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Plant dwarfing related protein GA2ox, and encoding gene and application thereof

A plant and gene technology, applied to the plant dwarfing-related protein GA2ox and its encoding gene and application fields, can solve the problems of long improvement period, high pruning frequency and low effect, and achieve the effect of reducing pruning frequency and being less prone to lodging.

Inactive Publication Date: 2012-10-31
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the large amount of vertical growth of tall fescue, it also caused a series of problems, the most prominent of which is: the high frequency of mowing makes it difficult to guarantee the quality of the lawn
Excessive pruning not only increases the cost of lawn management and reduces ornamental properties, but also accelerates the lack of nutrients in the grass body, leading to premature aging
[0005] It is difficult to obtain dwarf phenotypes through traditional breeding techniques, because conventional breeding methods have a long period of germplasm improvement for tall fescue and low effectiveness

Method used

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  • Plant dwarfing related protein GA2ox, and encoding gene and application thereof
  • Plant dwarfing related protein GA2ox, and encoding gene and application thereof
  • Plant dwarfing related protein GA2ox, and encoding gene and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1, the discovery of GA2ox protein and its coding gene

[0036] 1. The RNA of tall fescue was extracted by SDS method.

[0037] 2. Using RNA with good integrity and no contamination as a template, use M-MLV enzyme (purchased from Promage) to reverse transcribe the RNA into cDNA.

[0038] 3. Cloning of intermediate fragments

[0039] Primers (F1 and R1; M=A / C, S=G / C, V=A / G / C) were designed according to the homologous conserved sequence of GA2ox, and the cDNA of tall fescue was used as a template for PCR amplification. Recovered and sequenced to obtain a 244bp intermediate fragment.

[0040] F1 (5' primer): 5'-MTS GGG TGG VTC GAG TAC-3';

[0041] R1 (3' primer): 5'-GGT AGT GGT TCA SSC GGA-3'.

[0042] 4. Cloning of 3'-end

[0043] A primer 3'GSP I (gene-specific primer) was designed according to the sequencing results of the intermediate fragment. The primer sequence was 5'-CAC GTC CAC CGC GCT GAG AGA C-3'. In order to improve product specificity, another pri...

Embodiment 2

[0064] Embodiment 2, the acquisition and identification of transgenic plants

[0065] 1. Construction of recombinant plasmid pCAMBIA1302-GA2ox

[0066] See the construction flow chart figure 1 .

[0067] 1. Extract the total RNA of tall fescue and reverse transcribe it into cDNA.

[0068] 2. Using the cDNA in step 1 as a template, carry out PCR amplification with a primer pair consisting of GA2ox_5' and GA2ox_3' to obtain a PCR amplification product (the target sequence is the open reading frame of the GA2ox gene with the stop codon removed).

[0069] GA2ox_5': 5'-ACT AGT ATG GTG GTG CTC GCG AAG-3';

[0070] GA2ox_3': 5'-ACT AGT GGA CCG GTG GTG GTG GCC-3'.

[0071] The 1.0% agarose gel electrophoresis of the PCR amplification product is shown in figure 2 .

[0072] 3. Insert the PCR amplification product in step 2 into the pMD18-T simple vector (purchased from TaKaRa Bioengineering Company) to obtain the recombinant plasmid pMD18-Tsimple-GA2ox.

[0073] 4. The recombin...

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Abstract

The invention discloses a plant dwarfing related protein GA2ox, and an encoding gene and application thereof. The protein provided by the invention is a protein (a) or (b), wherein the protein (a) has an amino acid sequence shown in Sequence 1 in a sequence table, and the protein (b) is obtained through carrying out substitution and / or deletion and / or addition of one or more amino acid residues on the amino acid sequence shown in Sequence 1, related to plant dwarfing and derived from Sequence 1. In the invention, a recombinant plasmid containing a GA2ox gene is introduced to tobacco through an agrobacterium-mediated transformation method to obtain transgenic tobacco which has an obvious dwarfing phenotype. The plant dwarfing related protein GA2ox has a great value for plant dwarfing breeding.

Description

technical field [0001] The invention relates to a plant dwarfing related protein GA2ox and its encoding gene and application. Background technique [0002] Dwarf breeding is of great significance in improving crop quality. During the "Green Revolution" of the last century, dwarf varieties were introduced into rice and wheat, which greatly increased the yield of rice and wheat. Studies have shown that genes causing dwarf traits are related to gibberellin metabolism or signal transduction. [0003] Gibberellin (GA) is an important class of plant hormones. The biologically active GA plays an important role throughout the life cycle of higher plants. It can promote seed germination, stem and root elongation, leaf Stretching, epidermal hair development, pollen tube elongation, flower and fruit development, etc., and plants can also mediate their own responses to environmental factors by regulating the biosynthesis of endogenous gibberellins. Under the mediation of GA2-oxidase ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C12N15/11A01H5/00
Inventor 王涛刘松虎徐光远马磊董江丽
Owner CHINA AGRI UNIV
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