Development and evaluation of specific PCR and LAMP assays for the rapid detection of Phytophthora melonis

Phytophthora melonis is a widespread and devastating pathogen for the Cucurbitaceae family. Early and accurate detection of P. melonis is essential to control the disease in the field. To establish a simple, visual, and rapid detection system for P. melonis, we developed nested polymerase chain reac...

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Bibliographic Details
Published in:European journal of plant pathology 2013-11, Vol.137 (3), p.597-607
Main Authors: Chen, Qinghe, Li, Benjin, Liu, Peiqing, Lan, Chengzhong, Zhan, Zhixiong, Weng, Qiyong
Format: Article
Language:English
Subjects:
Agriculture
Biomedical and Life Sciences
cross reaction
cucumbers
Cucumis sativus
Cucurbitaceae
detection limit
disease control
DNA
Ecology
fungi
genes
isolation techniques
Life Sciences
Melonis
Oomycetes
Pathogens
Phytophthora
Plant diseases
Plant Pathology
Plant Sciences
Polymerase chain reaction
Proteins
risk
soil
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Summary:Phytophthora melonis is a widespread and devastating pathogen for the Cucurbitaceae family. Early and accurate detection of P. melonis is essential to control the disease in the field. To establish a simple, visual, and rapid detection system for P. melonis, we developed nested polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) systems based on the Ras-related protein (Ypt1) gene. All 36 isolates of P. melonis, from geographically distinct counties in China, yielded positive detection results on LAMP or nested PCR assays. No cross reaction was observed with other oomycetes or fungal pathogens. A sensitivity assay showed that both methods had a detection limit of 10 fg genomic DNA. We also detected P. melonis in diseased cucumber tissues and soils, and evaluated positive detection rates using LAMP, nested PCR, and conventional isolation methods. The results suggest that the LAMP assay has the greatest potential for active detection of P. melonis in regions that are at risk of contracting the disease, and for use in resource-poor settings.
ISSN:0929-1873
1573-8469
DOI:10.1007/s10658-013-0273-9