CN-116179665-B - Identification method of emamectin benzoate resistance gene in spodoptera frugiperda, dsRNA and application thereof

Abstract

The invention provides an identification method of emamectin benzoate resistance genes in spodoptera frugiperda, dsRNA and application thereof, and belongs to the technical field of molecular biology. The invention combines genome re-sequencing and transcriptome sequencing to identify key resistance genes aiming at emamectin benzoate in spodoptera frugiperda, designs dsRNA based on mRNA conserved sequences of the key resistance genes GSTD6 and CTP6A18, can effectively reduce the drug resistance of spodoptera frugiperda to emamectin benzoate through RNAi technology, and has important practical significance for developing effective spodoptera frugiperda resistance management strategies to delay the generation of resistance and identify new targets for pest control.

Inventors

• SUN ZHONGXIANG
• LU ZHIHUI
• GUI FURONG
• CHEN YAPING
• FU PENGFEI
• LI DONGGUI
• LI HAO

Assignees

• 云南农业大学

Dates

Publication Date

20260505

Application Date

20230214

Claims (4)

1. 1. A dsRNA that reduces resistance to emamectin benzoate of spodoptera frugiperda, characterized in that said dsRNA is dsRNA-1 or dsRNA-2; The nucleotide sequence of the synthesized template of the dsRNA-1 is shown as SEQ ID NO. 1; the sense strand of the dsRNA-1 is amplified by using primers with nucleotide sequences shown as SEQ ID NO. 3 and SEQ ID NO. 6, the antisense strand of the dsRNA-1 is amplified by using primers with nucleotide sequences shown as SEQ ID NO. 4 and SEQ ID NO. 5, and the sense strand and the antisense strand of the dsRNA-1 are annealed to obtain the dsRNA-1; the nucleotide sequence of the synthesized template of the dsRNA-2 is shown as SEQ ID NO. 2, the sense strand of the dsRNA-2 is amplified by using primers with nucleotide sequences shown as SEQ ID NO. 7 and SEQ ID NO. 10, the antisense strand of the dsRNA-2 is amplified by using primers with nucleotide sequences shown as SEQ ID NO. 8 and SEQ ID NO. 9, and the sense strand and the antisense strand of the dsRNA-2 are annealed to obtain the dsRNA-2.
2. 2. Use of the dsRNA of claim 1 for the preparation of a product that reduces resistance to emamectin benzoate.
3. 3. Use of the dsRNA of claim 1 for controlling spodoptera frugiperda.
4. 4. A method for reducing resistance to emamectin benzoate in Spodoptera frugiperda, comprising introducing the dsRNA of claim 1 into the body of Spodoptera frugiperda.

Description

Identification method of emamectin benzoate resistance gene in spodoptera frugiperda, dsRNA and application thereof Technical Field The invention relates to the technical field of molecular biology, in particular to an identification method of emamectin benzoate resistance genes in spodoptera frugiperda, dsRNA and application thereof. Background Spodoptera frugiperda (Spodoptera frugiperda) is a major cross-border migratory pest for global early warning of grain and agricultural organizations in the united nations, and larvae can be more than 350 plants such as corn, wheat, rice and the like. At present, the prevention and control means of spodoptera frugiperda at home and abroad still mainly adopts chemical prevention and control, and has the characteristics of quick response, low cost and the like. Emamectin benzoate (simply referred to as emamectin benzoate) is a semisynthetic high-efficiency low-toxicity biological source pesticide and has been used for controlling crop pests such as vegetables, fruit trees and cotton. The research shows that the emamectin benzoate has a stronger virulence effect on spodoptera frugiperda, the corrected mortality rate is more than 90%, and the field control effect test shows that the emamectin benzoate has good quick-acting property and lasting effect on spodoptera frugiperda, is superior to other medicaments, and can be used as one of the first-choice medicaments for controlling spodoptera frugiperda. However, spodoptera frugiperda has developed resistance to a number of traditional chemical pesticides, and by investigating the resistance of spodoptera frugiperda field populations in the province of bergamot, flourishing, and shaggy, it has been found that spodoptera frugiperda field populations have developed resistance to the novel biological source pesticide emamectin benzoate to varying degrees. The current research on the effect of emamectin benzoate on spodoptera frugiperda mainly focuses on virulence effect and resistance inheritance, and the research on resistance mechanism is still freshly reported. Disclosure of Invention In view of the above, the present invention aims to provide an identification, dsRNA and application thereof of emamectin benzoate resistance genes in spodoptera frugiperda, the present invention combines genome re-sequencing and transcriptome sequencing to identify key resistance genes for emamectin benzoate in spodoptera frugiperda, and the dsRNA for regulating the emamectin benzoate resistance of spodoptera frugiperda by carrying out RNA interference on the key resistance genes can effectively regulate the emamectin benzoate resistance of spodoptera frugiperda, and has important practical significance for developing an effective spodoptera frugiperda resistance treatment strategy to delay the generation of resistance and identify new targets for pest control. In order to achieve the above object, the present invention provides the following technical solutions: A method for identifying emamectin benzoate resistance genes in spodoptera frugiperda, comprising the steps of: s1, genome resequencing analysis of a emamectin benzoate drug-resistant population and a sensitive population, S2, transcriptome sequencing analysis of the drug-resistant population and the sensitive population; S3, identifying drug resistance related candidate genes, namely cross-linking the resistance related genome region obtained in the S1 with the differential expression genes obtained in the S2 to find candidate resistance related genes crossed with the resistance related genome region; s4, verifying the expression quantity difference of the candidate resistance related genes in the emamectin benzoate resistant strain and the sensitive strain spodoptera frugiperda by using fluorescent quantitative PCR. The identification method identifies the obtained dsRNA for regulating the resistance of the spodoptera frugiperda emamectin benzoate, wherein the dsRNA is dsRNA-1 or dsRNA-2, the dsRNA-1 is transcribed from a nucleotide sequence shown as SEQ ID NO.1, and the dsRNA-2 is transcribed from a nucleotide sequence shown as SEQ ID NO. 2. The invention also provides template DNA for synthesizing the dsRNA-1 or the dsRNA-2 in the scheme, wherein the nucleotide sequence of the template mRNA of the dsRNA-1 is shown as SEQ ID NO. 1, and the nucleotide sequence of the template mRNA of the dsRNA-2 is shown as SEQ ID NO. 2. The invention also provides an application of the dsRNA or the template mRNA in regulating the resistance of spodoptera frugiperda emamectin benzoate. The invention also provides an application of the dsRNA or the template mRNA in preparing a product for regulating spodoptera littoralis emamectin benzoate resistance. The invention also provides an application of the dsRNA or the template mRNA in preventing and controlling spodoptera frugiperda. The invention also provides an application of the dsRNA or the template mRNA in preparing spodoptera frugiperda product.