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Screening method of chilo suppressalis reference genes under stress of chlorantraniliprole and application of chilo suppressalis reference genes

A technology of chlorantraniliprole and internal reference genes, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., and can solve the problems of non-existence of internal reference genes

Inactive Publication Date: 2016-01-27
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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Problems solved by technology

Internal reference genes commonly used in insects include Rps3, Actin, GAPDH, Tublin, Rpl10, 18SrRNA, 28SrRNA, etc., but in fact, the mRNA expression level of any internal reference gene is constant under all external or internal conditions, ideal internal reference genes don't exist

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  • Screening method of chilo suppressalis reference genes under stress of chlorantraniliprole and application of chilo suppressalis reference genes
  • Screening method of chilo suppressalis reference genes under stress of chlorantraniliprole and application of chilo suppressalis reference genes
  • Screening method of chilo suppressalis reference genes under stress of chlorantraniliprole and application of chilo suppressalis reference genes

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Embodiment Construction

[0046] The present invention will be further described below in conjunction with specific embodiments and accompanying drawings. The use of the internal reference gene stability analysis software was carried out according to its instructions.

[0047] 1. Feeding of Chilo borer.

[0048] Chilo borer larvae collected from paddy fields were reared to the 2nd generation with artificial feed in an artificial climate chamber: temperature 25°C, humidity 75%, light: darkness = 14 hours: 10 hours, and then the 3rd instar larvae were used for breeding. Chlorantraniliprole stress treatment. 2. Chlorantraniliprole Stress Treatment

[0049] 1. Prepare chlorantraniliprole as a 500ppm mother solution with acetone, and then dilute the mother solution with 0.1% TritonX-100 distilled water into 50ppm, 100ppm and 200ppm working solutions.

[0050] 2. Invert the rice seedlings with a height of 20 cm and immerse them in the prepared medicinal liquid, dip them into the base of the rice seedlings...

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Abstract

The invention discloses a screening method of chilo suppressalis reference genes under the stress condition of insecticide of chlorantraniliprole and application of the chilo suppressalis reference genes. The method comprises the following steps of using chilo suppressalis which is not treated by medicine as reference materials; using the chilo suppressalis treated by the stress of the chlorantraniliprole as experiment materials to be subjected to fluorescent quantitative PCR (Polymerase Chain Reaction) detection; inputting Ct values of all optional reference genes into reference gene stability analysis software delta Ct, RefFinder, BestKeeper, GeNorm and NormFinder to be analyzed; and screening the reference genes TF4 with the highest stability in the chilo suppressalis under the stress condition of the chlorantraniliprole. Therefore the genes can be used as the reference genes for accurately studying the expression of relevant genes under the stress of the chlorantraniliprole.

Description

technical field [0001] The invention belongs to the technical field of expression stability of real-time fluorescent quantitative PCR internal reference genes in molecular biology, and in particular relates to a screening method and application of internal reference genes of Chilorrhizae medullaris under stress of different concentrations of chlorantraniliprole. Background technique [0002] Chilosuppressalis belongs to the family Lepidoptera and is an important pest of rice in my country. It causes symptoms such as withered sheath, dead heart, white ear, withered booting ear and insect damage to rice, causing huge economic losses. With the renewal of rice cultivation system and the change of rice varieties, the population of Chilo suppressalis increases year by year. [0003] In recent years, the control of rice stem borer largely depends on the chemical pesticide chlorantraniliprole. However, due to the long-term and large-scale use of this agent, Chilo borer developed se...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q2600/158C12Q2600/166
Inventor 鲁艳辉王国迪钟列权王国荣黄贤夫徐红星郑许松吕仲贤
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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