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Vitis vinifera abscisic acid degradation pathway key enzyme 8'-hydroxylase CYP707A protein and coding gene thereof, and application thereof

A technology encoding gene and abscisic acid, applied in the field of key enzymes and their encoding genes, the key enzyme 8'-hydroxylase CYP707A protein in the degradation pathway of grape abscisic acid, can solve the problem of less research, CYP707A gene cloning, and rare reports on expression analysis, etc. question

Pending Publication Date: 2020-04-17
ZHEJIANG WANLI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, this gene has been widely studied in plants such as Arabidopsis thaliana, tomato, and tobacco, but relatively few studies have been done in grapes, and there are few reports on the cloning and expression analysis of CYP707A gene

Method used

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  • Vitis vinifera abscisic acid degradation pathway key enzyme 8'-hydroxylase CYP707A protein and coding gene thereof, and application thereof
  • Vitis vinifera abscisic acid degradation pathway key enzyme 8'-hydroxylase CYP707A protein and coding gene thereof, and application thereof
  • Vitis vinifera abscisic acid degradation pathway key enzyme 8'-hydroxylase CYP707A protein and coding gene thereof, and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1, the cloning of grape fruit CYP707A gene

[0027] 1. Acquisition of plant material

[0028] The plant material used in this experiment is the fruit of a new grape variety - "Yinhong" grape cultivated by Wu Yueyan's scientific research team (formerly named: Yongyou No. 1, a commercially available Changshu variety; Yemawan Grape Farm, Dongqian Lake Tourism Resort, Ningbo, Wanli, Zhejiang College, Ningbo Yinzhou District Forestry Technology Management Service Station as the breeding unit, this variety is a European and American hybrid. The tree is strong, the leaves are thick, and the leaves are smooth and hairless. The average ear weight is 500-700 grams. The fruit is oval and the average grain size The weight is about 12 grams. The skin is thick and tough, purple red, and the fruit powder is medium. The flesh is hard, the flavor is sweet, and the soluble solids are more than 17%. The maturity period of greenhouse cultivation in Ningbo area is generally from ...

Embodiment 2

[0034] Example 2. Sequence information of grape fruit CYP707A gene

[0035] The obtained CYP707A gene was predicted by ORF, and there were 14 open reading frames in the sequence, the longest one starting from 97 to 1509 bases, with a total length of 1413bp and 470AA. According to the whole genome comparison of grapes, it can be seen that the CYP707A gene sequence is located on chromosome 2 of grapes, contains 7 exons and 6 introns, and can encode 653 amino acids, and after analysis, it can be known that it belongs to CYP707A1 in the CYP707A gene family.

Embodiment 3

[0036] Example 3. Physicochemical properties and functional analysis of grape fruit CYP707A protein

[0037] According to the subroutine analysis of Expasy, it can be seen that the protein molecular formula encoded by VvCYP707A is C3421H5336N874O931S25, the relative molecular mass is 74406.80, the number of amino acid residues is 654, the theoretical isoelectric point is 9.49, and the total number of negatively charged amino acid residues (Asp+Glu) is 54, positively charged. The total number of charged amino acid residues (Arg+Lys) is 82, the fat index (AI) is 87.40, the total average hydrophilicity (GRAVY) is -0.141, and the instability coefficient (II) is 42.85, showing that the protein is relatively stable (Stability protein standard around 40).

[0038] Hydrophilic and hydrophobic and transmembrane domain analysis shows that CYP707A protein belongs to hydrophobic and transmembrane proteins (see attached Figure 4 ). Using PredictProtein( https: / / www.predictprotein.org / ...

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Abstract

The invention relates to a vitis vinifera abscisic acid degradation pathway key enzyme 8'-hydroxylase CYP707A protein and a coding gene thereof, and application thereof. The amino acid sequence of theprotein is shown as SEQ ID No.2, and the sequence of the coding gene is shown as SEQ ID No.1. The invention also discloses a preparation method of the grape abscisic acid degradation pathway key enzyme 8'-hydroxylase CYP707A protein. Regulating factors in the vitis vinifera ripening process can be filled up, the plant fruit ripening mechanism is perfected, and the blank of cloning and expressionanalysis of key gene CYP707A family members of the abscisic acid degradation pathway and the blank of CYP707A protein and coding genes in new variety 'Yinhong' grape fruits are filled up.

Description

technical field [0001] The present invention relates to the key enzyme and its coding gene in the degradation pathway of the phytohormone abscisic acid (ABA) that regulates fruit ripening in "Yinhong" grapes, in particular to the key enzyme 8'-hydroxylase CYP707A protein of the degradation pathway of grape abscisic acid , coding gene and application thereof belong to the field of biotechnology. Background technique [0002] Among plants, the fruit not only has the function of plant reproduction and propagation, but also has certain economic and edible value, and is widely loved by people because of its unique taste and high nutritional value. Fruit growth and development is a complex and gradual process, which is mainly regulated by exogenous factors, endogenous hormones and transcription factors. Endogenous genes control the synthesis and degradation of a variety of endogenous hormones. It is known that the phytohormones related to promoting fruit ripening mainly include a...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12N15/53C12N15/82C12N5/10C12N15/10A01H5/08A01H6/88
CPCC12N9/0073C12N15/8243C12Y114/13093
Inventor 吴月燕杨洋贾永红
Owner ZHEJIANG WANLI UNIV
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