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Fast detection method of pine tree wilt disease pathogenic nematode

A pathogenic nematode and detection method technology, which is applied in the field of forest disease detection, can solve the problems of high price and application limitation, and achieve the effect of convenient operation, high accuracy, and favorable promotion and application

Inactive Publication Date: 2006-06-28
YUNNAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the completion of this method must rely on expensive fluorescent real-time PCR instruments, detection equipment and reagents, thus limiting its application

Method used

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  • Fast detection method of pine tree wilt disease pathogenic nematode
  • Fast detection method of pine tree wilt disease pathogenic nematode
  • Fast detection method of pine tree wilt disease pathogenic nematode

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Experimental program
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Effect test

Embodiment 1

[0020] 1. Design of specific primers for the detection of B. xylophilus and B. xylophilus: 2 populations of B. xylophilus and 5 populations of B. xylophilus from different sources were amplified using the general primers of the nematode ITS, and ITS1, 5.8 containing rDNA were obtained. S, the amplified fragment of ITS2 region; the DNA sequence of each amplified fragment was determined by the dideoxy termination method; the obtained sequence information was analyzed with DNASTAR 5.0 software to find out the difference sequence between the populations of B. xylophilus and B. xylophilus. According to the difference The sequence was designed using primer Premier 5.0 software to design pine xylophilus specific primers BX1: 5'-CGATGATGCGATTGGTGACTT-3' and BX25'-ATGCGAGACGACTGTCACAACG-3', and pine xylophilus specific primers BM1: 5'-TTCTCAAGTTTCTGCATTTGTAAG-3' and BM25' -GGCGCGGTCGCACAATCGAG-3', where BX1 is located in the ITS1 region of rDNA, BX2 is located in the ITS2 region of rDNA...

Embodiment 2

[0025]1. Design of specific primers for the detection of B. xylophilus and B. xylophilus: 2 populations of B. xylophilus and 5 populations of B. xylophilus from different sources were amplified using the general primers of the nematode ITS, and ITS1, 5.8 containing rDNA were obtained. S, the amplified fragment of ITS2 region; the DNA sequence of each amplified fragment was determined by the dideoxy termination method; the obtained sequence information was analyzed with DNASTAR 5.0 software to find out the difference sequence between the populations of B. xylophilus and B. xylophilus. According to the difference The sequence was designed using primer Premier 5.0 software to design pine xylophilus specific primers BX1: 5'-CGATGATGCGATTGGTGACTT-3' and BX25'-ATGCGAGACGACTGTCACAACG-3', and pine xylophilus specific primers BM1: 5'-TTCTCAAGTTTCTGCATTTGTAAG-3' and BM25' -GGCGCGGTCGCACAATCGAG-3', where BX1 is located in the ITS1 region of rDNA, BX2 is located in the ITS2 region of rDNA,...

Embodiment 3

[0030] 1. Design of specific primers for the detection of B. xylophilus and B. xylophilus: 2 populations of B. xylophilus and 5 populations of B. xylophilus from different sources were amplified using the general primers of the nematode ITS, and ITS1, 5.8 containing rDNA were obtained. S, the amplified fragment of ITS2 region; the DNA sequence of each amplified fragment was determined by the dideoxy termination method; the obtained sequence information was analyzed with DNASTAR 5.0 software to find out the difference sequence between the populations of B. xylophilus and B. xylophilus. According to the difference The sequence was designed using the primer Premier 5.0 software to design the specific primers BX1 of B. xylophilus: 5'-CGATGATGCGATTGGTGACTT-3' and BX25'-ATGCGAGACGACTGTCACAACG-3', and the specific primers BM1 of B. xylophilus: 5'-TTCTCAAGTTTCTGCATTTGTAAG-3' and BM2: 5'-GGCGCGGTCGCACAATCGAG-3', in which BX1 is located in the ITS1 region of rDNA, BX2 is located in the I...

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Abstract

A method for quickly detecting pathogenic threadworm of pine wilt disease includes designing each pair of specific primer for PCR detection of pine threadworm and pine imitate-threadworm, obtaining single threadworm DNA as template by mechanical grinding and filtering paper absorption, carrying out specific PCR amplification of pine threadworm and pine imitate-threadworm as well as detecting PCR product.

Description

Technical field: [0001] The invention belongs to the technical field of forest disease detection, in particular to a method capable of detecting pine wood nematodes and pseudo-pine wood nematodes. Background technique: [0002] Pine wilt disease caused by pine wood nematode (Bursaphelenchus xylophilus) is a devastating forest disease, which has the characteristics of extremely serious damage, rapid spread, and difficult control. Once it occurs, it will cause great losses to the national economy and the ecological environment , has now become the number one biological disaster in my country's forestry production, and its pathogenic pine xylophilus is an important quarantine object both internally and externally in my country. Due to the lack of good countermeasures after the occurrence of pine wilt disease, the current control of pine wilt disease is mainly to control the speed of its spread by removing the source of infection and cutting off the infection route. The diseased...

Claims

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Application Information

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IPC IPC(8): G01N21/78B01D57/02C12Q1/02
Inventor 王扬喻盛甫杨佩文胡先奇
Owner YUNNAN AGRICULTURAL UNIVERSITY
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