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Method for evaluating the influence of insecticidal protein on adults of propylaea japonica and feeding device

A technology of insecticidal protein and ladybug, which is applied in the field of environmental safety of transgenic plants, can solve the problems that the pollen of transgenic plants cannot be reached, has not been considered, and affects the risk of ladybugs, etc., and achieves simple structure, improved survival rate, The effect of increasing egg production

Inactive Publication Date: 2018-09-28
INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] When evaluating the impact of exogenous insecticidal proteins on the mole beetle, the exogenous insecticidal protein that is more than 10 times the maximum exposure to the mole beetle is generally used to feed the mole beetle, and the direct use of transgenic plant pollen cannot achieve the above-mentioned Require
In previous evaluations of the effects of exogenous insecticidal proteins on Coccinella mori, the risk of the interaction between secondary metabolites in plants and insecticidal proteins to affect Copa aureus was not considered

Method used

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  • Method for evaluating the influence of insecticidal protein on adults of propylaea japonica and feeding device
  • Method for evaluating the influence of insecticidal protein on adults of propylaea japonica and feeding device
  • Method for evaluating the influence of insecticidal protein on adults of propylaea japonica and feeding device

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: Fructose improves the survival rate and the amount of eggs laid by the adult beetle

[0032] Collect 100g of healthy non-genetically modified cotton leaves, add 200g of sterile distilled water after grinding, put it in a container, soak it in a shaker at 200rpm for 2 hours at room temperature, use a centrifuge at 12000g for 10min, extract the supernatant, and obtain it after filtration Cotton juice extract.

[0033] Weigh 0.4g fructose and 0.4g sucrose and place them in 2.0ml centrifuge tubes respectively, then add cotton juice extract, set the volume to 1mL, and mix well to obtain the prepared 0.4g / ml fructose solution and 0.4g / ml ml of sucrose solution.

[0034] Dispense 0.4g / ml fructose solution and 0.4g / ml sucrose solution into 1.5mL centrifuge tubes, 250μL per tube, cover the cap, pierce the cap of the centrifuge tube cap with a sharp object, and insert a capillary of appropriate length (inner diameter 0.53mm) to the bottom of the solution, drain th...

Embodiment 2

[0040] Example 2: Evaluation of the Effect of Exogenous Insecticidal Protein Cry1Ac Protein on Cod.

[0041] Collect 100g of healthy non-genetically modified cotton leaves, add 200g of sterile distilled water after grinding, put it in a container, soak it in a shaker at 200rpm for 2 hours at room temperature, use a centrifuge at 12000g for 10min, extract the supernatant, and obtain it after filtration Cotton juice extract.

[0042] Weigh 20 μg of exogenous insecticidal protein Cry1Ac and 400 mg of fructose into a 2.0ml centrifuge tube, then add cotton juice extract to make up to 1mL, and mix well to obtain the prepared exogenous insecticidal protein with a concentration of 20 μg / ml. Insect protein Cry1Ac solution, which contains 0.4g / ml fructose solution.

[0043] Weigh 400μg of protease inhibitor E-64 and 400mg of fructose into a 2.0ml centrifuge tube, add cotton juice extract to make the volume to 1mL, and mix well to obtain the prepared protease inhibitor containing 400μg / ...

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Abstract

The invention discloses a method for evaluating the influence of insecticidal protein on adults of propylaea japonica and a feeding device. The invention firstly designs a feeding device for simulating the plant gland, and the suction mode of cotton flower extrafloral nectaries is simulated, and then a cotton juice extract is used as a solvent to dissolve the exogenous insecticidal protein, and fructose is added to serve as a phagostimulant of the propylaea japonica adults; a single-pair of the propylaea japonica adults are put into the feeding device with a feeding solution, and enough pea aphids are supplemented to culture in an illumination cultivation box. According to the indexes of the to-be-detected solution on the death rate of the propylaea japonica adults, the egg laying early stage, the egg laying amount and the weight, whether the insecticidal protein has the influence on the propylaea japonica adults or not is judged. On the base of establishing the risk theory of evaluating that the secondary metabolites and the insecticidal protein in the plant interact to influence the propylaea javanica, the natural environment of the insecticidal protein threat which is faced by the propylaea ladyridis can be reflected more truly, the evaluation method is simple, and the applicability is high.

Description

technical field [0001] The invention relates to the field of environmental safety of transgenic plants, in particular to an evaluation method and a feeding device for the influence of exogenous insecticidal proteins on adults of the natural enemy insect, ladybird beetle. Background technique [0002] Bacillus thuringiensis is a Gram-positive bacterium and is one of the most widely used microbial insecticides in the world. The main source of its insecticidal activity is the parasporal crystal protein produced in the strain. Researchers have introduced Bt genes into conventional crops to obtain transgenic crops expressing exogenous insecticidal proteins. Since the first transgenic crops were planted in 1996, transgenic crops have been widely promoted around the world, creating a huge economy for agricultural production all over the world. , social and environmental benefits. [0003] Transgenic exogenous insecticidal protein gene crops can express exogenous insecticidal prote...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/033G01N33/50
CPCA01K67/033G01N33/5085
Inventor 张帅崔金杰雒珺瑜王丽
Owner INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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