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DNA bar code standard detection sequence for apocynum venetum aphids and application of DNA bar code standard detection sequence

A standard detection and barcode technology, applied in applications, microbial determination/inspection, enzymes, etc., can solve problems such as difficulty in identification of Apocynum aphid species, and achieve the effect of reducing costs, saving time, and filling gaps in the industry

Inactive Publication Date: 2020-10-13
LANZHOU UNIVERSITY
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Problems solved by technology

[0006] The purpose of the present invention is to provide a standard detection sequence of DNA barcode of apocynum acystis and its application in species identification, which can make up for the technical defects existing in identifying aphids by morphology at present, and aim at the difficult problem of species identification of apocynum apocynum, Provide a fast, simple and accurate molecular biological identification method

Method used

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Embodiment Construction

[0023] 1. Extraction of total DNA from Apocynum abaceae

[0024] (1) The DNA was extracted by SDS method. Add liquid nitrogen to a single insect in a mortar for thorough grinding, then add 500 μL of DNA extraction solution (500ml ddh20, 100ml 1M ph8.0 Tris-Hcl, 100ml 0.5M ph8.0 EDTA, 100ml 5M Nacl, 200ml 10% SDS), and all transferred to 2ml centrifuge tube.

[0025] (2) Wash the mortar with 150 μL of DNA extraction solution, transfer to the same centrifuge tube, shake well, and place in a water bath at 65°C for 30 minutes (shake well 2-3 times in the middle).

[0026] (3) After standing at room temperature, add 195 μL of potassium acetate, ice-bath for 5 minutes (-20°C); add 600 μL of chloroform / isoamyl alcohol (v:v=24:1), mix well; room temperature 12000 r / min, centrifuge for 10 minutes.

[0027] (4) Pipette 360 ​​μL supernatant into a new 1.5ml pointed centrifuge tube, add 36 μL 3M Nacl and 240 μL isopropanol to mix well, and place at room temperature for 10 min; after tak...

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Abstract

The invention discloses a DNA bar code standard detection sequence for apocynum venetum aphids and a molecular identification method based on the standard sequence. The DNA bar code standard detectionsequence is shown as SEQ ID NO. 1. According to the invention, by adoption of a reliable DNA molecular identification technology, the method aims to rapidly identify main pests, namely the apocynum venetum aphids in an apocynum venetum field, and has the characteristics of accuracy, rapidness, convenience and low cost; and compared with a traditional morphological characteristic identification method, the method provided by the invention saves time and reduces cost. The method provided by the invention has low requirements on used instruments and equipment, can be carried out in general molecular biology laboratories, is more accurate and reliable than traditional morphological identification, obtains the CO1 sequence of the apocynum venetum aphids for the first time, fundamentally solvesthe problems that the apocynum venetum aphids are difficult to accurately identify and easy to mix with cotton aphids in a conventional apocynum venetum aphid prevention and treatment process, fillsindustrial blank and has important significance.

Description

technical field [0001] The invention belongs to the technical field of species identification, and in particular relates to a DNA barcode standard detection sequence of Apocynum apocynum and its application in species identification. Background technique [0002] Apocynum apocynum is a species of the genus Aphididae in the order Homoptera. Apocynum apocynum has polymorphism, and the common types in the field are apterous viviparous females and winged viviparous females. The body length is 2mm and the width is 1-1.2mm. The common color is yellow-green and dark green. After death, the color becomes darker, mostly dark green. At the end of April and the beginning of May, the apocynum aphid begins to appear in the field, and the peak of development is from mid-July to the end of July. At the end of August and the beginning of September, the apocynum aphid gradually dies. Mainly distributed in the top 1-30 cm of the apocynum plant, the damage is concentrated vertically, and the ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6869C12N15/53
CPCC12N9/0053C12Q1/6869C12Q1/6888C12Y109/03001C12Q2531/113C12Q2563/185
Inventor 袁明龙冯润秋洛芳珍张莉君曹亚李敏
Owner LANZHOU UNIVERSITY
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