Rapid detection of potato late blight using a loop-mediated isothermal amplification assay
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 bas...
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| Format: | Article |
| Language: | English |
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KeAi Communications Co., Ltd.
2020-05-01
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| Series: | Journal of Integrative Agriculture |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2095311919628169 |
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| author | Liang KONG Hui-bin WANG Shuai-shuai WANG Ping-ping XU Ruo-fang ZHANG Suomeng DONG Xiao-bo ZHENG |
| author_facet | Liang KONG Hui-bin WANG Shuai-shuai WANG Ping-ping XU Ruo-fang ZHANG Suomeng DONG Xiao-bo ZHENG |
| author_sort | Liang KONG |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-e07ac6184fcb43a7bf5611d6ad4ce33a |
| institution | Kabale University |
| issn | 2095-3119 |
| language | English |
| publishDate | 2020-05-01 |
| publisher | KeAi Communications Co., Ltd. |
| record_format | Article |
| series | Journal of Integrative Agriculture |
| spelling | doaj-art-e07ac6184fcb43a7bf5611d6ad4ce33a2025-08-20T03:58:04ZengKeAi Communications Co., Ltd.Journal of Integrative Agriculture2095-31192020-05-011951274128210.1016/S2095-3119(19)62816-9Rapid detection of potato late blight using a loop-mediated isothermal amplification assayLiang KONG0Hui-bin WANG1Shuai-shuai WANG2Ping-ping XU3Ruo-fang ZHANG4Suomeng DONG5Xiao-bo ZHENG6Department of Plant Pathology, Nanjing Agricultural University, Nanjing 210095, P.R.ChinaDepartment of Plant Pathology, Nanjing Agricultural University, Nanjing 210095, P.R.ChinaDepartment of Plant Pathology, Nanjing Agricultural University, Nanjing 210095, P.R.ChinaNanjing Exit and Entry Inspection and Quarantine Bureau, Nanjing 210001, P.R.ChinaPotato Research Center, Inner Mongolia University, Hohhot 010021, P.R.ChinaDepartment of Plant Pathology, Nanjing Agricultural University, Nanjing 210095, P.R.China; The Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education, Nanjing 210095, P.R.China; Correspondence DONG Suo-mengDepartment of Plant Pathology, Nanjing Agricultural University, Nanjing 210095, P.R.China; The Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education, Nanjing 210095, P.R.ChinaPotato 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.http://www.sciencedirect.com/science/article/pii/S2095311919628169Phytophthora infestanspotato late diseaseLAMP assaydiagnostic detection |
| spellingShingle | Liang KONG Hui-bin WANG Shuai-shuai WANG Ping-ping XU Ruo-fang ZHANG Suomeng DONG Xiao-bo ZHENG Rapid detection of potato late blight using a loop-mediated isothermal amplification assay Journal of Integrative Agriculture Phytophthora infestans potato late disease LAMP assay diagnostic detection |
| title | Rapid detection of potato late blight using a loop-mediated isothermal amplification assay |
| title_full | Rapid detection of potato late blight using a loop-mediated isothermal amplification assay |
| title_fullStr | Rapid detection of potato late blight using a loop-mediated isothermal amplification assay |
| title_full_unstemmed | Rapid detection of potato late blight using a loop-mediated isothermal amplification assay |
| title_short | Rapid detection of potato late blight using a loop-mediated isothermal amplification assay |
| title_sort | rapid detection of potato late blight using a loop mediated isothermal amplification assay |
| topic | Phytophthora infestans potato late disease LAMP assay diagnostic detection |
| url | http://www.sciencedirect.com/science/article/pii/S2095311919628169 |
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