Sesame drought-resistant gene SiSAM1 and application thereof

A sesame and genetic technology, applied in the sesame drought-resistant gene SiSAM1 and its application field, can solve the problems of weak drought resistance of sesame and few molecular related studies, and achieve the effect of improving drought resistance, improving drought resistance, and improving drought resistance

Inactive Publication Date: 2019-02-05
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 drought resistance of sesame is relatively weak. The research reports mainly focus on the impact of drought on the morpholog

Method used

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  • Sesame drought-resistant gene SiSAM1 and application thereof
  • Sesame drought-resistant gene SiSAM1 and application thereof
  • Sesame drought-resistant gene SiSAM1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0028] Example 1 Obtaining the Drought Resistance Gene SiSAM1 Gene in Sesame

[0029] 1. Discovery of Drought Resistance Gene SiSAM1 in Sesame

[0030] 1. Based on the phenotype, geographic origin and genetic diversity test results of drought-related traits from 7910 domestic and foreign resources preserved in the National Sesame Medium-term Bank, a gradual sampling strategy was adopted, and 400 sesame materials with different drought resistance were selected for reconstitution. Sequencing analysis;

[0031] 2. Using the Illumina Hiseq2500 sequencing platform, 400 pieces of sesame material were re-sequenced with low coverage by the 2×76 paired-end sequencing method, and the genome sequence with 2.6 times coverage was obtained;

[0032] 3. Combining the drought-resistant traits data, genotype data and population structure of the drought-resistant populations of germplasm resources, using EMMAX software package and Peal program to perform genome-wide association analysis on sesame-relat...

Example Embodiment

[0038] Example 2 Construction and genetic transformation of SiSAM1 gene overexpression vector

[0039] 1. Construction of overexpression vector

[0040] The drought-resistant-related sesame gene SiSAM1 cloned in Example 1 was ligated with the plasmid pCAMBIA1301S (provided by our laboratory) by homologous recombination to construct a plant expression vector, named pCAMBIA1301S-SiSAM1( figure 1 ), the specific operation is as follows:

[0041] 1. First, use the double enzyme digestion (BamHI and KpnI) (Takara) method to obtain the linearized vector, and then purify it by agarose gel electrophoresis and gel recovery kit (Tiangen Biochemical Technology Co., Ltd.) to obtain the high-purity linearized vector.

[0042] 2. Add the target fragment DNA and linearized vector to a 1.5ml centrifuge tube at a molar ratio of 3:1 for recombination. After mixing, place it at 37°C for about 30 minutes, and add 10μl of the reaction solution to 50μl of DH5a competence. In the cells, mix gently with a pi...

Example

[0074] Implementation 3 Determination of Drought Resistance of Positive Plants of SiSAM1 Overexpression Transgenic T2 Generation Family

[0075] Three strains of SiSAM1 gene overexpression transgenic T2 generation family plants in Example 2 were selected for drought stress experiments. The specific steps are as follows: Plant 3 plants for each material in a nutrient bowl (the T2 generation transgenic plants will grow to 3 pairs of leaves) and 3 untransgenic wild-type Arabidopsis plants. When the plants start to boot, the water is cut off and drought stress is applied 17 Day, then observe the growth status of the transgenic plants.

[0076] The results showed that the growth of the three transgenic plants overexpressing the SiSAM1 gene cloned in the present invention was slightly better than that of the control plant under normal conditions, but the difference was not much, but after the stress treatment, the control non-transgenic plants were obviously The ground wilted before the...

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Abstract

The invention belongs to the technical field of gene engineering, and provides a sesame drought-resistant gene SiSAM1 and application thereof. The sesame drought-resistant gene SiSAM1 is a nucleotidesequence as shown by SEQ ID NO.1; or a nucleotide sequence with drought-resistant performance generated by adding, substituting, inserting or omitting on or more nucleotide sequences of the nucleotidesequence as shown by SEQ ID NO.1. The SiSAM1 gene is established onto an expression vector, the SiSAM1 gene is transferred into Arabidopsis by virtue of agrobacterium-mediated conversion method to obtain transgenic agrobacterium; the transgenic agrobacterium and non-transgenic agrobacterium are simultaneously treated in a drought stress manner, and comparing the transgenic agrobacterium with thenon-transgenic agrobacterium, the expression and drought-resistant capacity of SiSAM1 can be greatly improved. By adopting the sesame drought-resistant gene SiSAM1, the drought-resistant capacity of plants can be improved, and the sesame drought-resistant gene SiSAM1 can be used for the breeding of an oil crop drought-resistant variety, so that the drought-resistant performance of crops can be improved, and the high yield and stable yield of the crops can be ensured.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to the sesame drought-resistant gene SiSAM1 and its application. Background technique [0002] Sesamum (Sesamum indicum L.) is an important oil crop, mainly distributed in tropical and subtropical regions from 40°N to 40°S. Due to the high nutritional value of sesame seeds (oil content 40%-62.7%; protein content 19%-30%), sesame has the reputation of "queen of oil". Sesame is one of the five major oil crops in my country and is a traditional characteristic agricultural product. As a major producer and trading country, my country's annual consumption ranks first in the world, but its self-sufficiency is less than 50%, and its import rate is increasing year by year. [0003] Due to the combined effects of biotic and abiotic stresses, the annual yield of sesame is low and unstable, which is also a major bottleneck in the development of the sesame industry. Sesame is main...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82C07K14/415A01H5/00A01H6/20
CPCC07K14/415C12N15/8273
Inventor 黎冬华山姆张秀荣周瑢玛丽刘爱丽游均魏鑫张艳欣王林海
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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