Lai Tongfei, Chen Yong, Li Boqiang, Qin Guozheng, Tian Shiping
Key Laboratory of Plant Resources, Institute of Botany, The Chinese Academy of Sciences, Xiangshan Nanxincun 20, Haidian District, Beijing 100093, China.
Key Laboratory of Plant Resources, Institute of Botany, The Chinese Academy of Sciences, Xiangshan Nanxincun 20, Haidian District, Beijing 100093, China.
J Proteomics. 2014 May 30;103:47-56. doi: 10.1016/j.jprot.2014.03.012. Epub 2014 Mar 24.
Penicillium expansum is an important fungal pathogen, which causes blue mold rot in various fruits and produces a mycotoxin (patulin) with potential damage to public health. Here, we found that nitric oxide (NO) donor could significantly inhibit germinability of P. expansum spores, resulting in lower virulence to apple fruit. Based on two dimension electrophoresis (2-DE) and mass spectrometry (MS) analysis, we identified ten differentially expressed proteins in response to exogenous NO in P. expansum. Among of them, five proteins, such as glutamine synthetase (GS), amidohydrolase, nitrilases, nitric oxide dioxygenase (NOD) and heat shock protein 70, were up-regulated. Others including tetratricopeptide repeat domain, UDP-N-acetylglucosamine pyrophosphorylase, enolase (Eno), heat shock protein 60 and K homology RNA-binding domain were down-regulated. The expression of three genes associated with the identified proteins (GS, NOD, and Eno) was evaluated at the mRNA level by RT-PCR. Our results provide the novel evidence for understanding the mechanism, by which NO regulates growth of P. expansum and its virulence.
Crop diseases caused by fungal pathogens lead to huge economic losses every year in the world. Application of chemical fungicides to control diseases brings the concern about food and environmental safety. Screening new antimicrobial compounds and exploring involved mechanisms have great significance to development of new disease management strategies. Nitric oxide (NO), as an important intracellular signaling molecule, has been proved to be involved in many physiological processes and defense responses during plant-pathogen interactions. In this study, we firstly found that NO at high concentration could distinctly delay spore germination and significantly reduce virulence of P. expansum to fruit host, identified some important proteins in response to NO stress and characterized the functions of these proteins. These results provide novel evidence for understanding the mechanism of NO regulating virulence of the fungal pathogen, but are beneficial for screening new targets of antifungal compounds.
扩展青霉是一种重要的真菌病原体，它会在各种水果中引起青霉腐烂，并产生一种对公众健康有潜在危害的霉菌毒素（棒曲霉素）。在此，我们发现一氧化氮（NO）供体可显著抑制扩展青霉孢子的萌发能力，从而降低其对苹果果实的毒力。基于二维电泳（2-DE）和质谱（MS）分析，我们鉴定出扩展青霉中响应外源NO的十种差异表达蛋白。其中，谷氨酰胺合成酶（GS）、酰胺水解酶、腈水解酶、一氧化氮双加氧酶（NOD）和热休克蛋白70等五种蛋白上调。其他包括四肽重复结构域、UDP-N-乙酰葡糖胺焦磷酸化酶、烯醇化酶（Eno）、热休克蛋白60和K同源RNA结合结构域的蛋白则下调。通过RT-PCR在mRNA水平评估了与鉴定出的蛋白（GS、NOD和Eno）相关的三个基因的表达。我们的结果为理解NO调节扩展青霉生长及其毒力的机制提供了新证据。
由真菌病原体引起的作物病害每年在全球造成巨大的经济损失。应用化学杀菌剂控制病害引发了对食品和环境安全的担忧。筛选新的抗菌化合物并探索其相关机制对开发新的病害管理策略具有重要意义。一氧化氮（NO）作为一种重要的细胞内信号分子，已被证明参与植物与病原体相互作用期间的许多生理过程和防御反应。在本研究中，我们首先发现高浓度的NO可明显延迟孢子萌发并显著降低扩展青霉对果实寄主的毒力，鉴定了一些响应NO胁迫的重要蛋白并表征了这些蛋白的功能。这些结果为理解NO调节真菌病原体毒力的机制提供了新证据，且有利于筛选抗真菌化合物的新靶点。
