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- Title
Development and evaluation of specific PCR and LAMP assays for the rapid detection of Phytophthora melonis.
- Authors
Chen, Qinghe; Li, Benjin; Liu, Peiqing; Lan, Chengzhong; Zhan, Zhixiong; Weng, Qiyong
- Abstract
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.
- Subjects
CHINA; POLYMERASE chain reaction; PHYTOPHTHORA diseases; CUCURBITACEAE; PATHOGENIC microorganisms; PEST control; RAP1 proteins; GENOMICS; DISEASES
- Publication
European Journal of Plant Pathology, 2013, Vol 137, Issue 3, p597
- ISSN
0929-1873
- Publication type
Article
- DOI
10.1007/s10658-013-0273-9