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Protein used for controlling anthocyanidin content, coding gene thereof, and application thereof

An anthocyanin content and gene technology, applied in the direction of introducing foreign genetic material using a vector, cells modified by introducing foreign genetic material, plant peptides, etc. , Enhance the body's antioxidant capacity, reduce the effect of disease incidence

Active Publication Date: 2014-01-01
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For a long time, the comprehensive utilization of rice husk has been extensively studied at home and abroad, and many available ways have been obtained, but there are not many utilization ways that can really form large-scale production and consume a large amount of rice husk

Method used

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  • Protein used for controlling anthocyanidin content, coding gene thereof, and application thereof
  • Protein used for controlling anthocyanidin content, coding gene thereof, and application thereof
  • Protein used for controlling anthocyanidin content, coding gene thereof, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1, the acquisition of protein GH-1 and its coding gene for controlling anthocyanin content

[0048] 1. Phenotype analysis of rice golden glume mutant gh-1

[0049] The rice golden glume mutant gh-1 (International Rice Research Institute, Gramene Accession: GR: 0060365) is a near allele of ZF802 (WT) (preserved by the National Rice Data Center of China Rice Research Institute, registration number GS01003-1989) gene line. Compared with the control ZF802, the phenotype of the golden glume was brown-red from the heading stage, and the husk and stalks were brown-red when mature ( figure 1 ). The mutant phenotype does not affect the normal growth and development of rice under normal cultivation conditions. figure 1 The upper middle left is the seed of the control ZF802, which is light yellow; the upper right is the panicle of the golden glume mutant gh-1, which is reddish brown; the lower left is the stem of the control ZF802, which is light yellow; the lower rig...

Embodiment 2

[0071] Embodiment 2, Complementation experiment of gh-1 phenotype of golden glume mutant

[0072] 1. Construction of complementary vector pCGH and complementary control vector pCGHC

[0073] BAC OsJNBa0017N05 (purchased from Shanghai National Gene Research Center, Chinese Academy of Sciences, No. OsJNBa0017N05) was digested with PstI and BamHI to obtain 3005 bases upstream of the start codon ATG and 3360 bases after the stop codon TGA of GH-1 The DNA fragment (7996bp) of the full-length sequence of bases was cloned between the PstI and BamHI recognition sites of pCAMBIA1300 (DingGuo, MCV033), and the complementary expression vector pCGH was constructed. The constructed complementary vector pCGH was digested with KpnI, the promoter region, exon and part of the 3' end sequence of the GH-1 gene were removed, and part of the regulatory region at the 3' end was retained, and the complementary control vector pCGHC was constructed ( image 3 Middle C).

[0074] 2. Obtaining and phe...

Embodiment 3

[0076] Embodiment 3, the overexpression verification test of GH-1 gene

[0077] 1. Construction of overexpression vector pGHOX

[0078] According to the method of step 5 in Example 1, the ORF of GH-1 was obtained, and the vector was digested with Sal I and Xba I to obtain a fragment of about 700bp, and then this fragment was connected to Sal I and Xba I of the overexpression vector pCam13OX for recognition Between the sites, the overexpression vector pGHOX was constructed.

[0079] The construction method of pCam130X is as follows:

[0080] Using the primer pair 35S-F: (5'-AAGCTTCCCAGATTAGCCTTTTCAAT-3') and 35S-R: (5'-CTGCAGTCCCCCGTGTTTCTCTCAA-3') PCR amplified plasmid pBI121 (DingGuo, MCV032) to obtain a constitutive CaMV35S promoter fragment of about 850bp , the fragment was double digested with Pst I and Hind III and then ligated into the Pst I and Hind III recognition sites of the vector pCAMBIA1300 (DingGuo, MCV033) to form the intermediate vector pCam13OXM. Plasmid pB...

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Abstract

The invention discloses a protein used for controlling the content of anthocyanidin, a coding gene thereof, and an application thereof. The invention discloses a paddy rice chalcone isomerase (CHI), a coding gene thereof, and an application thereof. The protein is represented by the following (1) or (2): (1) a protein composed of an amino acid residue sequence of SEQ ID No.2 in a sequence list; (2) a protein with a same activity as that of the amino acid residue sequence of SEQ ID No.2, wherein the protein is derived from the amino acid residue sequence of SEQ ID No.2 through substitution and / or deletion and / or addition of one or more amino acid residues. The paddy rice CHI gene provided by the invention can be used as a molecular marker and assists in paddy rice seed breeding. The paddy rice CHI gene also has an important value on the study of reasonable utilization of rice hulls.

Description

technical field [0001] The invention relates to a protein for controlling anthocyanin content, its coding gene and application. Background technique [0002] Flavonoids generally refer to a large class of secondary metabolites formed by connecting two benzene rings (A- and B-rings) with phenolic hydroxyl groups through the central three carbon atoms. There are more than 6,000 kinds of flavonoids in nature. The types and quantities of these compounds are distributed differently in different types of plants, and are also affected by the developmental stages and growth conditions of plants. Flavonoids widely exist in the plant kingdom and are the most important pigment compounds in plants. They make the leaves, flowers and fruits of plants present various colors such as red, blue and purple. These gorgeous colors can attract pollinators and make Plants are pollinated and seed dispersal is facilitated. In addition to making the various tissues and organs of plants present diff...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/63C12N7/01C12N5/10C07K14/415C12N15/82
Inventor 程祝宽洪丽兰李明唐丁王克剑
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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