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Information × Registration Number 0216U003444, 0111U003406 , R & D reports Title To elaborate methodic grounds and improve the technologies for production of spring barley haploids in anther and isolated microspore culture in vitro 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 barley anther culture in vitro, technological conditions of isolated microspore suspension production, gelling and trophic properties of chemically modified and natural starches as solidifying agents of nutrient media. Study Purpose - characterization of genotypic variability of new spring barley varieties by capacity for androgenesis in vitro, determination of technical and technological parameters of isolated microspore suspension production technique based on the use of an experimental laboratory compact blender, evaluation of gelling and trophic properties and morphogenetic effects of chemically modified and natural starches, elucidation of responses of genotypes with different in vitro androgenic ability to auxin- and cytokinin-active growth regulator contents in inductive medium, improvement of cold pretreatment of plant material, DH-line production from hybrids using the improved technology. Study Methods -anther and isolated microspore cultures in vitro, culture of isolated embryos, light microscopy, methods of variation statistics and analysis of variances (ANOVA). Based on the results of spring barley genotypic diversity evaluation for in vitro androgenic ability, three donors with high culture ability were selected among new varieties of Ukrainian breeding. Genotypic dependence of response plant growth regulator content in nutrient medium for anther culture in vitro was proved. For the first time, it was ascertained that genotypes with high androgenic capacity did not require auxin for androgenic structure induction unlike genotypes with low expression of this trait. At the same time, regardless of auxin- and cytokinin-active growth regulator contents and their ratios, androgenic capacity ranks of genotypes remained unchanged. The technique of barley microspore suspension production using an experimental small volume laboratory blender (patent No 9500) was elaborated. However, only the initial stages of sporophytic development of microspores were induced in isolated microspore culture due to unfavorable conditions of donor plant growth. On the basis of studying gelling properties and morphogenetic effects of eight new chemically modified starches and natural pea starches (patent No 103426), nutrient media for spring barley anther cultivation were worked out. A method of amylose content increase in corn seed starch was elaborated (patent No 101781), and the suitability of starch from aesu2 genotype as a gelling agent for nutrient medium was proved. For the first time, trophic properties of chemically modified and natural corn (mutations su2 and ae) and pea (normal type and mutation rugosus) starches with various amylose content in anther culture in vitro and in embryo culture were assessed. A new method of spike pretreatment at low positive temperature ensuring long-term storage of plant material and a sufficient increase in androgenic structure and plant-regenerate induction rates was worked out. A technology of barley androgenic haploid production in anther culture in vitro was improved due to these innovative elements. The efficiency of this technology in model genotypes exceeded 60 % of plants-regenerates of the total amount of cultivated anthers. When it was applied to obtain haploids from breeding hybrid material, it gave the mean plant regeneration frequency close to 20 %, which was 2.5-fold in comparison to that obtained by the basic technology. Within the program for waxy-barley breeding, 1,264 plants were regenerated and 170 doubled haploid lines were produced from 22 F1 hybrid cross combinations. The best DH-lines will be registered in NCPGRU. Scope of Application - genetic and physiological investigations in plant cell, tissue and organ in vitro cultivation; breeding aimed at generation and reproduction of valuable breeding material, including lines with improved grain quality Product Description popup.authors popup.nrat_date 2020-04-02 Close