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Gene capable of reducing nicotine content of tobacco and application thereof

A tobacco and genetic technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve problems such as easy thinking and unclear molecular mechanism

Inactive Publication Date: 2013-04-03
BEIJING NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

The experimental results prove that jasmonin can regulate the production of nicotine, but its molecular mechanism is not clear
[0006] At present, for the research on JAZ proteins, a total of 12 members have been found in Arabidopsis. It is not difficult for us to think whether there are such JAZ proteins in tobacco, and whether these proteins are involved in the synthesis of jasmonin-induced tobacco nicotine

Method used

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  • Gene capable of reducing nicotine content of tobacco and application thereof
  • Gene capable of reducing nicotine content of tobacco and application thereof
  • Gene capable of reducing nicotine content of tobacco and application thereof

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Experimental program
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Embodiment Construction

[0022] The following experimental methods, unless otherwise specified, are conventional methods. The reagents used in the following experimental methods were purchased from conventional biochemical reagent companies unless otherwise specified.

[0023] Plant material: tobacco suspension cell line BY-2 (Nicotiana tabacum L.cv. Bright Yellow 2)

[0024] 1. Obtaining experimental materials and extracting RNA

[0025] ■MeJA-treated BY-2 cells at different times

[0026] Tobacco suspension cells BY-2 in MS (4.3g / l MS, 100mg / l inositol, 30g / l sucrose, 200mg / l potassium dihydrogen phosphate, 1mg / l VB1, 0.2mg / l 2,4-D, pH =5.8) in liquid culture medium at 24°C, 120rpm shaker suspension culture, 7 days a cycle.

[0027] Materials used in Real-time PCR experiments: When jasmonin (MeJA) was treated, the suspension cells grown for 4 days were transferred to 2,4-D-free MS medium, and after 24 hours, 50 μM jasmonin (final concentration ) were processed, and samples were collected at 0, 0...

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Abstract

The invention discloses a gene capable of reducing nicotine content of tobacco and an application thereof. The gene is a tobacco JAZ protein gene which is named as NtJAZ3, and protein encoded by the gene disclosed by the invention contains a typical TIFY structural domain and a jas structural domain and is induced and expressed by jasmonate. An RNAi (Ribonucleic Acid Interfere) transgenic carrier of the NtJAZ3 gene is constructed, and the tobacco is converted, so that expression of the NtJAZ3 in tobacco cells is successfully inhibited, and the nicotine content of the obtained transgenic tobacco cell line is obviously reduced compared with that of a wild type tobacco cell line. The gene disclosed by the invention provides an important basis and a technical support for low-nicotine-content tobacco breeding.

Description

technical field [0001] The invention relates to the synthesis of tobacco nicotine and related genes for regulation, in particular to the application of the gene for regulating the synthesis of tobacco nicotine in reducing the content of tobacco nicotine. Background technique [0002] Smoking seriously endangers people's health, so how to reduce the harm of smoking has become one of the topics that governments, medical experts, and botanists have paid close attention to and studied. Many of the world's top tobacco companies have invested heavily in research on the metabolic pathways and regulatory mechanisms of tobacco secondary metabolites. Nicotine is a pyridine alkaloid mainly found in Nicotiana plants of the family Solanaceae, and is an important secondary metabolite in tobacco. Current research on nicotine biosynthesis mainly focuses on cloning enzymes related to its metabolism and analyzing its physiological and biochemical properties. Nicotine is formed in the roots ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N15/113A01H5/00
Inventor 韩生成王英典赵和平杨玉萍
Owner BEIJING NORMAL UNIVERSITY
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