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A pine wood sample preparation method for high target and quality DNA extraction for detection of Esteya vermicola by PCR from living pine
Author(s) -
Yin Can,
Wang Yunbo,
Zhang Yongan,
Wang Haihua,
Tao Ran,
Li Yu,
Sung Chang Keun
Publication year - 2019
Publication title -
journal of basic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0233-111X
DOI - 10.1002/jobm.201800522
Subject(s) - dna extraction , sample preparation , extraction (chemistry) , fungus , chromatography , polymerase chain reaction , pulp and paper industry , chemistry , biology , botany , biochemistry , gene , engineering
Esteya vermicola is an endophytic fungus of pine wood nematode with demonstrated biocontrol efficacy. At present, the methods for detection of this fungus from pine wood is still inconvenient and inefficient such as thin‐section microscopic observations. In the present study, a simple protocol was developed for wood sample preparation for effective extraction of fungal DNA from wood samples for PCR detection. The protocol of preparation of the sample involves washing in sterile water overnight on a shaking table followed by filtration and centrifugation to obtain the extracted DNA. The result indicates that with this sample preparation protocol, any proper DNA extraction method can be effectively used for the rapid and reliable detection of E. vermicola from pine wood. This method can provide valuable support for follow‐up studies with practical applications, such as investigation of the growth rate of E. vermicola and how long it remains viable inside a pine tree.