Molecular-biological method for quickly distinguishing noctuidae pests

A technology of molecular biology and noctuidae, applied in the field of molecular biology, can solve the problems of lack of scientific identification methods, inaccurate morphological identification of Spodoptera litura, cotton bollworm and tobacco caterpillar

Inactive Publication Date: 2012-03-14
YUNNAN NORMAL UNIV
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Problems solved by technology

[0006] The purpose of the present invention is to provide a scientific, effective, simple, quick and accurate molecular biological identificatio

Method used

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Examples

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Embodiment 1

[0028] 1. Use the standard phenol-chloroform method or DNA extraction kit to extract the total DNA of Spodoptera litura, cotton bollworm and N. tabaci. In order to avoid contamination by fungi and nematodes, the present invention discards the abdomen, wings and antennae, and only extracts DNA from the head, chest and legs. DNA was extracted by referring to the standard phenol / chloroform protocol (the standard phenol / chloroform protocol) or directly using a DNA extraction kit (Shanghai Huashun Bioengineering Co., Ltd.).

[0029] 2. Using DNA barcode primers to PCR amplify mitochondrial COI gene fragments and sequence determination, specifically including the following steps:

[0030] ①PCR reaction system is 50μl, including about 25ng of template DNA, 1×PCR buffer, 2.5mM Mgcl 2 , 1mM dNTP, 2 μg / μl BSA, 2 pM each of forward and reverse primers, 1 unit of TaqDNA polymerase. Add deionized water to adjust to a final volume of 50 μl, and cover the system with paraffin oil. Reactio...

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Abstract

The invention discloses a molecular-biological method for quickly distinguishing prodenia litura and other two noctuidae pests (cotton bollworm and tobacco budworm). The method comprises the following steps of: extracting total DNA (Deoxyribonucleic Acid) by adopting a standard phenol-chloroform method or a DNA extraction kit; using a DNA bar code primer to amplify a sequence at the front part of mitochondria CO1 as a label; and comparing diagnostic characters of nucleotides. The molecular-biological method can be used for distinguishing different biotypes and larvae or incomplete individuals of the prodenia litura, the cotton bollworm and the tobacco budworm, has the characteristics of accuracy, quickness, convenience and quickness, and is more accurate and reliable compared with the traditional morphological identification.

Description

technical field [0001] The invention belongs to the field of molecular biology. Specifically, the invention relates to a molecular biology method capable of quickly identifying Spodoptera litura and other two Noctuidae pests (cotton bollworm and budworm). Background technique [0002] Spodoptera litura ( Spodoptera litura (Fabricius)) is a polyphagous and overeating worldwide agricultural pest, one of the top ten economic pests in my country, and endangers plants including important energy crops such as corn, cassava and taro. Due to Spodoptera litura, cotton bollworm ( Helicoverpa armigera ) and tobacco budworm ( H. assault ) belong to the family Lepidoptera Noctuidae. Eggs, larvae and pupae are very similar in size, color and pattern, so it is difficult to identify; insect taxonomists usually identify them based on the characteristics of the genitalia of the three adults. However, this traditional species identification mainly relies on professional insect taxonomist...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 李青青段焰青叶辉施伟刘晓飞曹能
Owner YUNNAN NORMAL UNIV
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