Rapid detection of potato late blight using a loop-mediated isothermal amplification assay

doi:10.1016/S2095-3119(19)62816-9

Journal of Integrative Agriculture ›› 2020, Vol. 19 ›› Issue (5): 1274-1282.DOI: 10.1016/S2095-3119(19)62816-9

收稿日期:2019-05-10 出版日期:2020-04-01 发布日期:2020-03-25

Rapid detection of potato late blight using a loop-mediated isothermal amplification assay

KONG Liang¹, WANG Hui-bin¹, WANG Shuai-shuai¹, XU Ping-ping², ZHANG Ruo-fang³, DONG Suo-meng^{1, 4}, ZHENG Xiao-bo^{1, 4}

¹ Department of Plant Pathology, Nanjing Agricultural University, Nanjing 210095, P.R.China
² Nanjing Exit and Entry Inspection and Quarantine Bureau, Nanjing 210001, P.R.China
³ Potato Research Center, Inner Mongolia University, Hohhot 010021, P.R.China
⁴ The Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education, Nanjing 210095, P.R.China

Received:2019-05-10 Online:2020-04-01 Published:2020-03-25
Contact: Correspondence DONG Suo-meng, E-mail: smdong@njau.edu.cn
About author:KONG Liang, E-mail: ankl2013@163.com;
Supported by:
This study was funded by the Fok Ying Tung Education Foundation of China (151025), the National Natural Science Foundation of China (31772144).

摘要/Abstract

Abstract:

Potato late blight caused by Phytophthora infestans is one of the most destructive plant diseases that threaten global food security. Early and effective diagnosis of P. infestans is required before disease management decisions are made. Here, we developed a quick protocol to detect P. infestans based on a loop-mediated isothermal amplification (LAMP) assay. The P. infestans specific multiple copy DNA sequences (PiSMC), a transposon-like element, provides an ideal target for molecular detection of this pathogen. We designed highly specific and sensitive primers allowing effective LAMP detection of the pathogen at 64°C in 70 min. In the validation assay, all 15 P. infestans isolates collected from China, Europe and South America could be positively detected, but results of the other 9 Phytophthora species infecting different plants, fungal and bacterial plant pathogens tested were negative. The detection limit of this assay is 1 pg P. infestans DNA. Moreover, the LAMP-PiSMC assay is able to detect P. infestans from infected leaves, tubers and soil. Taken together, this study reports the development of a specific and sensitive LAMP-PiSMC assay for early diagnosis of potato late blight.

Key words: Phytophthora infestans , potato late disease , LAMP assay , diagnostic detection

. [J]. Journal of Integrative Agriculture, 2020, 19(5): 1274-1282.

KONG Liang, WANG Hui-bin, WANG Shuai-shuai, XU Ping-ping, ZHANG Ruo-fang, DONG Suo-meng, ZHENG Xiao-bo.

Rapid detection of potato late blight using a loop-mediated isothermal amplification assay

[J]. Journal of Integrative Agriculture, 2020, 19(5): 1274-1282.

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Rapid detection of potato late blight using a loop-mediated isothermal amplification assay

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