Abstract
Aim. Creation of glyphosate-resistant canola plants expressing bifunctional hybrid desC::licBM3 gene. In the hybrid gene the sequence of DesC desaturase of cyanobacterium S. vulcanus without plastid targeting was fused with the sequence of thermostable lichenase reporter LicBM3 gene. Methods. Agrobacterium tumefaciensmediated transformation, PCR, quantitative and qualitative determination of lichenase activity, genetic analysis. Results. Transgenic canola plants, carring the enolpyruvat shikimat phosphate syntase gene (epsps), conferring on plants resistance to phosphonomethyl glycine herbicides (Roundup), as well as the desC::licBM3 gene, were selected. The presence of transgenes was confimed by multiplex PCR. The epsps gene expression in canola was shown at the transcription level, during in vitro growth and after greenhouse herbicide treatment. Activity of the licBM3 gene product as a part of hybrid protein allowed quantitative and qualiative estimation of the desaturase gene expression. Inheritance of heterologous genes and their expression in the first generation were investigated. Conclusions. Transgenic canola plants were obtained, the presence of trangenes in plant genome was proved and expression of the target genes was detected.
Highlights
Due to climate changes, the plant resistance to stress factors of various origin, including low temperatures and phytopathogens, becomes of great importance.One of the plant adaptive mechanisms to cold is an increase in the unsaturation of fatty acid residues in cellular membranes, sustaining the required membrane fluidity at low temperatures [1]
An important role in this process is attributed to fatty acid desaturases, catalyzing the transformation of a single bond between carbon atoms in acyl chains (C-C) into the double bond (C=C)
This work was aimed at canola plants, resistant to Roundup herbicide, which express the desaturase DesC of cyanobacterium S. vulcanus as a part of the bifunctional reporter gene for further testing of transgene lines for resistance to stress factors of different origin
Summary
One of the plant adaptive mechanisms to cold is an increase in the unsaturation of fatty acid residues in cellular membranes, sustaining the required membrane fluidity at low temperatures [1]. An important role in this process is attributed to fatty acid desaturases, catalyzing the transformation of a single bond between carbon atoms in acyl chains (C-C) into the double bond (C=C). The Agrobacterium tumefaciensmediated transformation resulted in the introduction of the gene coding for D9-acyl-lipid desaturase of cyanobacterium Anacystic nidulans into the Nicotiana tabacum L. genome. This enzyme catalyzes the formation of cys-double bond in position D9 in both 16- and SAKHNO L.
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