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Transcription factor gene for regulating synthesis of xanthophylls and application thereof

A transcription factor and lutein technology, applied in the field of plant genetic engineering, can solve the problem that the transcription factor gene has not been cloned and isolated

Active Publication Date: 2017-11-21
HEFEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the transcription factor genes that are specifically up-regulated or down-regulated during the development of marigold flowers to control the synthesis of anthocyanins have not been cloned and isolated

Method used

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  • Transcription factor gene for regulating synthesis of xanthophylls and application thereof
  • Transcription factor gene for regulating synthesis of xanthophylls and application thereof
  • Transcription factor gene for regulating synthesis of xanthophylls and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] Example 1: High-throughput transcriptome sequencing of marigold

[0012] Since the marigold genome has not yet been determined, in order to obtain the sequence of the functional gene transcripts of marigold, the tissue samples of the pigmented marigold cultivar Chrysanthemum King were used to extract 1 copy of RNA from leaves, immature flowers, and mature flowers of three individual plants, and perform High-throughput transcriptome sequencing and assembly annotation.

[0013] 1. Reagents

[0014] Plant RNA extraction reagent Trizol was purchased from Invitrogen Company, DNase I (Dnase I) was purchased from Takara Company, RNA Library Prep Kit was purchased from Beijing Biomike Biotechnology Co., Ltd., and other reagents were imported. packaged or domestically produced analytically pure products.

[0015] 2. Plant material

[0016] Pigment marigold (Tagetes erecta L.) cultivar Juwang was bred by Chifeng Xinhui Horticultural Company.

[0017] 3. Method

[0018] 3.1 R...

Embodiment 2

[0051] Example 2: Cloning of Tagetes TePTF1 Gene

[0052] According to Example 1, the tagetes transcription factor TePTF1 gene sequence was isolated. Total RNA was extracted from mature flowers of Marigold, and TePTF1 gene was cloned by reverse transcription-PCR (reverse transcription-PCR, RT-PCR).

[0053] 1. Reagents

[0054] Plant RNA extraction reagent Trizol was purchased from Invitrogen Company; DNase I (Dnase I) was purchased from Takara Company; reverse transcriptase (TransScript Reverse Transcriptase), Pfu high-fidelity DNA polymerase, and cloning vector pEASY-Blunt Simple Cloning Vector were purchased from Beijing Quanshijin Biotechnology Co., Ltd.; primers were synthesized by Shanghai Yingjun Biotechnology Co., Ltd., and the rest of the reagents were imported or domestic analytically pure products.

[0055] 2. E. coli strains and plant material

[0056] Escherichia coli (Escherichia coli) strain DH5α was purchased from Beijing Quanshijin Biotechnology Co., Ltd.; ...

Embodiment 3

[0105] Example 3: Expression Analysis of TePTF1 in Flower Tissues of Marigold at Different Stages

[0106] According to Example 2, the full-length sequence of the TePTF1 gene was obtained by cloning, and quantitative PCR primers were designed by using the primer design software Primer Premier 5.0, and total RNA was extracted from immature flowers and mature flowers of marigold respectively, and cDNA was obtained by reverse transcription, and TePTF1 gene expression was carried out level of quantitative analysis.

[0107] 1. Reagents

[0108] RNA extraction and reverse transcription reagents are as described in Example 1. Quantitative PCR kit TransStart Top GreenqPCR SuperMix was purchased from Beijing Quanshijin Biotechnology Co., Ltd.; primers were synthesized by Shanghai Yingjun Biotechnology Co., Ltd., and the rest of the reagents were imported or domestic analytically pure products.

[0109] 2. Method

[0110] Samples of immature flowers and mature flowers at the develop...

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Abstract

The invention discloses a transcription factor gene for regulating synthesis of xanthophylls and application thereof, wherein the transcription factor gene TePTF1 has a nucleotide sequence shown as in a sequence table SEQ ID NO. 1 and an amino acid sequence show as in the sequence table SEQ IN NO. 2. The gene TePTF1 is acquired by cloning from mature flower tissues of Tagetes erecta. Real-time quantitative analysis shows that the gene TePTF1 has significantly higher expression in mature flowers than immature flowers. In transient expression of the gene TePTF1 in tobacco leaves, the content of xanthophylls increases. The gene shown as SEQ ID NO. 1 plays a positive regulation role in the biosynthesis of plant xanthophylls. The transcription factor gene TePTF1 cloned from Tagetes erecta pigment is applicable to increasing the content of xanthophylls in plants.

Description

technical field [0001] The present invention relates to the technical field of plant genetic engineering, in particular to cloning lutein biosynthesis transcription factor TePTF1 from pigment marigold, the nucleotide sequence and protein sequence of the gene, and the use of TePTF1 gene in regulating plant lutein synthesis application. Background technique [0002] Marigold (Tagetes erecta L.) is a plant of the genus Tagetes in the Asteraceae family that is widely grown around the world. It is native to Mexico and other places in the Americas. It has large flower shape, various flower colors, long flowering period, strong plant adaptability, and short production cycle (Wang Dianbei et al., "Northern Horticulture", 2007, 1: 44-46). Marigold flowers can extract a variety of secondary metabolites. According to "Food Science" (Li Dajing et al., 2005, 9: 582-586), marigold flower extract has 93% available pigments, of which all-trans and cis lutein and lutein ester account for 8...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00
CPCC07K14/415C12N15/825
Inventor 牛向丽冯国栋黄胜雄马飞毛云张政刘永胜王洋王莹莹
Owner HEFEI UNIV OF TECH
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