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Gene capable of reducing nicotine content of tobacco and application thereof

A tobacco and gene 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
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  • Claims
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AI Technical Summary

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

[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 treatment of BY-2 cells at different times

[0026] Tobacco suspension cells BY-2 in MS (4.3g / lMS, 100mg / l inositol, 30g / l sucrose, 200mg / l potassium dihydrogen phosphate, 1mg / lVB1, 0.2mg / l2,4-D, pH=5.8) Suspension culture in the liquid medium at 24°C and 120rpm in a shaker for 7 days.

[0027] Materials used for Real-time PCR experiment: When jasminin (MeJA) is processed, the suspension cells grown for 4 days are transferred to 2,4-D-free MS medium. After 24 hours, 50μM jasmin is added (final concentration ) For processing, collect samples at 0, 0.5, 2, 6, 12, and 24 h...

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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 NtJAZ1, 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 NtJAZ1 gene is constructed, and the tobacco is converted, so that expression of the NtJAZ1 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 present invention relates to the synthesis of tobacco nicotine and the regulation of related genes, in particular to the application of genes regulating the synthesis of tobacco nicotine in reducing the content of tobacco nicotine. Background technique [0002] Smoking seriously harms people's health, so how to reduce the harm of smoking has become one of the issues that governments, medical experts, and botanists all over the world pay attention to and research. 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 kind of pyridine alkaloid, which mainly exists in Nicotiana plants of the family Solanaceae. It is an important secondary metabolite in tobacco. The current research on nicotine biosynthesis is mainly focused on cloning its metabolic-related enzymes and analyzing its physiological and biochemical characteristics. Nicoti...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/113C12N15/82A01H5/00
Inventor 韩生成王英典赵和平杨玉萍
Owner BEIJING NORMAL UNIVERSITY
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