柑桔基因转化新方法的研究

Agrobacterium mediated in vivo trsformation of lateral buds of citrus plants has been developed and been combined with in vivo cloning. This new approach has ingeniously detoured plant regeneration and the long juvenile stage of regenerated plants from protoplasts, which limits gene transformation of most ftuit trees including cirtus. It makes the elimination of remaining bacteria and the selection of transformants be easy and efficient by using the natural growth Of buds. In addition, this approach has combined the cloning of transformants in order to increase population rapidly. In demonstrating experiment, 21out of 112 treated buds of Shatian pulmmelo ( Citrus grandis Osb.) developed into primary branehes which gave 76 secondary buds. 4 in 76 secondary buds (5.26%) were prvod to be positive by GUS activity assay. Among 14 cluster buds differentiated from the in vitro culture of 4 positive buds, 4 showing highest GUS activity and being positive in Southern hybridization were regenerated into true plants. This approach could be used as a simple and effecient methed for the transformation of a wide range og fruit tree species.