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Application of nicotiana benthamiana NbSMG7 gene in regulation and control of plant virus resistance and transgenic plant cultivation method

A technology of transgenic plants and cultivation methods, applied in the field of cultivation of NbSMG7 transgenic plants, can solve the problems of low efficiency and instability of seed treatment measures

Active Publication Date: 2021-03-23
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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Problems solved by technology

However, at this stage, no pesticides and disease-resistant plants with control effects against CGMMV have been developed, and studies have shown that seed treatment measures are not efficient and unstable

Method used

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  • Application of nicotiana benthamiana NbSMG7 gene in regulation and control of plant virus resistance and transgenic plant cultivation method
  • Application of nicotiana benthamiana NbSMG7 gene in regulation and control of plant virus resistance and transgenic plant cultivation method
  • Application of nicotiana benthamiana NbSMG7 gene in regulation and control of plant virus resistance and transgenic plant cultivation method

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Embodiment Construction

[0031] A method for cultivating an NbSMG7 transgenic plant resistant to cucumber green mottle mosaic virus, specifically comprising the following steps:

[0032] (1) Obtain Nicotiana benthamiana plants, extract RNA from leaves of Nicotiana benthamiana plants by Trizol method, remove genomic DNA in RNA samples, and reverse transcribe to obtain cDNA;

[0033] (2) Utilize the primer pair NbSMG7-F and NbSMG7-R, as shown in SEQ ID NO:2-3, carry out PCR amplification with cDNA as template, the primer sequence used is:

[0034] NbSMG7-F: atgatgaccatccaatgga,

[0035] NbSMG7-R: cacaaacaaacggccttc;

[0036] (3) the amplified product is constructed on the n-YFP vector to obtain the YFP-NbSMG7 recombinant plasmid; as figure 1 as shown in (A);

[0037] (4) Mix 1 μl of the recombinant plasmid and 100 μl of Agrobacterium competent and transfer it into an electric shock cup, use an electric shock device for 2500V electric shock transformation, add LB medium for 2 hours of recovery, and th...

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Abstract

The invention discloses application of a nicotiana benthamiana NbSMG7 gene in regulation and control of plant virus resistance and a transgenic plant cultivation method, the virus is cucumber green mottle mosaic virus, and a transcript sequence of the NbSMG7 gene is shown as SEQ ID NO: 1. The NbSMG7 gene in nicotiana benthamiana is cloned and constructed into a YFP-NbSMG7 vector with a fluorescentlabel, the YFP-NbSMG7 vector is transferred into agrobacterium tumefaciens and then infects tobacco for genetic transformation of the tobacco, and a YFP-NbSMG7 transgenic plant is obtained. The obtained nicotiana benthamiana overexpression YFP-NbSMG7 plant can significantly hinder the infection of cucumber green mottle mosaic viruses.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a method for cultivating stably inherited NbSMG7 transgenic plants resistant to cucumber green mottle mosaic virus. Background technique [0002] Cucumber green mottle mosaic virus (CGMMV) belongs to the Tobamovirus genus of Virgaviridae, which can infect cucumbers, gourds, melons, watermelons and other Cucurbitaceous crops, causing plant growth. Slow, mottled leaf chlorosis, fruit discoloration and fibrosis, resulting in serious losses, is an important phytosanitary pest in my country. [0003] CGMMV is a positive-sense single-stranded RNA virus with a full length of about 6400nt and encodes four proteins, namely a replicase of 129kD, a replication-associated protein of 186kD, a motor protein of 29kD and a coat protein of 17.4kD. [0004] The harm of CGMMV to cucurbit crops is a worldwide problem, and there is currently no effective chemical agent to control CGMMV. ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/00A01H6/82
CPCC07K14/415C12N15/8283
Inventor 李方方陈雅琳周雪平
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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