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Agapanthus praecox gibberellin synthesis dioxygenase APGA20ox protein, and coding gene and probe thereof

A technology for synthesizing gibberellins and dioxygenases from Agapanthus agapanthus, applied in genetic engineering, oxidoreductase, plant genetic improvement, etc.

Inactive Publication Date: 2014-09-03
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Before the present invention was published, there was no literature report related to the sequence of Agapanthus APGA20ox protein mentioned in this patent application

Method used

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  • Agapanthus praecox gibberellin synthesis dioxygenase APGA20ox protein, and coding gene and probe thereof
  • Agapanthus praecox gibberellin synthesis dioxygenase APGA20ox protein, and coding gene and probe thereof
  • Agapanthus praecox gibberellin synthesis dioxygenase APGA20ox protein, and coding gene and probe thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Cloning of APGA20ox Gene of Gibberellin Synthetic Dioxygenase from Agapanthus chinensis

[0043] 1. Acquisition of plant material

[0044] Agapanthus was introduced into my country from South Africa in 2000, and the introduction test was carried out in Shanghai. In his master's thesis "Research on the Dwarfing of Agapanthus Potted Plants" published in 2011, Sun Bo introduced the basic biological characteristics and introduction history of Agapanthus in detail. The agapanthus material involved in this embodiment can be obtained through public channels. Callus, embryogenic callus, somatic embryos, and seedlings were cultivated and preserved in a tissue culture room for RNA extraction. Plant healthy Agapanthus plants of the same size in the greenhouse and perform routine management. When the flower buds are formed and the petals are about to crack, various organs and tissues are collected for RNA extraction.

[0045] 2. Extraction of RNA

[0046] Total RNA ...

Embodiment 2

[0073] Example 2 , Sequence information and homology analysis of the APGA20ox gene of Agapanthus chinensis

[0074] The new Agapanthus APGA20ox full-length CDS open reading frame sequence of the present invention is 1086bp, and the detailed sequence is shown in the first sequence of SEQ ID NO.1. According to the CDS open reading frame sequence, the amino acid sequence of Agapanthus APGA20ox was deduced, with a total of 361 amino acid residues, a molecular weight of 41050.8 Daltons, and an isoelectric point (pI) of 6.73. For the detailed sequence, see the sequence shown in SEQ ID NO.2;

[0075] The CDS open reading frame sequence of Agapanthus APGA20ox and the amino acid sequence of its encoded protein were nucleated in Non-redundant GenBank+EMBL+DDBJ+PDB and Non-redundant GenBank CDS translations+PDB+SwissProt+Superdate+PIR databases using BLAST program Nucleotide and protein homology search, it was found that it has 67% identity with T. villosa GA20ox gene (EU142949.1) at ...

Embodiment 3

[0076] Example 3 , Agapanthus APGA20ox gene expression differences in different developmental stages and in different tissues of Agapanthus

[0077] 1. Acquisition of materials: At different stages of Agapanthus somatic embryo development (callus; callus; somatic embryo differentiation; seedling formation), laboratory tissue culture materials were used as research objects, and adult The main root, lateral root, stem, scape, old leaf, new leaf, small pedicel, petal, filament, anther, ovary, and seed of the plant were wrapped in aluminum platinum paper and immediately put into liquid nitrogen, and then transferred to Store in a -80°C ultra-low temperature freezer until use. The leaves of Agapanthus plants sprayed with GA and Paclobutrazol (PBZ) were sampled for RNA extraction.

[0078] 2. RNA extraction: RNA prep pure Plant Total RNA Extraction Kit (RNA prep pure Plant Kit: Shanghai Laifeng Biotechnology Co., Ltd.) was used to extract RNA from different developmental stages a...

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Abstract

The invention relates to an agapanthus praecox gibberellin synthesis dioxygenase APGA20ox protein, and a coding gene and a probe thereof. The protein is a protein consisting of the amino acid sequence shown in SEQ ID No.2. The invention also provides a corresponding nucleic acid sequence and a corresponding detection probe. According to steps, the 3' and 5' terminals of the APGA20ox gene are obtained by respective amplification through an RACE technology; the gene full-length sequence splicing and homology analysis are performed; and the expression difference condition of the APGA20ox gene is analyzed through spatio-temporal expression mode analysis and exogenous regulation substance treatment so as to verify the gene functions. To verify the functions of the APGA20ox, an agapanthus praecox embryonic callus is transformed by an RNAi technology, a dwarfing agapanthus praecox plant can be obtained by APGA20ox gene silencing, and therefore effects of the APGA20ox nucleic acid sequence on improvement of the plant height of agapanthus praecox are shown more definitely. According to the agapanthus praecox gibberellin synthesis dioxygenase APGA20ox protein and the coding gene and the probe thereof, the spatio-temporal expression characteristic of the agapanthus praecox gibberellin synthesis dioxygenase APGA20ox gene can be regulated so as to change the plant height development of a plant.

Description

technical field [0001] The invention relates to a key enzyme in the synthetic pathway of Agapanthus gibberellin, its coding gene and probe, in particular to APGA20ox protein, its coding gene and probe. Background technique [0002] The plant type of ornamental plants is one of the important factors that determine the commodity value and ornamental value. Changing and directional regulation of plant type to create dwarf and compact plant type can increase the commercial value and ornamental value of ornamental plants, and good cut flower traits can be obtained by increasing plant height. Gibberellin (Gibberellin, GA) is the most important phytohormone that determines the elongation of plant stems. The synthesis and metabolism of GA directly regulate the development of plant architecture. There are various types of gibberellins, and the biosynthetic pathway is very complicated. Only a few types of GAs are biologically active, and most of the intermediate products are not biol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02C12N15/53C12Q1/68C12N15/11
CPCC12N9/0071C12N15/827C12Y114/11
Inventor 申晓辉岳建华张荻任丽
Owner SHANGHAI JIAO TONG UNIV
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