The plant defense and pathogen counterdefense mediated by Hevea brasiliensis serine protease HbSPA and Phytophthora palmivora extracellular protease inhibitor PpEPI10.
The plant defense and pathogen counterdefense mediated by Hevea brasiliensis serine protease HbSPA and Phytophthora palmivora extracellular protease inhibitor PpEPI10.
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DOI：
10.1371/journal.pone.0175795
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Churngchow N
中科院分区：
文献类型：
--
作者：
Ekchaweng K;Evangelisti E;Schornack S;Tian M;Churngchow N
橡胶树(Vevea brasilisis Muell.)Arg)是泰国重要的经济作物。由侵袭性卵菌病棕榈疫霉菌引起的落叶和黑条病害，对橡胶树的生长造成严重危害，导致胶乳产量下降。为了深入了解巴西烟草枯草杆菌类丝氨酸蛋白酶的分子功能，通过农业渗透在本氏烟草中瞬时表达了HbSPA、HbSPB和HbSPC基因。HbSPA编码的一种功能性蛋白水解酶已成功地在N.benthamiana叶片的质外体中表达。HbSPA的瞬时表达增强了植物对棕榈疫霉菌的抗性，表明HbSPA在植物防御中起着重要作用。棕榈假单胞菌Kazal样胞外蛋白酶抑制因子10(PpEPI10)是一种质外体效应因子，通过抑制巴氏假单胞菌蛋白水解酶参与致病。半定量RT-PCR结果表明，PpEPI10基因在棕榈疫霉菌侵染橡胶树的过程中显著上调。同时，HbSPA基因在感染过程中高度表达。为了研究HbSPA与PpEPI10之间可能的相互作用，在大肠杆菌中表达了重组PpEPI10蛋白(RPpEPI10)，并用亲和层析法进行了纯化。凝胶内酶谱和免疫共沉淀(co-IP)分析表明，rPpEPI10特异性地抑制HbSPA并与其相互作用。PpEPI10对HbSPA的靶向作用揭示了一种由植物蛋白水解酶和病原菌蛋白水解酶抑制剂介导的防御-反防御机制。掌食性动物之间的相互作用。
Rubber tree (Hevea brasiliensis Muell. Arg) is an important economic crop in Thailand. Leaf fall and black stripe diseases caused by the aggressive oomycete pathogen Phytophthora palmivora, cause deleterious damage on rubber tree growth leading to decrease of latex production. To gain insights into the molecular function of H. brasiliensis subtilisin-like serine proteases, the HbSPA, HbSPB, and HbSPC genes were transiently expressed in Nicotiana benthamiana via agroinfiltration. A functional protease encoded by HbSPA was successfully expressed in the apoplast of N. benthamiana leaves. Transient expression of HbSPA in N. benthamiana leaves enhanced resistance to P. palmivora, suggesting that HbSPA plays an important role in plant defense. P. palmivora Kazal-like extracellular protease inhibitor 10 (PpEPI10), an apoplastic effector, has been implicated in pathogenicity through the suppression of H. brasiliensis protease. Semi-quantitative RT-PCR revealed that the PpEPI10 gene was significantly up-regulated during colonization of rubber tree by P. palmivora. Concurrently, the HbSPA gene was highly expressed during infection. To investigate a possible interaction between HbSPA and PpEPI10, the recombinant PpEPI10 protein (rPpEPI10) was expressed in Escherichia coli and purified using affinity chromatography. In-gel zymogram and co-immunoprecipitation (co-IP) assays demonstrated that rPpEPI10 specifically inhibited and interacted with HbSPA. The targeting of HbSPA by PpEPI10 revealed a defense-counterdefense mechanism, which is mediated by plant protease and pathogen protease inhibitor, in H. brasiliensis-P. palmivora interactions.