Accelerated development of rice populations using anther culture in vitro method

Background. Inclusion of doubled haploids in the breeding process allows increasing the speed and reliability of selection of desired forms with smaller population volumes. To stabilize the genotype, anthers of hybrid combinations of F2, F3, F4 and F5 generations, developed in crossings of white-grained samples with traits of high nutritional value of grain, and anthers of combinations of F4 generation from crossings of varieties contrasting in pericarp color and amylose content, were introduced into the culture. The responsiveness to gamete technologies was studied in 21 hybrid combinations. New genetically stable material (DH - doubled haploids) was developed. Phenotyping was carried out in conditions of a vegetation experiment for economic and biological traits and elements of plant productivity in four populations, which included 45 DH lines. Variability was noted in a number of traits within the DH line populations. Based on the results of the biometric analysis of the regenerated populations, 6 sources with a “1000 grain mass” of more than 30 grams were identified.

The study was carried out with the financial support of the Kuban Science Foundation and the Russian Science Foundation within the framework of the scientific project No. 25-16-20103 “Application of the genomic approach in rice breeding for high technological grain quality”

Purpose. To study the response of hybrids obtained from crossing contrasting rice samples to in vitro pollen culture, to accelerate the creation of DH line populations based on the studied genotypes, and to phenotype them.

Materials and methods. The research was conducted at the Laboratory of Biotechnology and Molecular Biology at the Federal Research Center for Rice, using the culture of isolated anthers in vitro according to the generally accepted method of R.G. Butenko (1990).

Results. The genetic determinism of the “regeneration” trait in the donor plants used in the crossbreeding was noted. The genotypes of the crossbreeding involving the varieties Favorit and Azovsky proved to be the most productive in terms of the output of highly morphogenic calluses and androgenic lines. Phenotypic analysis revealed significant diversity among the plants in the individual DH populations in terms of the shape of the panicle, the angle of the flag leaf deviation, the length of the growing season, the weight of 1,000 grains, the height of the plants, and the length of the growing season.

Conclusion. Phenotypic analysis of the DH lines’ traits showed that their genesis originates from microspores, thus these lines are a valuable genetic resource. The inclusion of doubled haploids in the breeding process will help to facilitate the assessment of recombinant genotypes arising from the cross, will allow to detect rare recessive alleles, will increase the speed and reliability of the selection of desired forms with smaller population volumes.