Journal
EUROPEAN JOURNAL OF PLANT PATHOLOGY
Volume 148, Issue 4, Pages 785-793Publisher
SPRINGER
DOI: 10.1007/s10658-016-1132-2
Keywords
Colletotrichum truncatum; Rpb1; Soybean anthracnose; Loop-mediated isothermal amplification (LAMP) assay
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Funding
- National High-Tech R&D Program (program 863) [2012AA101501]
- National Department Public Benefit Research Foundation [200903004]
- Chinese National Science Foundation Committee [31225022]
- public sector research funding [201303018]
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A loop-mediated isothermal amplification (LAMP) assay that directly detects Colletotrichum truncatum in diseased soybean tissues is described, thus allowing rapid diagnosis of soybean anthracnose. Using the target gene Rpb1 (that codes for the large subunit of RNA polymerase II), we designed and screened a set of species-specific primers allowing amplification at 62 A degrees C over 70 min. After addition of SYBR Green I to the LAMP reaction products, a yellow-green color (visible to the unaided eye) developed only in the presence of C. truncatum. The detection limit of the LAMP assay was 100 pg (per mu L genomic DNA). The Rpb1-Ct-LAMP assay has been successfully used to diagnose soybean anthracnose in field samples collected from Jiangsu, Anhui and Hubei provinces of China, and to detect C. truncatum in soybean seeds from farmers' markets. Our results show that the Rpb1-Ct-LAMP assay is a useful and convenient method for detecting C. truncatum, and thus for diagnosis of soybean anthracnose.
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