大豆与胞囊线虫互作中GmPR的表达研究

Expression of pathogenesis-related proteins(GmPRs) in interactions between soybean and soybean cyst nematode

为探究大豆在大豆胞囊线虫(Heterodera glycines)胁迫下病程相关蛋白基因(GmPR)的表达情况,以大豆(Glycine max)感病品种辽豆15和抗病品种灰皮支黑豆为材料,通过实时荧光定量PCR和亚细胞定位,检测了大豆接种大豆胞囊线虫后,六种GmPR基因的表达水平,分析这些基因在抗病品种和感病品种中的表达情况,并且构建了融合表达载体以探究GmPR2和GmPR10在亚细胞水平的分布情况。结果表明,在抗病品种中,胞囊线虫强烈诱导GmPR2(β-1,3-葡聚糖酶基因)的表达,在线虫的侵染初期和发育期大幅度上调表达,在感病品种中表达量差异不显著,表明葡聚糖酶基因可能在大豆抗胞囊线虫中起到抗性作用;GmPR3和GmPR10在线虫发育期表达量达到最高值,表明这些基因可能与大豆抗胞囊线虫发育的调控有关。通过亚细胞定位研究,GmPR2主要分布于细胞膜,GmPR10主要分布于细胞质和细胞核,与预测结果相符。大豆胞囊线虫的侵染能够不同程度地提高病程相关蛋白基因转录水平,GmPR基因的表达特点表明其在大豆抗胞囊线虫过程中起不同的作用。

Pathogenesis-related proteins were originally discovered to accumulate in plants that were infected by pathogens,but were later found to accumulate in plants in response to a variety of biotic and abiotic stresses.It played a very important role in the process of plant disease. In order to investigate the expression of pathogenesis-related proteins （GmPRs） in soybean roots after inoculated by Heterodera glycines. Six GmPRs genes expression were examined after inoculated by Heterodera glycines in resistant cultivar Huipizhi heidou and susceptible cultivar Liaodou 15 by using real-time quantitative PCR （RT-PCR） technology and subcellular localization. Furthermore expression vector was constructed to research the subcellular localization of GmPR2 and GmPR10.The results showed that, in the resistant cultivar,the expression of GmPR2（β-1,3- glucanase）were significantly induced by inoculation treatment at early stage of inoculation and development.The difference of expression was not significant in susceptible cultivars, indicating that GmPR2 may also play an important role in soybean against Heterodera glycines;GmPR3 and GmPR10 reach the peak at the stage of development indicating that it could be related to prevent nematode development.Through subcellular localization, GmPR2 mainly located in cytomembrane, GmPR10 mainly distributed in the cytoplasm and nucleus, which was consistent with the predicted results. The transcript levels of GmPRs gene increased in different degrees after inoculated by Heterodera glycines, the expression of GmPRs proved that it played a different role in the defense of soybean against soybean cyst nematode.