Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (7): 1285-1292.doi: 10.3864/j.issn.0578-1752.2012.07.006

• PLANT PROTECTION • Previous Articles     Next Articles

Development of a RT-LAMP Assay for Rapid Detection of Rice black-streaked dwarf virus

 ZHOU  Tong, DU  Lin-Lin, FAN  Yong-Jian, ZHOU  Yi-Jun   

  1. 1.江苏省农业科学院植物保护研究所/江苏省植物病毒病诊断检测技术中心，南京 210014.
    2.南京农业大学植物保护学院，南京210095
  • Received:2011-07-29 Online:2012-04-01 Published:2011-12-16

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Abstract: 【Objective】The objective of this study is to develop a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid and sensitive detection of Rice black-streaked dwarf virus (RBSDV) from host plants and insect vector.【Method】Four primers matching a total of 6 sequences of the S10 of RBSDV were synthesized for the RT-LAMP assay. The concentration of the primer and MgSO4 were optimized, and the best temperature and reaction time for detecting the virus were found, respectively. Detection sensitivity comparisons were performed between the RT-LAMP and RT-PCR assay using a single extraction of total RNA from RBSDV-infected rice leaves which was serially diluted in ten-fold increments in DEPC-treated water. In order to determine the specificity of the RT-LAMP assay, total RNA of rice leaves infected with RBSDV or Southern rice black-streaked dwarf virus (SRBSDV) was applied separately to the reaction system. RBSDV-infected rice plants collected from rice field were detected.【Result】 This method demonstrated a high degree of specificity for RBSDV, which can distinguish RBSDV from SRBSDV. The method was also proved to be extremely sensitive, which was as much as the RT-PCR for RBSDV detection. The detection of amplified products was easily monitored.【Conclusion】The RT-LAMP assay is suitable for rapid detection of RBSDV in host plant and vectors.

Key words: Rice black-streaked dwarf virus, RT-LAMP, detection

[1]阮义理, 陈声祥, 林瑞芬, 蒋文烈, 金登迪. 水稻黑条矮缩病的研究. 浙江农业科学, 1984(4): 185-187, 192.

Ruan Y L, Chen S X, Lin R F, Jiang W L, Jin D D. Studies on Rice black-streaked dwarf virus. Journal of Zhejiang Agricultural Sciences, 1984(4): 185-187, 192. (in Chinese)

[2]周国辉, 温锦君, 蔡德江, 李  鹏, 许东林, 张曙光. 呼肠孤病毒科斐济病毒属一新种: 南方水稻黑条矮缩病毒. 科学通报, 2008, 53(20): 2500-2508.

Zhou G H, Wen J J, Cai D J, Li P, Xu D L, Zhang S G. A new proposed Fijivirus species in the family Reoviridae: Southern rice black-streaked dwarf virus. Chinese Science Bulletin, 2008, 53(20): 2500-2508. (in Chinese)

[3]Zhang H M, Yang J, Chen J P, Adams M J. A black-streaked dwarf disease on rice in China is caused by a novel Fijivirus. Archives of Virology, 2008, 153(10): 1893-1898.

[4]Wang Q, Yang J, Zhou G H, Zhang H M, Yang J, Chen J P, Adams M J. The complete genome sequence of two isolates of Southern rice black-streaked dwarf virus, a new member of the genus Fijivirus. Journal of Phytopathology, 2010, 158(11/12): 733-737.

[5]朱凤美, 肖庆璞, 王法明, 陈毓芩. 江南稻区新发生的几种稻病. 植物保护, 1964, 2(3): 100-102.

Zhu F M, Xiao Q P, Wang F M, Chen Y Q. The new occurred rice disease in South of the Yangtze. Plant Protection, 1964, 2(3): 100-102. (in Chinese)

[6]陈声祥, 张巧艳. 我国水稻黑条矮缩病和玉米粗缩病研究进展. 植物保护学报, 2005, 32(1): 97-103.

Chen S X, Zhang Q Y. Advance in researches on rice black-streaked dwarf disease and maize rough dwarf disease in China. Acta Phytophylacica Sinica, 2005, 32(1): 97-103. (in Chinese)

[7]羊  健, 张恒木, 陈剑平, 戴良英. 水稻黑条矮缩病毒p8蛋白的原核表达、抗血清制备及其特性. 植物保护学报, 2007, 34(3): 252-256.

Yang J, Zhang H M, Chen J P, Dai L Y. Prekaryotic expression antiserum preparation and some properties of p8 protein of Rice black-streaked dwarf Fijivirus. Acta Phytophylacica Sinica, 2007, 34(3): 252-256. (in Chinese)

[8]Wang Z H, Fang S G, Zhang Z Y, Han C G, Li D W, Yu J L. Development of an ID-ELISA for the detection of Rice black-streaked dwarf virus in plants. Journal of Virological Methods, 2006, 134(1/2): 61-65.

[9]欧阳元龙, 吴建祥, 熊如意, 周益军, 周雪平. 水稻黑条矮缩病病毒外壳蛋白基因S10的原核表达、多克隆抗体制备及应用. 中国水稻科学, 2010, 24(1): 25-30.

Ouyang Y L, Wu J X, Xiong R Y, Zhou Y J, Zhou X P. Prokaryotic expression of coat protein gene S10 of Rice black-streaked dwarf virus, and preparation and application of its polyclonal antibody. Chinese Journal of Rice Science, 2010, 24(1): 25-30. (in Chinese)

[10]吕永平, 雷娟利, 金登迪, 陈声祥. 水稻黑条矮缩病毒的RT-PCR检测. 浙江农业学报, 2002, 14(2): 117-119.

Lü Y P, Lei J L, Jin D D, Chen S X. Detect RBSDV with RT-PCR. Acta Agriculturae Zhejiangensis, 2002, 14(2): 117-119. (in Chinese)

[11]王朝辉, 周益军, 范永坚, 程兆榜, 张文荟. 从单头灰飞虱中检测水稻黑条矮缩病毒简单快速的方法. 上海交通大学学报: 农业科学版, 2002, 20(4): 340-343.

Wang Z H, Zhou Y J, Fan Y J, Cheng Z B, Zhang W H. Rapid methods for the detection of rice black streaked dwarf Fijivirus in a single planthopper. Journal of Shanghai Jiaotong University: Agricultural Science, 2002, 20(4): 340-343. (in Chinese)

[12]王朝辉, 周益军, 范永坚, 薛宝娣, 吴淑华, 程兆榜, 张文荟. 应用RT-PCR、斑点杂交法和SDS-PAGE检测水稻黑条矮缩病毒. 南京农业大学学报, 2001, 24(4): 24-28.

Wang Z H, Zhou Y J, Fan Y J, Xue B D, Wu S H, Cheng Z B, Zhang W H. Detection of rice black-streaked dwarf Fifivirus by RT-PCR, dot-blot hybridization and SDS-PAGE. Journal of Nanjing Agricultural University, 2001, 24(4): 24-28. (in Chinese)

[13]季英华, 高瑞珍, 张  野, 程兆榜, 周  彤, 范永坚, 周益军. 一种快速同步检测水稻黑条矮缩病毒和南方水稻黑条矮缩病毒的方法. 中国水稻科学, 2011, 25(1): 91-94.

Ji Y H, Gao R Z, Zhang Y, Cheng Z B, Zhou T, Fan Y J, Zhou Y J. A simplified method for quickly detection of Rice black-streaked dwarf virus and Southern rice black-streaked dwarf virus. Chinese Journal of Rice Science, 2011, 25(1): 91-94. (in Chinese)

[14]Le D T, Netsu O, Uehara-Ichiki T, Shimizu T, Choi I R, Omura T, Sasaya T. Molecular detection of nine rice viruses by a reverse- transcription loop-mediated isothermal amplification assay. Journal of Virological Methods, 2010, 170(1/2): 90-93.

[15]Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T. Loop-mediated isothermal amplification of DNA. Nucleic Acids Research, 2000, 28(12): e63.

[16]Fukuta S, Iida T, Mizukami Y, Ishida A, Ueda J, Kanbe M, Ishimoto Y. Detection of Japanese yam mosaic virus by RT-LAMP. Archives of Virology, 2003, 148: 1713-1720.

[17]Enomoto Y, Yoshikawa T, Ihira M, Akimoto S, Miyake F, Usui C, Suga S, Suzuki K, Kawana T, Nishiyama Y, Asano Y. Rapid diagnosis of herpes simplex virus infection by a loop-mediated isothermal amplification method. Journal of Clinical Microbiology, 2005, 43(2): 951-955.

[18]鑫  婷, 侯绍华, 贾  红, 郭晓宇, 丁家波, 李延鹏, 丁  敏, 朱鸿飞. 猪呼吸与繁殖综合症病毒RT-LAMP检测方法的建立. 中国农业科学, 2010, 43(1): 185-191.

Xin T, Hou S H, Jia H, Guo X Y, Ding J B, Li Y P, Ding M, Zhu H F. Development of a novel RT-LAMP assay for porcine reproductive and respiratory syndrome virus. Scientia Agricultura Sinica, 2010, 43(1): 185-191. (in Chinese)

[19]李  健, 陈  沁, 熊  炜, 方雪恩. 口蹄疫病毒RT-LAMP检测方法的建立. 病毒学报, 2009, 25(2): 137-142.

Li J, Chen Q, Xiong W, Fang X E. Establishment of RT-LAMP for rapid detection of foot-and-mouth disease virus. Chinese Journal of Virology, 2009, 25(2): 137-142. (in Chinese)

[20]Zhao K, Liu Y, Wang X F. Reverse transcription loop-mediated isothermal amplification of DNA for detection of Barley yellow dwarf viruses in China. Journal of Virological Methods, 2010, 169(1): 211-214.

[21]周  彤, 吴丽娟, 王  英, 程兆榜, 季英华, 范永坚, 周益军. 灰飞虱从冷冻病叶获得水稻黑条矮缩病毒方法的研究初报. 中国水稻科学, 2010, 24(4): 425-428.

Zhou T, Wu L J, Wang Y, Cheng Z B, Ji Y H, Fan Y J, Zhou Y J. Preliminary report on the transmission of Rice black-streaked dwarf virus from frozen infected leaves to rice by insect vector small brown planthopper (Laodelphax striatellus). Chinese Journal of Rice Science, 2010, 24(4): 425-428. (in Chinese)

[22]周  彤, 王  英, 吴丽娟, 范永坚, 周益军. 水稻品种抗黑条矮缩病人工接种鉴定方法. 植物保护学报, 2011, 38(4): 301-305.

Zhou T, Wang Y, Wu L J, Fan Y J, Zhou Y J. Method of artificial inoculation identification of rice cultivar resistance to rice black-streaked dwarf. Acta Phytophylacica Sinica, 2011, 38(4): 301-305. (in Chinese)

[23]张恒木, 雷娟利, 陈剑平, 吕永平, 陈声祥, 薛庆中, Adams M J. 浙江和河北发生的一种水稻、小麦、玉米矮缩病是水稻黑条矮缩病毒引起的. 中国病毒学, 2001, 16(3): 246-251.

Zhang H M, Lei J L, Chen J P, Lü Y P, Chen S X, Xue Q Z, Adams M J. A dwarf disease on rice, wheat and maize from Zhejiang and Hebei is caused by rice black-streaked dwarf virus. Virologica Sinica, 2001, 16(3): 246-251. (in Chinese)

[24]Zhou T, Wang Y, Fan Y J, Cheng Z B, Zhou Y J. First report of rice black-streaked dwarf virus infecting barley in Jiangsu, China. Journal of Plant Pathology, 2010, 92(S4): 118.

[25]周国辉, 张曙光, 邹寿发, 许兆伟, 周志强. 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(1): 144-146.

Zhou G H, Zhang S G, Zou S F, Xu Z W, Zhou Z Q. Occurrence and damage analysis of a new rice dwarf disease caused by Southern rice black-streaked dwarf virus. Plant Protection, 2010, 36(1): 144-146. (in Chinese)
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