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Tobacco recessive PVY (Potato Virus Y) resistance gene eIF4E-1 and application thereof

An eif4e-1, tobacco technology, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve the problem that the recessive anti-PVY gene has not been reported in the literature, etc., and achieve the effect of major application prospects.

Inactive Publication Date: 2014-05-28
YUNNAN ACAD OF TOBACCO AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is speculated that the eIF4E gene in tobacco is related to the sensitivity to PVY, but the recessive anti-PVY gene in tobacco has not been reported in the literature

Method used

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  • Tobacco recessive PVY (Potato Virus Y) resistance gene eIF4E-1 and application thereof
  • Tobacco recessive PVY (Potato Virus Y) resistance gene eIF4E-1 and application thereof
  • Tobacco recessive PVY (Potato Virus Y) resistance gene eIF4E-1 and application thereof

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Embodiment 1

[0032] The pCR-BluntII-TOPO vector, Gateway LR clonase Enzyme Mix kit, and pENTR 2B vector were purchased from Invitrogen, and the pK7GWIWG2 expression vector was purchased from VIB (www.gateway.psb.ugent.be). Agrobacterium C58C1 (pMP90) (see McKersie, etc., 1999, Plant Physiol 119:839-847), and Agrobacterium GV3101 were purchased from Invitrogen. Plasmid DNA extraction kit, agarose gel DNA recovery kit, and DNA fragment purification kit were purchased from QIAGEN. Escherichia coli DH5α; restriction endonuclease, Klenow Fragment; reverse transcription kit, DNA Marker, LA Taq DNA polymerase, T4 DNA polymerase and T4 DNA ligase, spectinomycin were purchased from Dalian Bao Biological Corporation and Roche Corporation. The RNA extraction kit Trizol was purchased from Invitrogen, and the ELISA kit and test strips for detecting PVY were purchased from Agdia (www.agdia.com).

[0033] 1. Extraction of tobacco total RNA

[0034] Take 100 mg of young leaves of China Tobacco 100 and ...

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Abstract

The invention discloses a tobacco recessive PVY (Potato Virus Y) resistance gene eIF4E-1 and application thereof. The tobacco recessive PVY resistance gene eIF4E-1 comprises the DNA (Deoxyribonucleic Acid) fragment of an eIF4E-1 gene coding region, wherein the DNA fragment is obtained by PCR (Polymerase Chain Reaction) amplification of a specific primer to a primer 4E-1F (SEQIDNO.3) and a primer 4E-1R (SEQIDNO.4); a nucleotide sequence SEQIDNO.1 of a tobacco PVY resistance gene eIF4E-1 coding region and the amino acid sequence SEQIDNO.2 of protein coded by the tobacco PVY resistance gene are obtained first; the eIF4E-1 gene is a recessive disease resistance gene; the eIF4E-1 gene of infected tobacco is lost by a conventional breeding means, so that the non-GMO (Genetically Modified Organism) disease resistance tobacco is obtained; an interference vector plasmid is built by using the eIF4E-1 gene and converted to the infected tobacco; expression of the eIF4E-1 gene of the infected tobacco is inhibited, so that genetically-modified tobacco with obviously improved disease resistance is obtained; the gene has an important application prospect in cultivating the PVY viral disease resistance tobacco.

Description

technical field [0001] The invention relates to a tobacco anti-PVY gene eIF4E-1 and its application. Background technique [0002] Potato virus Y (PVY) can infect more than 170 kinds of plants belonging to 34 genera, and is very harmful to Solanaceae crops (such as: tobacco, potato, pepper, tomato, etc.). The disease of PVY infecting tobacco, also known as tobacco vein spot disease, is a worldwide epidemic disease. Due to the lack of main resistant varieties and the difficulty in implementing comprehensive control measures, it is seriously harmful in my country's smoking areas. PVY infection can reduce tobacco yield by 7%-18%, and reduce output value by 11%-80%. In 1989, tobacco vein spot disease caused direct economic losses to the Canadian tobacco industry of $1 million. In 1993, vein spot disease became popular in tobacco-growing areas in Shandong Province of my country, and by 1998 it became the largest tobacco disease. PVY is seriously harmful in some tobacco areas o...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82C12N15/84A01H5/00
Inventor 刘勇王丙武宋中邦高玉龙李文正谢贺杨大海陈学军肖炳光
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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