Morphological and molecular combined detection method for pine wood nematodes

A combined detection technology for B. xylophilus, applied in biochemical equipment and methods, measurement/inspection of microorganisms, material analysis through optical means, etc., can solve the instability of pine xylophilus tail shape, increase in accurate identification, difficulty, etc. problems, to achieve reliable detection means and technical support, efficient and accurate detection needs, and accurate molecular detection

Pending Publication Date: 2020-04-24
CHONGQING NORMAL UNIVERSITY
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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.

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  • Morphological and molecular combined detection method for pine wood nematodes
  • Morphological and molecular combined detection method for pine wood nematodes
  • Morphological and molecular combined detection method for pine wood nematodes

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Embodiment

[0087] 1. Materials and methods

[0088] 1.1 Collection of samples

[0089] The materials used in the experiments provided by the embodiments of the present invention come from the 2017 autumn census of forest diseases and insect pests in Pengshui, Wuxi, Tongliang and other 14 districts and counties in Chongqing City. There are 276 samples in total, which are preserved in the life of Chongqing Normal University. Institute of Science Insect and Molecular Biology Research -20°C refrigerator. The worm samples used in the experiment were all fresh living specimens, which were identified through morphological identification, among which the specimens of B. xylophilus and B. xylophilus were the main ones. The collection time, collection location and host plants of the case samples were registered in detail (as shown in Table 1).

[0090] Table 1 Sources and hosts of samples isolated from S.

[0091]

[0092]

[0093] 1.2 Separation of samples

[0094] 1.2.1 Sample processi...

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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...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/686G01N21/84
CPCC12Q1/6888C12Q1/686G01N21/84C12Q2565/125
Inventor 陈斌袁缓司风玲闫振天
Owner CHONGQING NORMAL UNIVERSITY
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