小麦TaMlo基因的原核表达、抗体制备及细胞化学分析(简报)

白粉病菌(Blumeria graminis)是一类高度专化性的寄生真菌,可侵染650多种单子叶植物和 9000多种双子叶植物,能够引起多种麦类作物的白粉病,给农业生产带来巨大的损失。由于白粉病菌生理小种多、变异快,所以利用专化性抗病基因难以解决植物的持久抗病性问题。人们在研究大麦白粉病时．发现大麦Mlo基因的隐性突变可导致大麦对绝大多数白粉病菌生理小种的高效持久的广谱抗病性。Schulze-Lefert等多家实验室合作于1997年成功克隆了野生的 Mlo基因。进一步研究表明．该基因编码一种植物特有的具有7个跨膜区和羧基端长尾的膜蛋白(Mlo),它可能对植物细胞的坏死起负调控作用。但Mlo基因如何表达及其在白粉病菌发育中的作用机制尚不清楚。

PROKARYOTIC EXPRESSION, ANTISERUM PREPARATION AND CYTOCHEMICAL ANALYSIS OF TaMlo3 IN WHEAT

Mlo gene in wheat is critical to determine wheat broad spectrum disease resistance against the pathogenic powdery mildew fungus. According to wheat TaMlo3 cDNA sequence, the two DNA fragments encoding the intracellular loop 2 and C-terminus of wheat Mlo protein were cloned respectively into the vector pET-30a, and then the fusion polypeptides were expressed in E. coli BL21. The recombinant proteins were purified by Ni2 -NTA agarose column, and then were used to immunize the rabbits respectively. Two polyclonal antisera were obtained and they could be specifically bound on the extrahaustorial membrane and extrahaustorial matrix of the powdery mildew fungus by immuno-gold labeling.