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Method for suppressing Zoysia japonica endogenous gene expression efficiently and rapidly

A Zoysia, fast technology, applied in the field of genetic engineering and biology, can solve the problem of no zoysia virus-induced gene silencing technology, etc., and achieve the effect of improving the infection efficiency

Inactive Publication Date: 2016-06-22
INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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Problems solved by technology

So far, several virus-induced gene silencing vectors have been constructed and successfully used in gene silencing experiments of rice, Brachypodium distachyon, sorghum and other gramineous plants. Unfortunately, there is still no Zoysia virus-induced gene silencing vector. technical reports

Method used

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  • Method for suppressing Zoysia japonica endogenous gene expression efficiently and rapidly
  • Method for suppressing Zoysia japonica endogenous gene expression efficiently and rapidly

Examples

Experimental program
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Effect test

Embodiment 1

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

[0022] Zoysia cultivar ‘Lanyin No. 3’ approved by the state and Zoysia cultivar ‘Crowne’ 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: PurkayasthaA, MathurS, VermaV, SharmaS, DasguptaI. Virus-inducedgene silencing in rice using avector derived fromaDNAvirus. 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. Zoysia phytoene dehydrogenase gene (phytoenedesaturase, PDS ) clones.

[0024] The total RNA of 'Lanyin No. 3' Zoysias leaves was extracted using the RNAprep Plant Total RNA Extraction Kit of Tiangen Biochemical Technology (Beijing) Co., L...

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Abstract

The invention discloses a novel method for suppressing Zoysia japonica endogenous gene expression efficiently and rapidly. Since Zoysia japonica genetic transformation is extremely difficult, carrying out Zoysia japonica gene function research and molecular breeding is restricted seriously. According to the method, Zoysia japonica endogenous gene expression is reduced specifically by agrobacterium-medated rice tungro bacillifrom virus gene silencing vectors, so that an efficient rapid technological means can be provided for gene functions of Zoysia japonica. The novel method includes the steps of 1), cloning target gene fragments of the Zoysia japonica; 2), constructing virus-induced gene silencing vectors; 3), impregnating Zoysia japonica plants by agrobacterium; 4), performing molecular detection to obtain genetic expression abundance. By the method, Zoysia japonica endogenous gene expression can be reduced by more than 30% successfully.

Description

technical field [0001] The invention relates to a method for efficiently and rapidly inhibiting the expression of endogenous genes of Zoysia sativa by virus-induced gene silencing technology mediated by Agrobacterium, and belongs to the fields of genetic engineering and biotechnology. Background technique [0002] Zoysia ( ZoysiajaponicaL .) is a perennial plant of the genus Zoysia in the Poaceae Teffae subfamily. It is recognized as an excellent warm-season turfgrass in the world. It has a wide range of soil adaptability and strong resistance (cold resistance, drought resistance, salt tolerance It is widely used in the construction of various lawns such as park greening, golf courses and stadiums in warm temperate and subtropical zones, and has a very important role in the lawn industry. status. [0003] Functional research on important genes related to stress resistance and development of Zoysia spp. can provide important candidate genes and theoretical basis for genetic...

Claims

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Application Information

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IPC IPC(8): C12N15/84A01H5/00
CPCC12N9/001C12N15/8205C12N15/8218
Inventor 张兵刘建秀史经昂郭海林宗俊勤陈静波李丹丹李建建汪毅郭爱桂
Owner INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
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