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Salt tolerance gene hgs2 of halophytic grass and its application

A salt-growing grass and genetic technology, applied in the field of plant bioengineering and transgenics, can solve the problems of salt-tolerant gene mining and other reports that are rarely reported, and achieve the effect of easy genetic transformation operation

Active Publication Date: 2020-05-15
GANSU AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The halophytic plant Halogeton glomeratus is widely distributed in the saline lands in the arid regions of Northwest my country, and has strong adaptability to saline lands. It is an excellent source for excavating salt-tolerant genes. rarely reported

Method used

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  • Salt tolerance gene hgs2 of halophytic grass and its application
  • Salt tolerance gene hgs2 of halophytic grass and its application
  • Salt tolerance gene hgs2 of halophytic grass and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The preparation method of halophytic grass salt tolerance gene HgS2 described in embodiment 1, its main feature is that the steps are:

[0035] Using 200mM NaCl stress treatment for 7 days as the material, the total RNA was extracted by Trizol method, and the first strand of cDNA was synthesized by reverse transcription using the cDNA synthesis kit (Dalian Bao Biological Engineering Co., Ltd.), and designed according to the sequence of HgS2 The upstream and downstream primers F1 (5'-AAAAAATGGCACAATTTCAGCTCC-3') and R1 (5'-GTAGTAGAAATGCGTTGATGTCCA-3') (synthesized by Shanghai Sangon Bioengineering Co., Ltd.) amplify the gene fragment, and the PCR reaction system is 10×Ex Taq Buffer (Mg 2+ Plus) 2.5 μL, dNTP Mixture (each 2.5 mM) 2 μL, upstream primer (5'-AAAAAATGGCACAATTTCAGCTCC-3') (10 μM) 1 μL, downstream primer (5'-GTAGTAGAAATGCGTTGATGTCCA-3' (10 μM) 1 μL, cDNA 1 μL, Ex Taq Enzyme 0.2 μL, ultrapure water 17.3 μL, total volume 25 μL; the amplification program was 94°C...

Embodiment 2

[0042] Embodiment 2: the preparation method of the described halophytic grass salt-tolerant gene HgS2 is characterized in that the steps are:

[0043] Using 500mM NaCl stress treatment for 3-7 days as material, the total RNA was extracted by the Trizol method, and the first strand of cDNA was synthesized by reverse transcription using a cDNA synthesis kit, and the gene fragment was amplified. The PCR reaction system was as follows: 10×Ex Taq Buffer Mg 2+ Plus 2.5 μL, dNTP Mixture 2.5 mM each 2 μL, upstream primer F1 5'-AAAAAATGGCACAATTTCAGCTCC-3' 10 μM 1 μL, downstream primer R1 5'-GTAGTAGAAATGCGTTGATGTCCA-3' 10 μM 1 μL, cDNA 1 μL, Ex Taq enzyme 0.2 μL, ultrapure water 17.3 μL, The total volume is 25 μL; the amplification program is 94°C for 4 min, 94°C for 50 s, 60°C for 50 s, 72°C for 25 s, a total of 32 cycles, and 72°C for 7 min; use the gel recovery kit to recover the PCR product, and follow the steps in the manual: first, agarose Use the gel to detect the size of the ta...

Embodiment 3

[0044] Embodiment 3: the construction method of the expression vector of described halophytic grass salt tolerance gene HgS2, its steps are:

[0045] Two restriction sites, Xba I and Sma I, were respectively added to the upstream and downstream primers of the salt-tolerant gene HgS2 of the halophytic grass, the upstream primer F1 was 5'-GCTCTAGAAAAAAATGGCACAATTTCAGCTCC-3', and the downstream primer R1 was 5'- CGACCCGGGGTAGTAGAAATGCGTTGATGTCCA-3':

[0046] Using 200-500mM NaCl stress treatment for 3-7 days as material, the total RNA was extracted by the Trizol method, and the first strand of cDNA was synthesized by reverse transcription using a cDNA synthesis kit, and the gene fragment was amplified, and the PCR reaction system was used. 10×Ex Taq Buffer Mg 2+Plus 2.5 μL, dNTP Mixture 2.5 mM each 2 μL, upstream primer 5’-GCTCTAGAAAAAAATGGCACAATTTCAGCTCC-3’ 10 μM 1 μL, downstream primer 5’-CGACCCGGGGTAGTAGAAATGCGTTGATGTCCA-3’ 10 μM 1 μL, cDNA 1 μL, Ex Taq enzyme 0.2 μL, ultrapu...

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Abstract

The invention relates to a salt tolerant gene HgS2 derived from halophyte halogeton glomeratus, and application thereof. The invention aims at providing a novel salt tolerant gene HgS2, encoding protein of the salt tolerant gene HgS2, and application of the salt tolerant gene HgS2 to improve the plant salt tolerance and culture a salt tolerant new variety (strain). The salt tolerant gene HgS2 contains a nucleotide sequence of SEQ ID NO.1cDNA, and the molecular weight is 678bp; the cDNA coding sequence is the nucleotide sequence from the 30th site to the 290th site in the SEQ ID NO.1, and the molecular weight is 261bp; the amino acid sequence is shown as SEQ ID NO.3 and consists of 86 amino acids. The salt tolerant gene HgS2 can obviously improve the salt tolerance of arabidopsis thaliana plants. The salt tolerant gene provided by the invention is favorable for the culture of salt tolerant crops and plant new varieties (strains).

Description

technical field [0001] The invention belongs to the field of plant bioengineering and transgenic technology, and specifically relates to a salt-tolerant gene of a halophyte halophyte and an application thereof. Background technique [0002] Soil salinization has become a major factor limiting global food production. It is estimated that 360 million hectares of dryland arable land worldwide are suffering from soil erosion, soil degradation and salinization, and 230 million hectares of irrigated arable land are also affected by salinization. Moreover, the trend of global warming in the next few decades will cause annual precipitation in subtropical regions to decrease year by year, and the contradiction between supply and demand of water resources will further intensify. The brutal reality of brackish or salt water irrigation. [0003] Improving the salt tolerance of crops is the key to solving the problem of soil salinization. This requires crop breeding to make breakthrou...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/10C12N15/82C12N15/66A01H5/00A01H6/20
CPCC07K14/415C12N15/1096C12N15/66C12N15/8273C12Q2531/113
Inventor 王化俊汪军成姚立蓉李葆春孟亚雄马小乐任盼荣司二静杨轲邹兰闫栋张燕
Owner GANSU AGRI UNIV
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