Application of pine wood nematode SSR labeled primer in pine wood nematode detection and PCR detection method of pine wood nematode

A technology of pine wood nematodes and labeled primers, applied in biochemical equipment and methods, microbial measurement/inspection, DNA/RNA fragments, etc., can solve the problems of pine forest area destruction, little effect, strong infectivity, etc., and achieve reduction The effect of a huge loss

Inactive Publication Date: 2017-05-31
SICHUAN AGRI UNIV
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Problems solved by technology

Because pine wood nematode disease is not easy to be found and identified in the early stage of the disease, and it is highly contagious, the damage to the pine forest area is very serious. Therefore, although the relevant departments have invested a lot of manpower and material resources to control it, the effect is very little. The harm of pine wood nematode still spreading rapidly

Method used

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  • Application of pine wood nematode SSR labeled primer in pine wood nematode detection and PCR detection method of pine wood nematode
  • Application of pine wood nematode SSR labeled primer in pine wood nematode detection and PCR detection method of pine wood nematode
  • Application of pine wood nematode SSR labeled primer in pine wood nematode detection and PCR detection method of pine wood nematode

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Experimental program
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Embodiment Construction

[0042] 1 Materials and methods

[0043] 1.1 Materials

[0044] For the test nematode a. the pine wood nematode (hereinafter represented by pine xylophilus ①) of artificial purification and cultivation, b. the pine wood nematode ( The following is represented by B. xylophilus ②), c. purified and cultivated B. xylophilus, and the above materials are all provided by the forest pathology laboratory of Forestry College, Sichuan Agricultural University.

[0045] Taq enzyme Taq enzyme is a thermostable DNA polymerase used in PCR amplification experiments, purchased from Chengdu Qingke Zixi Biotechnology Co., Ltd.

[0046] Primers were synthesized by Chengdu Qingke Zixi Biotechnology Co., Ltd. (see Table 1).

[0047] Table 1 PCR primer sequences and their amplification procedures

[0048] Table1 Primer sequence and amplification procedure about PCR

[0049]

[0050]

[0051] 1.2 Cultivation and collection of pine xylophilus

[0052] Get preserved Botrytis cinerea (Botrytis ...

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Abstract

The invention belongs to the field of phytopathogen detection and discloses a high-sensitivity primer marking the screened-out pine wood nematode based on SSR and an application thereof and establishes a PCR method for detecting pine wood nematode in a diseased pine plant. The primer and detection method disclosed by the invention can provide a stable and effective detection way for quarantine inspection and disease diagnosis of pine wood nematode and plays a positive role in reducing forestry loss caused by the pine wood nematode disease.

Description

technical field [0001] The invention belongs to the field of plant pathogen detection, and in particular relates to a use of a pine wood nematode SSR marker primer for detection of pine wood nematode and a PCR detection method for detecting pine wood nematode. Background technique [0002] Bine wood nematode is a devastating disease of pine trees caused by the pine wood nematode (Bursaphelenchus xylophilus) [1] . First reported in 1934 [2] , introduced to China in 1982 [3] , with its characteristics of strong pathogenicity, wide transmission route, fast spread, and difficult prevention and treatment [4-6] , bringing huge losses to the world's forestry. Because pine wood nematode disease is not easy to be found and identified in the early stage of the disease, and it is highly contagious, the damage to the pine forest area is very serious. Therefore, although the relevant departments have invested a lot of manpower and material resources to control it, the effect is very ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2565/125
Inventor 韩珊王方方朱天辉刘应高谯天敏李姝江刘裕峰张博阳李鸿朋黄焱焱阮若玉余鹏举何俊尧
Owner SICHUAN AGRI UNIV
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