硅介导番茄青枯病抗性的土壤定量蛋白质组学研究
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国家自然科学基金项目(31370456)、广东省自然科学基金项目(S2012010010331)和教育部博士点基金项目(20124404110010)资助


Soil quantitative proteomic analysis of silicon-mediated resistance of tomato (Solanum lycopersicum) to Ralstonia solanacearum
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    摘要:

    番茄( Solanum lycopersicum)是一种重要的经济作物。由青枯菌( Ralstonia solanacearum)侵染所产生的青枯病是一种严重影响茄科类作物的细菌性土传病害。传统的防治方法,如培育抗性品种、轮作、药剂防治等均存在一定的局限。而硅作为一种作物生长的有益元素,在提高植物适应生物胁迫和非生物胁迫中均起重要作用。本研究以易感青枯病的番茄品种“台湾红圣女”作为试验材料,利用iTRAQ(isobaric tags for relative and absolute quantitation)定量蛋白质组学技术,研究硅和/或青枯菌接种对番茄青枯病抗性及土壤蛋白质组的影响。结果表明,2.0 mmol L-1硅处理能显著降低番茄青枯病的病情指数,增强抗病性。基于iTRAQ定量蛋白质组学技术,硅和/或青枯菌处理对土壤蛋白质组的研究表明,在鉴定的30个土壤蛋白质中,有29个蛋白差异表达显著(上调≥1.2倍,下调≤0.8倍差异)。青枯菌侵染诱发22个土壤蛋白下调表达,5个上调表达。在不接菌的条件下,硅处理有5个土壤蛋白上调,19个蛋白下调;而接菌条件下则有8个蛋白上调,14个蛋白下调。将这29个差异蛋白进行GO基因功能分类结果显示,硅和/或青枯菌处理主要影响生理代谢过程以及核酸结合蛋白。硅对番茄青枯病的作用机理主要体现在:改变微生物的代谢能力,调控与抗性代谢、蛋白质的合成与翻译、信号转导以及免疫系统过程和胁迫响应有关蛋白的表达,影响钙离子信号转导等。

    Abstract:

    Tomato (Solanum lycopersicum) is an important economic crop. Bacterial wilt caused by Ralstonia solanacearum is a serious soil-borne disease of Solanaceae crops. Traditional controlling methods, such as cultivation of resistant varieties, crop rotation and use of chemicals, all have certain limitations. Silicon, as beneficial element to plant growth, plays an important role in enhancing plant adaptability to biotic and abiotic stresses. Proteomics is a new branch of science in post-genome era, focusing on proteomics as object to explore the full extent properties of various proteins, and further on pathogenesis, cell model and functional links of diseases at the protein level. In this study, the variety of tomato “Taiwan red cherry” which is susceptible to R. solanacearum was used. The experiment was designed as the following treatments: CK, Treatment Si (Si addition only), Treatment Rs (R. solanacearum inoculation only) and Treatment Si+Rs (Si addition followed by R. solanacearum inoculation), and the technique of iTRAQ (isobaric tags for relative and absolute quantitation) was used to analyze effects of Si addition on resistance of the tomato to R. solanacearum and soil proteome. Results showed that silicon addition significantly reduced disease index of the tomato crop by 19.4%, increased available silicon content in the soil and enhanced resistance of the tomato to R. solanacearum. ITRAQ analysis revealed that out of the 30 soil proteins identified, 29 were differentially expressed (up-regulation ≥1.2 fold and down-regulation ≤0.8 fold); isoelectric point of differentiated proteins varied between 4.1 and 10.7 and relative molecular weight in the range of 9 ~ 392 kDa; and confidence coefficient was ≥95%. Compared with CK, Treatment Si had five proteins up-regulated and nineteen down-regulated and Treatment Rs, five up-regulated and twenty-two down-regulated. Compared with Treatment Si, Treatment Si Rs had eight proteins up-regulated and fourteen down-regulated. Classification based on GO (gene ontology) function shows that of the twenty-nine differentially expressed proteins, 21.88% are involved in metabolic process, 15.63% in cellular process, 12.50% in cell communication, 3.13% in immune system process and another 3.13% in the process of response to stimulus, and classification in light of molecular function reveals that 25.64% are related to structural molecule activity, 25.64% to binding and 20.51% to catalytic activity, the three categories making up a major proportion. Classification according to protein class exposes that nucleic acid binding protein, hydrolase protein and cytoskeletal protein are the three major categories of proteins, accounting for 19.51%, 14.63% and 9.76%, respectively. To better understand effects of Treatment Si Rs and Treatment Rs on soil proteins, further analysis was done of the 22 differentially expressed proteins obtained from Treatments Si Rs/Rs. It was found compared with Treatment Rs, Treatment Si Rs had eight proteins up-regulated by as high as 1.6 folds and fourteen proteins down-regulated by 0.5 fold. All the findings indicate that Si could affect R. solanacearum by altering metabolic and cellular processes in biological process, structural molecule active protein and bindin protein in molecular function, and nucleic acid binding protein and cytoskeletal protein in protein classification. In a word, Si addition can reduce the disease index of tomato bacterial wilt significantly, thus enhancing the plant resistance to the disease. Infection of R. solanacearum down-regulates metabolic-related proteins, influences degradation of misfolded proteins, hinders signal communication and Ca2 signal transduction between microorganisms and microorganisms or plants, blocks synthesis of proteins, while Si helps tomato build up its resistance to R. solanacearum infection by affecting soil microbial metabolic capability, regulating expression of resistance and metabolism related proteins, improving signal communication and transduction between microorganisms and between microorganisms and plants, adjusting synthesis of soil proteins and expression of proteins involved in immune system process and stress response and influencing signal transduction of Ca2 .

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陈玉婷,林威鹏,范雪滢,蔡昆争.硅介导番茄青枯病抗性的土壤定量蛋白质组学研究[J].土壤学报,2015,52(1):162-173. DOI:10.11766/trxb201405200240 Chen Yuting, Lin Weipeng, Fan Xueying, Cai Kunzheng. Soil quantitative proteomic analysis of silicon-mediated resistance of tomato (Solanum lycopersicum) to Ralstonia solanacearum[J]. Acta Pedologica Sinica,2015,52(1):162-173.

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  • 收稿日期:2014-05-20
  • 最后修改日期:2014-07-22
  • 录用日期:2014-08-20
  • 在线发布日期: 2014-10-22
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