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Method for efficiently and rapidly inhibiting endogenous-gene expression of cynodon dactylon

A bermudagrass and fast technology, applied in the fields of genetic engineering and biology, can solve the problem of lack of bermudagrass virus-induced gene silencing technology, etc., and achieve the effect of improving the infection efficiency

Inactive Publication Date: 2016-06-15
INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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Problems solved by technology

However, up to now, there is still no report on bermudagrass virus-induced gene silencing technology

Method used

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  • Method for efficiently and rapidly inhibiting endogenous-gene expression of cynodon dactylon
  • Method for efficiently and rapidly inhibiting endogenous-gene expression of cynodon dactylon
  • Method for efficiently and rapidly inhibiting endogenous-gene expression of cynodon dactylon

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 The gene silencing of Bermudagrass phytoene dehydrogenase leads to albinism of plant leaves.

[0022] State-approved bermudagrass variety 'Yangjiang' bermudagrass and 'common' bermudagrass imported from the United States: Grass Industry Research Center, Institute of Botany, Chinese Academy of Sciences, Jiangsu Province. pRTBV-MVIGS plasmid: Department of Plant Molecular Biology, South Campus, University of Delhi, India (References: Purkayastha A, Mathur S, Verma V, Sharma S, Dasgupta I. Virus-induced gene silencing in rice using avector derived from a DNA virus. Planta, 2010, 232: 1531-1540). Escherichia coli DH5α and Agrobacterium EHA105: Grassland Research Center, Institute of Botany, Chinese Academy of Sciences, Jiangsu Province.

[0023] 1. Cloning of Bermudagrass phytoene dehydrogenase gene (phytoenedesaturase, PDS).

[0024] The total RNA of 'Yangjiang' bermudagrass leaves was extracted using the RNAprep plant total RNA extraction kit from Tiangen Bioch...

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Abstract

The invention discloses a method for efficiently and rapidly inhibiting endogenous-gene expression of cynodon dactylon. As genetic transformation of cynodon dactylon is quite difficult, the development of gene-function research and molecular breeding work of cynodon dactylon is seriously restricted. According to the new method, expression of endogenous genes of cynodon dactylon is specifically reduced through agrobacterium tumefaciens mediated transformation rice-tungro-bacilliform-virus-gene silencing vectors, and efficient and rapid technical means are provided for researching the gene function of cynodon dactylon. The method includes the following steps of 1, cloning of cynodon-dactylon target gene segments; 2, construction of the virus-induced gene silencing vectors; 3, cynodon-dactylon plant agrobacterium infection; 4, gene-expression-abundance molecular detection. By means of the method, expression of the endogenous genes of the cynodon dactylon can be successfully reduced by 50% or above.

Description

technical field [0001] The invention relates to a method for efficiently and rapidly inhibiting endogenous gene expression of bermudagrass by virus-induced gene silencing technology mediated by Agrobacterium, and belongs to the fields of genetic engineering and biotechnology. Background technique [0002] Bermudagrass (Cynodondactylon (L.) Pers.) is a plant of the genus Bermudagrass in the Poaceae Sansevieriae subfamily. It is an important warm-season turfgrass, forage and soil and water conservation plant. It is resistant to drought, salt, trampling, It has many advantages such as few pests and diseases, fast growth, thick grass layer, etc. It is widely used in the construction of slope protection lawns and grazing grasslands such as warm temperate, subtropical and tropical lawns, parks and road green belts, rivers and hillsides. very important position. [0003] In-depth research on the functions of important genes related to stress resistance and development of bermudagr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/53C12N15/84A01H5/00C12Q1/68
CPCC07K14/415C12N9/001C12N15/8218C12N15/8261C12N15/8271C12Q1/6895C12Y103/05005
Inventor 张兵刘建秀史经昂陈静波李丹丹郭海林宗俊勤李建建汪毅郭爱桂
Owner INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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