New wheat gene TaMYB3 for regulating synthetization and metabolization of anthocyanin

An anthocyanin and gene technology, applied in genetic engineering, plant genetic improvement, angiosperms/flowering plants, etc., can solve the problems of grape embryogenic callus cells producing anthocyanins, etc.

Inactive Publication Date: 2012-11-14
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI +1
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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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  • New wheat gene TaMYB3 for regulating synthetization and metabolization of anthocyanin
  • New wheat gene TaMYB3 for regulating synthetization and metabolization of anthocyanin
  • New wheat gene TaMYB3 for regulating synthetization and metabolization of anthocyanin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Cloning of TaMYB3 gene

[0031] All experimental materials were purchased from the Institute of Genetics and Developmental 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 Beijing Aoke Biotechnology 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). The plastids were extracted using a kit from Beijing Biotech Co., Ltd. (China). Nucleotide and protein sequences were aligned using the software Vector NTIAdvance 10.0 (http: / / www.invitrogen.com / ). The protein phylogenetic tree was constructed using software MEGA 4.0 (http: / / www.megasoftware.net / ). The gene map was drawn using GSDS software (http: / / gsds.cbi.pku.edu.cn / help.php).

[0032] Through the MYB-like transcription factors known to regulate anthoc...

Embodiment 2

[0033] Example 2: TaMYB3 bioinformatics analysis

[0034] In order to understand the relationship between TaMYB3 and other similar genes, so as to predict the biological function of TaMYB3, we conducted bioinformatics analysis on TaMYB3. Bioinformatics analysis showed that the predicted TaMYB3 protein (SEQ ID NO: 12) has two conserved domains ( figure 2 ). The gene structure of TaMYB3 is different from other homologous genes. TaMYB3 has only one intron, while the MYB gene regulating anthocyanin synthesis and metabolism in maize, rice, Arabidopsis and apple has two introns ( image 3 ). We selected the full-length sequences of MYB transcription factors known to regulate anthocyanin synthesis and metabolism to construct a phylogenetic tree. From the perspective of homology, TaMYB3 protein belongs to the MYB class of proteins that regulate anthocyanin synthesis and metabolism. It is divided into the same branch as ZmC1 and ZmPL of maize, and all proteins under this branch h...

Embodiment 3

[0035] Example 3: Chromosomal loci of the TaMYB3 gene

[0036]TaMYB3 gene-specific primers TaMYB3-F and TaMYB3-R were used to perform PCR amplification on various cytogenetic materials to determine the TaMYB3 chromosomal locus. TaMYB3 could not be amplified in Uraltu wheat IE29-1 (with common wheat A chromosome set, without B and D chromosome sets) and Azalea As69 (with common wheat D chromosome set, without A and B chromosome sets). However, TaMYB3 can be stably amplified in materials with B chromosome groups such as China Spring and Langdon. This shows that TaMYB3 is not on the A and D chromosomes, but on the B chromosome ( Figure 5 .B). TaMYB3 could not be amplified in the genomic DNA of tetraploid wheat Langdon (AABB) lacking chromosome 4B but adding chromosome 4D from Chinese Spring. TaMYB3 can be stably amplified in other substitution lines with 4B chromosomes, and it is inferred that TaMYB3 is located on the 4B chromosome of wheat ( Figure 5 .B). In the Chinese S...

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Abstract

The invention provides a transcription factor TaMYB3 of a MYB class, which is separated from the plateau 115 of a purple kernel wheat variety and is used for regulating the synthetization and the metabolization of anthocyanin. TaMYB3 is positioned between 0.62 and 0.95 of a long arm physical map of wheat 4B chromosome. The subcellular fraction of a protein product is positioned on a cell nucleus.Shown by a derived amino acid sequence, the TaMYB3 codes a MYB class transcription factor regulating the synthetization and the metabolization of anthocyanin. The expression quantity of the TaMYB3 inthe plateau 115 of a purple kernel wheat variety increases under the induction of light. The TaMYB3 can promote skin cells of the plateau 115 kernels after dark treatment to synthetize anthocyanin under the existence of transcription factors ZmR of corns bHLH during transient expression. Single TaMYB3 genes and single ZmR genes can not induct the synthetization of anthocyani, which shows that theTaMYB3 genes have the transcriptional activity of regulating the synthetization and the metabolization of anthocyanin. barly strip mosaic virus (BSMV) mediates the expression quantity decrease of theTaMYB3 in the plateau 115 kernels, and reduces the quantity of anthocyanin in kernels. This shows that the TaMYB3 participates in the biosynthesis of anthocyanin in the plateau 115 kernels.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and relates to a new wheat gene TaMYB3 for regulating the synthesis and metabolism of wheat anthocyanin, including a recombinant plasmid of the gene, a host and application thereof. 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). ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10C07K14/415A01H5/00
Inventor 王道文张怀刚刘宝龙安学丽秦焕菊刘昕
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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