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Molecular detection method for resistance gene of spodoptera frugiperda to bacillus thuringiensis Bt Cry1F toxin

A technology of Spodoptera frugiperda and Bacillus aureus, applied in the direction of genetic engineering, plant genetic improvement, botanical equipment and methods, etc.

Inactive Publication Date: 2021-08-24
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can be seen that the loss of function of the protein encoded by the ABCC2 gene will lead to the resistance of Spodoptera frugiperda to Cry1F toxin

Method used

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  • Molecular detection method for resistance gene of spodoptera frugiperda to bacillus thuringiensis Bt Cry1F toxin
  • Molecular detection method for resistance gene of spodoptera frugiperda to bacillus thuringiensis Bt Cry1F toxin
  • Molecular detection method for resistance gene of spodoptera frugiperda to bacillus thuringiensis Bt Cry1F toxin

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Embodiment 1

[0032] In this example, 14 individuals of Spodoptera frugiperda collected from Brazil with a high level of resistance to Cry1F insecticidal protein were selected for molecular detection, and the preferred technology of the present invention was used to detect that they carried exon 12 of ABCC2 gene No. 15 - Allele frequency of the bp insertion mutation. Among them, ss indicates that the site is wild homozygous, rs indicates that the site is heterozygous for the mutation, and rr indicates that the site is homozygous for the mutation. The specific detection information is as follows:

[0033] Table 1 Mutation detection results of exon 15 of ABCC2 gene in samples of Spodoptera frugiperda collected in Brazil

[0034]

[0035] According to the preferred molecular detection technology of the present invention, its specific implementation steps include:

[0036] 1. Use AXYprep TMThe Multisource Genomic DNA Miniprep Kit Genomic Kit extracts whole genome DNA from Spodoptera frugi...

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Abstract

The invention discloses a molecular detection method for resistance genes of spodoptera frugiperda to bacillus thuringiensis Bt Cry1F toxin. The method comprises the following steps: carrying out PCR amplification on a genome DNA template of the spodoptera frugiperda by utilizing a specific forward primer F as shown in SEQ ID NO.1 and a specific reverse primer R as shown in SEQ ID NO.2, and directly sequencing a PCR purified product; according to a sequencing chromatogram, identifying whether 12-bp base insertion occurs on a No.15 exon of the individual adenosine triphosphate (ATP) binding cassette transporter C2 gene of the spodoptera frugiperda; and distinguishing the sensitive homozygote, the resistant heterozygote and the resistant homozygote of the resistant gene at one time. The method has the advantages of rapidness, simplicity, convenience and high accuracy, and is specially used for molecular detection and early warning of the resistance of the spodoptera frugiperda to the Bt Cry1F toxin.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a molecular detection method for the mutation of the resistance gene of Spodoptera frugiperda to Bt Cry1F toxin of Spodoptera frugiperda, which is specially used for No. 15 of Spodoptera frugiperda adenosine triphosphate (ATP) binding box transporter C2 gene Highly sensitive and rapid detection of exonic 12-bp insertion mutations. [0002] technical background [0003] Spodoptera frugiperda (J.E.Smith) is a worldwide pest native to tropical and subtropical regions of America, and mainly damages crops such as corn, sorghum, cotton and soybean. Biological characteristics such as polyphagia, high fecundity, strong migratory ability and easy resistance to insecticides strengthen the pest status of F. frugiperda. Spodoptera frugiperda is divided into corn type and rice type. The corn type mainly harms corn, sorghum, cotton and other crops, and the rice type mainly damages rice and f...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11C12N15/12
CPCC07K14/43563C12Q1/6888C12Q2600/124C12Q2600/156
Inventor 吴益东王兴亮管放杨亦桦武淑文
Owner NANJING AGRICULTURAL UNIVERSITY
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