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Method of breeding non-flying Harmonia axyridis

A technology of ladybug and ladybug wings, applied in the field of bioengineering, can solve the problems of decreased viability and fecundity, unsuitable for field release, low flight ability, etc., to ensure large-scale cultivation, obvious social benefits and economic value, The effect of reducing economic costs

Inactive Publication Date: 2018-10-16
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, French scholars have screened ladybugs with low flying ability and non-flying male and female adults from the natural population of ladybug heterochromia, which are used for pest control in closed environments such as greenhouses. It takes several generations to obtain a stable strain, and the viability and fecundity will drop significantly. It is easy to be assimilated by the wild type in the natural environment and is not suitable for field release.

Method used

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  • Method of breeding non-flying Harmonia axyridis
  • Method of breeding non-flying Harmonia axyridis
  • Method of breeding non-flying Harmonia axyridis

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Experimental program
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Embodiment 1

[0046] Embodiments of the present invention provide a method for cultivating flightless ladybugs, specifically as follows:

[0047] 1. Raw materials

[0048] dsRNA: a vestigial dsRNA that interferes with the key genes of the wing development of the ladybug heterochromia.

[0049] According to the standard method for in vitro synthesis of dsRNA provided by Promega, dsRNA was synthesized using the T7RiboMAXExpress RNAi System kit provided by the company. The nucleotide sequence of dsRNA is shown in SEQ ID No.1.

[0050] 2. Method

[0051] The method for cultivating flightless ladybugs, the specific steps include:

[0052] (1) The dsRNA is prepared into a dsRNA solution with a concentration of 50.0 ng / ul;

[0053] (2) Take the larvae on the first day of the fourth instar, and inject 1 ul of the above-mentioned dsRNA solution from the third and fourth intersegmental membranes of the abdomen;

[0054] (3) Feed the larvae with aphids or artificial diet until it emerges, and obta...

Embodiment 2

[0056] Embodiments of the present invention provide a method for cultivating flightless ladybugs, specifically as follows:

[0057] 1. Raw materials

[0058] (1) Nano-carrier: The embodiment of the present invention takes the fluorescent nano-carrier molecule disclosed by CN103172776A as an example. The fluorescent nucleus inside the carrier has a high fluorescence quantum yield, good light, heat and chemical stability, which is beneficial to biological For in vivo tracking, a good cavity structure is formed between each side chain and a large number of specific functional groups are conducive to the binding and assembly of dsRNA molecules, and a stable dsRNA / nanocarrier complex is formed. The whole complex has good insect cell affinity. Harmony, which is conducive to the rapid entry of dsRNA molecules into cells to target target genes.

[0059] (2) dsRNA: a vestigial dsRNA that interferes with the key genes of the wing development of the heterochromia.

[0060] According to...

Embodiment 3

[0071] Embodiments of the present invention provide a method for cultivating flightless ladybugs, specifically as follows:

[0072] 1. Raw materials

[0073] (1) Nano-carrier: The embodiment of the present invention takes the fluorescent nano-carrier molecule disclosed by CN103172776A as an example. The fluorescent nucleus inside the carrier has a high fluorescence quantum yield, good light, heat and chemical stability, which is beneficial to biological For in vivo tracking, a good cavity structure is formed between each side chain and a large number of specific functional groups are conducive to the binding and assembly of dsRNA molecules, and a stable dsRNA / nanocarrier complex is formed. The whole complex has good insect cell affinity. Harmony, which is conducive to the rapid entry of dsRNA molecules into cells to target target genes.

[0074] (2) dsRNA: a vestigial dsRNA that interferes with the key genes of the wing development of the heterochromia.

[0075] According to...

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Abstract

The invention discloses a method for breeding non-flying Harmonia axyridis. The method uses gene interference methods to interfere with expressions of vestigial or ultrabithorax, key genes of the wingdevelopment of the Harmonia axyridis, to breed the non-flying Harmonia axyridis. The breeding of the non-flying Harmonia axyridis includes three specific methods: injecting dsRNA, feeding dsRNA / nano-carriers, or soaking the body surface of the Harmonia axyridis in dsRNA / nano-carriers / surface active agents, or spraying or dripping the dsRNA / nano-carriers / surface active agents onto the body surfaceof the Harmonia axyridis. The method breeds the non-flying Harmonia axyridis by three potential gene interference methods, and the three potential gene interference methods are highly-efficient and stable and capable of thoroughly solving a problem of difficult colonization of artificially released natural enemies Harmonia axyridis in the control area, and has important theoretical and practicalsignificance. By applying the feeding method and the body wall penetration method to gene interference, the current situation of time-consuming and labor-consuming traditional methods is completely changed, efficiency and success rates are greatly improved, and costs and operation difficulty are reduced. The method for breeding the non-flying Harmonia axyridis has obvious social and economic benefits and no potential ecological risk.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method for cultivating flightless ladybugs. Background technique [0002] There are many kinds of ladybugs, which belong to the Coleoptera Polyphagae in the insect classification system. Currently, there are about 500 genera and 5,000 known species of ladybugs, among which predatory ladybugs account for about 80% of the species of ladybugs, mainly preying on scale insects, aphids, whiteflies, spider mites and other arthropods, mostly An important natural enemy of agricultural and forestry pests, among which Harmonia axyridis is one of the predatory ladybugs that has received widespread attention. Compared with other predatory ladybugs, it has the advantages of easy feeding, strong predation ability, and large food intake characteristics, strong environmental adaptability makes the heterochromia a dominant natural enemy of predating aphids. [0003] Although ladybugs ha...

Claims

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Application Information

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IPC IPC(8): A01K67/033C12N15/85
CPCA01K67/033C12N15/8509
Inventor 沈杰马中正王丹刘小侠尹梅贞
Owner CHINA AGRI UNIV
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