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Specificity molecule marker of penicillum IPC strain and its application

A kind of Paecilomyces lilacinus, molecular marker technology, applied in the direction of determination/inspection of microorganisms, sugar derivatives, biochemical equipment and methods, etc. , Bio-control bacteria detection sensitivity and accuracy are not high, to achieve the effect of solving legal disputes

Inactive Publication Date: 2004-07-21
YUNNAN ACAD OF TOBACCO AGRI SCI
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Problems solved by technology

The main reason is that the detection sensitivity and accuracy of bio-control bacteria are not high, and it cannot accurately and objectively reflect the colonization, survival, growth and decline of bio-control bacteria in the natural environment

Method used

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  • Specificity molecule marker of penicillum IPC strain and its application
  • Specificity molecule marker of penicillum IPC strain and its application
  • Specificity molecule marker of penicillum IPC strain and its application

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Embodiment Construction

[0021] The following examples are used to further describe the present invention, but the content of the present invention is not limited thereto.

[0022] 1. Acquisition of specific DNA fragments

[0023] By cultivating the pure culture mycelia of the M-14 strain and the control strain, extracting their genomic DNA respectively, carrying out PCR amplification with universal primers, and screening the primers with DNA polymorphisms for all tested strains. The amplified DNA polymorphisms were compared, and primer OPC-11 (5'-AAA GCT GCG G 3') amplified a DNA fragment unique to the M-14 strain with a size of about P850bp, which was named P850; primer OPD-05 (5'-TGA GCG GAC A-3') amplified a DNA fragment unique to the M-14 strain with a size of about P1400bp, and the fragment was named P1400.

[0024] 2. Determination of specific DNA fragment sequence

[0025] The specific DNA fragments P850 and P1400 on the agarose gel after electrophoresis were excised under the ultraviolet vi...

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Abstract

A specific molecular marker of lilacinus pseudopanicillum sp. M-14 strain and its application are disclosed. The randomly amplifying DNA polymorphism technique is used for molecular fingerprint analysis to M-14 strain and the frequently seen fungus in soil to obtain the RAPD specific DNA fragments P850 and P1400 of M-14 strain, which are recovered, purified, cloned and sequenced. The specific SCAR-PCR primer is designed for PCR amplification to the DNA extracted from the soil where M-14 stain is applied and the amplified fragments P850 and P1400 are marked by digoxin to obtain the hybridizing probe for spot hybridization to said extracted DNA, so specifically detecting if there is M-14 strain in natural soil and its quantity.

Description

Technical field: [0001] The invention relates to a specific molecular marker of Paecilomyces lilacinus M-14 strain and an application thereof, belonging to the field of biotechnology. technical background: [0002] Soybean cyst nematode (Heterodera glycines) is one of the important groups of plant parasitic nematodes. In my country, nematodes mainly harm major economic crops such as tobacco, flowers, vegetables, cotton, soybeans, peanuts, Panax notoginseng, American ginseng, etc., and become the key to the development of these crops. important limiting factor. In Heilongjiang Province, the annual loss caused by soybean cyst nematode alone is as much as 800 million yuan. What's more serious is that the crop wounds caused by the nematode infestation cause serious root diseases of the crops. According to incomplete statistics, nematode diseases cause an average loss of 10-15% of the yield of various crops every year. Nematodes have been controlled by chemical pesticides for a...

Claims

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Application Information

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IPC IPC(8): C07H21/04C12P19/34C12Q1/68
Inventor 祝明亮夏振远张克勤刘杏忠
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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