为了培育出无筛选标记基因的转基因水稻,试验将loxp-hpt-loxp基因与Bt基因连锁在一起转化水稻方法,得到loxp-hpt-loxp-Bt转基因水稻植株,再与同质的带有cre基因的水稻杂交,以定向删除潮霉素抗性筛选标记。试验表明以水稻品种"皖粳97"为供试材料,将成熟胚来源的愈伤组织用根癌农杆菌EHA105/pCAMBIA1305.1感染后,筛选出抗性愈伤组织并获得再生植株。经PCR验证,得到20棵转基因水稻植株。

Xa23,Pi9 and Bt genes which possess wide resistance spectrum and high resistance level respectively to bacterial blight(BB),fungi blast,stemborers and leaffolders of rice were pyramided by molecular marker-assisted selection(MAS).Two homozygous lines HB1471 and HB1473 carrying the three resistant genes were obtained.Resistance identification showed that the pyramided lines HB1471 and HB1473 simultaneously possessed high resistance level to BB,blast and leaffolders(Cnaphalocrocis medialis).Compared with the Xa23 and Pi9 genes donor parent L10,the pyramided lines had the similar resistance spectrum and level to BB and blast.Meanwhile,they exhibited equivalent resistance to leaffolders with Bt donor parent MH63-Bt.The homozygous lines carrying Xa23,Pi9 and Bt genes should be very useful in rice breeding as donor of multiple resistance genes.

以转Bt基因的"克螟稻"为研究材料,通过使用特异的引物和荧光标记探针,以已知转基因成份含量的水稻样品为模板建立标准曲线,对转基因水稻的NOS和Bt外源基因进行了荧光定量检测分析.初步建立了转基因水稻定量检测的技术方法.