Rapid molecular detection of Esteya vermicola based on specific primers and the FTA-DNA extraction method

Abstract

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.

Keywords:

• Esteya vermicola
• molecular detection
• specific primers
• FTA-DNA extraction

Funding

This work was supported by the State 863 Project funded by the Ministry of Science and Technology [grant no. 2012AA101503] of P.R. China.

Additional information

Funding

Funding: This work was supported by the State 863 Project funded by the Ministry of Science and Technology [grant no. 2012AA101503] of P.R. China.