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对番茄叶霉病菌不同分离株进行比较蛋白质组学分析,以发掘对番茄植株致病力起作用的差异表达蛋白。

Comparative Proteomic Analysis of Different Isolates of f.sp. to Exploit the Differentially Expressed Proteins Responsible for Virulence on Tomato Plants.

作者信息

Manikandan Rajendran, Harish Sankarasubramanian, Karthikeyan Gandhi, Raguchander Thiruvengadam

机构信息

Department of Plant Pathology, Centre for Plant Protection Studies, Tamil Nadu Agricultural University, Coimbatore, India.

出版信息

Front Microbiol. 2018 Mar 6;9:420. doi: 10.3389/fmicb.2018.00420. eCollection 2018.

Abstract

The vascular wilt of tomato caused by f.sp. is an important soil borne pathogen causes severe yield loss. The molecular characterization and their interaction with its host is necessary to develop a protection strategy. 20 isolates of f.sp. (FOL) were isolated from wilt infected tomato plants across Tamil Nadu. They were subjected to cultural, morphological, molecular and virulence studies. The results revealed that all the isolates produced both micro and macro conidia with different size, number of cells. The colors of the culture and growth pattern were also varied. In addition, chlamydospores were observed terminally and intercalary. The PCR analysis with species-specific primer significantly amplified an amplicon of 600 bp fragment in all the isolates. Based on the above characters and pathogenicity, isolate FOL-8 was considered as virulent and FOL-20 was considered as least virulent. Proteomics strategy was adopted to determine the virulence factors between the isolates of FOL-8 and FOL-20. The 2D analyses have showed the differential expression of 17 different proteins. Among them, three proteins were down regulated and 14 proteins were significantly up regulated in FOL-8 than FOL-20 isolate. Among the 17 proteins, 10 distinct spots were analyzed by MALDI-TOF. The functions of the analyzed proteins, suggested that they were involved in pathogenicity, symptom expression and disease development, sporulation, growth, and higher penetration rate on tomato root tissue. Overall, these experiments proves the role of proteome in pathogenicity of f.sp. in tomato and unravels the mechanism behinds the virulence of the pathogen in causing wilt disease.

摘要

由番茄枯萎病菌(Fusarium oxysporum f.sp. lycopersici)引起的番茄维管束萎蔫病是一种重要的土传病原菌,会导致严重的产量损失。开展保护策略研究,有必要对其进行分子特征分析以及研究它与宿主的相互作用。从泰米尔纳德邦各地感染枯萎病的番茄植株中分离出20株番茄枯萎病菌(FOL)。对它们进行了培养、形态、分子和毒力研究。结果显示,所有分离株均产生了大小和细胞数量不同的微菌核和大分生孢子。培养物的颜色和生长模式也各不相同。此外,在顶端和中间观察到厚垣孢子。用物种特异性引物进行PCR分析,在所有分离株中均显著扩增出一个600 bp片段的扩增子。基于上述特征和致病性,分离株FOL-8被认为是高毒力的,而FOL-20被认为是低毒力的。采用蛋白质组学策略来确定FOL-8和FOL-20分离株之间的毒力因子。二维分析显示有17种不同蛋白质存在差异表达。其中,与FOL-20分离株相比,FOL-8中有3种蛋白质表达下调,14种蛋白质显著上调。在这17种蛋白质中,通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF)分析了10个不同的斑点。对所分析蛋白质功能的研究表明,它们参与了致病性、症状表达和疾病发展、孢子形成、生长以及对番茄根组织的较高穿透率。总体而言,这些实验证明了蛋白质组在番茄枯萎病菌致病性中的作用,并揭示了该病原菌导致萎蔫病的毒力背后的机制。

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