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A method of identifying whether and what effect a substance has on insect behavior

A technology of insects and substances, applied in the field of identifying whether and what kind of influence substances have on insect behavior, can solve the problem of lack of relatively systematic research methods, and achieve the effect of guaranteeing effectiveness

Active Publication Date: 2020-02-18
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 functions of these compounds have been further identified through behavioral and field trapping experiments. response, there is no systematic research method to determine

Method used

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  • A method of identifying whether and what effect a substance has on insect behavior
  • A method of identifying whether and what effect a substance has on insect behavior
  • A method of identifying whether and what effect a substance has on insect behavior

Examples

Experimental program
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Effect test

Embodiment 1

[0042] Identification and Analysis of Glandular Components of Cotton Bollworm

[0043] 1. Take 1-4 day-old virgin female moths as test insects, extract sex pheromones 6-7 hours after entering the hidden weapon, gently squeeze the abdomen of female moths to force them to stretch out sex pheromone glands, and use surgery to Cut off the gland with a blade, place it in a micro glass tube, add 10 μl of n-hexane solvent to soak for 30 minutes, and then analyze it by gas chromatography.

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

[0045] 3. The sex pheromone components are qualitatively compared with th...

Embodiment 2

[0047] CRISPR / Cas9 Knockout of HarmOR16 (OR16) Receptor

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

[0049] For the flow chart of CRISPR / Cas9 knockout of OR16, see figure 2 .

[0050] 1. Design and in vitro synthesis of sgRNA: According to the design criteria of sgRNA target site (5'-GG(18N)NGG-3'), select a small fragment sequence 5'-ggatcggtctaagcaagccatgg- located 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 in vitro transcription of the sgRNA. The double-stranded DNA template for synthesizing sgRNA was obtained by long-chain PCR amplification. The PCR reaction system: 25 μL of 2×Premix PrimeSTAR Hs, 1.5 μL of HarmOR16-F1 and HarmOR16-R1 (10 μM), 22 μL of nuclease-free water. PCR reaction conditions:...

Embodiment 3

[0056] Single sensilla recorder records sensilla electrophysiological responses to sex pheromone components

[0057] 1. First, select 3-4 day-old unmated male moths and place them in an Axygen 1.5mL centrifuge tube. The head of the male moth is exposed from the port of the centrifuge tube. Fix the moth's lower lip whiskers on the edge of the port with dental wax, exposing one of the moths. Root antennae were fixed on coverslips stained with double-sided adhesive.

[0058] 2. The fixed male moth is used for single sensory recording. We use a ground tungsten wire electrode to insert into 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 obviously higher than the noise signal). The operations recorded above were performed under a Le...

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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 present invention relates to methods of identifying whether and / or what effect a substance has on insect behaviour. 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 has not yet been matured. [0003] For example, in the sex pheromone communication of moth insects, the female insects mainly release a certain specific (specific composition and ratio) sex pheromone, while the male insects transmit tens of thousands of sex information on the antennae over a long distance. The sensilla senses it and produces tendency behavior, finds the female and mates with it. With the development of chemical analysis techniques, a large number of sex pheromone components of moth insects have been identified, and the behavioral functions of these compounds have been further identified through behavioral and field ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/12A01K67/033
CPCA01K67/033
Inventor 王桂荣常贺坦刘杨艾东
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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