Glufosinate Ammonium Doses for In Vitro Selection of Bread Wheat Explants

Abstract

One of the problems of cereal biotechnology is the low efficiency of genetic transformation. The process of creating transgenic plants consists of three stages: transfer of foreign genetic material into the plant genome; obtaining of regenerated plants; selection of transgenic plants. Each stage is important and requires well-developed techniques. A widely used method of making transgenic plant is selection by including a selective antibiotic or herbicide resistance gene in DNA constructs. Effective breeding of transgenic plants requires knowledge of the exact concentrations and exposure duration of selective agents for the specific tissue culture species used. Our study aimed to identify the optimal concentration and exposure duration to glufosinate ammonium (Basta®) for the selection of resistant regenerates of bread wheat in vitro. Several options for the selection of resistant shoots were proposed based on the study results: induction of shoot regeneration on medium with 7.5 mg/L of glufosinate ammonium for 7 days after callus formation in the immature embryos culture on medium with Basta (concentration of at least 7.5 mg/L); constant cultivation of explants on media with glufosinate ammonium (minimum concentration—5 mg/L), when working with the culture of mature embryos on maternal grains; selection at the organogenesis stage should be carried out on the media with glufosinate ammonium concentration of 15 mg/L for 14 days or 10 mg/L for 28 days. The results of the work represent a significant contribution to accelerating work in the field of genetic and genomic studies of bread wheat—an important agricultural crop.