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Overexpression transcription transcription factor SlBBX20 for improving the content of tomato anthocyanin

A tomato anthocyanin, transcription factor technology, applied in the field of plant genetic engineering

Active Publication Date: 2019-01-04
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, zinc finger protein transcription factors found in many plants can regulate the accumulation of anthocyanins have not been reported.

Method used

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  • Overexpression transcription transcription factor SlBBX20 for improving the content of tomato anthocyanin
  • Overexpression transcription transcription factor SlBBX20 for improving the content of tomato anthocyanin
  • Overexpression transcription transcription factor SlBBX20 for improving the content of tomato anthocyanin

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Example 1: Cloning and genetic transformation of SlBBX20 gene

[0027] 1) Cloning and analysis of SlBBX20 gene

[0028] The SlBBX20 gene cloned in the present invention is based on the results of whole genome association analysis and the transgenic function verification of a large number of candidate genes. It is found that the SlBBX20 transcription factor gene can significantly improve the ability of tomato to synthesize and accumulate anthocyanins. According to the gene information annotated in the database (Http: / / solgenomics.net / tomato / ), the primers for amplifying the SlBBX20 gene are designed, the forward primer is SlBBX20For: 5`-CAGAGACTTTAGGTGTGAGCCC-3`, and the reverse primer is SlBBX20Rev: 5` -GGAAAAAACTACACATCTGGTCG-3`, amplify the full-length SlBBX20 gene cDNA sequence from TS-93 (quoted from the Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences) by PCR (see Table 1 for details). The amplification method is to first extract the RNA of...

Embodiment 2

[0041] Example 2: Phenotype identification of overexpression of SlBBX20 gene

[0042] 1) Tissue expression profile analysis of SlBBX20 gene

[0043] Using Q-RT-PCR technology to analyze the expression of SlBBX20 gene in different tissues and organs (roots, stems, leaves, flowers, fruits) of tomato, the results showed that under normal growth conditions, the SlBBX20 gene is in the detected It is expressed in tissues and organs, and the expression is higher in flowers (see figure 2 Figure A in the middle), we further analyzed the different structures of flowers and found that the expression level is the highest in the petals. The structural expression of other floral organs is relatively low (see figure 2 Figure B in).

[0044] 2) Phenotype analysis of overexpression of SlBBX20

[0045] After overexpression of SlBBX20, the leaf color changed significantly compared with the transgenic recipient material, that is, the color became dark purple, while the interference plant was green ( ...

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Abstract

The invention belongs to the technical field of plant genetic engineering, and particularly relates to an overexpression transcription factor SlBBX20 for improving the content of tomato anthocyanin. The method of the invention clones a key gene SlBBX20 for controlling anthocyanin synthesis in tomato leaves, and performs biological function identification. The cDNA sequence of the gene is shown inSEQ ID NO: 1 of the sequence table and the sequence of the encoded protein is shown as SEQ ID NO: 3. The transgenic function verification shows that the overexpression of the gene in tomato can significantly regulate the function of anthocyanin content in tomato leaves.

Description

Technical field [0001] The invention belongs to the field of plant genetic engineering. Specifically, it relates to overexpression of the transcription factor SlBBX20 to increase the content of tomato anthocyanins. In the present invention, a reverse genetics method is used to isolate and clone a functional gene SlBBX20 that can increase the content of tomato anthocyanins, and overexpress the gene in tomatoes, which can significantly improve the synthesis and accumulation of anthocyanins in tomato leaves. The invention can effectively increase the anthocyanin content in tomato leaves. Background technique [0002] Anthocyanin, also known as anthocyanin, is the most important component of flavonoids. It mainly imparts various colors to plant fruits, leaves, flowers and other tissues and organs. It is a water-soluble pigment. As an important product of plant secondary metabolism, anthocyanins participate in a variety of biological processes. When plants are subjected to external...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82
CPCC07K14/415C12N15/825
Inventor 叶志彪王涛涛林爱华张春丽罗丹李汉霞张俊红熊程
Owner HUAZHONG AGRI UNIV
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