23 Jul Efficient transgene method for inducing caespitose shoots of soybean at high frequency
Abstract
The invention discloses an efficient transgene method for inducing caespitose shoots of soybean at a high frequency. The method comprises the following steps of: removing two cotyledons, primary leaves and redundant hypocotyls from an aseptic seedling along hypocotyls and growing point meristematic tissues of cotyledons to prepare an explant; analyzing pure tiny particles by adopting quartz sand; fully and uniformly mixing the pure tiny particles with engineering agrobacterium solution; performing intense vortex concussion on the explant on a vortex concussion instrument; and uniformly manufacturing a wound. According to the method, the dip dyeing cycle is shortened, the co-culture time is reduced, the toxic action on the plant explant is lightened and simultaneously the infected area of agrobacterium tumefaciens is increased; compared with cotylcdonary node transformation mediated by the agrobacterium tumefaciens, more caespitose shoot (mbt) tissues/tissue blocks of bud-leaf cushions are formed on a bud-induced culture medium, and transgene bud primordial can be detected about three weeks after the caespitose shoot (mbt) tissues/tissue blocks of the bud-leaf cushions are infected by the agrobacterium tumefaciens; besides, the regeneration frequency is high, so the genetic transformation efficiency of the soybean is greatly improved.
Year: 2011
Country: CN
Doc No: 102124947