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Molecular evidence for the involvement of a polygalacturonase-inhibiting protein GhPGIP1 in enhanced resistance to Verticillium and Fusarium wilts in cotton

机译:聚半乳糖醛酸酶抑制蛋白GhPGIP1参与棉花抗黄萎病和枯萎病抗性增强的分子证据

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摘要

Polygalacturonase-inhibiting protein (PGIP), belonging to a group of plant defence proteins, specifically inhibits endopolygalacturonases secreted by pathogens. Herein, we showed that purified GhPGIP1 is a functional inhibitor of Verticillium dahliae and Fusarium oxysporum f. sp. vasinfectum, the two fungal pathogens causing cotton wilt. Transcription of GhPGIP1 was increased in cotton upon infection, wounding, and treatment with defence hormone and H2O2. Resistance by GhPGIP1 was examined by its virus-induced gene silencing in cotton and overexpression in Arabidopsis. GhPGIP1-silenced cotton was highly susceptible to the infections. GhPGIP1 overexpression in transgenic Arabidopsis conferred resistance to the infection, accompanied by enhanced expression of pathogenesis-related proteins (PRs), isochorismate synthase 1 (ICS1), enhanced disease susceptibility 1 (EDS1), and phytoalexin-deficient 4 (PAD4) genes. Transmission electron microscopy revealed cell wall alteration and cell disintegration in plants inoculated with polygalacturonase (PGs), implying its role in damaging the cell wall. Docking studies showed that GhPGIP1 interacted strongly with C-terminal of V. dahliae PG1 (VdPG1) beyond the active site but weakly interacted with C-terminal of F. oxysporum f. sp. vasinfectum (FovPG1). These findings will contribute towards the understanding of the roles of PGIPs and in screening potential combat proteins with novel recognition specificities against evolving pathogenic factors for countering pathogen invasion.
机译:聚半乳糖醛酸酶抑制蛋白(PGIP),属于一组植物防御蛋白,特异性抑制病原体分泌的内聚半乳糖醛酸酶。在这里,我们表明纯化的GhPGIP1是黄萎病菌和尖孢镰刀菌f的功能抑制剂。 sp。 vasinfectum,引起棉花枯萎的两种真菌病原体。感染,受伤和用防御激素和过氧化氢处理后,棉花中GhPGIP1的转录增加。 GhPGIP1的抗性通过其病毒诱导的棉花基因沉默和拟南芥中的过表达来检测。 GhPGIP1沉默的棉花极易感染。 GhPGIP1在转基因拟南芥中的过度表达赋予了对感染的抵抗力,并伴随着发病机制相关蛋白(PRs),等渗硫酸合酶1(ICS1),疾病易感性1(EDS1)和植物抗毒素缺乏4(PAD4)基因的表达增强。透射电子显微镜显示接种聚半乳糖醛酸酶(PGs)的植物中细胞壁的改变和细胞的分解,表明其在破坏细胞壁中的作用。对接研究表明,GhPGIP1与活性位点以外的大丽花PG1 PG1(VdPG1)的C末端强烈相互作用,但与oxysporum f。的C末端弱相互作用。 sp。血管感染(FovPG1)。这些发现将有助于理解PGIP的作用,并筛选具有对抗发展中的致病因子的新型识别特异性的潜在战斗蛋白,以对抗病原体的入侵。

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