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Ziziphus jujuba SOD (superoxide dismutases) gene and application thereof

A technology of superoxide and dismutase, applied in the field of genetic engineering to achieve the effect of promoting understanding

Inactive Publication Date: 2016-11-16
山西省农业科学院农业资源与经济研究所 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] So far, there is no report on isolating jujube superoxide dismutase gene from jujube for plant variety improvement

Method used

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  • Ziziphus jujuba SOD (superoxide dismutases) gene and application thereof
  • Ziziphus jujuba SOD (superoxide dismutases) gene and application thereof
  • Ziziphus jujuba SOD (superoxide dismutases) gene and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment 1: the biological information analysis of jujube superoxide dismutase gene ZjSOD

[0051] The present invention constructs pot jujube ( Ziziphus jujuba Mill hupingzao) screened the complete sequence of jujube superoxide dismutase gene from the cDNA library of fruiting branch (namely jujube tree), and named it as wxya ( Z iziphus j ujuba superoxide dismutases ), the analysis on the NCBI website shows that the protein structure of ZjSOD1 has a conserved SOD domain ( figure 1 ), belonging to the Fe-SOD type. By looking for protein sequences with high homology and high sequence similarity with ZjSOD protein from different species, the phylogenetic relationship analysis was carried out, and the results showed that ZjSOD has a close relationship with rubber tree CAB53458.1 and CAC13961.1, and is located in the evolutionary tree On the same branch of the peach tree Q9SM64.1 in the same sub-branch ( figure 2 ). Sequences with higher homology to the...

Embodiment 2

[0055] Embodiment 2: Construction, amplification and Agrobacterium-mediated expression vector of jujube superoxide dismutase gene ZjSOD wxya Gene Transformation of Arabidopsis

[0056] According to the sequence encoded by jujube superoxide dismutase gene cDNA, the design contains EcoR I and Sma Ⅰ Specific primers for restriction sites, the primers were synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd. Its upstream primer is the sequence of P1: 5'—ac gaattc agatggcaatgggaagtg—3', containing EcoR Ⅰ Restriction enzyme cutting site (that is, the underlined part). The downstream primer is the sequence of P2: 5'—at cccggg tagcagactctctctcata—3', contains the Sma Ⅰ Restriction enzyme cutting site (that is, the underlined part). restriction endonuclease EcoR I and Sma ⅠAfter modification, it was connected to the plant expression vector PEZR(K)-LNY carrying the yellow fluorescent protein gene YFP, and then the transformants were screened on th...

Embodiment 3

[0057] Example 3: wxya Gene expression analysis under abiotic stress (high salt, osmotic) conditions

[0058] 1, wxya Genes are induced by high salt at the transcriptional level

[0059] Wild-type jujube seedlings were treated with 50 mM, 100 mM, 200 mM and 300 mM NaCl, respectively ( Figure 4 ), followed by extraction of RNA, after reverse transcription into cDNA, the wxya Gene expression analysis, the results showed that: compared with the control not treated with NaCl, wxya The induction of genes by salt stress is mainly manifested in the treatment of different concentrations of NaCl for the same time or the same concentration of NaCl for different times wxya The expression levels of the genes were significantly increased, and the expression level was the highest after treatment at 100 mM NaCl concentration for 48 h ( Figure 5 ).

[0060] , wxya Genes are induced by drought at the transcriptional level

[0061] The wild type jujube seedlings were treated wit...

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Abstract

The invention relates to the technical field of genetic engineering, particularly to a ziziphus jujuba SOD (superoxide dismutase) gene and application thereof. A nucleotide sequence of the ziziphus jujuba SOD gene is indicated in the SEQ ID (sequence identifier) NO (number):1, and an encoded amino acid sequence of the nucleotide sequence is indicated in the SEQ ID:2. The ziziphus jujuba SOD gene and the application thereof have the advantages that the ziziphus jujuba SOD gene is segregated from a ziziphus jujuba tree for the first time, a plant expression vector is constructed, a transgenic arabidopsis plant is obtained, transferring the ziziphus jujuba SOD gene as a target gene into the transgenic arabidopsis plant is practically significant to plant breed improvement, in-depth study of a mechanism of resistance of a high-resistance plant is benefited, molecular structures, expressions, functions and regulations of stress-related genes are defined beneficially, certain experimental data are provided and theoretical bases are established to further improve stress resistance of the plants by means of fully making use of high-resistance genes, understanding of functions of SOD in ROS (reactive oxygen species) signal transaction of the plants is facilitated, and people are helped to understand reactive oxygen serving as signal molecules.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a jujube superoxide dismutase gene and its application. Background technique [0002] Plants will produce a large amount of reactive oxygen species (such as H 2 o 2 , OH — , O 2 — ) cause protein denaturation, DNA mutation and membrane lipid oxidation, etc., which endanger normal physiological activities. In order to minimize the damage effect of reactive oxygen species, plants have evolved a series of enzymatic and non-enzymatic mechanisms to remove reactive oxygen species. Superoxide dismutases (SODs, EC1.15.1.1.) are a class of metalloenzymes widely present in plants, which can specifically remove superoxide anion radicals in biological oxidation, and disproportionate them to generate O 2 and H 2 o 2 . According to the metal cofactors in the active site of the enzyme, SOD can be divided into three categories: Cu / ZnSOD, FeSOD and Mn-SOD, Cu / ZnSOD protein ma...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12N9/02C12N15/82A01H5/00
Inventor 聂园军曹秋芬李建军肖蓉邓舒董艳辉薄晓峰李亚莉侯丽媛侯富恩
Owner 山西省农业科学院农业资源与经济研究所
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