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Formula for improving dsRNA insecticidal effect

A technology of formula and butanobuterol, which is applied in the field of biopesticide, can solve the problem of resistance, insecticidal efficiency and other problems

Active Publication Date: 2021-07-13
SHANGHAI PLANT SCI BIOTECHNOLOGY LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still many problems to be solved in pest control based on RNAi technology, for example, 1. How to quickly and high-throughput discover target genes; 2. How to easily apply to field production; 3. Resistance issues; 4. Safety issues ; 5. Insecticidal efficiency, etc.

Method used

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  • Formula for improving dsRNA insecticidal effect
  • Formula for improving dsRNA insecticidal effect
  • Formula for improving dsRNA insecticidal effect

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Embodiment 1 prepares liquid formula

[0059] Prepare the liquid formula according to the composition and ratio in Table 1 below.

[0060] Table 1. Liquid formula of dsRNA auxiliary agent

[0061]

[0062] After preparation, store at room temperature for later use. When spraying on farmland, temporarily dilute 1000 times.

Embodiment 2

[0063] Embodiment 2 prepares the dsRNA of killing cotton aphid

[0064] 2.1 Extraction of total RNA from cotton aphid

[0065] The RNA was extracted by the conventional Trizol method, purified by the conventional method, and treated with DNase to obtain a total RNA sample with a concentration ≥ 300 ng / ul, a total amount ≥ 6 μg, and an OD260 / 280 of 1.8-2.2.

[0066] 2.2 mRNA isolation and cDNA synthesis

[0067] PolyA-bearing mRNA was isolated using magnetic beads with oligo-dT, and cDNA first-strand synthesis was then performed using random hexamers and Invitrogen's Superscript II reverse transcriptase kit.

[0068] 2.3 Gene amplification and sequencing

[0069] Utilize the target gene-specific primers shown in Table 2, use the above-mentioned synthetic cDNA as a template to amplify the genes listed in the table, purify the obtained gene fragments, connect to the pMD18-T vector (Takara company), and transform into Escherichia coli Top10 strains, blue-white screening, positi...

Embodiment 3

[0076] Example 3 The Stability Investigation of Liquid Formula Preservation dsRNA

[0077] The dsRNA (ds_hemocytin) synthesized in Example 2 was dissolved in the liquid formulation prepared in Example 1 to a final concentration of 50 mg / mL. The synthetic dsRNA liquid and the aqueous solution of dsRNA dissolved in water with a concentration of 50 mg / mL were used as controls to investigate the stability of dsRNA.

[0078] After the three samples were exposed to the natural environment for 15 days, the bands were compared by agarose gel electrophoresis, and the results were as follows: figure 1 shown. The dyeing brightness of dsRNA in the strip represents the concentration of dsRNA, that is, the content, and the higher the content of dsRNA, the more stable the dsRNA in the sample. figure 1 M is Mark, A is the synthetic dsRNA solution, B is the dsRNA solution in the auxiliary agent, and C is the dsRNA aqueous solution. From figure 1 It can be seen from the comparison of the ba...

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Abstract

The invention discloses a formula for improving the insecticidal effect of dsRNA (double-stranded ribonucleic acid). The formula comprises the following components in parts by weight: 2 parts of potassium oleate; 0.1 to 0.5 part of geraniol; 0.005 to 0.05 part of synergistic ether; 0.05 to 0.3 part of diatomite; 0.5 to 2 parts of SDS; 0.05 to 0.3 part of sodium lauroyl sarcosinate; 0.5-2 parts of alkali metal chloride; and 0.1-0.5 part of alkaline earth metal chloride. According to the formula, the lasting time of dsRNA in the natural environment can be remarkably prolonged, and the prevention and treatment effect on cotton aphids is improved.

Description

technical field [0001] The invention belongs to the field of biological pesticides, relates to a formula for improving the insecticidal effect of dsRNA and its application in pest control, in particular to a formula for maintaining or improving the insecticidal effect of dsRNA on cotton aphids and its application in the control of cotton aphids application. Background technique [0002] The phenomenon of RNA interference (RNAi) has developed rapidly since it was discovered in 1991. Studies have shown that the use of RNAi to inhibit the expression of key genes in the process of insect growth and development can lead to insect developmental deformities and even death (BettencourtR, Terenius O, Faye I.2002.Insect Mol.Biol., 11(3):267-271.Whitten Miranda, M.A., et al,. Proceedings of the Royal Society B: Biological Sciences 283, 20160042.). This characteristic of RNAi inspired entomology researchers to apply it to the exploration of pest control. Through years of research, a ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N37/44A01N57/16A01P7/04
CPCA01N37/44A01N57/16A01P7/04C12N15/113A01N61/00C12N15/82C12N15/63
Inventor 苗雪霞李海超关若冰
Owner SHANGHAI PLANT SCI BIOTECHNOLOGY LTD
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