转PvPGIP2基因小麦的获得与纹枯病抗性鉴定

Development and Characterization of PvPGIP2 Transgenic Wheat Plants with Enhanced Resistance to Rhizoctonia cerealis

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。

Polygalacturcuase-inhibing proteins （PGIPs） are defense proteins produced in plants,which can in- hibit the growth of pathogenic fungi. In this study, the coding sequence of the bean PGIP gene PvPGIP2 was cloned, and its transformation vector, pA25-PvPGIP2 was constructed, in which the expression of PvPGIP2 was drived by the maize ubiquitin promoter. A total of 4000 embryo callus of wheat variety Yangmai 18 were bombarded by the particle containing pA25-PvPGIP2,and 203 regenerated plants were obtained. By PCR test,65 positive transgenic plants were identified in the To generation, thus the transformation frequency was 1. 625%. The transgenic wheat plants in T1- T2 generations were subjected to PCR, RT-PCR, and Q-RT-PCR analysis, and disease resistance evalu- ation following inoculations with Rhizoctonia cereali. The results showed that the introduced alien PvPGIP2 gene in these transgenic wheat plants could be inherited,and expressed in the wheat plants. The transgenic wheat plants ex- pressing PvPGIP2 showed enhanced resistance to R. cerealis compared with untransformed Yangmai 18.