Method for extracting symbiotic microorganisms in coleopteran insects

A technology for coleopteran insects and microorganisms, applied in the biological field, can solve the problems of unclear coevolution of coleopteran insects and symbiotic bacteria, lack of research methods, etc.

Pending Publication Date: 2022-03-29
SOUTH CHINA AGRI UNIV
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Problems solved by technology

This kind of symbiotic bacteria spreads vertically in coleopteran insects by smearing symbiotic bacteria near them after laying eggs, and passes them from mother to offspring, so that larvae can hatch and assist their growth under more suitable environmental conditions, but At present, due to the lack of suitable research methods, the co-evolution of coleopteran insects and symbiotic bacteria has been unclear

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  • Method for extracting symbiotic microorganisms in coleopteran insects

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Embodiment

[0016] The method proceeds as follows:

[0017] (1) Preparation: Select a few sexually mature female glass stag beetles, turn them upside down, gently squeeze the middle and back of their abdomen with your fingers to make them protrude from the female sexual organs, repeat the operation several times, and squeeze again Press the end of the abdomen of the female to make it protrude more tissue. At this time, an oval-shaped small cyst-like structure (mostly yellow or white tissue inside) will protrude from the front of the sex organ. This tissue is the coleopteran symbiotic fungal sac.

[0018] (2) Obtain coleopteran insect fungus: sterilize the head of the tweezers by burning them with a flame. In order to avoid the deactivation of the insect tissue due to the high temperature of the tweezers, immerse the sterilized tweezers into 1.5ml of PBS solution containing a certain concentration. Cool down in the separation tube, and then use the cooled tweezers to remove the symbiotic b...

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Abstract

The invention relates to a method for extracting symbiotic microorganisms in coleopteran insects, which comprises the following steps of: selecting a plurality of sexually mature female coleopteran insects, enabling the bellies of the female coleopteran insects to face upwards, and extruding the middle and rear sections of the bellies to extend out of symbiotic bacterium sacs (one ellipse has a small cyst type structure, and the interiors of the symbiotic bacterium sacs are mostly yellow or white tissues); the method comprises the following steps: completely picking off the tissue with tweezers under a microscope, immersing the tissue into a vessel containing a PBS (Phosphate Buffer Solution) with a certain concentration, grinding the tissue with a grinding rod to fully dissolve the content in the solution, carrying out concentration dilution, transferring the tissue into a culture dish containing YNB, placing the culture dish in a constant temperature box with a proper temperature (25 DEG C), culturing for 2-3 days, and if white dot-shaped or strip-shaped tissue appears, taking out the tissue. And comparing with the known symbiotic bacteria gene sequence, and if the symbiotic bacteria belong to the same clade, indicating that the symbiotic microorganisms in the coleopteran insects are successfully extracted.

Description

technical field [0001] The invention belongs to the field of biological technology, in particular to a method for extracting mycocyst symbiotic microorganisms in insects. Background technique [0002] As the insect with the largest population in nature, it faces various threats in the environment, such as natural enemies, pathogenic microorganisms or adversity. Studies have found that insect symbiotic bacteria can protect the host from predators and parasitism by natural enemies, resist the invasion of pathogenic microorganisms, and assist them to tolerate poor environments. Among them, protosymbiotic bacteria (i.e., endosymbiotic bacteria) are necessary for the survival of host insects, and there is a long-term co-evolutionary relationship between them, and they are mainly distributed in specialized cell-bacteria-containing tissues of host insects (for example: Coleoptera fungi) within, and strictly vertically propagated. The role played by endosymbionts is closely relate...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N1/02
CPCC12N1/14C12N1/02
Inventor 朱雪姣温秀军久保田耕平
Owner SOUTH CHINA AGRI UNIV
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