Tobacco transcription repressor protein OFP1 and application thereof

A repressor protein and tobacco technology, applied in the field of tobacco genetic engineering, can solve problems such as poor appearance quality, affecting tobacco grade and quality, and producing harmful substances

Active Publication Date: 2019-08-23
ZHENGZHOU TOBACCO RES INST OF CNTC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

"Green-yellow smoke" not only has poor appearance, but also produces harmful substances when burned, which in turn affects the grade and quality of tobacco

Method used

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  • Tobacco transcription repressor protein OFP1 and application thereof
  • Tobacco transcription repressor protein OFP1 and application thereof
  • Tobacco transcription repressor protein OFP1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The present embodiment is about the coding gene of tobacco transcription repressor protein OFP1 OFP1 The cloning and construction process of the silencing vector are briefly introduced as follows.

[0032] (1) Tobacco NtOFP1 gene cloning

[0033] According to the previous research on the tobacco genome and related OFP1 genes, the specific coding sequence was selected as the target fragment, and the primer sequences for PCR amplification were designed as follows:

[0034] NtOFP1-F: 5'-ACCGAATTCTCTGCTAACTTGCCTAAA-3',

[0035] NtOFP1-R: 5'-ACCGGATCCAAGCCTCTTGTATAAAACTATG-3';

[0036] Using the cDNA of tobacco K326 leaves as a template, PCR amplification was carried out to obtain NtOFP1 Gene;

[0037] The PCR amplification program was: pre-denaturation at 95°C for 3 min; denaturation at 95°C for 15 s, annealing at 55°C for 15 s, extension at 72°C for 30 s, and after 34 cycles, complete extension at 72°C for 5 min;

[0038] The PCR amplification products were detect...

Embodiment 2

[0049] On the basis of Example 1, using Agrobacterium-mediated VIGS technology, the inventors further transformed the constructed recombinant TRV2-NtOFP1 vector into tobacco plants, and verified and analyzed the phenotypic changes of related plants. The specific experimental process is briefly introduced as follows.

[0050] (1) Transformation of Agrobacterium

[0051] It should be noted that, referring to the operation of Example 1 and the prior art, the inventor simultaneously prepared TRV2-GFP and TRV2-PDS recombinant vectors as controls, and the specific transformation process is as follows:

[0052] The positive cloning plasmids of TRV2-GFP (vector control), TRV2-PDS (VIGS efficiency control) and TRV2-NtOFP1 were transformed into Agrobacterium GV3101 competent cells by electric shock transformation respectively, using 50mg / L Kan and 50mg / L Kan and 50mg / L The YEB plate of LRif was cultured and screened, and after 2 days of inverted culture at 28°C, colony PCR was used to ...

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Abstract

The invention belongs to the technical field of tobacco genetic engineering, and particularly relates to a tobacco transcription repressor protein OFP1 and application thereof. The tobacco transcription repressor protein OFP1 is composed of 87 amino acid residues, wherein the amino acids from 34 locus to 83 <rd> locus are at a conserved structure domain. The protein is relevant to the contentof pigment substances in plant leaves, and after the expression of the protein is lowered, the content of the pigment substances in the leaves is significantly lowered, wherein the pigment substancesare neoxanthin, violaxanthin, xanthophyll, chlorophyll a/b and beta-carotene. According to the tobacco transcription repressor protein OFP1 and the application thereof, through the preliminary studyon genes of the tobacco transcription repressor protein OFP1, it is found that the genes are highly relevant to the content of pigment substances in tobacco, and after the genes are silenced, the content of the pigment substances in the tobacco is significantly lowered. By utilizing the feature, a new reference can be provided for cultivation of new varieties of tobacco plants.

Description

technical field [0001] The invention belongs to the technical field of tobacco genetic engineering, and specifically relates to a tobacco transcription repressor protein OFP1 and a patent application for its application. Background technique [0002] The tobacco that people usually smoke belongs to the genus Nicotiana of the genus Nicotiana ( Nicotiana ) There are more than 60 species, and the tobacco used for the preparation of smoking is mainly two cultivars, of which common tobacco (also known as safflower tobacco, Nicotiana tabacum ) occupies the main area, while Nicotiana chrysalis ( Nicotiana rustica ) is relatively small. [0003] Cultivated tobacco can be divided into six types: flue-cured tobacco, sun-cured tobacco, air-dried tobacco, burley tobacco, oriental tobacco and yellow flower tobacco according to the characteristics of tobacco leaf quality, biological traits, and cultivation methods, among which flue-cured tobacco is the most widely cultivated common to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82C12N15/11A01H5/00A01H6/82
CPCC07K14/415C12N15/825
Inventor 卢鹏陈千思许亚龙李泽锋刘萍萍郑庆霞金静静曹培健周会娜
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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