Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (18): 3939-3945.doi: 10.3864/j.issn.0578-1752.2013.18.025

• RESEARCH NOTES • Previous Articles     Next Articles

Detection of Sweet potato chlorotic stunt virus Strain WA by Reverse Transcription Loop-Mediated Isothermal Ampli?cation Reaction

 QIAO  Qi, ZHANG  Zhen-Chen, QIN  Yan-Hong, ZHANG  De-Sheng, TIAN  Yu-Ting, WANG  Shuang, WANG  Yong-Jiang   

  1. Institute of Plant Protection, Henan Academy of Agricultural Sciences/Henan Key Laboratory of Crop Pest Control/IPM Key Laboratory in Southern Part of North China for Ministry of Agriculture, Zhengzhou 450002
  • Received:2013-03-26 Online:2013-09-15 Published:2013-06-17

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Abstract: 【Objective】The objective of this study is to develop a one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid and sensitive detection of Sweet potato chlorotic stunt virus strain WA (SPCSV-WA). 【Method】Four RT-LAMP primers, designed on the basis of the coat protein (CP) gene sequences of SPCSV-WA, and template RNA from infected leaves were used for one-step RT-LAMP which were carried out under isothermal conditions at 65℃ for 1 h. RT-LAMP products were analyzed by electrophoresis in agarose gels followed by staining with SYBR green I. A fragment from the RT-LAMP product was then cloned and sequenced to con?rm the spect?city of the assay. The RT-LAMP, and meanwhile compared with the conventional RT-PCR assay, was performed to detect SPCSV-WA from 14 sweet potato samples. 【Result】The RT-LAMP showed a high specificity and the detection limit was 101 copies/µL RNA template. A fragment cloned from the RT-LAMP product indicated that the nucleotide sequence cloned was SPCSV-WA CP. Fourteen sweet potato samples testing results of RT-LAMP were consistent with that of RT-PCR. 【Conclusion】 The RT-LAMP described in this study represents a very sensitive, speci?c and rapid assay for the detection of SPCSV-WA.

Key words: Ipomoea batatas , Sweet potato chlorotic stunt virus strain WA , RT-LAMP , detection

[1]Clark C A, Davis J A, Abad J A, Cuellar W J, Fuentes S, Kreuze J F, Gibson R W, Mukasa S B, Tugume A K, Tairo F D, Valkonen J P T. Sweetpotato viruses: 15 years of progress on understanding and managing complex diseases. Plant Disease, 2012, 96(2): 168-185.

[2]Mukasa S B, Rubaihayo P R, Valkonen J P T. Interactions between a crinivirus, an ipomovirus and a potyvirus in coinfected sweetpoato plants. Plant Pathology, 2006, 55: 458-467.

[3]Untiveros M, Fuentes S, Salazar L F. Synergistic interaction of Sweet potato chlorotic stunt virus (Crinivirus) with carla-, cucumo-, ipomo-, and potyviruses infecting sweet potato. Plant Disease, 2007, 91(6): 669-676.

[4]Gutiérrez D L, Fuentes S, Salazar L F. Sweetpotato virus disease (SPVD): Distribution, incidence, and effect on sweetpotato yield in Peru. Plant Disease, 2003, 87(3): 297-302.

[5]Njeru R W, Mburu M W K, Cheramgoi E, Gibson R W, Kiburi Z M, Obudho E, Yobera D. Studies on the physiological effects of viruses on sweet potato yield in Kenya. Annals of Applied Biology, 2004, 145: 71-76.

[6]张振臣, 乔奇, 秦艳红, 张德胜, 田雨婷. 我国发现由甘薯褪绿矮化病毒和甘薯羽状斑驳病毒协生共侵染引起的甘薯病毒病害. 植物病理学报, 2012, 42(3): 328-333.

Zhang Z C, Qiao Q, Qin Y H, Zhang D S, Tian Y T. First evidence for occurrence of Sweet potato virus disease (SPVD) caused by dual infection of Sweet potato feathery mottle virus and Sweet potato chlorotic stunt virus in China. Acta Phytopathologica Sinica, 2012, 42(3): 328-333. (in Chinese)

[7]Qin Y H, Zhang Z C, Qiao Q, Zhang D S, Tian Y T, Wang Y J. Molecular variability of Sweet potato chlorotic stunt viruses (SPCSV) and five potyviruses infecting sweet potato in China. Archives of Virology, 2013, 158(2): 491-495.

[8]Kokkinos C D, Clark C A. Real-time PCR assays for detection and quantification of sweetpotato viruses. Plant Disease, 2006, 90(6): 783-788.

[9]Mukasa S B, Rubaihayo P R. Valkonen J P T. Incidence of viruses and viruslike diseases of sweetpotato in Uganda. Plant Disease, 2003, 87(4): 329-335.

[10]Aritua V, Bua B, Barg E, Vetten H J, Adipala E, Gibson R W. Incidence of ?ve viruses infecting sweetpotatoes in Uganda; the ?rst evidence of Sweet potato caulimo-like virus in Africa. Plant Pathology, 2007, 56: 324 -331.

[11]Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanebe K, Amino N, Hase T. Loop-mediated isothermal ampli?cation of DNA. Nucleic Acids Research, 2000, 28(12): e63.

[12]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(9): 1713-1720.

[13]Zhao L, Cheng J, Hao X, Tian X, Wu Y. Rapid detection of tobacco viruses by reverse transcription loop-mediated isothermal amplification. Archives of Virology, 2012, 157(12): 2291-2298.

[14]Peng J, Shi M, Xia Z, Huang J, Fan Z. Detection of cucumber mosaic virus isolates from banana by one-step reverse transcription loop-mediated isothermal amplification. Archives of Virology, 2012, 157(11): 2213-2217.

[15]周彤, 杜琳琳, 范永坚, 周益军. 水稻黑条矮缩病毒RT-LAMP快速检测方法的建立. 中国农业科学, 2012, 45(7): 1285-1292.

Zhou T, Du L L, Fan Y J, Zhou Y J. Development of a RT-LAMP assay for rapid detection of Rice black-streaked dwarf virus. Scientia Agricultura Sinica, 2012, 45(7): 1285-1292. (in Chinese)

[16]Yang L, Li J, Bi Y, Xu L, Liu W. Development and application of a reverse transcription loop-mediated isothermal ampli?cation method for rapid detection of Duck hepatitis A virus type 1. Virus Genes, 2012, 45(3): 585-589.

[17]Komiyama C, Suzuki K, Miura Y, Sentsui H. Development of loop-mediated isothermal amplification method for diagnosis of bovine leukemia virus infection. Journal of Virological Methods, 2009, 157(2): 175-179.

[18]Rovira A, Abrahante J, Murtaugh M, Munoz-Zanzi C. Reverse transcription loop-mediated isothermal ampli?cation for the detection of Porcine reproductive and respiratory syndrome virus. Journal of Veterinary Diagnostic Investigation, 2009, 21: 350-354. 

[19]Hoyer U, Maiss E, Jelkmann W, Lesemann D E, Vetten H J. Identification of the coat protein gene of a sweet potato sunken vein closterovirus isolate from Kenya and evidence for a serological relationship among geographically diverse closterovirus isolates from sweet potato. Phytopathology, 1996, 86(7): 744-750.

[20]Aritua V, Barg E, Adipala E, Gibson R W, Vetten H J. Further evidence for limited genetic diversity among East African isolates of Sweet potato chlorotic stunt virus. Journal of Phytopathology, 2008, 156: 181-189.

[21]Tairo F, Mukasa S B, Jones R A C, Kullaya A, Rubaihayo P R, Valkonen J P T. Unravelling the genetic diversity of the three main viruses involved in sweet potato virus disease (SPVD), and its practical implications. Molecular Plant Pathology, 2005, 6(2): 199-211.

[22]Qiao Q, Zhang Z C, Qin Y H, Zhang D S, Tian Y T, Wang Y J. First report of Sweet potato chlorotic stunt virus infecting sweet potato in China. Plant Disease, 2011, 95(3): 356.

[23]Hong T C, Mai Q L, Cuong D V, Parida M, Minekawa H, Notomi T, Hasebe F, Morita K. Development and evaluation of a novel loop-mediated isothermal amplification method for rapid detection of severe acute respiratory syndrome coronavirus. Journal of Clinical Microbiology, 2004, 42(5): 1956-1961.
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