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Sesame drought resisting and salt-tolerant gene SiMYB75 as well as encoded protein and application thereof

A salt-tolerant gene and sesame technology, applied in the field of genetic engineering, can solve the problems of weak abiotic stress research and few molecular-related research on sesame, improve drought resistance and salt tolerance, ensure high and stable yield, and improve drought resistance and salt tolerance. Effect

Inactive Publication Date: 2019-02-15
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The research on the abiotic stress of sesame is relatively weak, and there are few molecular related studies

Method used

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  • Sesame drought resisting and salt-tolerant gene SiMYB75 as well as encoded protein and application thereof
  • Sesame drought resisting and salt-tolerant gene SiMYB75 as well as encoded protein and application thereof
  • Sesame drought resisting and salt-tolerant gene SiMYB75 as well as encoded protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Acquisition of Sesame Drought Resistance and Salt Tolerance Gene SiMYB75 Gene

[0032] 1. Discovery of the drought-resistant and salt-tolerant gene SiMYB75 in sesame

[0033] 1. A pair of sesame drought-resistant material ZZM0635 and drought-sensitive material ZZM4728 were selected, potted and planted in an arid shed, and subjected to drought stress treatment at the initial flowering stage (47 days after sowing). Take d0 (soil moisture content 35%), d1 (soil moisture content 15%), d2 (soil moisture content 9%), d3 (soil moisture content 6%) and d4 (soil moisture content 35% after rehydration)5 The leaves of two materials at each time point were used for drought-resistant transcriptome analysis;

[0034] 2. Use the Illumina Hiseq4000 sequencing platform to perform transcriptome sequencing on 30 samples, and each sample produces an average of 6.86Gb of data. After aligning the sequencing reads to the sesame reference genome and reconstructing the transcripts, ...

Embodiment 2

[0041] Example 2 Construction and genetic transformation of SiMYB75 gene expression vector

[0042] 1. Construction of overexpression vector

[0043] The drought-resistant and salt-tolerant related sesame gene SiMYB75 that embodiment 1 is cloned utilizes the method for homologous recombination and pCAMBIA1301S (provided by this laboratory) plasmid connection construction plant expression vector, named after pCAMBIA1301S-SiMYB75 ( figure 1 ), the specific operations are as follows:

[0044] 1. First, the linearized vector was obtained by double enzyme digestion (BamHI and KpnI) (Takara), and then purified by agarose gel electrophoresis and gel recovery kit (Tiangen Biochemical Technology Co., Ltd.) to obtain a high-purity linearized vector.

[0045] 2. Add the DNA of the target fragment and the linearized carrier to a 1.5ml centrifuge tube at a molar ratio of 3:1 for recombination reaction, mix well and place it at 37°C for about 30min, add 10μl of the reaction solution to 50μ...

Embodiment 3

[0082]Implementation 3 Determination of Drought Resistance and Salt Tolerance of Positive Plants of SiMYB75 Overexpression Transgenic T2 Generation Families

[0083] 1. Determination of drought resistance of T2 generation positive Arabidopsis plants

[0084] After the transgenic plants of the T2 generation grew to the stage of three pairs of leaves, the transgenic Arabidopsis plants and the non-transgenic wild-type Arabidopsis plants were subjected to drought stress treatment for 17 days. The results showed that compared with the wild-type Arabidopsis control, the drought resistance of the SiMYB75-transgenic Arabidopsis lines obtained in this study was significantly higher than that of the wild-type Arabidopsis control ( Figure 8 ), indicating that the overexpression of SiMYB75 gene in sesame can improve the drought resistance of plants.

[0085] 2. Determination of Salt Tolerance of T2 Generation Positive Arabidopsis Plants

[0086] After the transgenic plants of the T2 ge...

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Abstract

The invention belongs to the technical field of genetic engineering and provides a sesame drought resisting and salt-tolerant gene SiMYB75 as well as encoded protein and application thereof. The sesame drought resisting and salt-tolerant gene SiMYB75 is a nucleotide sequence shown as SEQ ID NO:1 or a nucleotide sequence with drought resistance and salt tolerance, generated by adding, displacing, inserting or deleting one or more nucleotides. The gene SiMYB75 disclosed by the invention is constructed to an expression vector and is transferred into arabidopsis by a agrobacterium tumefaciens medicated transformation method to obtain transgenic arabidopsis; by overexpression of the drought resisting and salt-tolerant gene SiMYB75 in the arabidopsis, the gene is found to remarkably improve thedrought resistance and the salt tolerance of plants. Therefore, the sesame drought resisting and salt-tolerant gene SiMYB75 has a good application prospect in improving the properties of the plants, such as the drought resistance and the salt tolerance.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to the sesame drought-resistant and salt-tolerant gene SiMYB75 and its coded protein and application. Background technique [0002] The use of high-quality oils in the daily human diet is an important part of overall health. Sesame (Sesamumindicum L.) is one of the oldest oil crops, widely grown in tropical and subtropical regions. Sesame oil is a high-quality vegetable oil rich in important minerals and vitamins, and is rich in unique classes of substances such as phytosterols, polyunsaturated fatty acids, tocopherols and lignin, such as sesamin, which has been identified as a compound that has beneficial effects on human health. In addition, among the main oil crops, sesame seeds have the highest oil content (about 55%), and also contain higher protein content (about 27%). The world's population is growing rapidly, and the demand for vegetable oil is also increasing ...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/10C12N15/82A01H5/00A01H6/20
CPCC07K14/415C12N15/1096C12N15/8273C12Q2531/113
Inventor 黎冬华山姆游均玛丽周瑢刘爱丽张秀荣
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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