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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 and other issues

Inactive Publication Date: 2011-01-05
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 the 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 snap-frozen in liquid nitrogen and stored in a -80 freezer after sampling. All nucleotide sequence determinations were performed at Beijing Aoke Biotechnology Co., Ltd. DNA was extracted by CTAB method (Murray and Thompson, 1980), and RNA was extracted by modified Trizon method (Li and Trick, 2005). Plastid extraction was performed using a kit from Beijing Bodatech Bio-Gene Technology 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 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).

[0032] By targeting the MYB-like transcripti...

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 performed bioinformatics analysis on TaMYB3. Bioinformatics analysis showed that the predicted TaMYB3 protein (SEQ ID NO: 12) had two conserved domains of R2 and R3 with high homology compared with other MYB transcription factors that regulate anthocyanin synthesis ( figure 2 ). The gene structure of TaMYB3 is different from other homologous genes. TaMYB3 has only one intron, while the MYB gene that regulates anthocyanin synthesis and metabolism in maize, rice, Arabidopsis and apple has two introns ( image 3 ). We selected the protein full-length sequences of MYB-like transcription factors known to regulate anthocyanin synthesis and metabolism to construct a phylogenetic tree. In terms of homology, TaMYB3 protein belongs to the MYB class of proteins that regulate anthocyanin synthe...

Embodiment 3

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

[0036]The TaMYB3 chromosomal locus was determined by PCR amplification with TaMYB3 gene-specific primers TaMYB3-F and TaMYB3-R in various cytogenetic materials. TaMYB3 could not be amplified in Urartu wheat IE29-1 (with common wheat A chromosome set, but without B and D chromosome sets) and Junjie wheat As69 (with common wheat D chromosome set, without A and B chromosome sets). TaMYB3 can be stably amplified in materials with B genomes such as Chinese Spring and Langdon. This means that TaMYB3 is not on the A and D chromosome sets, but on the B chromosome set ( Figure 5 .B). TaMYB3 could not be amplified in the genomic DNA of tetraploid wheat Langdon (AABB) which lacked chromosome 4B and added chromosome 4D in Chinese spring. TaMYB3 can be stably amplified in other substitution lines with chromosome 4B, and it is inferred that TaMYB3 is located on chromosome 4B of wheat ( Figure 5 .B). In the Chinese spring 4B chro...

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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 in the 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 the TaMYB3 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 the TaMYB3 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 that regulates the synthesis and metabolism of wheat anthocyanins, including a recombinant plasmid of the gene, a host and applications 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-heart rate disorders, 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...

Claims

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Application Information

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Patent Type & Authority Applications(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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