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Encoding genes of key enzyme, namely dihydroflavonol-4-reductase in metabolic pathway of plant anthocyanin

A technology encoding gene and gene, applied in the direction of plant gene improvement, oxidoreductase, genetic engineering, etc., can solve the problems of low reduction efficiency, inability to produce anthocyanin-like anthocyanins, inability to reduce DHK, etc.

Inactive Publication Date: 2009-11-18
BEIJING UNIV OF AGRI +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In all dicotyledonous plants that use DHK as a substrate except petunia, the E (glutamic acid) at position 145 is highly conserved. The best substrate of petunia DFRA is DHM, and the reduction efficiency of DHQ Very low, can not reduce DHK (ForkmannG., Ruhnau B. Distinct substrate specificity of dihydroflavonol 4-reductase from flowers of Wtunia hybrida. Z. Narturforsch, 1987, 42C: 1146-1148.), so can not produce geranoid anthocyanins

Method used

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  • Encoding genes of key enzyme, namely dihydroflavonol-4-reductase in metabolic pathway of plant anthocyanin
  • Encoding genes of key enzyme, namely dihydroflavonol-4-reductase in metabolic pathway of plant anthocyanin
  • Encoding genes of key enzyme, namely dihydroflavonol-4-reductase in metabolic pathway of plant anthocyanin

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Embodiment Construction

[0021] The methods used in the following examples are conventional methods unless otherwise specified, and the primer synthesis and sequencing work are all completed by Shanghai Sangon Biotechnology Service Co., Ltd. Restriction enzymes EcoR I, HindIII and Taq enzymes, T4 DNA ligase were purchased from Dalian Bao Biological Engineering Co., Ltd., pUC18-T Vector was purchased from Bao Biological Engineering Company, BD SMART TM RACE cDNA Amplification Kit was purchased from Clontech Company.

[0022] Amp antibiotics, Tris saturated phenol, water saturated phenol, DEPC, DL2000 DNA Marker, agarose, yeast extract, and tryptone were purchased from Beijing Dingguo Biotechnology Co., Ltd.

[0023] 1. Acquisition of the 3' end sequence of MrDFR gene

[0024] Total RNA was extracted from leaves of Malus Royalty (Malus Royalty) (Malus Royalty, Beijing Agricultural University Germplasm Resource Garden), dissolved in 50 μl of DEPC water, and detected by electrophoresis. Analyzing by fo...

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Abstract

The invention discloses encoding genes of key enzyme in a metabolic pathway of plant anthocyanin and application thereof. Protein is: 1) protein consisting of amino acid sequences shown in a sequence 2 in a sequence table, or 2) protein which is derived from the 1) protein, obtained by performing substitution and / or deletion and / or addition of one or more amino acid residues on the amino acid sequence shown in the sequence 2 in the sequence table and can change flower color or leaf color of a plant.

Description

technical field [0001] The invention relates to an enzyme coding gene related to the formation of plant anthocyanin and its application. Background technique [0002] Flower color is one of the important factors that determine the ornamental value of flowers. Flower color is mainly determined by three major substances: flavonoids, carotenoids, and alkaloids (Tanaka Y, S Tsuda, T Kusumi. Metabolic Engineering to modify flowers color. Plant Cell Physioloy, 1998(11): 1119-1126.). [0003] Anthocyanins (anthocyanins) are a class of main pigments in flavonoids, which control the pink, red, brick red, blue, purple and other flower colors of flowers. There are three basic anthocyanins in plants, namely anthocyanin, Anthocyanins, delphinidins, and methylated derivatives of these anthocyanins (Xu Jizun, Wang Lihui, Pan Qingyu. Research progress in flower color gene transformation of ornamental plants [J]. China Agricultural Science and Technology Herald, 2006, 8(5): 56~60.). [00...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12N15/53C12N15/11
Inventor 姚允聪沈红香宋婷婷田佶
Owner BEIJING UNIV OF AGRI