Homozygous Transgenic Barley (Hordeum vulgare L.) Plants by Anther Culture.

Ludmila Ohnoutková,Tomáš Vlčko

doi:10.3390/plants9070918

Abstract

Production of homozygous lines derived from transgenic plants is one of the important steps for phenotyping and genotyping transgenic progeny. The selection of homozygous plants is a tedious process that can be significantly shortened by androgenesis, cultivation of anthers, or isolated microspores. Doubled haploid (DH) production achieves complete homozygosity in one generation. We obtained transgenic homozygous DH lines from six different transgenic events by using anther culture. Anthers were isolated from T0 transgenic primary regenerants and cultivated in vitro. The ploidy level was determined in green regenerants. At least half of the 2n green plants were transgenic, and their progeny were shown to carry the transgene. The process of dihaploidization did not affect the expression of the transgene. Embryo cultures were used to reduce the time to seed of the next generation. The application of these methods enables rapid evaluation of transgenic lines for gene function studies and trait evaluation.

Highlights

The plant life cycle includes the switch between the haploid gametophytic phase and the diploid sporophyte phase
Verified transgenic plants were grown in the greenhouse until the optimal stage for spike collecting
We addressed the question of how to and quickly obtain homozygous lines from T0 transgenic barley plants

Summary

Introduction

The plant life cycle includes the switch between the haploid gametophytic phase and the diploid sporophyte phase. This change is considered so critical that it is not possible to interfere with it. Since 1964, when the development of haploid embryos and plantlets from microspores of Datura inoxia was performed using anther cultures, there has been extensive research on the process and its use This process of inducing haploid or doubled haploid (DH) plants from meiotic spores (microspores) through anther culture or isolated microspores cultivated in vitro is called androgenesis [1]. The production of pure, homozygous plants is an important step in biological and genetic studies and in crop breeding. Populations derived from pure lines allow the study of phenotypic traits of selected genotypes in different environmental conditions and developmental stages [3]

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Discussion

Conclusion