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Novel wheat gene TaMYB7D capable of adjusting and controlling synthesis and metabolism of anthocyanin

A kind of anthocyanin and gene technology, applied in the direction of genetic engineering, plant gene improvement, recombinant DNA technology, etc., can solve the problem of anthocyanin produced by grape embryogenic callus cells

Inactive Publication Date: 2015-09-09
CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Maize ZmC1 and ZmB genes can induce maize suspension cells to synthesize anthocyanins, but cannot induce grape embryogenic callus cells to produce anthocyanins

Method used

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  • Novel wheat gene TaMYB7D capable of adjusting and controlling synthesis and metabolism of anthocyanin
  • Novel wheat gene TaMYB7D capable of adjusting and controlling synthesis and metabolism of anthocyanin
  • Novel wheat gene TaMYB7D capable of adjusting and controlling synthesis and metabolism of anthocyanin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Cloning of TaMYB7D gene

[0035] All experimental materials were purchased from Northwest Institute of Plateau Biology, Chinese Academy of Sciences. All samples for DNA and RNA extraction were quickly frozen in liquid nitrogen and stored in a -80 freezer after sampling. All nucleotide sequences were determined at Shenzhen Huada Gene Co., Ltd. DNA was extracted using the CTAB method (Murray and Thompson, 1980), and RNA was extracted using the improved Trizon method (Li and Trick, 2005). Plasmids were extracted using a kit from Beijing Biotech Co., Ltd. (China). Nucleotide and protein sequences were aligned using the software Vector NTI Advance 10.0 (http: / / www.invitrogen.com / ). The protein phylogenetic tree was constructed using the software MEGA 4.0 (http: / / www.megasoftware.net / ). The gene map was drawn using GSDS software (http: / / gsds.cbi.pku.edu.cn / help.php).

[0036] In the present invention, the MYB gene AtTT2 and ZmC1 sequences related to anthocyani...

Embodiment 2

[0040] Example 2: TaMYB7D bioinformatics analysis

[0041] The full-length CDS sequence of TaMYB7D is 759bp, encoding 252 amino acids ( Figure 4 ). There is a typical R2R3-MYB domain at its N-terminal, 3 tryptophan residues W are regularly distributed in the R2 domain, 2 tryptophan residues W are regularly distributed in the R3 domain, and at the same time in the R3 There is a non-identical D / ELx in the domain that interacts with bHLH protein 2 R / Kx 3 Lx 6 Lx 3 R structural element; a possible transcriptional activation region exists at the C-terminus. This suggests that TaMYB-7D may be a positive regulator of R2R3-MYB transcription factor.

[0042] The known and functionally verified MYB genes were selected for phylogenetic tree analysis, including snapdragon, Arabidopsis, pepper, carrot, strawberry, gerbera, tomato, apple, tobacco, petunia, grape, Phylogenetic tree of MYB transcription factors in maize and wheat ( Figure 5 ). On the phylogenetic tree, TaMYB-7D is cl...

Embodiment 3

[0044] Example 3: Subcellular localization of TaMYB3 protein

[0045] Using the pk7FWG2-R vector with GFP and 35S promoter, the TaMYB-7D gene was constructed on the pk7FWG2-R vector, and the constructed pk7FWG2-R-TaMYB-7D fusion expression vector was transformed into onion by gene gun transformation method Epidermal cells. The results showed that in the onion epidermal cells transformed with empty vector pk7FWG2-R, the green fluorescence excited by GFP expression was distributed throughout the onion epidermal cells; when the recombinant plasmid pk7FWG2-R-TaMYB-7D was transformed into the onion epidermal cells, only in Green fluorescence was observed on the nucleus and cell membrane ( Figure 7 ). This indicates that TaMYB-7D may localize to the cell membrane in addition to the nucleus.

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Abstract

The invention discloses a novel wheat gene TaMYB7D capable of adjusting and controlling the synthesis and the metabolism of anthocyanin, and belongs to the field of plant genetic engineering, wherein the novel wheat gene TaMYB7D is obtained through separation of the purple-grain wheat variety highland 115. The TaMYB7D is located on a wheat chromosome 7D, and a protein product subcell is located in a cell nucleus. A deduced amino acid sequence shows that TaMYB3 encodes an MYB transcription factor capable of adjusting and controlling the synthesis and the metabolism of anthocyanin. The expression quantity of the TaMYB7D in the coleoptiles of a purple wheat variety highland 115 grain can be increased under the induction of light. During transient expression, the TaMYB7D and the pericarpic cell of the highland 115 grain synthetize anthocyanin. Moreover, coseparation occurs between the color of the coleoptiles of the highland 115 grain and the expression of the TaMYB-7D, that is, the TaMYB7D has transcriptional activity used for adjusting and controlling the synthesis and the metabolism of anthocyanin, and is the major gene of the purple coleoptiles of the TaMYB7D highland 115 grain.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, in particular to a new wheat gene TaMYB7D that regulates anthocyanin synthesis and metabolism. Background technique [0002] Anthocyanins are important secondary metabolites in plants and are involved in a wide range of biological processes (Beckwith et al., 2004; Downs and Siegelman, 1963; Lo and Nicholson, 1998; Vinterhalter et al., 2007; Zhou and Singh , 2002). As a food, anthocyanins are not only excellent food additives, but also have anti-arrhythmia, anti-cancer functions, and reduce the incidence of cardiovascular disease, obesity, and diabetes (Lietti et al., 1976). Therefore, people need more anthocyanins from fruits, vegetables and grains (Boyd, 2000), but due to dietary habits and economic reasons, fruits and vegetables cannot provide enough anthocyanins for people (Butelli et al. , 2008). [0003] The control of anthocyanin synthesis and metabolism in plants is mainly achie...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10C12N15/82A01H5/00
Inventor 刘宝龙张怀刚刘登才张波陈文杰刘瑞娟魏乐李建民王延谦
Owner CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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