Chitin-synthesized inhibitory pesticide and preparing method and application thereof

A technology of chitin synthase and chitin, applied in pesticides, botany equipment and methods, applications, etc., can solve problems such as difficulty in developing new structures, increasing insect resistance, and limiting the use of pesticides , to achieve the effect of hindering normal growth and reproduction, significant control effect, and no damage to the ecological environment

Active Publication Date: 2016-06-22
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the increase in insect resistance and the difficulty in developing new structures, this greatly limits the use of such insecticides. In-depth research on the mechanism of insect growth and development based on CHSA will help us find new Chitin synthesis inhibitor insecticide

Method used

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  • Chitin-synthesized inhibitory pesticide and preparing method and application thereof
  • Chitin-synthesized inhibitory pesticide and preparing method and application thereof
  • Chitin-synthesized inhibitory pesticide and preparing method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Prepare dsRNA as follows:

[0051] 1. Select the nymphs of different ages of brown planthopper

[0052] 2. Extraction of total RNA: 20 nymphs were taken, ground in liquid nitrogen, and total RNA was extracted by TRIzol method.

[0053] 3. Synthesis of the first strand of cDNA: 1 μg of total RNA was reverse-transcribed into cDNA according to the instructions of the PrimeScript RTreagent Kit with gDNA Eraser kit from Takara Company.

[0054] 4. The preparation process of the NlCHSA intermediate fragment is described below.

[0055] (1) Polymerase chain reaction (PCR) reagent (GenestarTaq DNA polymerase HiFi) and reaction conditions:

[0056]

[0057] The specific steps of the above polymerase chain reaction are as follows: pre-denature the above mixture at 95°C for 5 minutes, and then enter the following cycle: denaturation at 95°C for 30 s, annealing at 55°C for 30 s, extension at 72°C for 60 s, a total of 35 cycles, and finally 72°C for 35 cycles. Extend for 10 mi...

Embodiment 2

[0069] (1) Design specific primers with T7 promoter to synthesize dsRNA of CHSA in vitro, and use primers of green fluorescent protein gene to synthesize dsGFP as a control.

[0070] dsCHSA-F: 5'-GAAGACAAGGCTGAGAAGG-3'

[0071] dsCHSA-R: 5'-GGAATCGTTTGCGGAAGG-3'

[0072] dsCHSA-T7F: 5'-GGATCCTAATACGACTCACTATAGGGAAGACAAGGCTGAGAAGG-3'

[0073] dsCHSA-T7R: 5'-GGATCCTAATACGACTCACTATAGGGGAATCGTTTGCGGAAGG-3'

[0074] dsGFP-F: 5'-AAGGGCGAGGAGCTGTTCACCG-3'

[0075] dsGFP-R: 5'-CAGCAGGACCATGTGATCGCGC-3'

[0076] dsGFP-T7F:

[0077] 5'-GGATCCTAATACGACTCACTATAGGAAGGGCGAGGAGCTGTTCACCG-3'

[0078] dsGFP-T7R:

[0079] 5'-GGATCCTAATACGACTCACTATAGGCAGCAGGACCATGTGATCGCGC-3'

[0080] (2) Using the plasmid named pMDT-CHSA obtained in Example 1 as a template, use the primers designed in step 1 to amplify according to the following reaction conditions to obtain the target fragment.

[0081] A reaction system is as follows:

[0082]

[0083] B reaction system is as follows:

[0084] ...

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Abstract

The invention provides a chitin-synthesized inhibitory pesticide.The chitin-synthesized inhibitory pesticide is prepared from double-stranded RNA serving as an active ingredient, wherein the nucleotide sequence of a positive-sense strand of a transcription template of the double-stranded RNA is shown as SEQ ID NO:1.The chitin-synthesized inhibitory pesticide has a remarkable prevention and treatment effect on brown planthopper; in addition, pesticide resistance and environment pollution can be avoided, the ecological environment is not destroyed, and safety and environment friendliness are achieved.

Description

technical field [0001] The invention belongs to the field of insect control, and in particular relates to a chitin synthesis inhibiting insecticide, a preparation method and an application of the insecticide. Background technique [0002] Chitin, also known as chitin, is a polysaccharide of N-acetylglucosamine, which widely exists in fungi and arthropods, and is the core of insect exoskeleton, trachea and midgut peritrophic matrix (PM). Important components (Cohen, 2001; Merzendorfer, 2006). Current studies have also shown that chitin is also an important component of some insect eggs, such as Aedes aegypti, Tribolium castaneum and Rhodnius prolixus (Arakane et al., 2008; Mansure et al., 2014 ; Moreira et al., 2007). The chitin biosynthetic pathway of insects begins with trehalose and ends with chitin (Candy et al., 1962), which involves the participation of 8 enzymes, and the last enzyme is chitin synthase (Chitinsynthase, CHS). Under its action, the chitin precursor uri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N57/16A01P7/04C12N15/10
CPCA01N57/16C12N15/102
Inventor 张文庆李腾超陈洁范晓斌邱洁琪
Owner SUN YAT SEN UNIV
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