DEVELOPMENT OF AN EFFECTIVE TECHNIQUE FOR IN VITRO Agrobacterium - MEDIATED GENETIC TRANSFORMATION OF WINTER RAPE Brassica napus L.

摘要

The aim of the research was to improve the genetic transformation and adaptation conditions of commercial winter rapeseed line Brassica napus L. with subsequent production of transgenic seeds.It was optimized the technique of Agrobacterium-mediated genetic transformation of winter rapeseed Bn1 line (Brassica napus L.) using hypocotyls of 6- day-old seedlings as explants. GUS activity histochemical analysis showed a positive reaction in calli cultures and regenerant plant leaves obtained after Agrobacterium tumefaciens GV3101 transformation, which carried pCB203 plasmid with gus and bar genes. The optimal transformation conditions were determined as follows: the bacterial suspension optical density at a 600 nm wavelength - OD = 0.5, 10 min of inoculation and 48 h of co-cultivation. Stable integration of marker genes was confirmed by both histochemical and PCR analysis.