抗根腐病的转GmPGIP3基因小麦扬麦18的获得与鉴定

Development and Characterization of GmPGIP3 Transgenic Yangmai 18 with Enhanced Resistance to Wheat Common Root Rot

GmPGIP3是大豆的一种多聚半乳糖醛酸酶抑制蛋白,能够特异性地抑制部分病原真菌内切多聚半乳糖醛酸活性,从而减弱病原菌对植株的侵害。利用基因重组技术构建了GmPGIP3基因的单子叶植物表达载体pA25-GmPGIP3,通过基因枪介导法将pA25-GmPGIP3转入小麦品种扬麦18中。对转GmPGIP3基因扬麦18的T0至T2代植株进行PCR、Southern杂交、半定量RT-PCR和荧光定量Q-RT-PCR分析,并对根腐病进行抗性鉴定。结果表明,GmPGIP3已转入扬麦18,并在转基因小麦中遗传、转录和表达;与受体材料相比,5个GmPGIP3过表达的转基因小麦株系对根腐病的抗性有明显提高。

GmPGIP3 is a polygalacturonase-inhibiting protein from soybean, which can reduce the infection of fungal pathogen through inhibiting the endo-polygalacturonase activity of pathogen fungi. Using genetic recombination technique, we constructed the transformation vector of GmPGIP3 gene expressing highly in monocot plants, pA25-GmPGIP3, in which GmPGIP3 gene was driven by maize ubiquitin promoter. Embryo callus of Yangmai 18 was bombarded by the particle containing pA25-GmPGIP3 vector DNA. The GmPGIP3 transgenic wheat plants from T 0 to T 2 generations were detected by PCR, Southern blot, RT-PCR, and Q-RT-PCR analyses. The alien GmPGIP3 proved to be introduced into the transgenic wheat lines with heritability and expression events. We also evaluated the disease resistance in these GmPGIP3 transgenic plants through inoculating the common root rot pathogen, Bipolaris sorokiniana. Compared to untransformated Yangmai 18, five GmPGIP3 transgenic lines showed significantly-enhanced resistance to Bipolaris sorokiniana.