, and 14 from \u003cem\u003eA. orientalis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e. These embryos were cultured on suitable MS medium and incubated in the dark for germination. The hybrid plant regeneration frequency was 61.64%, with 45 of the 73 embryos germinating within 7\u0026ndash;10 days. Of these, 43 embryos developed into seedlings with 3\u0026ndash;4 leaves.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\n\u003ch2\u003e3.3 Evaluation of BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e derivatives\u003c/h2\u003e\n\u003cp\u003eThe BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e derivatives were assessed for a range of agro-morphological and quality traits during the \u003cem\u003eRabi\u003c/em\u003e season of 2020-21, alongside parent lines. The evaluation revealed significant variations in key traits among the derivatives (Tables\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e and \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). Days to 50% flowering ranged from 117 to 142 days, with several plants, including P\u003csub\u003e1\u003c/sub\u003e of \u003cem\u003eA. orientalis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e (124 days) and P\u003csub\u003e2\u003c/sub\u003eand P\u003csub\u003e3\u003c/sub\u003e of \u003cem\u003eA. longiglumis\u003c/em\u003e \u0026times; \u003cem\u003eA. Sativa\u003c/em\u003e (122 and 124 days, respectively), flowering earlier than the overall BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e mean (130.30 days). Whereas, \u003cem\u003eA. strigosa\u003c/em\u003e (123 days) was significantly superior for this trait among the parents.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab5\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003ePromising BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e plants selected for important agro-morphological and quality traits.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eSr. No.\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eCross\u003c/p\u003e\n\u003c/th\u003e\n\u003cth colspan=\"4\" align=\"left\"\u003e\n\u003cp\u003ePlant number\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eSeed yield per plant (g)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eBiological yield per plant (g)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eDays to 75% maturity\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eBeta-glucan content (%)\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eA. orientalis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e1\u003c/sub\u003e, P\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e2.\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eA. longiglumis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e3\u003c/sub\u003e, P\u003csub\u003e4,\u003c/sub\u003e P\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e1\u003c/sub\u003e, P\u003csub\u003e2\u003c/sub\u003e, P\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e1\u003c/sub\u003e, P\u003csub\u003e2\u003c/sub\u003e, P\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eA. strigosa\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e1\u003c/sub\u003e, P\u003csub\u003e2\u003c/sub\u003e, P\u003csub\u003e5\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e1\u003c/sub\u003e, P\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e1\u003c/sub\u003e, P\u003csub\u003e2\u003c/sub\u003e, P\u003csub\u003e3\u003c/sub\u003e, P\u003csub\u003e5\u003c/sub\u003e, P\u003csub\u003e6\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e1\u003c/sub\u003e, P\u003csub\u003e3\u003c/sub\u003e, P\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eA. sativa\u003c/em\u003e \u0026times; \u003cem\u003eA. byzantina\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e1\u003c/sub\u003e, P\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5.\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eA. sativa\u003c/em\u003e \u0026times; \u003cem\u003eA. sterilis\u003c/em\u003e cv. HFO-508\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e1\u003c/sub\u003e, P\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e1\u003c/sub\u003e, P\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eA. sativa\u003c/em\u003e \u0026times; \u003cem\u003eA. sterilis\u003c/em\u003e cv. HFO-878\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eP\u003csub\u003e2\u003c/sub\u003e, P\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003ePlant height among the BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e derivatives varied from 58 cm to 95 cm. Plants P\u003csub\u003e1\u003c/sub\u003e of \u003cem\u003eA. orientalis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e (80.00 cm) and P\u003csub\u003e3\u003c/sub\u003e of \u003cem\u003eA. longiglumis\u003c/em\u003e \u0026times; \u003cem\u003eA. Sativa\u003c/em\u003e (95.00 cm) were notably taller than the overall mean (74.0 cm). \u003cem\u003eA. strigosa\u003c/em\u003e (82.00 cm) and \u003cem\u003eA. sterilis\u003c/em\u003e cv. HFO-508 (83.00 cm) showed significant superiority for plant height. Similarly, leaf number ranged from 22 to 60, with \u003cem\u003eA. strigosa\u003c/em\u003e (48.00), \u003cem\u003eA. byzantina\u003c/em\u003e (52.00), and \u003cem\u003eA. sterilis\u003c/em\u003e cv. HFO-878 (52.00) exhibiting superior performance.\u003c/p\u003e\n\u003cp\u003eThe number of tillers per plant varied from 5 to 14, with plants P\u003csub\u003e3\u003c/sub\u003e of \u003cem\u003eA. longiglumis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e (14.00) and P\u003csub\u003e1\u003c/sub\u003e and P\u003csub\u003e5\u003c/sub\u003e of \u003cem\u003eA. strigosa\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e (12.00 and 10.00, respectively) showing significant superiority. Leaf area ranged from 13.87 to 56.00 cm\u0026sup2;, with plants P\u003csub\u003e3\u003c/sub\u003e of \u003cem\u003eA. orientalis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e (26.50 cm\u003csup\u003e2\u003c/sup\u003e), and P\u003csub\u003e4\u003c/sub\u003e and P\u003csub\u003e5\u003c/sub\u003e of \u003cem\u003eA. longiglumis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e (56.00 and 37.33 cm\u003csup\u003e2\u003c/sup\u003e, respectively) demonstrating superior values. Number of spikelets per plant varied from 28 to 76, with several plants across different crosses showing significant superiority.\u003c/p\u003e\n\u003cp\u003eGreen fodder yield per plant ranged from 72 to 118 g. \u003cem\u003eA. strigosa\u003c/em\u003e (110.00 g) and \u003cem\u003eA. longiglumis\u003c/em\u003e (106.00 g) were exhibited superiority among the parents. The leaf:stem ratio ranged from 0.22 to 0.65, with significant superiority in several plants including P\u003csub\u003e3\u003c/sub\u003e of \u003cem\u003eA. orientalis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e (0.39), P\u003csub\u003e2\u003c/sub\u003e of \u003cem\u003eA. strigsa\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e (0.40). Dry matter yield per plant varied from 15.54 to 32.96 g, with \u003cem\u003eA. byzantina\u003c/em\u003e (24.09 g), \u003cem\u003eA. strigosa\u003c/em\u003e (29.77 g) and \u003cem\u003eA. longiglumis\u003c/em\u003e (31.10 g) showing significant superiority.\u003c/p\u003e\n\u003cp\u003eDays to 75% maturity ranged from 155 to 178 days, with several BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e derivatives showing earlier maturity than the overall mean. Biological yield per plant varied from 27.06 to 42.90 g, with \u003cem\u003eA. sterilis\u003c/em\u003e cv. HFO-508 (36.87 g) and \u003cem\u003eA. sterilis\u003c/em\u003e cv. HFO-878 (36.34 g) showing significant superiority. Seed yield per plant ranged from 3.11 to 7.30 g, with several plants, including those from \u003cem\u003eA. orientalis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e, showing superior values.\u003c/p\u003e\n\u003cp\u003eHarvest index ranged from 8.64 to 17.97%, with significant superiority in several plants from cross \u003cem\u003eA. orientalis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e and \u003cem\u003eA. longiglumis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e. Thousand-grain weight varied from 28.30 to 35.40 g, with \u003cem\u003eA. orientalis\u003c/em\u003e (32.70 g), \u003cem\u003eA. byzantina\u003c/em\u003e (34.40 g), and \u003cem\u003eA. longiglumis\u003c/em\u003e (35.70 g) showing significant superiority. Seed length ranged from 11.95 to 14.15 mm, with \u003cem\u003eA. orientalis\u003c/em\u003e (15.12 mm), \u003cem\u003eA. byzantina\u003c/em\u003e (14.60 mm), \u003cem\u003eA. longiglumis\u003c/em\u003e (10.40 mm), and \u003cem\u003eA. strigosa\u003c/em\u003e (10.15 mm) showing superior values.\u003c/p\u003e\n\u003cp\u003eProtein content ranged from 7.3\u0026ndash;11.00%, and all wild accessions except \u003cem\u003eA. sterilis\u003c/em\u003e cv. HFO-878 demonstrating superior values than overall mean and cultivated species \u003cem\u003eA. sativa\u003c/em\u003e. Beta-glucan content varied from 2.89\u0026ndash;5.42%, with \u003cem\u003eA. longiglumis\u003c/em\u003e (4.43%) and \u003cem\u003eA. strigosa\u003c/em\u003e (4.62%) showing significant superiority. Plant P\u003csub\u003e3\u003c/sub\u003e (3.97%) of cross \u003cem\u003eA. orientalis \u0026times; A. sativa\u003c/em\u003e; P\u003csub\u003e1\u003c/sub\u003e (5.42%), P\u003csub\u003e2\u003c/sub\u003e (4.31%) and P\u003csub\u003e5\u003c/sub\u003e (4.70%) of cross \u003cem\u003eA. longiglumis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e; P\u003csub\u003e1\u003c/sub\u003e (4.74%), P\u003csub\u003e3\u003c/sub\u003e (4.00%) and P\u003csub\u003e4\u003c/sub\u003e (4.95%) of \u003cem\u003eA. strigosa\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e; P\u003csub\u003e3\u003c/sub\u003e (3.94%) of \u003cem\u003eA. byzantina\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e; P\u003csub\u003e1\u003c/sub\u003e (4.10%) of \u003cem\u003eA. sativa\u003c/em\u003e \u0026times; \u003cem\u003eA. sterilis\u003c/em\u003e cv. HFO-508 showed significant superiority for this trait over overall mean.\u003c/p\u003e\n\u003cp\u003eOverall, the BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e derivatives exhibited promising traits for introgression into cultivated oat genotypes, potentially enhancing their performance. These findings align with previous research highlighting the benefits of interspecific crosses in improving plant traits (Murphy, 1981).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\n\u003ch2\u003e3.4 Molecular Screening\u003c/h2\u003e\n\u003cp\u003eMolecular characterization of all \u0026lsquo;\u003cem\u003eAvena\u003c/em\u003e\u0026rsquo; species accessions utilized 25 SSR primers. Among these, five SSR primers displayed polymorphism between wild species and the cultivated oat variety, \u003cem\u003eA. sativa\u003c/em\u003e cv. HJ-8. Each wild species exhibited unique polymorphic primers, except for MAMA_4, which was polymorphic in both \u003cem\u003eA. orientalis\u003c/em\u003e and \u003cem\u003eA. sterilis\u003c/em\u003e cv. HFO-878 (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e). The polymorphic primers confirmed the hybridity of F\u003csub\u003e1\u003c/sub\u003e plants from interploidy crosses, demonstrating the presence of alleles from both parent species (Plate 2). For intraploidy crosses, hybridity was verified in five plants of \u003cem\u003eA. sativa\u003c/em\u003e \u0026times; \u003cem\u003eA. byzantina\u003c/em\u003e, four plants of \u003cem\u003eA. sativa\u003c/em\u003e \u0026times; \u003cem\u003eA. sterilis\u003c/em\u003e HFO-508, and four plants of \u003cem\u003eA. sativa\u003c/em\u003e \u0026times; \u003cem\u003eA. sterilis\u003c/em\u003e HFO-878, all showing alleles from both parent species.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003ctable id=\"Tab6\" style=\"width: 514px;\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eList of SSR primers polymorphic for different \u003cem\u003eAvena\u003c/em\u003e species\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth style=\"width: 36px;\" align=\"left\"\u003e\n\u003cp\u003eS. No.\u003c/p\u003e\n\u003c/th\u003e\n\u003cth style=\"width: 47px;\" align=\"left\"\u003e\n\u003cp\u003ePrimer\u003c/p\u003e\n\u003c/th\u003e\n\u003cth style=\"width: 187.549px;\" align=\"left\"\u003e\n\u003cp\u003eCross\u003c/p\u003e\n\u003c/th\u003e\n\u003cth style=\"width: 133.451px;\" align=\"left\"\u003e\n\u003cp\u003eAnnealing Temp. (\u0026ordm;C)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth style=\"width: 81px;\" align=\"left\"\u003e\n\u003cp\u003eNo. of Alleles\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 36px;\" align=\"left\"\u003e\n\u003cp\u003e1.\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 47px;\" align=\"left\"\u003e\n\u003cp\u003eAM87\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 187.549px;\" align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eA. sativa \u0026times; A. byzantina\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 133.451px;\" align=\"left\"\u003e\n\u003cp\u003e52.59\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\" align=\"left\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 36px;\" align=\"left\"\u003e\n\u003cp\u003e2.\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 47px;\" align=\"left\"\u003e\n\u003cp\u003eMAMA_4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 187.549px;\" align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eA. orientalis \u0026times; A. sativa\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 133.451px;\" align=\"left\"\u003e\n\u003cp\u003e52.30\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\" align=\"left\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 36px;\" align=\"left\"\u003e\n\u003cp\u003e3.\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 47px;\" align=\"left\"\u003e\n\u003cp\u003eMAMA_5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 187.549px;\" align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eA. longiglumis \u0026times; A. sativa\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 133.451px;\" align=\"left\"\u003e\n\u003cp\u003e53.09\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\" align=\"left\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 36px;\" align=\"left\"\u003e\n\u003cp\u003e4.\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 47px;\" align=\"left\"\u003e\n\u003cp\u003eAM115\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 187.549px;\" align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eA. strigosa \u0026times; A. sativa\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 133.451px;\" align=\"left\"\u003e\n\u003cp\u003e51.17\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\" align=\"left\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 36px;\" align=\"left\"\u003e\n\u003cp\u003e5.\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 47px;\" align=\"left\"\u003e\n\u003cp\u003eAstavea\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 187.549px;\" align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eA. sativa \u0026times; A. sterilis\u003c/em\u003e cv. HFO 508\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 133.451px;\" align=\"left\"\u003e\n\u003cp\u003e54.35\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\" align=\"left\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 36px;\" align=\"left\"\u003e\n\u003cp\u003e6.\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 47px;\" align=\"left\"\u003e\n\u003cp\u003eMAMA_4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 187.549px;\" align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eA. sativa \u0026times; A. sterilis\u003c/em\u003e cv. HFO 878\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 133.451px;\" align=\"left\"\u003e\n\u003cp\u003e52.30\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\" align=\"left\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eIn the BC\u003csub\u003e1\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e generation, all plants from intraploidy crosses presented amplicon segments from both parent species, whereas in interploidy crosses, all plants of \u003cem\u003eA. orientalis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e possessed alleles from both parents. However, in the cross \u003cem\u003eA. longiglumis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e, plants 4, 7, and 8 out of 9 lacked the amplicon region from the donor parent P\u003csub\u003e1\u003c/sub\u003e.\u003c/p\u003e\n\u003cp\u003eIn the BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e generation, the cross \u003cem\u003eA. sativa\u003c/em\u003e \u0026times; \u003cem\u003eA. byzantina\u003c/em\u003e yielded four plants, three of which exhibited the amplified region of primer AM87 from both parents (Plate 3). Similarly, two plants each in the crosses \u003cem\u003eA. sativa\u003c/em\u003e \u0026times; \u003cem\u003eA. sterilis\u003c/em\u003e cv. HFO-508 and \u003cem\u003eA. sativa\u003c/em\u003e \u0026times; \u003cem\u003eA. sterilis\u003c/em\u003e cv. HFO-878 demonstrated amplicon regions from the donor parent. These plants were notably superior in seed yield per plant, biological yield per plant, and plant height. In the interploidy cross \u003cem\u003eA. orientalis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e, only one plant showed the allelic region for SSR marker MAMA_4 from both parents, and this plant excelled in days to 50% flowering, seed yield per plant, and tillers per plant compared to the recurrent parent \u003cem\u003eA. sativa\u003c/em\u003e cv. HJ-8. Similarly, in the cross \u003cem\u003eA. longiglumis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e, only plant P\u003csub\u003e1\u003c/sub\u003e exhibited the allelic region from the donor parent, and it was significantly superior in beta-glucan content, protein content, and days to 75% maturity over the recurrent parent. In the cross \u003cem\u003eA. strigosa\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e, three plants showed the allelic region from the donor parent and were also significantly superior in beta-glucan, protein content, and days to 75% maturity.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eThe successful introgression of desirable chromatin from wild \u0026lsquo;\u003cem\u003eAvena\u003c/em\u003e\u0026rsquo; species into cultivated oat underscores the potential of interspecific and interploidy hybridization in enhancing the genetic diversity and agronomic traits of cultivated oats (Tinker et al., \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). CWRs can significantly expand the genetic diversity of crops and introduce beneficial traits (Brozynska et al., 2016; Zhang et al., 2017). However, their direct use in breeding programs is often challenging due to the presence of undesirable traits like, linkage drag (peng et al., 2014) and common breeding barriers (Kashyap et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) with cultivated crops.Cross compatibility is notably low in hexaploid \u0026times; diploid \u0026lsquo;Avena\u0026rsquo; hybrids (Latta et al., \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). A significant challenge in transferring genes from wild diploid and tetraploid oat species into hexaploid cultivars is successfully producing healthy, vigorous inter-ploidy F\u003csub\u003e1\u003c/sub\u003e hybrids (Aung et al., 2010). These hybrids must maintain all chromosomes from both the cultivated and wild parents, ensuring that the gene of interest is effectively expressed (Aung et al., 2010).Few studies have focused on transferring traits from wild relatives to cultivated oats, particularly for agronomic characteristics (Hermann et al., 2014), nutritional traits (Ahokas and Manninen, \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2000\u003c/span\u003e), crown rust resistance (Rines et al., \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Aung et al., 2010), and powdery mildew resistance (Yu and Hermann, 2006) and they got positive result by successful introgression.\u003c/p\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003e4.1 Molecular Screening\u003c/h2\u003e \u003cp\u003eThis study generated a significant number of F\u003csub\u003e1\u003c/sub\u003e hybrids from various interspecific crosses, although the cross between \u003cem\u003eA. barbata\u003c/em\u003e and \u003cem\u003eA. sativa\u003c/em\u003e was unsuccessful due to the absence of embryo formation. The subsequent backcrossing efforts yielded promising BC\u003csub\u003e1\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e and BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e generations, indicating the feasibility of incorporating wild \u0026lsquo;Avena\u0026rsquo; chromatin into cultivated backgrounds.\u003c/p\u003e \u003cp\u003eEmbryo culture proved crucial in rescuing hybrid embryos from interploidy crosses, with a notable regeneration frequency of 61.64%. The development of 43 viable seedlings from 73 germinated embryos highlights the effectiveness of this technique in overcoming barriers associated with interploidy hybridization. These results align with previous studies, which suggest that using species with lower ploidy levels as females in crosses increases the likelihood of success. The hybrids demonstrated a broad range of agro-morphological variations, with several BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e derivatives exhibiting significant improvements in traits such as plant height, number of tillers, leaf area, green fodder yield, seed yield, and other agronomic characteristics.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003e4.2 Evaluation of BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e derivatives\u003c/h2\u003e \u003cp\u003eThe evaluation of BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e derivatives in our study revealed significant variations in key agro-morphological and quality traits, highlighting the potential of introgressing desirable traits from wild \u0026lsquo;Avena\u0026rsquo; species into cultivated oat (\u003cem\u003eA. sativa\u003c/em\u003e L.). The observed variability in days to 50% flowering, ranging from 117 to 142 days, demonstrates the potential for modifying flowering times through interspecific crosses. Notably, plants derived from \u003cem\u003eA.orientalis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e and \u003cem\u003eA.longiglumis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e exhibited earlier flowering compared to the overall BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e mean, suggesting that these crosses can be used to accelerate flowering times in cultivated oats, an important trait for adapting to varying climatic conditions (Smith et al., \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe significant variation in plant height among the derivatives, with values ranging from 58 cm to 95 cm, underscores the potential for introducing height traits from wild species such as \u003cem\u003eA. strigosa\u003c/em\u003e and \u003cem\u003eA. sterilis\u003c/em\u003e cv. HFO-508. Taller plants may offer advantages in forage yield and mechanical harvesting efficiency, consistent with findings from previous studies (Semchenko and Zobel, \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Rosser, 2014; Coblentz et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Similarly, the variation in leaf number and leaf area suggests that wild species can contribute to improved canopy architecture and productivity, aligning with the observations of leaf trait improvement in other interspecific crosses (Carlson et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe variation in number of tillers per plant and green fodder yield per plant further emphasizes the potential for improving these traits through wild species introgression. The observed superiority of certain plants for these traits, such as those from \u003cem\u003eA.longiglumis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e and \u003cem\u003eA. strigosa\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e, indicates that these wild species can enhance forage yield and quality, which is critical for sustainable oat production (Humphreys, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Kumar et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Chand et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Sood et al., \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn terms of yield components, the significant differences in biological yield, seed yield, harvest index, thousand-grain weight, and seed length suggest that wild species introgression can positively impact yield and grain quality. The superior values observed in plants from crosses involving \u003cem\u003eA. orientalis\u003c/em\u003e, \u003cem\u003eA. byzantina\u003c/em\u003e, and \u003cem\u003eA. longiglumis\u003c/em\u003e highlight the potential for developing high-yielding oat cultivars with desirable agronomic characteristics (Beavis and Frey, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e1987\u003c/span\u003e; Sharma et al., \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Additionally, the higher protein and beta-glucan content in certain wild accessions, particularly \u003cem\u003eA. longiglumis\u003c/em\u003e and \u003cem\u003eA. strigosa\u003c/em\u003e, aligns with findings from previous research that emphasize the nutritional benefits of integrating wild species into cultivated crops (Frison et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Kahane et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Ebert, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe promising traits observed in the BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e derivatives underscore the value of utilizing wild \u0026lsquo;\u003cem\u003eAvena\u003c/em\u003e\u0026rsquo; species for improving cultivated oat performance. This study's findings support previous research highlighting the benefits of interspecific crosses in enhancing plant traits and emphasize the potential for developing improved oat cultivars through continued exploration of wild species (Murphy, 1981; Johnson et al., 2004).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003e4.3 Molecular Screening\u003c/h2\u003e \u003cp\u003eThe molecular characterization of \u0026lsquo;\u003cem\u003eAvena\u003c/em\u003e\u0026rsquo; species accessions using 25 SSR primers provided valuable insights into the genetic introgression from wild species into cultivated oat (\u003cem\u003eA. sativa\u003c/em\u003e L.). Notably, five SSR primers exhibited polymorphism between the wild species and the cultivated variety, \u003cem\u003eA. sativa\u003c/em\u003e cv. HJ-8. This finding underscores the potential of SSR markers in distinguishing between wild and cultivated accessions, offering a robust tool for genetic analysis in oat breeding programs (Gunnaiah et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Thapa et al., \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Liu et al., 2022).\u003c/p\u003e \u003cp\u003eThe distinct polymorphic patterns observed in wild species, except for the MAMA_4 primer which was shared between \u003cem\u003eA. orientalis\u003c/em\u003e and \u003cem\u003eA. sterilis\u003c/em\u003e cv. HFO-878, highlight the genetic diversity present within the wild \u0026lsquo;\u003cem\u003eAvena\u003c/em\u003e\u0026rsquo; species. Such variability is crucial for the introgression of beneficial traits from wild relatives into cultivated varieties. Previous studies have similarly demonstrated the utility of SSR markers in revealing genetic differences between species and in assessing genetic diversity (Yarvaan et al., 2020; Arora et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Koroluk et al., 2022; Kaur et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe confirmation of hybridity in F\u003csub\u003e1\u003c/sub\u003e plants from interploidy crosses, as indicated by the presence of alleles from both parent species, reflects successful genetic transfer. This aligns with findings from other research where SSR markers were used to validate hybridization and the introgression of alien chromatin (Boutin et al., 2006; Zhang et al., 2009). Specifically, the detection of polymorphic alleles in intraploidy crosses, such as \u003cem\u003eA. sativa\u003c/em\u003e \u0026times; \u003cem\u003eA. byzantina\u003c/em\u003e and \u003cem\u003eA. sativa\u003c/em\u003e \u0026times; \u003cem\u003eA. sterilis\u003c/em\u003e, further supports the effectiveness of these markers in tracking genetic introgression and verifying the presence of alien genes.\u003c/p\u003e \u003cp\u003eHowever, the cross between \u003cem\u003eA. longiglumis\u003c/em\u003e and \u003cem\u003eA. sativa\u003c/em\u003e revealed a notable exception. In this cross, plants 4, 7, and 8 out of 9 lacked the amplicon region from the donor parent (P\u003csub\u003e1\u003c/sub\u003e). This observation highlights the challenges associated with introgressing genetic material from certain wild relatives. It suggests that the introgression from \u003cem\u003eA. longiglumis\u003c/em\u003e may be less efficient or that specific genetic barriers might be present (Jones et al., 2022). Such variability in successful introgression has been reported in other studies, where the presence or absence of donor chromatin in progeny varied significantly (Lee et al., 2020; Kim \u0026amp; Park, 2021).\u003c/p\u003e \u003cp\u003eThe differential success in introgression observed in this study underscores the need for a tailored approach when selecting wild relatives for breeding programs and highlights the importance of comprehensive molecular screening to ensure the retention of desirable chromatin segments. The successful integration of traits from wild \u0026lsquo;\u003cem\u003eAvena\u003c/em\u003e\u0026rsquo; species into cultivated oat demonstrates the value of molecular techniques in enhancing the genetic base of crop varieties. This approach aligns with other studies that emphasize the role of wild relatives in broadening genetic diversity and improving traits such as disease resistance and stress tolerance (Perrino et al., 2020; Bohra et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Kashyap et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Future research should focus on understanding the underlying genetic mechanisms influencing successful introgression and developing strategies to overcome barriers associated with specific wild species.\u003c/p\u003e \u003cp\u003eThe overall study's findings emphasize the importance of utilizing wild relatives and landraces to broaden the genetic base of cultivated oat crop. The introgressed lines developed in this study not only possess enhanced agronomic traits but also offer potential for further genetic improvement through conventional breeding and molecular techniques. The observed transgressive segregation and superior performance of certain BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e plants underscore the potential for creating new oat cultivars with improved yield, quality, and resilience to biotic and abiotic stresses.\u003c/p\u003e \u003cp\u003eThe introgression of desirable alien chromatin from wild \u0026lsquo;Avena\u0026rsquo; species into cultivated oat has yielded promising results, demonstrating the value of wild species in oat improvement programs (Boczowska et al., 2015; 2016). Several past research studies indicates that genetic diversity within modern oat gene pools is limited. Over time, valuable alleles have been lost through selection, introduction, and domestication making it essential to explore obsolete cultivars, wild ancestors, and landraces for new germplasm to advance oat breeding efforts (Okoń et al., \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Koroluk et al., 2022).\u003c/p\u003e \u003cp\u003eThe combination of agro-morphological evaluation and molecular characterization has provided a robust framework for selecting superior lines (Ghazy et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Bayahi and Rezgui, 2015; Guizani et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; khan et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), paving the way for the development of new oat varieties that meet the challenges of modern agriculture (Kumar et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Kebede et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Further research should focus on the continued evaluation and utilization of these introgressed lines in breeding programs, with particular attention to their performance under diverse environmental conditions.\u003c/p\u003e \u003c/div\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003eThis study successfully demonstrated the potential of interspecific and interploidy hybridization in \u0026lsquo;\u003cem\u003eAvena\u003c/em\u003e\u0026rsquo; species for enhancing cultivated oat (\u003cem\u003eA. sativa\u003c/em\u003e L.) through the introgression of desirable alien chromatin. A total of 73 F\u003csub\u003e1\u003c/sub\u003e hybrids were generated, and subsequent backcrossing produced BC\u003csub\u003e1\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e and BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e generations, revealing significant agro-morphological and quality trait variations. Key traits such as plant height, leaf number, green fodder yield, and protein and β-glucan content showed notable improvements in several BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e derivatives compared to the recurrent parent. These newely developed interspecific derivatives will be screened for ploidy level and alien introgressions using cytological techniques such as Genomic In situ Hybridization, after two more successive bakcrossings to achieve the stable useful introgressions. Molecular screening using SSR markers confirmed the introgression of chromatin from wild species into cultivated oat, validating the hybridity and the presence of alleles from both parent species. This integration of wild genetic material into cultivated oat shows promise for improving yield, stress tolerance, and nutritional quality, aligning with the benefits observed in previous research. The findings highlight the potential for utilizing wild \u0026lsquo;Avena\u0026rsquo; species to enhance the genetic base and performance of cultivated oats.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eAuthors contribution statement:\u003c/p\u003e\n\u003cp\u003eConceptualization, Methodology, Software, Validation, Formal analysis, Resources, Data curation, Writing-original draft- SK, VKS, SKS,\u003c/p\u003e\n\u003cp\u003eWriting-review and editing, Visualization, Supervision- GS, VR, RK.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAhokas H, Manninen ML (2000) Introgressive hexaploid oats from the \u003cem\u003eAvena abyssinia\u003c/em\u003e (♀) \u0026times; A. sativa hybrid: performance, grain lipids and proteins. Euphytica 113:153\u0026ndash;160\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eArora A, Sood VK, Chaudhary HK et al (2021) Genetic diversity analysis of oat (\u003cem\u003eAvena sativa\u003c/em\u003e L.) germplasm revealed by agro-morphological and SSR markers. Range Manage Agrofor 42:38\u0026ndash;48\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAung T, Zwer P, Park R et al (2010a) Hybrids of Avena sativa with two diploid wild oats (CIav6956) and (CIav7233) resistant to crown rust. Euphytica 174:189\u0026ndash;198. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10681-009-0111-5\u003c/span\u003e\u003cspan address=\"10.1007/s10681-009-0111-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAung T, Zwer P, Park R et al (2010b) Hybrids of Avena sativa with two diploid wild oats (CIav6956) and (CIav7233) resistant to crown rust. Euphytica 174:189\u0026ndash;198. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10681-009-0111-5\u003c/span\u003e\u003cspan address=\"10.1007/s10681-009-0111-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBayahi K, Rezgui S (2017) Genetic Study of Nutritional and Physicochemical Characters of Chickpea Lines and Cultivars (\u003cem\u003eCicer arietinum\u003c/em\u003e L). J Agric Sci 9:151. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.5539/jas.v9n3p151\u003c/span\u003e\u003cspan address=\"10.5539/jas.v9n3p151\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBeavis WD, Frey KJ (1987) Expression of nuclear-cytoplasmic interactions and heterosis in quantitative traits of oats (\u003cem\u003eAvena\u003c/em\u003e spp). Euphytica 36:877\u0026ndash;886. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/bf00051873\u003c/span\u003e\u003cspan address=\"10.1007/bf00051873\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBoczkowska M, Harasimiuk M, Onyśk A (2015) Studies on genetic variation within old Polish cultivars of common oat. Cereal Res Commun 43:12\u0026ndash;21. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1556/crc.2014.0025\u003c/span\u003e\u003cspan address=\"10.1556/crc.2014.0025\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBoczkowska M, Łapiński B, Kordulasińska I et al (2016) Promoting the Use of Common Oat Genetic Resources through Diversity Analysis and Core Collection Construction. PLoS ONE 11:e0167855. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1371/journal.pone.0167855\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0167855\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBohra A, Kilian B, Sivasankar S et al (2022) Reap the crop wild relatives for breeding future crops. Trends Biotechnol 40:412\u0026ndash;431. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.tibtech.2021.08.009\u003c/span\u003e\u003cspan address=\"10.1016/j.tibtech.2021.08.009\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBrozynska M, Furtado A, Henry RJ (2015) Genomics of crop wild relatives: expanding the gene pool for crop improvement. Plant Biotechnol J 14:1070\u0026ndash;1085. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/pbi.12454\u003c/span\u003e\u003cspan address=\"10.1111/pbi.12454\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCarlson CH, Stack GM, Jiang Y et al (2021) Morphometric relationships and their contribution to biomass and cannabinoid yield in hybrids of hemp (\u003cem\u003eCannabis sativa\u003c/em\u003e). J Exp Bot 72:7694\u0026ndash;7709. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/jxb/erab346\u003c/span\u003e\u003cspan address=\"10.1093/jxb/erab346\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChand S, Indu, Singhal RK, Govindasamy P (2022) Agronomical and breeding approaches to improve the nutritional status of forage crops for better livestock productivity. Grass Forage Sci 77:11\u0026ndash;32. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/gfs.12557\u003c/span\u003e\u003cspan address=\"10.1111/gfs.12557\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChhuneja P, Kaur S, Dhaliwal HS (2016) Introgression and Exploitation of Biotic Stress Tolerance from Related Wild Species in Wheat Cultivars. Sustainable Development and Biodiversity. Springer International Publishing, Cham, pp 269\u0026ndash;324\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCoblentz WK, Jokela WE, Bertram MG (2014) Cultivar, Harvest Date, and Nitrogen Fertilization Affect Production and Quality of Fall Oat. Agron J 106:2075\u0026ndash;2086. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2134/agronj13.0436\u003c/span\u003e\u003cspan address=\"10.2134/agronj13.0436\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCurto M, Winter S, Seiter A et al (2019) Application of a SSR-GBS marker system on investigation of European Hedgehog species and their hybrid zone dynamics. Ecol Evol 9:2814\u0026ndash;2832. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/ece3.4960\u003c/span\u003e\u003cspan address=\"10.1002/ece3.4960\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEbert A (2014) Potential of Underutilized Traditional Vegetables and Legume Crops to Contribute to Food and Nutritional Security, Income and More Sustainable Production Systems. Sustainability 6:319\u0026ndash;335. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/su6010319\u003c/span\u003e\u003cspan address=\"10.3390/su6010319\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEng W-H, Ho W-S (2019) Polyploidization using colchicine in horticultural plants: A review. Sci Hort 246:604\u0026ndash;617. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.scienta.2018.11.010\u003c/span\u003e\u003cspan address=\"10.1016/j.scienta.2018.11.010\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFrison EA, Cherfas J, Hodgkin T (2011) Agricultural Biodiversity Is Essential for a Sustainable Improvement in Food and Nutrition Security. Sustainability 3:238\u0026ndash;253. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/su3010238\u003c/span\u003e\u003cspan address=\"10.3390/su3010238\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGhazy AI, Ali MA, Ibrahim EI et al (2024) Characterization of Improved Barley Germplasm under Desert Environments Using Agro-Morphological and SSR Markers. Agronomy 14:1716. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/agronomy14081716\u003c/span\u003e\u003cspan address=\"10.3390/agronomy14081716\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGuizani A, Babay E, Askri H et al (2024) Screening for drought tolerance and genetic diversity of wheat varieties using agronomic and molecular markers. Mol Biol Rep 51. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s11033-024-09340-9\u003c/span\u003e\u003cspan address=\"10.1007/s11033-024-09340-9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGunnaiah R, Jagadeesha RC, Cholin S et al (2021) Genetic diversity assessment and population structure analysis of pomegranate cultivars from different countries and Himalayan wild accessions. J Hortic Sci Biotechnol 96:614\u0026ndash;623. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/14620316.2021.1899854\u003c/span\u003e\u003cspan address=\"10.1080/14620316.2021.1899854\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHerrmann MH, Yu J, Beuch S, Weber WE (2014) Quantitative trait loci for quality and agronomic traits in two advanced backcross populations in oat (\u003cem\u003eAvena sativa\u003c/em\u003e L). Plant Breeding 133:588\u0026ndash;601. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/pbr.12188\u003c/span\u003e\u003cspan address=\"10.1111/pbr.12188\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHisano H, Abe F, Hoffie RE, Kumlehn J (2021) Targeted genome modifications in cereal crops. Breed Sci 71:405\u0026ndash;416. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1270/jsbbs.21019\u003c/span\u003e\u003cspan address=\"10.1270/jsbbs.21019\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHuang X-Y, Shang J, Zhong Y-H et al (2022) Disaggregation of Ploidy, Gender, and Genotype Effects on Wood and Fiber Traits in a Diploid and Triploid Hybrid Poplar Family. Front Plant Sci 13. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fpls.2022.866296\u003c/span\u003e\u003cspan address=\"10.3389/fpls.2022.866296\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHumphreys MO (2005) Genetic improvement of forage crops \u0026ndash; past, present and future. J Agricultural Sci 143:441\u0026ndash;448. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1017/s0021859605005599\u003c/span\u003e\u003cspan address=\"10.1017/s0021859605005599\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eImam AG, Allard RW (1965) Population studies in predominantly self-pollinated species. Vi. Genetic variability between and within natural populations of wild oats from differing habitats in california. Genetics 51:49\u0026ndash;62. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/genetics/51.1.49\u003c/span\u003e\u003cspan address=\"10.1093/genetics/51.1.49\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKahane R, Hodgkin T, Jaenicke H et al (2013) Agrobiodiversity for food security, health and income. Agron Sustain Dev 33:671\u0026ndash;693. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s13593-013-0147-8\u003c/span\u003e\u003cspan address=\"10.1007/s13593-013-0147-8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKashyap A, Garg P, Tanwar K et al (2022) Strategies for utilization of crop wild relatives in plant breeding programs. Theor Appl Genet 135:4151\u0026ndash;4167. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s00122-022-04220-x\u003c/span\u003e\u003cspan address=\"10.1007/s00122-022-04220-x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKaur R, Kapoor R, Vikal Y, Amandeep A (2024) Molecular characterization of oats (Avena sativa L.) diversity: Implications for dual-purpose breeding. Genetika 56:103\u0026ndash;120. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2298/gensr2401103k\u003c/span\u003e\u003cspan address=\"10.2298/gensr2401103k\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKebede G, Worku W, Feyissa F, Jifar H (2023) Agro-morphological traits-based genetic diversity assessment on oat (\u003cem\u003eAvena sativa\u003c/em\u003e L.) genotypes in the central highlands of Ethiopia. All Life 16. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/26895293.2023.2236313\u003c/span\u003e\u003cspan address=\"10.1080/26895293.2023.2236313\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKhan T, Rashid R, Shah L et al (2024) Genetic and phenotypic diversity in \u003cem\u003eSolanum lycopersicum\u003c/em\u003e genotypes: insights from morpho-molecular and biochemical analyses. Plant Biotechnol Rep 18:207\u0026ndash;221. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s11816-024-00894-5\u003c/span\u003e\u003cspan address=\"10.1007/s11816-024-00894-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKoroluk A, Paczos-Grzęda E, Sowa S et al (2022a) Diversity of Polish Oat Cultivars with a Glance at Breeding History and Perspectives. Agronomy 12:2423. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/agronomy12102423\u003c/span\u003e\u003cspan address=\"10.3390/agronomy12102423\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKoroluk A, Paczos-Grzęda E, Sowa S et al (2022b) Diversity of Polish Oat Cultivars with a Glance at Breeding History and Perspectives. Agronomy 12:2423. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/agronomy12102423\u003c/span\u003e\u003cspan address=\"10.3390/agronomy12102423\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumar R, Kumar D, Datt C et al (2018) Forage Yield and Nutritional Characteristics of Cultivated Fodders as Affected by Agronomic Interventions: A Review. Indian J Anim Nutr 35:373. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.5958/2231-6744.2018.00057.9\u003c/span\u003e\u003cspan address=\"10.5958/2231-6744.2018.00057.9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumar R, Varghese S, Jayaswal D et al (2023) Agro-morphological and genetic variability analysis in oat germplasms with special emphasis on food and feed. PLoS ONE 18:e0280450. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1371/journal.pone.0280450\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0280450\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLatta RG, Bekele WA, Wight CP, Tinker NA (2019) Comparative linkage mapping of diploid, tetraploid, and hexaploid Avena species suggests extensive chromosome rearrangement in ancestral diploids. Sci Rep 9. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/s41598-019-48639-7\u003c/span\u003e\u003cspan address=\"10.1038/s41598-019-48639-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLiu Q, Lin L, Zhou X et al (2017) Unraveling the evolutionary dynamics of ancient and recent polyploidization events in Avena (Poaceae). Sci Rep 7. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/srep41944\u003c/span\u003e\u003cspan address=\"10.1038/srep41944\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLiu R, Huang Y, Yang T et al (2021) Population genetic structure and classification of cultivated and wild pea (\u003cem\u003ePisum\u003c/em\u003e sp.) based on morphological traits and SSR markers. J Syst Evol 60:85\u0026ndash;100. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/jse.12710\u003c/span\u003e\u003cspan address=\"10.1111/jse.12710\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265\u0026ndash;275. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/s0021-9258(19)52451-6\u003c/span\u003e\u003cspan address=\"10.1016/s0021-9258(19)52451-6\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMaluszynski M, Kasha K, Forster BP, Szarejko I (2013) Doubled Haploid Production in Crop Plants: A Manual. Springer Science \u0026amp; Business Media\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMohler V, Paczos-Grzęda E, Sowa S (2023) Loving the Alien: The Contribution of the Wild in Securing the Breeding of Cultivated Hexaploid Wheat and Oats. Agriculture 13:2060. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/agriculture13112060\u003c/span\u003e\u003cspan address=\"10.3390/agriculture13112060\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMurashige T, Skoog F (1962) A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures. Physiol Plant 15:473\u0026ndash;497. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.1399-3054.1962.tb08052.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1399-3054.1962.tb08052.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNelson DR, Adger WN, Brown K (2007) Adaptation to Environmental Change: Contributions of a Resilience Framework. Annu Rev Environ Resour 32:395\u0026ndash;419. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1146/annurev.energy.32.051807.090348\u003c/span\u003e\u003cspan address=\"10.1146/annurev.energy.32.051807.090348\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOkoń SM, Chrząstek M, Kowalczyk K, Koroluk A (2014) Identification of new sources of resistance to powdery mildew in oat. Eur J Plant Pathol 139:9\u0026ndash;12. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10658-013-0367-4\u003c/span\u003e\u003cspan address=\"10.1007/s10658-013-0367-4\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePeng T, Sun X, Mumm RH (2013) Optimized breeding strategies for multiple trait integration: I. Minimizing linkage drag in single event introgression. Mol Breeding 33:89\u0026ndash;104. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s11032-013-9936-7\u003c/span\u003e\u003cspan address=\"10.1007/s11032-013-9936-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePerrino EV, Perrino P (2020) Crop wild relatives: know how past and present to improve future research, conservation and utilization strategies, especially in Italy: a review. Genet Resour Crop Evol 67:1067\u0026ndash;1105. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10722-020-00930-7\u003c/span\u003e\u003cspan address=\"10.1007/s10722-020-00930-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRey E, Moln\u0026aacute;r I, Doležel J (2015) Genomics of Wild Relatives and Alien Introgressions. Alien Introgression in Wheat. Springer International Publishing, Cham, pp 347\u0026ndash;381\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRines HW (2003) Oat haploids from wide hybridization. Doubled Haploid Production in Crop Plants. Springer Netherlands, Dordrecht, pp 155\u0026ndash;159\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRines HW, Porter HL, Carson ML, Ochocki GE (2007) Introgression of crown rust resistance from diploid oat Avena strigosa into hexaploid cultivated oat \u003cem\u003eA. sativa\u003c/em\u003e by two methods: direct crosses and through an initial 2x\u0026middot;4x synthetic hexaploid. Euphytica 158:67\u0026ndash;79. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10681-007-9426-2\u003c/span\u003e\u003cspan address=\"10.1007/s10681-007-9426-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRogo U, Fambrini M, Pugliesi C (2023) Embryo Rescue in Plant Breeding. Plants 12:3106. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/plants12173106\u003c/span\u003e\u003cspan address=\"10.3390/plants12173106\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRosser CL, Beattie AD, Block HC et al (2016) Effect of maturity at harvest for whole-crop barley and oat on dry matter intake, sorting, and digestibility when fed to beef cattle1. J Anim Sci 94:697\u0026ndash;708. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2527/jas.2015-0063\u003c/span\u003e\u003cspan address=\"10.2527/jas.2015-0063\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSemchenko M, Zobel K (2005) The effect of breeding on allometry and phenotypic plasticity in four varieties of oat (\u003cem\u003eAvena sativa\u003c/em\u003e L). Field Crops Res 93:151\u0026ndash;168. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.fcr.2004.09.019\u003c/span\u003e\u003cspan address=\"10.1016/j.fcr.2004.09.019\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSemedo MC, Karmali A, Fonseca L (2015) A high throughput colorimetric assay of β-1,3-d-glucans by Congo red dye. J Microbiol Methods 109:140\u0026ndash;148. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.mimet.2014.12.020\u003c/span\u003e\u003cspan address=\"10.1016/j.mimet.2014.12.020\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSharma D, Tiwari A, Sood S et al (2018) Genome wide association mapping of agro-morphological traits among a diverse collection of finger millet (\u003cem\u003eEleusine coracana\u003c/em\u003e L.) genotypes using SNP markers. PLoS ONE 13:e0199444. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1371/journal.pone.0199444\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0199444\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSmith EG, Neil Harker K, O\u0026rsquo;Donovan JT et al (2018) The profitability of diverse crop rotations and other cultural methods that reduce wild oat (\u003cem\u003eAvena fatua\u003c/em\u003e). Can J Plant Sci 98:1094\u0026ndash;1101. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1139/cjps-2018-0019\u003c/span\u003e\u003cspan address=\"10.1139/cjps-2018-0019\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSood VK, Sanadya SK, Kumar S et al (2022) Health benefits of oat (\u003cem\u003eAvena sativa\u003c/em\u003e) and nutritional improvement through plant breeding interventions. Crop Pasture Sci 74:993\u0026ndash;1013. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1071/cp22268\u003c/span\u003e\u003cspan address=\"10.1071/cp22268\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTadsse W, Sanchez-Garcia M (2019) Genetic Gains in Wheat Breeding and Its Role in Feeding the World. Crop Breeding, Genetics and Genomics. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.20900/cbgg20190005\u003c/span\u003e\u003cspan address=\"10.20900/cbgg20190005\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eThapa R, Edwards M, Blair MW (2021) Relationship of Cultivated Grain Amaranth Species and Wild Relative Accessions. Genes 12:1849. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/genes12121849\u003c/span\u003e\u003cspan address=\"10.3390/genes12121849\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTinker NA, Wight CP, Bekele WA et al (2022) Genome analysis in Avena sativa reveals hidden breeding barriers and opportunities for oat improvement. Commun Biology 5. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/s42003-022-03256-5\u003c/span\u003e\u003cspan address=\"10.1038/s42003-022-03256-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ewood PJ, Paton D, Siddiqui R (2006) Determination of beta-glucan in oats and barley. Cereal Chem 54:524\u0026ndash;533\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYarvaan M, Zongwen Z (2020) Study of genetic diversity in oat accessions identified using of SSR markers. Mongolian J Agricultural Sci 29:120\u0026ndash;124. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.5564/mjas.v29i1.1380\u003c/span\u003e\u003cspan address=\"10.5564/mjas.v29i1.1380\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhang H, Mittal N, Leamy LJ et al (2016) Back into the wild\u0026mdash;Apply untapped genetic diversity of wild relatives for crop improvement. Evol Appl 10:5\u0026ndash;24. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/eva.12434\u003c/span\u003e\u003cspan address=\"10.1111/eva.12434\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003eTables 3 and 4 are available in the Supplementary Files section.\u003c/p\u003e"},{"header":"Plates","content":"\u003cp\u003ePlates 1 and 3 are available in the Supplementary Files section\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":true,"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":"Avena species, Interspecific crosses, Embryo culture, Oat","lastPublishedDoi":"10.21203/rs.3.rs-5057298/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5057298/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe genetic enhancement of polyploid oats poses a significant challenge for crop production improvement. This study aimed to introgress desirable agronomic and quality traits from wild \u0026lsquo;\u003cem\u003eAvena\u003c/em\u003e\u0026rsquo; species into cultivated \u003cem\u003eA. sativa\u003c/em\u003e L. We utilized three diploid species (\u003cem\u003eA. strigosa\u003c/em\u003e L., \u003cem\u003eA. longiglumis\u003c/em\u003e L., and \u003cem\u003eA. orientalis\u003c/em\u003e L.), one tetraploid species (\u003cem\u003eA. barbata\u003c/em\u003e L.), and three hexaploid species (\u003cem\u003eA. sterilis\u003c/em\u003e L., \u003cem\u003eA. byzantina\u003c/em\u003e L., and \u003cem\u003eA. sativa\u003c/em\u003e L.) for interspecific and interploidy crosses. Interspecific crosses were advanced to BC\u003csub\u003e1\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e and BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e generations over three \u003cem\u003eRabi\u003c/em\u003e seasons at CSK Himachal Pradesh Agricultural University, India. Phytohormone application, embryo culture, and colchicine treatment facilitated interploidy crosses, while intraploidy crosses were performed without manipulation. We developed 41 embryos from \u003cem\u003eA. strigosa\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e, 18 from \u003cem\u003eA. longiglumis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e, and 14 from \u003cem\u003eA. orientalis\u003c/em\u003e \u0026times; \u003cem\u003eA. sativa\u003c/em\u003e, with 15 plants surviving colchicine treatment (34.88% survival rate). SSR primers confirmed hybridity, and molecular screening of BC\u003csub\u003e1\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e and BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e generations showed donor alleles in 16 of 26 BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e plants. The BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e derivatives exhibited significant improvements over the recurrent parent, including up to 75% higher beta-glucan content, 41% higher protein content, 11.28% reduced maturity days, and 70.96% higher seed yield. These advancements underscore the potential of BC\u003csub\u003e2\u003c/sub\u003eF\u003csub\u003e1\u003c/sub\u003e derivatives in oat breeding programs for trait introgression and crop improvement.\u003c/p\u003e","manuscriptTitle":"Introgression of Desirable Alien Chromatin from Wild Avena Species into Cultivated Oat (Avena sativa L.) and Its Identification through Agro-Morphological and Molecular Techniques","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-11-21 16:35:59","doi":"10.21203/rs.3.rs-5057298/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":"5daca783-45cf-4229-be37-7253529a059b","owner":[],"postedDate":"November 21st, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-09-25T14:05:15+00:00","versionOfRecord":[],"versionCreatedAt":"2024-11-21 16:35:59","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5057298","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5057298","identity":"rs-5057298","version":["v1"]},"buildId":"cBFmMYwuxLRRLfASyISRj","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}