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mir-novel-0287-3p and its application in pest control

A technology for pest control and pest control, which is applied to miR-novel-0287-3p and its application in pest control, can solve problems such as environmental pollution, pest resistance, and low efficiency.

Active Publication Date: 2021-10-08
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, agricultural control and physical control have a large workload and low efficiency, and chemical control is likely to make pests resistant to pesticides and pollute the environment
However, in biological control, there are very few studies involving the molecular mechanism and the role of miRNA

Method used

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  • mir-novel-0287-3p and its application in pest control
  • mir-novel-0287-3p and its application in pest control

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Effect of novel-m0287-3p on the growth of Spodoptera litura

[0045] Select Spodoptera litura larvae from the first day of the 5th instar (L5D1) and the third day of the 6th instar (L6D3) with the same body size and good health, weigh each larva, and take worms with similar weights for the experiment. They were randomly divided into novel-m0287-3p treatment group and NC control group, with 20 rats in each group for L5D1 and 30 rats in each group for L6D3, repeated 3 times. Along the direction of blood circulation, use a micro-syringe to inject synthetic novel-m0287-3pagomir and NC agomir from the penultimate abdominal segment of the flank of Spodoptera litura larvae; the injection volume of L5D1 is 4 μg per worm, and the injection volume of L6D3 6 μg per worm, fed with normal culture medium after injection. After the injection, the morphological changes of the worms were observed every day and photographed for evidence, and the molting rate 48 hours after the...

Embodiment 2

[0047] Example 2 Effect of novel-m0287-3p on the protein expression of EcR

[0048] In order to find the possible reason why novel-m0287-3p affects the development of Spodoptera litura. Through the prediction, novel-m0287-3p may bind to the 3' non-coding region of the ecdysone receptor EcR, and the binding site was predicted, and the 3' non-coding region of EcR was constructed into the luciferase activity assay vector, and the vector and novel -m0287-3p mimic was transfected into cells, and the effect of novel-m0287-3p on EcR was verified by dual luciferase activity (according to the kit method of Promega Company). The gene number of EcR in NCBI is: HM046618.2.

[0049] The result is as image 3 As shown, (a) and (b), (c) and (d), (e) and (f) are the three predicted sites and the corresponding dual-luciferase activity verification results, respectively, proving that novel-m0287- 3p can bind these sites.

[0050] In order to verify whether novel-m0287-3p regulates the expre...

Embodiment 3

[0051] Example 3 Effects of indole-3-carbinol and Novel-m0287-3p on the growth and development of Spodoptera litura

[0052] Spodoptera litura likes to feed on cruciferous plants, and indole-3-carbinol (I3C) is the main secondary substance of mustard (Brassicaceae). This experiment studied the growth and development of Spodoptera litura treated with Novel-m0287-3p injection after eating artificial medium containing plant secondary substance indole-3-carbinol (I3C).

[0053] Select worms with the same size and good health on the first day of the 4th instar, inject 4 μg novel-m0287-3p agomir per worm, and inject the same amount of NC agomir as the control group, 20 worms in each group, and continue to feed the worms containing or Artificial medium without 0.5% I3C.

[0054] The weight change of Spodoptera litura is as follows: Figure 5 As shown, the body weights of NC (NC agomir+artificial medium not containing I3C) group and novel-m0287-3p (novel-m0287-3p agomir+artificial m...

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Abstract

The invention relates to the field of biotechnology, in particular to a miR-novel-0287-3p and its application in pest control. Provided is the application of the miR-novel-0287-3p or its mimic or its agonist in controlling pests or preparing products for controlling pests or preparing GSTe1 inhibitors. miR‑novel‑m0287‑3p can inhibit the expression of detoxification enzymes required to resist plant secondary substances and the ecdysone receptor that regulates molting in the worm, thereby slowing down the growth and development of the worm body, and ultimately causing the worm body to delay molting and pupation , and even produce deformed pupae. Therefore, insect miRNA novel-m0287-3p can be used for the preparation of insecticides, or transgenic plants expressing novel-m0287-3p can be prepared for pest control.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a miR-novel-0287-3p and its application in pest control. Background technique [0002] MicroRNA (miRNA) is a class of endogenous non-coding RNA discovered in the 1990s (Lee RC, Feinbaum RL, Ambros V. The C. elegans heterochronic gene lin-4 encodes smallRNAs with antisense complementarity to lin-14. Cell , 1993,75(5):843-54.), a single-stranded small RNA of about 22 nucleotides, processed from hairpin transcripts produced endogenously in cells. The transcription of most miRNAs is mediated by RNA polymerase II, and a few are mediated by RNA polymerase III, forming primary transcripts (pri-miRNAs) with a length of hundreds or even thousands of bases. has a cap structure at the 5' end and a poly(A) tail with a stem-loop structure inside. pri-miRNA is cleaved by RNase III in the nucleus to generate miRNA precursor (pre-miRNA). Precursor miRNA is transported into the cytoplasm through R...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113
CPCA01N63/60
Inventor 郑思春曹晓聪马康
Owner SOUTH CHINA NORMAL UNIVERSITY
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