Method of authenticating whether substance influences insect behavior or not and what kind of influence

A technology for insects and substances, which is applied in the field of identifying whether substances have an impact on insect behavior and what kind of impact it has, and can solve problems such as the absence of more systematic research methods to achieve the effect of ensuring effectiveness

Active Publication Date: 2017-06-20
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With the development of chemical analysis techniques, a large number of sex pheromone components of moth insects have been identified, and the behavioral funct...

Method used

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  • Method of authenticating whether substance influences insect behavior or not and what kind of influence
  • Method of authenticating whether substance influences insect behavior or not and what kind of influence
  • Method of authenticating whether substance influences insect behavior or not and what kind of influence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Identification and Analysis of Glandular Components of Helicoverpa armigera

[0043] 1. Take 1-4 days old virgin female moths as test insects, extract sex pheromone at 6-7 hours after entering the hidden object, gently squeeze the female moth’s abdomen with your hand to force it to extend the sex pheromone gland, and use surgery The glands were cut off with a blade, placed in a miniature glass tube, and 10μl of n-hexane solvent was added to soak for 30 minutes and then analyzed by gas chromatography.

[0044] 2. GC-MS analysis: The GC-MS used were HP-6890 capillary gas chromatograph and Agilent 5973 mass spectrometer, and the capillary column model was Agilent J&W DB-1MS GC column, 30m, 0.25mm, 0.25μm. The working conditions of the chromatogram were as follows: the initial temperature was 40°C, kept for 4 minutes, and then increased to 280°C at a temperature increase rate of 10°C / min, and then kept for 20 minutes.

[0045] 3. The sex pheromone component is compared with the r...

Embodiment 2

[0047] CRISPR / Cas9 knockout HarmOR16 (OR16) receptor

[0048] Through the Xenopus oocyte expression system and the two-electrode voltage clamp technology, this laboratory identified the target receptor for sensing Z11-16:OH as the sex pheromone receptor OR16.

[0049] See the flowchart of CRISPR / Cas9 knockout OR16 figure 2 .

[0050] 1. Design and in vitro synthesis of sgRNA: According to the design criteria of the sgRNA target site (5'-GG(18N)NGG-3'), select a small fragment sequence 5'-ggatcggtctaagcaagccatgg- in the third exon region of OR16 3'is the target site. At the same time, the front end of the sgRNA sequence is the T7 promoter sequence, which is used for sgRNA in vitro transcription. The double-stranded DNA template for the synthesis of sgRNA is amplified by long-strand PCR to obtain the PCR reaction system: 2×Premix PrimeSTAR Hs 25μL, HarmOR16-F1 and HarmOR16-R1 (10μM) 1.5μL, and nuclease-free water 22μL. PCR reaction conditions: pre-denaturation at 98°C for 1min; den...

Embodiment 3

[0056] Single-sensor recorder records the electrophysiological response of the sensilla to sex pheromone components

[0057] 1. First select the unmated male moths 3-4 days old and put them into the Axygen 1.5mL centrifuge tube. The head of the male moth exposes the port of the centrifuge tube, and the moth's lower lip must be fixed on the edge of the port through dental wax, exposing one of them. The root tentacles are fixed on a cover glass coated with double-sided tape.

[0058] 2. The fixed male moth is used for single-sensor recording. We use the ground tungsten wire electrode to insert the compound eye of the male moth as a reference electrode. The recording electrode connected to the olfactory amplifier (sytech) is also inserted into the base of the single sensor through the ground tungsten wire electrode until a stable electrical signal can appear (the frequency of the signal is significantly higher than the noise signal). The operations recorded above are performed under ...

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Abstract

The invention relates to a method of authenticating whether a substance influences insect behavior or not and/or what kind of influence. The method comprises the steps of 1, determining the substance; 2, after the substance is utilized to stimulate an insect, analyzing an electrophysiological and/or behavioral reaction of the insect. The substance is preferably a substance generated by the insect itself; the substance more preferably comprises pheromone and/or a pheromone antagonist; the pheromone is most preferably selected from (z)-11-hexedecanal and/or (z)-9-hexedecanal; the pheromone antagonist is selected from (z)-11-hexadecenol and/or (z)-9-fulure.

Description

Technical field [0001] The invention relates to a method for identifying whether and / or what kind of influence a substance has on insect behavior. Background technique [0002] Insects can synthesize thousands of small molecular compounds or macromolecular substances in their bodies. How to determine the functions of these substances in their bodies is not yet a mature method to determine them. [0003] For example, in the sex pheromone communication of moth insects, females mainly release certain specific sex pheromones (specific components and proportions), while males pass tens of thousands of sexual information on their antennae at long distances. The sensilla feels it and produces a trending behavior, finding and mating with females. With the development of chemical analysis technology, the sex pheromone composition of a large number of moth insects has been identified, and the behavioral functions of these compounds have been further identified through behavioral studies and...

Claims

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Application Information

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IPC IPC(8): A01K67/033
CPCA01K67/033
Inventor 王桂荣常贺坦刘洋艾东
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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