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Agapanthus praecox gibberellin receptor APGID1b protein, and encoding gene and probe thereof

A technology of Agapanthus gibberellin and coding genes, which can be used in genetic engineering, plant genetic improvement, recombinant DNA technology, etc., and can solve problems such as unexamined

Inactive Publication Date: 2013-10-09
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Before the publication of the present invention, there was no literature report related to the Agapanthus Agapanthus APGID1b protein sequence mentioned in this patent application

Method used

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  • Agapanthus praecox gibberellin receptor APGID1b protein, and encoding gene and probe thereof
  • Agapanthus praecox gibberellin receptor APGID1b protein, and encoding gene and probe thereof
  • Agapanthus praecox gibberellin receptor APGID1b protein, and encoding gene and probe thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Cloning of gibberellin receptor APGID1b gene from Agapanthus chinensis

[0042] 1. Acquisition of plant material

[0043] 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.

[0044] 2. Extraction of RNA

[0045] Total RNA was extracted wit...

Embodiment 2

[0070] Example 2 , Sequence Information and Homology Analysis of Agapanthus Agapanthus APGID1b Gene

[0071] The new full-length CDS open reading frame sequence of the Agapanthus agapanthus APGID1b of the present invention is 1029bp, 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 APGID1b was deduced, with a total of 342 amino acid residues, a molecular weight of 38255.7 Daltons, and an isoelectric point (pI) of 6.11. For the detailed sequence, see the sequence shown in SEQ ID NO.2;

[0072] The CDS open reading frame sequence of Agapanthus agapanthus APGID1b 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 searches showed that it has 68% identity with upland cotton GID1b gene ...

Embodiment 3

[0073] Example 3 , Expression differences of Agapanthus agapanthus APGID1b gene at different developmental stages and in different tissues

[0074] 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.

[0075] 2. Extraction of RNA: 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...

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Abstract

The invention relates to an agapanthus praecox gibberellin receptor APGID1b protein, and an encoding gene and a probe thereof. The protein is the following protein of (a) or (b): (a) a protein composed by an amino acid sequence as shown in SEQ ID NO.2; and (b) a protein derived from the protein of (a), having agapanthus praecox gibberellin receptor activity, and composed by a sequence obtained by substituting, deleting or adding one or a plurality of amino acids of the amino acid sequence as shown in the SEQ ID NO.2. Furthermore, the invention provides a nucleic acid sequence encoding the protein, and the detection probe. Applications comprise the following steps: amplifying to obtain a 3' end and a 5' end of an APGID1b gene by an RACE technique, carrying out gene full-length sequence splicing and homology analysis, through spatial and temporal expression mode analysis and external source regulation substance processing, analyzing different conditions of APGID1b gene expression, so as to verify gene functions. Spatial and temporal expression characteristics of the agapanthus praecox gibberellin receptor APGID1b gene are regulated by utilizing a gene engineering technology, and thus the plant height of a plant can be changed.

Description

technical field [0001] The invention relates to a receptor protein and its encoding gene and probe in the process of gibberellin signal transduction from Agapanthus abaciens, in particular to an Agapanthus gibberellin receptor APGID1b protein and its encoding gene and probe. Background technique [0002] The plant type of ornamental plants is one of the important factors to determine its ornamental value and commercial value. Changing and directional regulation of the plant type to create a dwarf and compact plant type can increase the ornamental value and commercial value of ornamental plants, and good cut flower traits can be obtained by increasing the plant height. The morphogenesis of plants is regulated by a variety of hormones. Gibberellin (Gibberellin, GA) is the most important hormone that determines the elongation of plant stems. The synthesis and metabolism of gibberellin directly affect the plant type development. GID1, the receptor for gibberellin signal transdu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12Q1/68C12N15/11A01G7/06
Inventor 申晓辉岳建华张荻任丽
Owner SHANGHAI JIAO TONG UNIV
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