小白杏自交不亲和S-RNase基因RNAi表达载体的构建及遗传转化  

Construction of RNAi expression vector and genetic transformation of self-incompatibility S-RNase gene from Xiaobaixing apricot(Prunus armeniaca L.)

作　　者：刘小芳[1] 冯建荣[1] 刘海楠[1] 罗明[2] 郭明星[1] 谢晓婷[1] 

机构地区：[1]石河子大学农学院,特色果蔬栽培生理与种质资源利用兵团重点实验室,新疆石河子832003 [2]西南大学生物技术中心,农业部生物技术与作物品质改良重点开放实验室,重庆北碚400715

出　　处：《石河子大学学报（自然科学版）》2016年第4期409-414,共6页Journal of Shihezi University(Natural Science)

基　　金：国家自然科学基金项目(31272129)

摘　　要：为了应用RNAi技术调控杏自交不亲和性状,根据自交不亲和S66-RNase基因序列(Gen Bank登录中)设计特异引物,通过RT-PCR扩增S66-RNase基因(Gen Bank登录中)高变区及下游(63 bp)作为靶基因,构建RNAi表达载体的ihp RNA。以植物表达载体p CAMBIA为骨架载体,利用农杆菌介导的方法转化本氏烟,利用子房转化法转化小白杏。结果显示:获得了长度为439 bp的ihp RNA。表达载体酶切检验表明S66-RNase基因的RNAi表达载体构建和转入农杆菌LBA4404成功。通过抗性筛选和GUS染色检测,获得了16株阳性植株。结论:获得转基因烟草植株证明了本研究构建的RNAi植物表达载体有效,为诱导杏S-RNase基因转录后基因沉默、获得自交亲和的杏提供参考。We aimed to regulate self-incompatibility in apricot by RNAi technology in this study. The hypervariable region and downstream（63 bp） of S66-RNase gene based on the S66-RNase gene sequence of ＇Xiaobaixing＇ apricot was selected as target gene to construct ihp RNA of RNAi expression vector, which was amplified with the specific primers by PCR method. The expression vector p CAMBIA was used as backbone vector. The result showed that RNAi vector was transformed into Nicotiana benthamiana by Agrobacterium-mediated transformation. The length of ihp RNA was 439 bp, and RNAi expression vector of S66-RNase gene was constructed and transferred into LBA4404 successfully. 16 transgenic plants were obtained by means of resistance screening and GUS staining detection. This study provided a basis for inducing posttranscriptional gene silencing in the transformed plants and obtaining self-compatibility apricot.

关 键 词：杏 S-RNASE基因 ihpRNA RNAi表达载体 烟草 

分 类 号：S662.2[农业科学—果树学]