Journal
BIOCONTROL SCIENCE AND TECHNOLOGY
Volume 24, Issue 8, Pages 872-881Publisher
TAYLOR & FRANCIS LTD
DOI: 10.1080/09583157.2014.900736
Keywords
Esteya vermicola; molecular detection; specific primers; FTA-DNA extraction
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Funding
- State 863 Project - Ministry of Science and Technology of P.R. China [2012AA101503]
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Esteya vermicola is an endoparasitic fungus of the pinewood nematode and thus has great biocontrol value. At present, the detection of this fungus is still based on microscopic observations and morphological identification, and the sampling is notably inconvenient and inefficient. In the present study, a pair of specific primers (upstream primer, 5'-GTGCCTCTACCAAGACTCGC-3'; downstream primer, 5'-CGCCAAATGTCAAGATCCGC-3') was designed to detect E. vermicola. The analysis of the PCR amplification and the agarose gel electrophoresis results led to the establishment of a new method for the detection of E. vermicola through the presence of a 176-bp specific fragment. In addition, the use of a FTA-DNA direct extraction method for the detection of E. vermicola was explored. The results suggest that the proposed method can be effectively used for the rapid detection of E. vermicola and may provide important technical support for follow-up studies of the fungus in field experiments.
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