A kind of microinjection method of whitefly small insect adult

A technique of microinjection and microinjection, applied in the field of entomology, to achieve the effects of reducing experimental errors, simple operation, and reducing mortality

Active Publication Date: 2021-11-19
SHENYANG AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For example, in the study of gene interference experiments, especially the injection of worms with different treatments at the same time period, so as to reduce the error, the worms are required to survive for a long time after injection, and the current injection method and feeding method of injected worms cannot meet the demand

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  • A kind of microinjection method of whitefly small insect adult
  • A kind of microinjection method of whitefly small insect adult

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1 whitefly class small insect adult microinjection method

[0021] Injection objects: Bemisia tabaci, greenhouse whitefly, brown planthopper, and brown planthopper.

[0022] Injection method:

[0023] (1) Adjust the microscope platform: Uncover the dust cover of the microscope and needle grinder, take off the lens cover; turn on the light source; turn on the air pump switch, and confirm that the balance pressure (pc) has been adjusted to 0;

[0024] The microinjection platform includes: Olympus stereo microscope, eppendorf air pump injection system, Japanese Chengmo micromanipulator, Sutter needle pulling instrument and needle grinding instrument, Liansheng brand shockproof table;

[0025] (2) Fix the ice box on the mobile platform of the base of the microscope, put in a 14.5×10×2cm ice box, put the fixed medium on the ice box; adjust the microscope manipulator to an angle of 45° with the horizontal;

[0026] (3) Production method of injection needles: inje...

Embodiment 2

[0036] Effects of Different Microinjection Methods on Injection Efficiency and Survival Rate in PGRP Gene Silencing Test of Bemisia tabaci

[0037] Process 1: adopt the microinjection method of embodiment 1

[0038] Treatment 2: Injection using conventional techniques, conventional injection methods reference Tissue-specific genesilencing by RNA interference in the whitefly Bemisia tabaci (Gennadius). Ghanim et al. BMC Genomics 2013, 14:160

[0039] Injection Efficiency Survey Results:

[0040] The number of worms injected within 1 hour is 80;

[0041] The number of worms injected within 2 hours of treatment was 20.

[0042] Survival rate survey results:

[0043] At 24 hours after injection, the survival rate of Bemisia tabaci in treatment 1 was about 80%-100%, and that in treatment 2 was about 50%-60%. The survival rate of treatment 1 was 30%-40% higher than that of treatment 2;

[0044] At 48 hours after injection, the survival rate of Bemisia tabaci was about 65%-75%...

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Abstract

The invention relates to a microinjection method for small whitefly insect adults. The microinjection of whitefly adult insects uses a glass microinjection needle to inject on a microscopic platform, comprising the following steps: preparing a glass microinjection needle; Insect anesthesia; place the anesthetized insect in the V-shaped groove of the special insect fixed medium plate for injection, and perform microinjection; put the injected insect on the top of the culture medium in the leaf culture dish, After being placed in the light incubator and raising for 10-14 hours, the leaf raising culture dish is turned upside down, so that the leaves are facing upwards, and the feeding is continued; the operation link of the microinjection of whitefly insects in this method is scientific and reasonable, and the injection efficiency is significantly improved. To reduce the experimental error, 80 whitefly adults and 40 planthopper adults can be injected per hour, and the injection speed is increased by about 3-4 times compared with conventional injection methods; the mortality rate of insects after injection is greatly reduced, and the experimental error is reduced.

Description

technical field [0001] The invention relates to the field of entomology, in particular to a microinjection method for whitefly small insect adults. Background technique [0002] Microinjection technology is a practical technology widely used in the field of life sciences, mainly used in microbiology, medicine, entomology and other research fields. Microinjection technology is the operation technique of injecting solution into animals, plants and microorganisms by using injection needles through microscope observation. Microinjection is widely used for the injection of adults or nymphs and many eggs of small insects. The operation of microinjection technology is complex, especially for small insects, the operation requirements are high. At present, most of the microinjection techniques disclosed by various research institutions are aimed at cells or large and medium-sized animals and plants, and there is no microinjection technique specifically for whitefly insect adults or...

Claims

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Application Information

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Patent Type & AuthorityPatents(China)
IPC IPC(8): C12N15/89A01K67/033
CPCA01K67/033C12N15/89
Inventor栾军波闫金阳
OwnerSHENYANG AGRI UNIV