MiRNA stimulant, chitin synthesis inhibition type biopesticide and application thereof

A technology of biological pesticides and simulants, applied in the directions of DNA/RNA fragments, applications, plant growth regulators, etc., can solve the problems of low efficiency of agricultural control methods, poor control effects, and difficulty in sustainable development of brown planthopper control work. Effective insecticidal effect

Inactive Publication Date: 2017-01-04
SUN YAT SEN UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Agricultural control methods are inefficient and effective, and chemical drugs often lead to problems such as insect resistance and environmental pollution, making it difficult to achieve sustainable development of brown planthopper control

Method used

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  • MiRNA stimulant, chitin synthesis inhibition type biopesticide and application thereof
  • MiRNA stimulant, chitin synthesis inhibition type biopesticide and application thereof
  • MiRNA stimulant, chitin synthesis inhibition type biopesticide and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Step 1: Synthesize miR-2703 mimics according to the nucleotide sequence (SEQ ID NO: 1) (commissioned by Guangzhou Ruibo Company).

[0025] The miR-2703 mimic is a chemically synthesized mature miRNA duplex, which can enhance the function of endogenous miRNA, including a sequence consistent with the target miRNA mature body sequence (as shown in SEQ ID NO: 1, 5'-TAGCTTTGATCTGTAGCTAGC-3' ), and a sequence complementary to the above miRNA mature body sequence. The synthetic miRNA mimic dry powder was diluted with RNase-free water to store 1 μg / μL, aliquoted and stored at -20°C.

[0026] The nucleotide sequences of the forward and reverse primers for the synthesis of miR-2703 mimics are as follows:

[0027] Forward primer: 5'-UAGCUUUGAUCUGUAGCUAGC-3'; (SEQ ID NO: 2)

[0028] Reverse primer: 5'-GCUAGCUACAGAUCAAAAGCUA-3'. (SEQ ID NO: 3)

[0029] Step 2: miR-2703 was mixed separately in the artificial diet of brown planthopper (Nilaparvata lugens), and fed to brown plantho...

Embodiment 2

[0037] Step 1: Count the hatching rate of eggs.

[0038] Select normal brachypterous females that have been eclosion for one day, inject 50ng of miR-2703 simulants per head, and then pair with untreated single males to lay eggs, and count the hatching rate of eggs.

[0039] The result is as Figure 4 shown. Figure 4 is a graph of the hatchability of offspring of N. lugens after injection of miRNA mimics. Injection of miR-2703 mimics resulted in a significant decrease in the hatching rate of offspring of N. lugens compared to controls (no injection of miR-2703 mimics).

[0040] Dissected female ovaries after processing, the results are as follows: Figure 5 shown. Figure 5 It is the change of ovary of N. lugens planthopper after injection of miRNA mimics. The left picture is the morphology of female ovaries treated with miR-2703 mimics, and the right picture is the morphology of untreated female ovaries. Depend on Figure 5 It can be seen that the shape of the ovary aft...

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Abstract

The invention provides an miRNA stimulant and a chitin synthesis inhibition type biopesticide. The nucleotide sequence of the miRNA stimulant is shown as SEQ ID NO:1. The miRNA stimulant and the chitin synthesis inhibition type biopesticide have the advantages of low dosage, high efficiency and obvious insect killing effect.

Description

technical field [0001] The invention belongs to the field of insect control, and specifically relates to a miRNA simulant, a chitin synthesis inhibiting biological pesticide, and an application of the biological pesticide. Background technique [0002] Chitin is a polysaccharide of N-acetylglucosamine, which widely exists in fungi and arthropods, and is an important component of insect exoskeleton, trachea and midgut peritrophic matrix (PM) (Cohen, 2001; Merzendorfer, 2006). The chitin biosynthetic pathway of insects starts from trehalose and forms chitin through the participation of eight enzymes (Candy et al., 1962). Currently the most studied are trehalase (Trehalase, Tre) and chitin synthase (CHS), the first and last enzymes of chitin synthesis (Zhu et al., 2016). Insects usually have two chitin synthetase genes, chitin synthetase A (CHSA) and chitin synthetase B (CHSB) (Zhu et al., 2016). The chitin synthase A gene is mainly expressed in the epidermis and trachea, an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A01N43/90A01P7/04
CPCA01N43/90C12N15/113C12N2310/141
Inventor 张文庆陈洁李腾超
Owner SUN YAT SEN UNIV
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