Method for micro-injection of adults of small-sized aleyrodid insects

A technology of microinjection and microinjection, applied in the field of entomology, to achieve the effects of improving injection efficiency, simple operation, and scientific and reasonable operation links

Active Publication Date: 2020-06-12
SHENYANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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

Method used

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  • Method for micro-injection of adults of small-sized aleyrodid insects

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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 method for micro-injection of adults of small-sized aleyrodid insects. According to the method for micro-injection of adults of small-sized aleyrodid insects, a glass micro-injection needle is adopted for injection on a microscopic platform, and the method includes the following steps: preparing the glass micro-injection needle, narcotizing the adults before injection, placing the narcotized adults in an injection-special V-shaped groove of an insect-fixing medium plate, performing micro-injection, placing the injected adults on a breeding medium in a leaf feeding culture dish, placing the culture dish in a lighting incubator, performing culture for 10-14 hours, placing the leaf feeding culture dish upside down so as to make leaves face up, and performing culturecontinuously. The method for micro-injection of adults of small-sized aleyrodid insects has scientific and reasonable operation steps, the injection efficiency is improved significantly, the test error is reduced, and 80 aleyrodid adults and 40 planthopper adults can be injected every hour; and when the method is compared with a conventional injection method, the injection speed is increased by about 3-4 times, the mortality of the injected insects is reduced greatly, and the test 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 & AuthorityApplications(China)
IPC IPC(8): A01K67/033C12N15/89
CPCA01K67/033C12N15/89
Inventor栾军波闫金阳
OwnerSHENYANG AGRI UNIV