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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。
Polygalacturonase inhibitory protein (PGIP) is a plant defensive protein, can prevent some pathogenic fungi. In this study, we cloned the coding sequence of PvP-GIP2 gene from lentils and constructed the expression vector pA25-PvPGIP2 of PvPGIP2 gene under the control of ubiquitin promoter. The pA25-PvPGIP2 gene was transformed into the progeny of wheat variety Yangmai 18 by particle bombardment Injured tissue 4,000, obtained 203 regenerated plants. PCR detected 65 positive plants, the conversion rate was 1.625%. The transgenic wheat plants T1 ~ T2 were subjected to PCR, RT-PCR, Q-RT-PCR analysis and wheat sheath blight resistance identification. The results showed that the transferred PvPGIP2 gene could be inherited, transcribed and expressed in transgenic wheat. The expression of PvPGIP2 gene increased the resistance of transgenic plants to wheat sheath blight.