Speakers - 2026

Title: Application of anther cuiture in vitro for production of introgression dh-lines from hybrids between hordeum vulgare L. and hordeum spontaneum C. koch

Abstract

Despite great achievements in the fields of genetic transformation and gene edition, wild hybridization remains and probably will remain the important alternative for development of crop cultivars with improved adaptability to biotic and abiotic stresses as well as with high grain quality. In most cases, breeding process based on wide hybridization is aimed at production of introgression lines possessing small segments of wild genome after elimination of undesirable traits of wild parent by repeating backcrossing and self-pollination. Anther culture in vitro as universal method of experimental haploidy opens the possibility to speed up breeding process in comparison to backcrosses. Additional advantages may be provided by gametic selection occurred in anther culture in vitro. This phenomenon seems to be underestimated in regard to determination of genetic diversity of DH-lines. At the same time preferential sporophytic development (formation of callus, somatic embryos and regenerated plants instead of pollen grains) of microspores with favorable for androgensesis in vitro chromosome sets can induce distorted segregation in DH-populations depending on androgenetic response of parental components of hybrids. The present research work was carried out to produce doubled haploid lines from hybrids between cultivated barley Hordeum vulgare and Hordeum spontaneum via anther culture in vitro. This latter species is considered to be wild relative, progenitor of cultivated barley and a source of valuable genes. Ten crosses were used as plant material. Previous screening of parent forms showed that all four H. spontaneum accessions were characterized by low androgenetic response. In contrast, among five spring barley genotypes one line of androgenetic origin and one variety had high androgenetic capacities. Donor plants were grown in field. Cut tillers were cold pretreated for 5 days at 4 ºC in water. According to another protocol, isolated spikes were subjected to long-term cold pretreatment for 28 days at the same temperature. Anthers were cultivated on agar solidified nutrient media containing N6 macro- and MS micronutrients, physiologically active substances and maltose (9.0 %). It was found that the number of morphogenic anthers varied from 13.2 to 64.0 % depending on cross combination and cold pretreatment. The yield of green plans ranged from 1.0 to 39.4 per 100 cultivated anthers. In model genotype DH00-126 the percentage of morphogenic anthers reached 76.1 %, and the number of green plants exceeded 100. More than 500 green plants were obtained in the experiment. In cross combinations developed by involving DH0-126 which is a model spring barley genotype with the highest androgenetic ability, introgression DH-lines of vulgare-type were prevailed. When hybrids created by crossing less contrast parents were used as plant material, most of DH-lines had intermediate spike morphology and grain size. Several plants of spontaneum-type with nonbrittle rachises and spring barley growth habit as distinct from H. spontaneum were found.