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Halogeton glomeratus salt-tolerant gene HgS3 and applications thereof

A salt-tolerant gene and salt-growing grass technology, applied in the field of plant bioengineering and transgenics, to achieve the effect of excellent salt-tolerant characteristics

Active Publication Date: 2017-10-24
GANSU AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are few studies on the excavation of salt-tolerant genes of native halophytes in the arid regions of Northwest China, especially the salt-tolerant pioneer plant Halogeton glomeratus. The discovery of excellent salt-tolerant genes is of great importance for the cultivation of salt-tolerant plant (crop) materials. use value

Method used

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  • Halogeton glomeratus salt-tolerant gene HgS3 and applications thereof
  • Halogeton glomeratus salt-tolerant gene HgS3 and applications thereof
  • Halogeton glomeratus salt-tolerant gene HgS3 and applications thereof

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Embodiment 1

[0033] The preparation method of halophytic grass salt tolerance gene HgS3 described in embodiment 1, its main feature is that the steps are:

[0034] Using 200mM NaCl treatment of 3-7d leaf tissue of Saltwort seedlings as material, total RNA was extracted by Trizol method, the first strand of cDNA was synthesized by reverse transcription using a cDNA synthesis kit (Dalian Bao Biological Engineering Co., Ltd.), and the amplified Gene fragment, PCR reaction system: 5×PrimeSTAR Buffer 5 μL, dNTP Mixture (2.5 mM each) 2 μL, upstream primer F15’-AAAAGACACTCCATAATCTTGTGTT-3’ (10 μM) 1 μL, downstream primer R1 5’-TTTTATTGATGCAAATAAGCTACTA-3’ (10 μM) 1 μL (Synthesized by Shanghai Sangon Bioengineering Co., Ltd.), 1 μL of cDNA, 0.25 μL of PrimeSTAR HS DNA polymerase, 14.75 μL of ultrapure water, and a total volume of 25 μL; the amplification program was pre-denaturation at 94°C for 4 min, 94°C for 50 s, 60°C for 20 s, 72°C for 1min, a total of 32 cycles, 72°C for 8min, the PCR amplifi...

Embodiment 2

[0040] Embodiment 2: the preparation method of the described halophytic grass salt tolerance gene HgS3 is characterized in that the steps are:

[0041] Using 500mM NaCl treatment of 3-7d leaf tissue of Saltwort seedlings as material, total RNA was extracted by Trizol method, the first strand of cDNA was synthesized by reverse transcription using a cDNA synthesis kit (Dalian Bao Biological Engineering Co., Ltd.), and the amplified Gene fragment, PCR reaction system: 5×PrimeSTAR Buffer 5 μL, dNTP Mixture (2.5 mM each) 2 μL, upstream primer F15’-AAAAGACACTCCATAATCTTGTGTT-3’ (10 μM) 1 μL, downstream primer R1 5’-TTTTATTGATGCAAATAAGCTACTA-3’ (10 μM) 1 μL (synthesized by Shanghai Sangon Bioengineering Co., Ltd.), 1 μL of cDNA, 0.25 μL of PrimeSTAR HS DNA polymerase, 14.75 μL of ultrapure water, and a total volume of 25 μL; the amplification program was pre-denaturation at 94°C for 4 min, 94°C for 50 s, 60.4°C for 15 s, 72°C for 1min, a total of 35 cycles, 72°C for 8min, the PCR ampl...

Embodiment 3

[0042] Embodiment 3: the construction method of the expression vector of described halophytic grass salt tolerance gene HgS3, its steps are:

[0043] Two restriction sites, BamHI and SacI, were added to the upstream and downstream primers of the salt-tolerant gene HgS3 of the halophytic grass, respectively. The upstream primer F1 was 5'-CGGGATCCAAAAGACACTCCATAATCTTGTGTT-3', and the downstream primer R1 was 5'-CGAGCTCTTTTATTGATGCAAATAAGCTACTA- 3' (synthesized by Shanghai Sangon Bioengineering Co.):

[0044] Using 200mM NaCl treatment of 3-7d leaf tissue of Saltwort seedlings as material, total RNA was extracted by Trizol method, the first strand of cDNA was synthesized by reverse transcription using a cDNA synthesis kit (Dalian Bao Biological Engineering Co., Ltd.), and the amplified Gene fragment, PCR reaction system: 5×PrimeSTAR Buffer 5 μL, dNTP Mixture (2.5 mM each) 2 μL, upstream primer F1 5’-CGGGATCCAAAAGACACTCCATAATCTTGTGTT-3’ (10 μM) 1 μL, downstream primer R1 5’-CGAGCT...

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Abstract

The present invention relates to a salt-tolerant gene HgS3 isolated from a halophyte halogeton glomeratus, and applications thereof. A purpose of the present invention is to provide a new salt-tolerant gene HgS3 and an encoding protein thereof, and applications of the salt-tolerant gene HgS3 in improvement of the salt tolerance of plants and cultivation of new salt-tolerant varieties (lines). According to the present invention, the salt-tolerant gene HgS3 comprises the cDNA nucleotide sequence represented by SEQ ID NO.1, and has the molecular weight of 468 bp, the cDNA coding sequence is a site 208-site 336 nucleotide sequence represented by SEQ ID NO.1, the molecular weight is 129 bp, and the amino acid sequence is represented by SEQ ID NO.3, and comprises 42 amino acids; the salt-tolerant gene HgS3 can significantly improve the salt tolerance of transgenic Arabidopsis seedlings, and can contribute to the cultivation of new salt-tolerant plant (crop) varieties (lines).

Description

technical field [0001] The invention belongs to the field of plant bioengineering and transgenic technology, and in particular relates to a salt-tolerant gene of a halophyte halophyte and its application in improving plant salt tolerance. Background technique [0002] Soil salinization is one of the most important abiotic stresses affecting crop growth and yield. With the gradual warming of the global climate, the process of soil salinization has been exacerbated, especially for the vast arid and semi-arid areas in Northwest my country. The originally extremely limited rainfall has been further reduced and evaporation has been further strengthened, resulting in soil The situation of salinization is even more serious, and due to the excessive irrigation of traditional agriculture, only irrigation without drainage has caused large areas of valuable arable land resources to be swallowed up by salinization, and eventually had to be abandoned by agricultural production and gradual...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/10C12N15/82C12N15/66A01H5/00
CPCC07K14/415C12N15/1096C12N15/66C12N15/8273
Inventor 汪军成王化俊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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