水稻草矮病毒侵染的水稻根系差异蛋白鉴定
收稿日期: 2015-12-29
修回日期: 2016-05-23
网络出版日期: 2016-07-10
基金资助
国家自然科学基金资助项目(31301640)； 国家科技支撑计划资助项目(2012BAD19B03)。
Identification of Differentially Expressed Proteins in Roots of Rice（Oryza sativa） Infected with Rice grassy stunt virus
Received date: 2015-12-29
Revised date: 2016-05-23
Online published: 2016-07-10
为鉴定与水稻草矮病毒（Rice grassy stunt virus， RGSV）侵染引起的水稻根系发育不良的相关蛋白，解析根系发育不良症状的形成机理以及水稻草矮病毒与水稻互作的分子机制，采用双向荧光差异凝胶电泳（2DDIGE）结合基质辅助激光解析串联飞行时间质谱技术（MALDITOFMS）对RGSV侵染水稻后根系的差异表达蛋白进行分离和鉴定，并对鉴定的差异表达蛋白进行GO聚类分析和KEGG生物通路注释。结果表明，差异倍数大于2的蛋白质点有56个，其中，表达丰度升高的点有34个，表达丰度下降的点有22个；质谱成功鉴定出27个蛋白质点，分属于25种蛋白质。GO聚类分析表明差异表达蛋白涉及14个生物学过程，在生物功能上分属10类。细胞组件分析显示差异表达蛋白定位于不同的细胞部位。KEGG通路分析显示差异蛋白参与了12个生物通路。
关键词： 水稻草矮病毒; 水稻根系; 差异表达蛋白; 双向荧光差异凝胶电泳; 基质辅助激光解析串联飞行时间质谱技术
丁新伦,谢荔岩,张洁,吴祖建* . 水稻草矮病毒侵染的水稻根系差异蛋白鉴定[J]. 中国水稻科学, 2016 , 30(4) : 356 -362 . DOI: 10.16819/j.1001-7216.2016.5193
In order to find the proteins related to root maldevelopment caused by Rice grassy stunt virus(RGSV), elucidate the mechanism of root maldevelopment and better understand the molecular basis of interaction between RGSV and host rice, 2DDIGE coupled with MALDITOFMS was used to screen and identify differentially expressed proteins between RGSVinfected and RGSVuninfected rice roots. The acquired proteins were further analyzed using GoMiner programs and KEGG pathway database. The results showed that 56 protein spots (differential ratio>2) were differentially expressed, including 34 upregulated and 22 downregulated protein spots. Among them, 27 differentially expressed protein spots which belonged to 25 proteins were identified, and these proteins were mainly involved in 14 biological processes, 10 molecular functions and 12 KEGG pathways.
\[1\]周国辉, 张曙光, 邹寿发, 等. 水稻新病害南方水稻黑条矮缩病发生特点及危害趋势分析. 植物保护, 2010, 36(2): 144146.
Zhou G H, Zhang S G, Zou S F, et al. Occurrence and damage analysis of a new rice dwarf disease caused by Southern rice blackstreaked dwarf virus. Plant Prot, 2010, 36(2): 144146. (in Chinese with English abstract)
\[2\]周雪平, 濮祖芹, 方中达. 黄瓜花叶病毒(CMV)土壤非介体传播研究. 南京农业大学学报, 1994, 17(2): 3942.
Zhou X P, Pu Z Q, Fang Z D. Nonvectored soil transmission of cucumber mosaic virus. J Nanjing Agric Univ, 1994, 17(2): 3942. (in Chinese with English abstract)
\[3\]Keeling B L. Effect of soybean mosaic virus on root volume and dry weight of soybean plants. Crop Sci, 1982, 22(3): 629630.
\[4\]陈德鑫, 王凤龙, 李多川, 等. 山东烟草病毒病发生特点及防治对策. 中国烟草科学, 2007, 28(1)： 2528.
Chen D X, Wang F L, Li D C, et al. Epidemic characteristics of tobacco virus disease and control measures in Shandong Province. Chin Tob Sic. 2007, 28(1):2528. (in Chinese with English abstract)
\[5\]王煜炜, 冯俊丽, 林若虹, 等. 病毒侵染对番茄根茎组织中miRNAs调控通路的影响. 北京： 中国科技论文在线\[20120223\]. http: //www.paper.edu.cn/releasepaper/content/201202928.
Wang Y W， Feng J L， Lin R H, et al. The interference of miRNAs pathway during tomato root and stem tissues development upon virus infections. Beijing： Sciencepaper Online\[20120223\]. http://www.paper.edu.cn/releasepaper/content/201202928.
\[6\]林奇英, 谢联辉, 谢荔岩, 等. 中菲两种水稻病毒病的比较研究: Ⅱ.水稻草状矮化病的病原学//谢联辉. 水稻病毒：病理学与分子生物学. 福州： 福建科学技术出版社， 2001：176180.
Lin Q Y, Xie L H, Xie L Y, et al. Comparative study on two kinds of rice virus disease between China and Philippines: Ⅱ. Pathogen of rice grassy stunt virus disease//Xie L H. Rice Virus:Pathology and Molecular Biology. Fuzhou: Fujian Science & Technology Publishing House, 2001：176180.
\[7\]丁新伦, 谢荔岩, 潘贤, 等. 水稻草状矮化病毒侵染下水稻根发育相关基因的表达分析. 中国农学通报, 2014, 30(36): 120123.
Ding X L, Xie L Y, Pan X, et al. Transcriptional analysis of the root development related genes in host rice (Oryza sativa) infected with Rice grassy stunt virus. Chin Agric Sci Bull, 2014, 30(36)：120123. (in Chinese with English abstract)
\[8\]Carpentier S C, Witters E, Laukens K, et al. Preparation of protein extracts from recalcitrant plant tissues: An evaluation of different methods for twodimensional gel electrophoresis analysis. Proteomics, 2005, 5: 24972507.
\[9\]丁新伦, 谢荔岩, 吴祖建. 水稻草状矮化病毒侵染寄主水稻差异表达蛋白的鉴定和分析. 中国农业科学, 2014, 47(9)：17251734.
Ding X L, Xie L Y, Wu Z J. Identification and analysis of differentially expressed proteins of host rice (Oryza sativa) infected with Rice grassy stunt virus. Sci Agric Sin, 2014, 47(9): 17251734. (in Chinese with English abstract)
\[10\]Chomchan P, Miranda G J, Shirako Y. Detection of rice grassy stunt tenuivirus nonstructural proteins p2, p5 and p6 from infected rice plants and from viruliferous brown planthoppers. Arch Virol, 2002, 147(12): 22912300.
\[11\]Kakutani T, Hayano Y, Hayashi T, et al. Ambisense segment 4 of rice stripe virus: Possible evolutionary relationship with phleboviruses and uukuviruses (Bunyaviridae). J Gen Virol, 1990, 71: 14271432.
\[12\]Zhu Y, Hayakawa T, Toriyama S. Complete nucleotide sequence of RNA4 of rice stripe virus isolate T and comparison with another isolate and with maize stripe virus. J Gen Virol, 1992, 73: 13091312.
\[13\]Sangster TA, Queitsch C. The HSP90 chaperone complex, an emerging force in plant development and phenotypic plasticity. Curr Opin Plant Biol, 2005, 8: 8692.
\[14\]Liu Y, BurchSmith T, Schiff M, et al. Molecular chaperone Hsp90 associates with resistance protein N and its signaling proteins SGT1 and Rar1 to modulate an innate immune response in plants. J Biol Chem, 2004, 279: 21012108.
\[15\]Wang G F, Wei X N, Fan R C, et al. Molecular analysis of common wheat genes encoding three types of cytosolic heat shock protein 90 (Hsp90): Functional involvement of cytosolic Hsp90s in the control of wheat seedling growth and disease resistance. New Phytol, 2011, 191(2): 418431.
\[16\]Velazhahan R, Chencole K, Anuratha C S, et al. Induction of thaumatinlike protein(TLPs)in Rhizoctoniasolaniinfected rice and characterization of two new cDNA clones. Physiol Plantarum, 1998, 102: 2128.
\[17\]Fu D, Tisserat N A, Xiao Y, et al. Overexpression of rice TLPD34 enhances dollarspot resistance in transgenic bentgrass. Plant Sci, 2005, 168(3): 671680.
\[18\]Qi X, Bao F S, Xie Z. Small RNA deep sequencing reveals role for Arabidopsis thaliana RNAdependent RNA polymerases in viral siRNA biogenesis. PLoS ONE, 2009, 4(3): e4971.
\[19\]Xie Z, Fan B, Chen C, et al. An important role of an inducible RNAdependent RNA polymerase in plant antiviral defense. PNAS, 2001, 98(11): 65166521.
\[20\]Yu D, Fan B, MacFarlane S A, et al. Analysis of the involvement of an inducible Arabidopsis RNAdependent RNA polymerase in antiviral defense. Mol Plant Microbe Interact, 2003, 16(3): 206216.
\[21\]Jiang L, Qian D, Zheng H, et al. RNAdependent RNA polymerase 6 of rice (Oryza sativa) plays role in host defense against negativestrand RNA virus, Rice stripe virus. Virus Res, 2012, 163(2): 512519.
\[22\]LaCamera S, Gouzerh G, Dhondt S, et al. Metabolic reprogramming in plant innate immunity: The contributions of phenylpropanoid and oxylipin pathways. Immunol Rev, 2004, 198(1): 267284.
\[23\]Vogt T. Phenylpropanoid biosynthesis. Mol Plant, 2010, 3(1): 220.
\[24\]Gruner K, Griebel T, Návarová H, et al. Reprogramming of plants during systemic acquired resistance. Front Plant Sci, 2013, 4: 128.
\[25\]Malamy J, Carr J P, Klessig D F, et al. Salicylic acid : A likely endogenous signal in the resistance response of tobacco to viral infection. Science, 2014, 80(250): 10021004.
\[26\]Tonnessen B W, Manosalva P, Lang J M, et al. Rice phenylalanine ammonialyase gene OsPAL4 is associated with broad spectrum disease resistance. Plant Mol Biol, 2015, 87(3): 273286.
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