Abstract
Using Agrobacterium-mediated transformation procedure, a total of 204 primary transformants of the Elizaveta, Lugovskoi and Nevski potato varieties carrying the cry IIIa gene were obtained. A number of biomolecular tests, including Southern hybridization, insert copy number screening, immunofluorescence analysis for assessment of the target gene expression levels, and PCR control for testing of the insert integrity, was carried out for these transformants, as well as the biosafety field trials for the transgenic lines selected during the biomolecular analysis. As a result, 3 transgenic lines (E2, N1, and L5) carrying one insert copy per genome with target gene expression level over 10 ppm were selected.
Highlights
The estimated losses that the Colorado beetle (Leptinotarsa decemlineata Say) brings to potato crops vary from 18% of the potential yield for large state potato producers to 40–90% for private producers
The common strategy to create genetically modified plants resistant to insect pests is the usage of natural proteinaceous insecticides (δ-endotoxins); the genes encoding these proteins were extracted from various strains of the bacterium Bacillus thuringiensis [1,2,3,4,5]
The Agrobacterium tumefaciens GV 3101 strain [12,13] carrying the pMON38943 plasmid kindly provided by Monsanto Co was used for plant transformation
Summary
The estimated losses that the Colorado beetle (Leptinotarsa decemlineata Say) brings to potato crops vary from 18% of the potential yield for large state potato producers to 40–90% for private producers. The common strategy to create genetically modified plants resistant to insect pests is the usage of natural proteinaceous insecticides (δ-endotoxins); the genes encoding these proteins were extracted from various strains of the bacterium Bacillus thuringiensis [1,2,3,4,5] Such transgenic plants were named Bt-protected plants. It was found that foodstuffs and their components derived from Bt-plants were identical to the same products obtained from regular plants in virtually all respects [10,11] All these findings were the inspiration for the onset and successful realization of the program on creating genetically modified varieties of Russian selection resistant to Colorado beetle. This work is dedicated to creation of new biotechnological potato varieties by means of genetic engineering via Agrobacteria-mediated transformation by inserting a genetic vector carrying the cryIIIa gene into the genome of Elizaveta, Lugovskoi and Nevski potato plant varieties
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