转PvPGIP2基因小麦的获得与纹枯病抗性鉴定

多聚半乳糖醛酸酶抑制蛋白(PGIP)是一种植物防卫蛋白,可阻止一些病原真菌的侵害。本研究克隆出扁豆PvP-GIP2基因编码序列,构建了受玉米泛素(ubiquitin)启动子控制的PvPGIP2基因表达载体pA25-PvPGIP2;采用基因枪法将pA25-PvPGIP2转化小麦推广品种扬麦18幼胚愈伤组织4000块,获得了203株再生植株。PCR检测出阳性植株65株,转化率为1.625%。对转PvPGIP2基因小麦T1～T2植株,进行外源基因的PCR、RT-PCR、荧光定量RT-PCR(Q-RT-PCR)分析和小麦纹枯病抗性鉴定。结果表明,转入的PvPGIP2能够在转基因小麦中遗传、转录与表达;PvPGIP2基因的表达提高了转基因植株对小麦纹枯病的抗性。

Development and Characterization of PvPGIP2 Transgenic wheat Plants with enhanced resistance to Rhizoctonia cerealis

Polygalacturcuase-inhibing proteins (PGIPs) are defense proteins produced in plants, which can inhibit the growth of pathogenic fungi. In this study, pA25-PvPGIP2, the transformation vector of a bean PGIP gene PvPGIP2, which can be highly expressed in monocot plants, was correctly constructed. In the pA25-PvPGIP2 vector, PvPGIP2 gene was driven by maize ubiquitin promoter. 4000 of embryo callus of Yangmai 18 were bombarded by the particle containing pA25- PvPGIP2. Among the regenerated 203 plants, 65 positive transgenic individuals were identified through PCR assay in the T0 generation with a transformation frequency of 1.625 %. The transgenic wheat plants from T0 to T2 generations were subjected to PCR, RT-PCR, and Q-RT-PCR, and inoculations with Rhizoctonia cerealis as well as the disease resistance tests. Results showed that the alien PvPGIP2 gene was introduced into these transgenic wheat plants, can be inherited from T0 to T2, and expressed in the wheat background. The transgenic wheat plants expressing PvPGIP2 showed enhanced resistance to R. cerealis compared with untransformed Yangmai 18.