Method for determining influence of exogenous dsRNA on toxicity of ladybugs

A ladybug, exogenous technology, applied in the field of insect molecular biology research, can solve the problem of no direct toxic effects of ladybugs, etc., and achieve the effect of simple and feasible method, good effectiveness and sensitivity

Active Publication Date: 2018-03-30
SOUTH CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

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  • Method for determining influence of exogenous dsRNA on toxicity of ladybugs
  • Method for determining influence of exogenous dsRNA on toxicity of ladybugs
  • Method for determining influence of exogenous dsRNA on toxicity of ladybugs

Examples

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Embodiment 1

[0038] In this example, taking the ladybug Heterochromia as the research object, taking its v-ATPaseA gene as an example, the method of the present invention was used to determine the direct toxicity of exogenous dsRNA on ladybugs.

[0039] A simple and sensitive assay for detecting the direct toxic effect of exogenous dsRNA on ladybugs, by mixing in vitro synthesized dsHA or dsCS or dsCM evenly into sucrose solution and feeding them to ladybugs for 2 days, and then providing sufficient pea aphids, Taking the sucrose solution mixed with potassium arsenate as the positive control and the sucrose solution mixed with dsGUS as the negative control, by comparing the changes in the expression of v-ATPaseA gene in ladybugs between the treatment group and the control group, as well as the survival rate and developmental history of ladybugs. The potential toxicity of dsRNA can be evaluated by the changes of important biological parameters such as period.

Embodiment 2

[0041] The present embodiment is the research object with the seven-spot ladybug, and all the other are researched with embodiment 1.

Embodiment 3

[0043] The present embodiment takes the 12-spot ladybug as the research object, and all the other are researched with embodiment 1.

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Abstract

The invention discloses a method for determining influence of exogenous dsRNA on toxicity of ladybugs. The method comprises the following specific steps: uniformly mixing the exogenous dsRNA into a sucrose solution, directly feeding the ladybugs for 2-3 days and then feeding with pea aphids; then detecting and analyzing the expression changes of target genes in the ladybugs, and observing the biological changes of the ladybugs so as to evaluate the toxicity of the exogenous dsRNA to the ladybugs, wherein the ladybugs include harmonia axyridis, coccinella septempunctata or henosepilachna pusillanima. By the method, direct influence of the exogenous dsRNA on the toxicity of the harmonia axyridis, the coccinella septempunctata or the henosepilachna pusillanima or other ladybugs can be effectively determined; in addition, the method is simple, feasible and high in effectiveness and sensitivity, and has a great significance and an application prospect in study on functions of related genesas well as environmental risk assessment on related RNAi transgenic crops.

Description

technical field [0001] The invention belongs to the technical field of insect molecular biology research. More specifically, it relates to a method for determining the toxic effects of exogenous double-stranded RNA (dsRNA) on ladybugs (Lady beetle, Coccyx chinensis or Ladybug todeca), including exogenously synthesized dsRNA or expression of RNAi transgenic crops Determination of the toxic effects of dsRNA on ladybugs (Lady beetles, Ladybugs, or Ladybugs). Background technique [0002] As the core technology of the second green revolution, agricultural genetically modified technology has rapidly reshaped the pattern of modern agriculture. In 1996, insect-resistant crops transfected with the Bacillus thuringiensis (Bt) insecticidal protein gene began to be commercially planted. By 2015, the number of countries planting Bt crops had reached 28, with an annual planting area of ​​nearly 2.7 billion mu (http: / / www .isaaa.org / ). Ranking the countries planting GM crops by the pla...

Claims

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Application Information

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IPC IPC(8): C12Q1/6851
CPCC12Q1/6851C12Q2531/113
Inventor 潘慧鹏杨春晓邱宝利吴建辉桑文王兴民金丰良
Owner SOUTH CHINA AGRI UNIV
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