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
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Embodiment 1
[0028] Example 1 Acquisition of Sesame Drought Resistance Gene SiSAM1 Gene
[0029] 1. Discovery of the drought resistance gene SiSAM1 in sesame
[0030] 1. From the 7910 domestic and foreign resources preserved in the National Sesame Medium-term Bank, according to the phenotype, geographical origin and genetic diversity detection results of drought resistance-related traits, a step-by-step sampling strategy was adopted, and 400 sesame materials with different drought resistance were selected for re-analysis. Sequencing analysis;
[0031] 2. Using the Illumina Hiseq2500 sequencing platform, the 2×76 double-end sequencing method was used to resequence the whole genome of 400 sesame materials with low coverage, and obtained a genome sequence with 2.6 times coverage;
[0032] 3. Combining the data of drought-resistance-related traits, genotype data and population structure in the drought-resistant population of germplasm resources, the EMMAX software package and Peal program wer...
Embodiment 2
[0038] Example 2 Construction and genetic transformation of SiSAM1 gene overexpression vector
[0039] 1. Construction of overexpression vector
[0040]The drought-resistance related sesame gene SiSAM1 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-SiSAM1 ( figure 1 ), the specific operations are as follows:
[0041] 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.
[0042] 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 50μl of DH5a compete...
Embodiment 3
[0074] Implementation 3 Determination of Drought Resistance of Positive Plants of SiSAM1 Overexpression Transgenic T2 Generation Families
[0075] Three transgenic T2 generation family plants overexpressing the SiSAM1 gene in Example 2 were selected to carry out the drought stress experiment. The specific steps are as follows: plant 3 plants of each material in the nutrient pot (when the T2 generation transgenic plants grow to the stage of 3 pairs of leaves) and 3 plants of non-transgenic wild-type Arabidopsis thaliana plants, when the plants start to boot, the water is cut off to carry out drought stress for 17 days, and then observe the growth status of the transgenic plants.
[0076] The results show that the growth of the 3 transgenic plants overexpressed by the SiSAM1 gene cloned by the present invention is slightly better than that of the control plant under normal conditions but the difference is not very large, but after the adversity stress treatment, the growth of th...
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