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Molecular detection method of panonychus citri Mc Gregor response abamectin stress protein encoding gene

A technology of citrus panonychus and abamectin, which is applied in the field of molecular biology, can solve the problems of decreased ovary weight, adult mite egg production, average life expectancy, and egg hatching rate, and achieves high detection sensitivity and high detection results. Accurate, reliable and specific effects

Inactive Publication Date: 2017-07-21
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is no report about the effect of abamectin on Vg, but abamectin can affect the growth and development of acarids. After injecting abamectin, the weight of the ovary of the long-horned blood tick Haemaphysalis longicornis was found to be significantly reduced; The study on the sublethal effect of mycocin on Tetranychus turkestan showed that the egg production, average lifespan and egg hatching rate of adult mites were significantly reduced after treatment

Method used

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  • Molecular detection method of panonychus citri Mc Gregor response abamectin stress protein encoding gene
  • Molecular detection method of panonychus citri Mc Gregor response abamectin stress protein encoding gene
  • Molecular detection method of panonychus citri Mc Gregor response abamectin stress protein encoding gene

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Embodiment 1

[0020] Example 1. Differential protein analysis of Panonychus citrus in response to abamectin stress

[0021] 1. Trial mite treatment

[0022] Choose Abamectin LC 30 (0.091mg.L -1 ) concentration, compared with double distilled water, the leaf disc dipping method was used to treat the test mites (see patent application CN201510038090.8 for the leaf disc dipping method). The treated test mites were placed in an artificial climate box, set temperature at 25±1°C, relative humidity at 75±5%, light-dark ratio L:D=14:10h, took them out after 24h, examined under microscope, and picked out dead ones. For Panonychus citrus, the surviving test mites were collected into a 1.5mL centrifuge tube and stored at -80°C for later use. Each treatment and control were repeated 4 times, and each treatment had 500 mites (commonly used mites in the experiment: 500×4×2=4000).

[0023] 2. Total protein extraction

[0024] Take out the frozen lysate stored at ‐20℃, dissolve at room temperature, ad...

Embodiment 2

[0045] The protein spots whose expression levels changed by more than 2 times were significantly less than the protein spots with little fluctuation. The reasons for the analysis may be as follows: (1) There are many kinds of proteins in the organism. The stress proteins that act are only a small part compared to the whole; (2) the mechanism of action of abamectin determines that its lethal effect is slow, and there are not many types of stress proteins induced in a short period of 24 hours; (3) the used The treatment concentration was low, resulting in little change in the fold of the induced protein. The low treatment concentration may be the main reason. This reason has also been confirmed in other studies. For example, Aedes aegypti larvae were fed with biotoxin Cry11Aa for 5 hours, and their midguts were dissected for two-dimensional electrophoresis. It was found that when the feeding concentration was LC10 , only ATP synthase subunit beta and serine‐type endopeptidase ch...

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Abstract

The invention discloses a molecular detection method of panonychus citri Mc Gregor response abamectin stress protein encoding gene. The molecular detection method comprises the following steps: firstly, extracting total RNA (Ribonucleic Acid) of panonychus citri Mc Gregor treated by abamectin, and then transcribing the RNA into cDNA (complementary Deoxyribonucleic Acid); secondly, carrying out real-time quantitative detection on RPSA, EEF2, ATPsyn-alpha, FAH, V-ATPsyn-B, CALR, sHsp, CarE-B, Actin, CRY, Fer, Vg, ATPsyn-D, TPX, TCP-1-eta and GAPDH genes by using a qPCR (quantitative Polymerase Chain Reaction) method, wherein upstream and downstream sequences of qPCR detecting primer pairs of 16 genes are shown as SEQ ID No: 1 to 32; thirdly, analyzing qPCR results to obtain expression results of the 16 genes treated by the abamectin with different concentrations at different time periods.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a molecular detection method for a gene coding for a protein encoding a response to abamectin stress by Panonychus citrus. Background technique [0002] Citrus whole claw mite (Panochus citri Mc Gregor) belongs to the order of Acariles, Tetranychidae, and it is distributed in every citrus producing area in China. Hosts in citrus, saffron, and sweet-scented osmanthus. It pierces the epidermis of host leaves with its mouthparts and sucks the juice. The damaged leaves show countless small off-white spots. Branches affect tree vigor and yield, and are the number one pest in citrus production in my country. It is a worldwide harmful mite, which has a great impact on the yield and quality of citrus. At present, chemical control is still an important measure to control Panonychus citrus. [0003] After a biological individual is stimulated by a drug, the response ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q2600/158
Inventor 豆威沈晓敏钟锐李刚王进军
Owner SOUTHWEST UNIV
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