Genes used for regulating and controlling synthesis and transport of nicotine in tobaccos and application thereof

A nicotine and gene technology, applied in the field of gene technology and botany, can solve problems affecting nicotine content, achieve good content traits, and improve content traits

Active Publication Date: 2014-04-30
浙江圣庭医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] The synthesis and transport of tobacco nicotine are regulated by many factors. At present, the research on the anabolic pathway and transport of nicotine from the molecular point of view has not been completely thorough. Although some key genes in the nicotine synthesis pathway have been identified and cloned, such as QPT , PMT, MPO, etc. (Dwey and Xie, 2013, Phytochemistry), but there is no report on the research on the regulation of nicotine synthesis and transport key genes through miRNA, thereby affecting nicotine content

Method used

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  • Genes used for regulating and controlling synthesis and transport of nicotine in tobaccos and application thereof
  • Genes used for regulating and controlling synthesis and transport of nicotine in tobaccos and application thereof
  • Genes used for regulating and controlling synthesis and transport of nicotine in tobaccos and application thereof

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Effect test

Embodiment 1

[0035] Based on high-throughput sequencing to obtain the small RNA sequences of tobacco roots and leaves after topping and leaf damage, as well as the genome sequences of two ancestral species of tobacco, a large number of tobacco gene sequences can be found, which can form a stable hairpin structure, and its stem can Production or expression of small RNA sequences (mature sequences). The hairpin structures formed by 6 gene sequences are shown in Figure 1-6 . These predicted hairpin structures were stable (all MEF values ​​were less than −24.82). The expression sequences of these genes were detected, especially in the roots and leaves of topping and leaf damage tobacco, which indicated that the expression of these genes was probably induced by these treatments.

Embodiment 2

[0037] The small RNA sequences generated by the expression of the above six genes are predicted by the target genes, and all target the key protein-coding genes in the tobacco nicotine synthesis and transport pathway, including QPT (quinolinate phosphirobosyltransferase), PMT (putrescine N-methyltransferase) and MATE (multi antimicrobial extrusion family protein) genes (Table 1). A large number of studies have shown that these genes are the key functional genes of nicotine synthesis (QPT and PMT) and transport (MATE), respectively (Dwey and Xie, 2013, Phytochemistry). The predicted values ​​of the 6 genes and their target genes found in the present invention are all between 0-2.0 (generally, the close value is 3.0, exceeding this value indicates that the binding result is unreliable), and all of them regulate the target genes by cleavage. At the same time, the analysis results of the degradome data obtained in the experiment showed that 6-105 cut fragments were detected (the c...

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Abstract

The invention relates to the fields of genetic technology and botany and aims to provide related non-coding microRNA (ribonucleic acid) (miRNA) genes for regulating and controlling synthesis and transport of nicotine in tobaccos. The sequence of the genes is any one of SEQ ID NO:1-6. The genes have the beneficial effects that the obtained six genes can form a stable hairpin structure; analysis of the obtained tobacco non-coding miRNA sequence indicates that part of miRNA comes from stem sequences of the tobaccos; the miRNA predicts that target genes of the miRNA are respectively targeted at key protein coding genes for synthesis and transport of nicotine in tobaccos; the obtained miRNA regulation and control mechanism can provide a deeper understanding for the roles played by the key genes in synthesis and transport pathways of nicotine in tobaccos, so that the content character of nicotine in tobaccos can be better improved.

Description

technical field [0001] The invention belongs to the field of gene technology and botany; the invention relates to regulating non-coding microRNA (miRNA) genes related to the synthesis and transport of tobacco nicotine. Background technique [0002] The synthesis and transport of tobacco nicotine are regulated by many factors. At present, the research on the anabolic pathway and transport of nicotine from the molecular point of view has not been completely thorough. Although some key genes in the nicotine synthesis pathway have been identified and cloned, such as QPT , PMT, MPO, etc. (Dwey and Xie, 2013, Phytochemistry), but there is no report on the research on the regulation of nicotine synthesis and transport key genes through miRNA, thereby affecting nicotine content. Therefore, this patent, as an important part of the nicotine synthesis and transport pathway, plays an important role in improving tobacco nicotine content and the like. Contents of the invention [0003]...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A01H5/00
Inventor 樊龙江王卫娣肖炳光卢秀萍李永平叶楚玉王煜
Owner 浙江圣庭医学检验实验室有限公司
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