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Method for infecting peony seedling against TRV vector-based virus-induced gene silencing system

A technology of gene silencing and peony, applied in the field of plant genetic engineering, can solve the problems such as difficulty in obtaining mutants, limitation of peony, immature genetic transformation system, etc., and achieve the effects of good infection effect and convenient operation process.

Pending Publication Date: 2018-12-18
NORTHWEST A & F UNIV
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Problems solved by technology

However, as a woody plant, peony is not easy to obtain mutants, and its genetic transformation system is still immature, which limits the research on the gene function of woody plants such as peony, and brings a lot of inconvenience to the molecular breeding of such plants

Method used

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  • Method for infecting peony seedling against TRV vector-based virus-induced gene silencing system
  • Method for infecting peony seedling against TRV vector-based virus-induced gene silencing system
  • Method for infecting peony seedling against TRV vector-based virus-induced gene silencing system

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Embodiment Construction

[0026] The present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments, the purpose of which is to better understand the content of the present invention rather than limit the protection scope of the present invention.

[0027] Based on the VIGS system, the present invention proposes an infection method for TRV vectors to mediate whole plant virus-induced gene silencing of tree peony seedlings

[0028] (1) Sources of raw materials and reagents

[0029] 1) 2 to 3-year-old peony seedlings: the peony variety is Fengdan (seeds purchased from Shaanxi Zhongzi Guoye Peony Industry Development Co., Ltd.), planted in the same environment and grown for 2-3 years (from sowing) Seedlings (planting location: Northwest Agriculture and Forestry University Resource Garden).

[0030] 2) EcoR I and BamH I were purchased from Thermo Fisher Scientific.

[0031] 3) T4-DNA ligase, DH5α competent cells, and pUCm-T Vector were purchased ...

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Abstract

The invention discloses a method for infecting a peony seedling against a TRV vector-based virus-induced gene silencing system. TRV1 vectors and vectors of TRV-PDS, TRV-GFP and the like constructed onthe basis of TRV2 vectors are used for conducting vacuum infection on the peony seedling, symptoms of bleached peony leave, fluorescent leaves and root tissues, and the like are developed, gene silencing of a peony plant is achieved, and a simple fast low-cost method is provided for studying functions of various tissue related genes in a peony growth and development process.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and relates to a method for virus-induced gene silencing directed at whole peony seedlings mediated by TRV vectors. Background technique [0002] Virus-induced gene silencing (VIGS) is a post-transcriptional gene silencing technology, which has been widely used in many plants as an effective reverse genetics technology. The principle of VIGS is that plants will produce specific endonuclease Dicer in the process of resisting virus infection, which will cut the double-stranded RNA produced during virus replication into small interfering RNA (siRNA) of 21-24 nucleotides. ). After siRNA is further amplified in plant cells, it binds to Agronaute 1 (AGO1) protein in a single-stranded form to form an RNA-induced silencing complex (RISC). RISC can specifically cause the degradation of mRNA homologous to siRNA in the plant cytoplasm, resulting in the silencing of the target gene. [0...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/65
CPCC12N9/001C12N15/65C12N15/8218
Inventor 张延龙谢力行张庆雨孙道阳牛立新胡佳媛
Owner NORTHWEST A & F UNIV
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