Editing of genes coding protein involved in maize gamete membrane interaction and fusion

An agrobacterial transformation of maize was performed using a CRISPR/Cas9 vector for knockout the Zm_gex2 and Zm_gsc1 genes.

Agrobacterial transformation of maize by in planta and in vitro methods: problems and decisions

Problems of agrobacterial transformation of maize by methods in planta and in vitro are described. Experimental data on optimization of regeneration from transformed embryogenic maize callus of the KM line are presented.

Experssion vectors for editing kafirin (k1C5, gKAF1) and ameiotic 1 genes in sorghum by agrobacterial transformation

One of the effective approach to improve the nutritional value of sorghum grains is the production of mutants with partially or completely blocked synthesis or the altered amino acid composition of spare proteins - kafirins. The usage of the genomic editing method allows to solve this problem by introducing mutations into the nucleotide sequences of the genes α- and γ-kafirins (k1C5 and gKAF1). Another important area in the genetics and selection of agricultural plants is the development and use of apomixis technologies to fix heterosis in F1 hybrids. The DYAD arabidopsis gene and its homolog ameiotic 1 are the potential apomixis candidate genes involved in the normal course of meiosis, the induction mutations in which is one of the way to solve this problem. In the course of research, the genomic targets (20 bp sequences) were selected to introduce mutations into the genes of α- and γ-kafirins of sorghum and ameiotic 1 gene through CRISPR/Cas genomic editing technology. Guide RNA design is performed using bioinformatic programs CRISPROR and CHOPCHOP. Cloning of the α- and γ- kafirins and ameiotic 1 targets of sorghum into vector pBAtC was carried out at sites AarI. Validation of the success of target cloning was performed by DNA sequencing. The created constructions were transferred by electroporation to the agrobacterial strain AGL0. Currently, the created vectors are used for the agrobacterial transformation of sorghum plants.

Optimization of the parameters of Agrobacterium-mediated transformation of bar-shaped millet

An efficient Agrobacterium-mediated transformation method has been developed and optimized for switchgrass (Panicum virgatum L.). The aim of this study is to optimize the conditions for agrobacterial transformation of upland switchgrass cultivars Alamo, Forestburg, Pathfinder, Shawnee, and Trailblazer. For genetic transformation, the authors have used embryogenic calli of switchgrass cultivars. As a result of the study the effect of the virulence gene inducer, surfactant, optical density of bacterial suspension and co-cultivation time, it has been revealed that the presence of acetosyringone with silwet-L77 in the medium, bacterial suspension at an optical density of 0.6 and 8 days of co-cultivation with bacteria is optimal for Agrobacterium– mediated transformation of embryogenic calli of switchgrass. The results obtained can serve as a basis for the creation of transgenic plants of switchgrass.

Copper and nickel resistance at ipt-transgenic potato plants

Using agrobacterial transformation collection of /pr-transgenic plants based on cv. Adretta was obtained. Analysis of transgenic plants susceptibility to the exogenous phytohormones auxin (IAA) and cytokinin (kinetin) for 18 from 22 analyzed forms reveals altered reaction to hormones. Among 18 plants with altered reaction to phytohormones for 16 forms enhanced resistance to copper sulfate and/or nickel chloride was observed