Abstract
Anthracnose caused by Colletotrichum gloeosporioides is an economic disease that affects soybean production worldwide. This study developed a rapid, sensitive method for the detection of C. gloeosporioides using a loop-mediated isothermal amplification (LAMP) assay. By targeting a glutamine synthetase (GS) gene sequence, the GS-Cg-LAMP assay works most efficiently at 64°C and allows the detection of C. gloeosporioides DNA within 70 min based on a color change from orange to yellow-green following the addition of SYBR Green I to the LAMP reaction products. The detection limit was 1 pg μL−1 of genomic DNA per reaction. In specificity tests, the positive reaction (yellow-green color by the naked eye) was observed only in the presence of C. gloeosporioides, and none of other Colletotrichum spp. or fungal isolates produced a color change. Moreover, the GS-Cg-LAMP assay was successfully used to diagnose anthracnose caused by C. gloeosporioides in diseased soybean samples obtained from the field and detected the pathogen in soybean seeds bought from farmers’ markets. Our study provides a simple, sensitive tool for the rapid diagnosis of soybean anthracnose caused by C. gloeosporioides.
Highlights
Colletotrichum gloeosporioides is an economically important pathogen that infects a wide range of crops causing enormous production losses each year (Jeffries 1990)
We used the glutamine synthetase (GS)-Cg-loopmediated isothermal amplification (LAMP) assay to detect C. gloeosporioides in samples suspected of having soybean anthracnose collected from the field in the Jiangsu, Anhui, and Hubei Provinces
The results show that our GS-Cg-LAMP assay was both rapid and accurate compared with traditional isolation methods
Summary
Colletotrichum gloeosporioides is an economically important pathogen that infects a wide range of crops causing enormous production losses each year (Jeffries 1990). In addition to Colletotrichum gloeosporioides, C. truncatum, C. capsici, C. dematium, C. destructivum and C. coccodes cause soybean anthracnose (Ghosh et al 2016; Lou et al 2009; Hartman et al 1988; Roy 1982). These species are difficult to differentiate by microscopic examination because of their morphological similarity. The disease symptoms on soybean plants caused by the Colletotrichum spp. listed above and C. gloeosporioides are similar and difficult to differentiate. New techniques for the rapid diagnosis of anthracnose caused by C. gloeosporioides are needed
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