Morphological and molecular combined detection method for pine wood nematodes

Abstract

The invention belongs to the technical field of morphological and molecular combined detection, and discloses a morphological and molecular combined detection method for pine wood nematodes. The method comprises the steps of sample separation, morphological detection of the pine wood nematodes and molecular detection of the pine wood nematodes. A simple pine wood nematode separation device is designed in the invention; saprophytic nematodes and aphelenchoides nematodes are distinguished by adopting the presence or absence of stylets, and morphological identification is carried out according tothe shape difference of the tails and the tips of the female adults of pine wood nematodes and the quasi pine wood nematodes; molecular identification is carried out on the aphelenchoides nematodes,the pine wood nematodes and bursaphelenchus mucronatus by adopting a specific PCR technology; and complete detection technology system details are designed. The same pine wood sample is repeatedly sampled for three times during DNA extraction, so that accidental detection errors during sampling are avoided. The method meets requirements of forestry workers for rapid, efficient and accurate pine wood nematode detection, and provides timely and reliable detection means and technical support for general investigation of pine wood nematode diseases of forestry departments.

Description

technical field [0001] The invention belongs to the technical field of morphological and molecular combined detection, in particular to a morphological and molecular combined detection method of pine xylophilus. Background technique [0002] Currently, the closest prior art: [0003] Pine wood nematodes belong to the phylum Aschelminthes, class Nematoda, order Aphelenchina, family Aphelenchoididae, and genus Bursaphelenchus. Currently, there are more than 80 species of genus Bursaphelenchus . Most of the nematodes of the genus Umbrella parasitize in insects, and the species related to the death of pine trees are mainly B. xylophilus and B. xylophilus. The morphological characteristics of B. xylophilus and B. xylophilus are very similar. The main difference lies in the shape of the tail of the female. The tail of the female adult B. xylophilus is broad and round, finger-shaped, and the tail tip protrudes less than 2 microns, while the female adult B. xylophilus has a tapere...

AI Technical Summary

Problems solved by technology

[0007] In summary, the problem existing in the prior art is that according to the relatively stable characteristics of the shape of the female tail and tail tip, B. xylophilus and B. xylophilus can be basically distinguished. There is a certain degree of difference between them; and the relative instability of the tail shape of B. xylophilus increases the difficulty of accurate identification; and the identification of morphology is limited to female adults and is not suitable for the identification of males and larvae

The DNA extraction method of the existing PCR identification technology is complicated, and cannot detect all the nematodes of the genus S.

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