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Establishment and evaluation of a TEF1-α based loop-mediated isothermal amplification assay for detection of Phomopsis longicolla

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Abstract

Phomopsis longicolla is an important seed-borne pathogen affecting soybean. In this research, transcription elongation factor 1-α (TEF1-α) was identified as a suitable target for detecting P. longicolla using Loop-mediated isothermal amplification (LAMP) method. The specificity of the method was tested against 54 P. longicolla isolates, 13 oomycetes isolates and 28 other fungal isolates. With the addition of hydroxynaphthol blue (HNB) prior to amplification, a sky-blue color was only observed in the presence of P. longicolla, whereas other isolates showed no color change. The method was sensitive enough to detect as little as 100 pg/uL fungal DNA. In addition, the assay also detected P. longicolla from diseased soybean tissues and residues from different origins. The results demonstrated that the LAMP assay provides a rapid and sensitive tool for detecting P. longicolla from plant tissues.

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Acknowledgments

This work was supported by National Natural Science Foundation of China under Grant 31500526, the National ‘863’ Program under Grant 2012AA101501, China Postdoctoral Science Foundation under Grant 2014 M561657, Jiangsu Postdoctoral Sustentation Fund under Grant 1402159c and the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

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Authors and Affiliations

  1. Co-Innovation Center of the Sustainable Forestry in Southern China, College of Forestry, Nanjing Forestry University, Nanjing, Jiangsu, 210037, China

    Ting-ting Dai

  2. Department of Plant Pathology, Nanjing Agricultural University, Nanjing, Jiangsu, 210095, China

    Hao Shen & Xiao-Bo Zheng

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  1. Ting-ting Dai
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  2. Hao Shen
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  3. Xiao-Bo Zheng
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Correspondence to Ting-ting Dai or Xiao-Bo Zheng.

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Dai, Tt., Shen, H. & Zheng, XB. Establishment and evaluation of a TEF1-α based loop-mediated isothermal amplification assay for detection of Phomopsis longicolla . Australasian Plant Pathol. 45, 335–337 (2016). https://doi.org/10.1007/s13313-016-0415-6

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  • DOI: https://doi.org/10.1007/s13313-016-0415-6

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