Chowrira G M, Akella V, Fuerst P E, Lurquin P F
Department of Genetics and Cell Biology, Washington State University, Pullman 99164-4234, USA.
Mol Biotechnol. 1996 Apr;5(2):85-96. doi: 10.1007/BF02789058.
Electroporation-mediated gene transfer into intact plant tissues was demonstrated in pea, cowpea, lentil, and soybean plants. Transient expression of a chimeric gus reporter gene was used to monitor the uptake and expression of the introduced DNA in electroporated nodal axillary buds in vivo. The branches that grew out of the nodal meristems were chimeric and expressed the introduced gene up to 20 d after electroporation. Transgenic R1 pea, lentil, and cowpea plants were recovered from seeds originating on these chimeric branches as shown by Southern blot hybridization and GUS expression. Transgenic R2 soybean and lentil plants were also obtained. Segregation ratios in these populations showed a strong bias against transgene presence or expression.
在豌豆、豇豆、小扁豆和大豆植株中证明了通过电穿孔介导的基因转移到完整植物组织中。使用嵌合gus报告基因的瞬时表达来监测体内电穿孔的节间腋芽中导入DNA的摄取和表达。从节间分生组织长出的枝条是嵌合体,并且在电穿孔后长达20天表达导入的基因。通过Southern印迹杂交和GUS表达表明,从这些嵌合枝条上产生的种子中获得了转基因R1豌豆、小扁豆和豇豆植株。还获得了转基因R2大豆和小扁豆植株。这些群体中的分离比例显示出对转基因存在或表达的强烈偏向。