狼尾草农杆菌转化体系的优化和转基因植株的获得

The Optimization of Pennisetum Agrobacterium-Mediated Transformation System and Obtaining of Transgenic Plants

以狼尾草[Pennisetum alopecuroides(L.)Spreng.]愈伤组织为遗传转化受体,利用农杆菌介导法导入黑麦草抗冻蛋白基因LpAFP,初步建立了狼尾草的遗传转化体系,获得了转化植株并利用PCR对14株抗性植株进行初步检测,检测呈阳性的有2株,PCR阳性转化率为14.28%;对2株阳性狼尾草进行RT-PCR检测,2株狼尾草RNA均进行了逆转录,对抗性植株进行了进一步的筛选。结果表明,LpAFP基因已整合到狼尾草基因组中。

The LpAFP gene was introduced into the Pennisetum alopecuroides （L.） Spreng. callus by agrobacterium-mediated method and the genetic transformation systerm was established initially. Fourteen transgenic plants were obtained,and two were identified as positive transgenic plants by PCR assay,PCR transform rate is 14.28% ; In 2 strains of positive RT-PCR test of 2 strains of Pennisetum americanum, RNA were reverse transcribed, antagonism plant for further screening. The results demonstrated the LpAFP gene was successfully integrated the genome of Pennisetum.