Identification method and special primer for trogoderma glabrum and trogoderma variabile

The technology of the black beetle and the black beetle, which is applied in the biological field, can solve the problems of no rapid molecular biology detection method, time-consuming and high cost, shorten the time required for the detection, improve the detection accuracy, and achieve credible results. high degree of effect

Inactive Publication Date: 2012-04-04
CHINA AGRI UNIV
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Problems solved by technology

[0005] Molecular biological techniques have been used in the identification and classification of insects. At present, the identification of closely related species of the genus Dermatomus mainly adopts traditional morphological taxonomy. Although there are also methods that use PCR amplification fragments and sequence identification, but Time-consuming and expensive, there is no relevant rapid molecular biology detection method

Method used

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  • Identification method and special primer for trogoderma glabrum and trogoderma variabile
  • Identification method and special primer for trogoderma glabrum and trogoderma variabile
  • Identification method and special primer for trogoderma glabrum and trogoderma variabile

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Embodiment 1

[0042] Embodiment 1, the design of primer

[0043] According to the sequence (gene sequence number: FJ58973) of the existing Trogoderma zebra, design primers:

[0044] Tgla-F: 5-CAACATTTATTTTTGATTT-3 (SEQ ID NO: 1)

[0045] Tgla-R: 5-TCCAATGCACTAATCTGCCA-3 (SEQ ID NO: 2)

Embodiment 2

[0046] Embodiment 2, identification of Dermoderma melanogaster and Dermoderma versicolor

[0047] 1. Extraction of DNA from dermatomid samples

[0048] The two dermatomid beetles numbered 1 and 2 were respectively provided by Huangpu Entry-Exit Inspection and Quarantine Bureau (collected in Huangpu, Guangzhou) and Shandong Agricultural University (collected in Tai’an, Shandong). Tiangen Tissue Genomic DNA Extraction Kit (TIANamp Genomic DNA Kit ), follow the instructions to extract the total DNA of the sample to be tested. Specifically: Grind live worms in 200 μl of GA buffer, mix well, add 20 μl of proteinase K, and place at 37° C. for 4 hours. Then add 200 μl of GB buffer, mix well, and place at 70° C. for 10 minutes. Add 200ul absolute ethanol and mix well for 15s. Transfer the obtained solution into the CB3 adsorption column, centrifuge at 12,000rpm for 30s, pour off the waste liquid, and put the adsorption column back into the collection tube. Add 0.5ml of GD buffer s...

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Abstract

The invention discloses an identification method and a special primer for trogoderma glabrum and trogoderma variabile. A primer pair provided by the identification method consists of a primer 1 and a primer 2. In the primer pair, nucleotide sequences of the primer 1 and the primer 2 are respectively a sequence 1 and a sequence 2 in a sequence table. Experiments prove that species specificity variation sites of different species of trogoderma are found according to mitochondrion gene sequence differences between the species of the trogoderma, a molecular identification system of the trogodermaspecies is built, and finally, the fast, accurate and efficient identification of the quarantine type trogoderma is realized.

Description

technical field [0001] The invention relates to the field of biological technology, in particular to an identification method of Dermoderma melanogaster and Dermoderma versicolor and special primers thereof. Background technique [0002] Trogoderma pests are important storage pests. At present, the identification of Trogoderma is mainly based on morphological characteristics. However, because the adults of Trogoderma are generally small and the morphological differences between species are not obvious, it often causes confusion between species in identification. [0003] Dermatomid beetles are small, and the adult body length is 2.0-3.5mm, and the shape is similar and difficult to distinguish. The identification of adults is mainly based on the number of antennal rod segments, the depth of antennal fossae, the color of elytra and the characteristics of markings, etc.; the body length of larvae is 4.0-6.0mm, generally based on the number and type of setae attached, and the ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 班丽萍蒋湘刘聪樊武疆王新国林惠娇吕英姿刘二龙
Owner CHINA AGRI UNIV
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