CN-122012508-A - Application of double-stranded RNA molecule and related biological material thereof in prevention and treatment of soybean thrips

Abstract

The invention discloses a double-stranded RNA molecule and application of a related biological material thereof in preventing and controlling soybean thrips, and relates to the technical field of agricultural pest prevention and control. The double-stranded RNA molecule provided by the invention targets Muhsp genes of soybean thrips, and the coding sequence of the Muhsp genes is SEQ ID NO. 1. The double-stranded RNA molecule provided by the invention can effectively interfere the gene expression of the soybean thrips Muhsp so as to cause death of the soybean thrips, and the double-stranded RNA molecule is further applied together with a biological source pesticide, so that the death rate of the soybean thrips is further improved. The invention can realize the accurate control of soybean thrips, improve the control efficiency of biological source pesticides and has good application prospect and agricultural sustainable development value.

Inventors

• WANG DAN
• SHEN JIE
• JIA KEHAN

Assignees

• 中国农业大学

Dates

Publication Date

20260512

Application Date

20260415

Claims (10)

1. 1. The double-stranded RNA molecule is characterized in that the double-stranded RNA molecule targets Muhsp genes of soybean thrips, and the coding sequence of the Muhsp genes is SEQ ID NO. 1.
2. 2. The double-stranded RNA molecule of claim 1, wherein the double-stranded RNA molecule consists of a sense strand and an antisense strand, the nucleotide sequence of the sense strand being SEQ ID No. 2 and the nucleotide sequence of the antisense strand being SEQ ID No. 3.
3. 3. A biological material associated with the double stranded RNA molecule of claim 1 or 2, characterized in that said biological material is selected from at least one of A1) -A5): A1 A DNA molecule encoding the double stranded RNA molecule; a2 An expression cassette comprising A1) said DNA molecule; A3 A) a recombinant vector comprising the DNA molecule of A1) or a recombinant vector comprising the expression cassette of A2); a4 A) a recombinant microorganism comprising the DNA molecule of A1) or a recombinant microorganism comprising the expression cassette of A2) or a recombinant microorganism comprising the recombinant vector of A3); A5 A transgenic cell line comprising A1) said DNA molecule or a transgenic cell line comprising A2) said expression cassette or a transgenic cell line comprising A3) said recombinant vector.
4. 4. A product comprising the double stranded RNA molecule of claim 1 or 2.
5. 5. The product of claim 4, further comprising a double stranded RNA molecule delivery vehicle and/or an insecticide.
6. 6. The product of claim 5, wherein the double stranded RNA molecule delivery vehicle is a polymeric vehicle represented by formula 1: ; In formula 1, R 1 and R 2 are each independently selected from H, CH 3 、CH 2 CH 3 or CH 2 CH 2 CH 3 ;n 1 、n 2 、n 3 、n 4 and each independently takes a value of 1-100.
7. 7. The product of claim 5 or 6, wherein the pesticide comprises at least one of emamectin benzoate, spinetoram.
8. 8. Use of the double stranded RNA molecule of any one of claims 1-2 or the biomaterial of claim 3 or the product of any one of claims 4-7, wherein the use is selected from at least one of B1) -B6): B1 Application in reducing gene expression quantity of bean thrips Muhsp; B2 Application in preparing products for reducing gene expression quantity of bean thrips Muhsp; B3 Application in preventing and controlling the invasion of plants by soybean thrips; B4 The application of the composition in the preparation of products for preventing and treating the damage of plants by soybean thrips; b5 Application in reducing survival rate of soybean thrips; b6 The application of the soybean thrips survival agent in the preparation of products for reducing soybean thrips survival rate.
9. 9. A method for reducing survival rate of soybean thrips, comprising feeding or contacting the double-stranded RNA molecule of claim 1 or 2 or the product of any one of claims 4 to 7 with soybean thrips, and reducing survival rate of soybean thrips after the double-stranded RNA molecule enters the soybean thrips.
10. 10. A method for controlling soybean thrips infestation of plants, comprising applying the double stranded RNA molecule of claim 1 or 2 or the product of any one of claims 4-7 to a plant surface, and after feeding or contacting the double stranded RNA molecule with soybean thrips, feeding the double stranded RNA molecule into the soybean thrips, and controlling soybean thrips infestation of plants by reducing soybean thrips survival.

Description

Application of double-stranded RNA molecule and related biological material thereof in prevention and treatment of soybean thrips Technical Field The invention relates to the technical field of agricultural pest control, in particular to a double-stranded RNA molecule and application of related biological materials thereof in controlling soybean thrips. Background Thrips sojae (Megalurothrips usitatus) belonging to the Thysanoptera thrips family (THRIPIDAE) is a destructive pest of leguminous plants. The soybean thrips directly eat leaf blade, flower device and pod juice mainly through the file suction type mouth gag, so that the leaf blade is shrunken, the flower device falls off, and the pod is black head and black tail. It can also transmit various plant viruses, seriously affecting crop yield and quality. The insect body is small, the concealment is strong, the propagation is quick, and the prevention and control difficulty is high. Under the large background of chemical pesticide reduction and synergy, biological source pesticides such as emamectin benzoate and spinetoram which are commonly used at present have drug resistance, and the resistance is enhanced year by year, and development of a green and efficient novel control technology is needed to cope with resistance pressure. RNA interference (RNA INTERFERENCE, RNAI) technology can utilize double-stranded RNA (double-STRANDED RNA, DSRNA) to mediate target mRNA specific degradation, thereby achieving gene silencing. In the field of pest control, the technology can effectively interfere the normal growth and development or physiological functions of the dsRNA aiming at the key genes of pests and even lead to the death of the pests by designing the dsRNA. In particular, at the end of 2023, commercialization of the first spray type RNA biopesticide Ledprona in the world has been formally approved internationally, marking the entry of pest control into the nucleic acid era. RNA pesticide based on artificial synthesized dsRNA is gradually developed into an important pest green prevention and control strategy, and has wide application prospect. Therefore, the mining of highly efficient functional genes and the construction of stable double-stranded RNA (dsRNA) delivery systems are of great importance for increasing the potential of application of RNAi technology. Disclosure of Invention The invention provides an application of double-stranded RNA molecules and related biological materials thereof in preventing and controlling soybean thrips, which are used for reducing the survival rate of soybean thrips, thereby effectively protecting plants from being damaged by soybean thrips. In a first aspect, the invention provides a double-stranded RNA molecule, wherein the double-stranded RNA molecule targets Muhsp (heat shock protein) genes of soybean thrips, and the coding sequence of the Muhsp genes is SEQ ID NO. 1. A double-stranded RNA molecule as described above, which consists of a sense strand and an antisense strand, wherein the nucleotide sequence of the sense strand is SEQ ID NO. 2, and the nucleotide sequence of the antisense strand is SEQ ID NO. 3. The double-stranded RNA molecule can be transcribed in vitro by using a coding sequence fragment of Muhsp gene (the nucleotide sequence shown at positions 61-567 of SEQ ID NO: 1) as a template. In a second aspect, the present invention provides a biological material associated with the double stranded RNA molecule of any one of the above, said biological material being selected from at least one of A1) -A5): A1 A DNA molecule encoding the double stranded RNA molecule; a2 An expression cassette comprising A1) said DNA molecule; A3 A) a recombinant vector comprising the DNA molecule of A1) or a recombinant vector comprising the expression cassette of A2); a4 A) a recombinant microorganism comprising the DNA molecule of A1) or a recombinant microorganism comprising the expression cassette of A2) or a recombinant microorganism comprising the recombinant vector of A3); A5 A transgenic cell line comprising A1) said DNA molecule or a transgenic cell line comprising A2) said expression cassette or a transgenic cell line comprising A3) said recombinant vector. The biological material as described above, wherein in A1) said DNA molecule comprises the nucleotide sequence shown in positions 61-567 of SEQ ID NO. 1. In a third aspect, the invention provides a product comprising a double stranded RNA molecule as defined in any one of the preceding claims. A product as described above, said product further comprising a double stranded RNA molecule delivery vehicle and/or an insecticide. It will be appreciated that the delivery route of the double stranded RNA molecule may be by injection, feeding, epidermis etc. and that the delivery vehicle used will vary depending on the delivery route and further that the product is suitable for delivery by the feeding route. Further, the double-stranded RNA molecule delivery carrier is