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Information × Registration Number 0216U003443, 0111U003405 , R & D reports Title To substantiate optimal parameters of the technology for production of winter rape haploids in anther culture in vitro and create lines of doubled haploids for heterotic breeding popup.stage_title Head Bilynska Olena Volodymyrivna, Кандидат біологічних наук Registration Date 03-03-2016 Organization PLANT PRODUCTION INSTITUTE ND. A. V. YA. YURYEV NATIONAL ACADEMY OF AGRАRIAN SCIENCES OF UKRAINE popup.description2 Study Object - the main factors of morphogenesis in oilseed rape anther, leaf and cotyledon explants cultures in vitro, micropropagation and long-term cultivation of regenerated plants. Study Purpose - determination of effects growth and pretreatment conditions of plant-anther donors on the efficiency of experimental androgenesis in vitro in oilseed rape; evaluation of spring and winter oilseed rape capacity for haploid formation in anther culture and direct morphogenesis in leaf and cotyledon explant cultures; improvement of nutrient media to induce embryogenesis in anther culture in vitro, to regenerate plants, and to perform micropropagation, finishing-growing, and long-term cultivation; improvement of the method for ploidy determination in regenerated androgenic plants; rape haploid and doubled haploid production from hybrid material. Study Methods - anther, shoot, plant, leaf and cotyledon explant cultures in vitro, light microscopy, methods of variation statistics. Effects of sowing time and plant growth temperature on the yield of androgenic embryoids and regenerated plants were investigated. The feasibility of investigations of rape doubled haploid production in anther culture in vitro in autumn-winter period and growing anther donor plants in climate chambers or greenhouses owing to a strong negative impact of high temperature and drought during the budding stage on the efficiency of haploid production at the traditional sowing time was experimentally rationalized. A technique of combined growing of donor plants (field/greenhouse), which ensured 50 %energy saving, was worked out. In experiments on the improvement of regimes and modes of pretreatment, the traditional methodic approaches (treatment of buds for several days at low positive temperature, including application of osmotic active substances) were showed to be ineffective, at least for plants obtained in stress growth conditions. An efficient technique for aseptic anther culture in vitro production was developed. Inductive nutrient medium containing normal pea starch instead of agar (patent No 103426) and medium for plant regeneration from embryoids, which increased the percentage of plants with normal morphology by 30 %, were proposed. Methods for clonal micropropagation of androgenic plants, growing and long-term in vitro cultivation on media containing chemically modified starches were elaborated. A technique for determination of plant ploidy by chloroplast counting in leaf stomatal closing сells was developed. For the first time, androgenic potential in 11 winter rape varieties and 11 spring rape varieties bred in Ukraine and Russia was evaluated. Two sourсes with high ability for culture formation (varieties 'Arion' and 'Chornyy Veleten' with the plant regeneration frequency of 68.7% and 8.9%, respectively), which will be registered in the NCPGRU, were identified. For the first time, the capacity for direct morphogenesis in cotyledon explants culture was described for 7 varieties bred in Ukraine, and varieties 'Arion' and 'Mariya' with high frequency of shoot regeneration (80 %) were identified. 34 plants of androgenic origin were regenerated from F1 hybrids in 5 cross combinations of spring rape and 7 cross combinations of winter rape. Scope of Application - genetic and physiological investigations in plant cell, tissue and organ cultivation in vitro, breeding aimed at creation and reproduction of valuable breeding material, elaboration of methods for long-term in vitro storage of plants in gene banks of plant genetic resources Product Description popup.authors Глухова Наталія Анатоліївна popup.nrat_date 2020-04-02 Close