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Application of TT1 gene in improving saline alkali tolerance of plants

A gene and plant technology, applied in the field of TT1 gene in improving plant salt-alkaline tolerance, to achieve the effect of improving plant salt-alkali tolerance, the method is simple and effective, and the content of proline is increased

Active Publication Date: 2013-12-04
SICHUAN BIODESIGN GENE ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still few reports on genes that improve plant salt-alkaline tolerance

Method used

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  • Application of TT1 gene in improving saline alkali tolerance of plants
  • Application of TT1 gene in improving saline alkali tolerance of plants
  • Application of TT1 gene in improving saline alkali tolerance of plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Cloning and acquisition of TT1 gene

[0035] Using the atp6 (genebank gi: 89279377) gene in rapeseed as the bait protein, an EST sequence (shown in SEQ ID NO: 3, encoded The amino acid sequence is shown in SEQ ID NO: 4), and then according to the sequence screened in this section, the method for improving plant and microbial resistance described in the present invention is obtained by 5' RACE (seeing the 5' RACE method data disclosed by Takara Company). The nucleotide sequence of the thermal gene is shown in SEQ ID NO: 1 in the sequence listing. Primers were designed according to the nucleotide sequence shown in SEQ ID NO: 1,

[0036] Upstream primer (SEQ ID NO: 7): 5'-ATGTCGGATCATTTGAGTTTATG-3',

[0037] Downstream primer (SEQ ID NO: 8): 5'-TCAGACTGGTGTTGGGTTGGATAT-3'.

[0038] Then the nucleotide sequence shown in SEQ ID NO: 1 is amplified from the rapeseed cDNA by PCR.

[0039] The PCR program is as follows:

[0040] 1.95℃ for 4min (pre-denaturation) ...

Embodiment 2

[0064] Example 2, Preparation of Transgenic Arabidopsis Plants and Obtaining of Seeds

[0065] 1. Obtaining transgenic Arabidopsis plants and seeds

[0066] Primers were designed according to the nucleotide sequence shown in SEQ ID NO: 1,

[0067] Upstream primer (SEQ ID NO: 13): 5'-CGC GGATCCATGTCGGATCATTTGAGTTTATG-3',

[0068] Downstream primer (SEQ ID NO: 14): 5'-CCGGAGC TCTCAGACTGGTGTTGGGTTGGATAT-3'.

[0069] Through PCR, the complete nucleotide sequence shown in SEQ ID NO: 1 is amplified from rapeseed cDNA,

[0070] The PCR program is as follows:

[0071] 1.95℃ for 4min (pre-denaturation)

[0072] 2.95℃ 30s (denaturation)

[0073] 3.53℃ 30s (refolding)

[0074] 4.72℃ 50s (extension)

[0075] 5.2~4 steps cycle 30 times

[0076] 6.72℃ for 5min (final extension)

[0077] Store at 7.4°C.

[0078] Purify the PCR product (see Qiagen’s PCR Product Purification Kit Instructions), then digest it with BamH1 and Sac1, recover from the gel, and connect it to the vector pBI1...

Embodiment 3

[0097] Embodiment 3, the influence of different concentrations of NaCl on the germination rate of Arabidopsis thaliana

[0098] Salts in saline-alkaline soils are usually NaCl, Na2SO, Na2CO, and NaHCO3, etc. Salt stress can not only reduce the water potential, but also cause ion stress caused by the increase of Na ions, which affects the absorption of nutrients such as K ions and Ca ions by plants. causing damage to plants. Therefore, NaCl was used to simulate salt stress conditions in this experiment.

[0099] The prepared MS medium (recipe in Table 1, pH adjusted to 5.8 with KOH) was added with NaCl before sterilization, so that the final NaCl concentrations were 0mmol / L (control group), 50mmol / L, 100mmol / L , 150mmol / L, 200mmol / L, 250mmol / L, 300mmol / L, after high-pressure steam sterilization, put them into petri dishes. After the medium is solidified, suspend the seeds with 2 mL of sterile water and transfer them to the medium. After the seeds are evenly sown, remove the e...

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Abstract

The invention relates to the technical field of organisms, in particular to application of a TT1 gene in improving the saline alkali tolerance of plants, aiming to solve the technical problem on how to provide a new effective selection for the technical field of transgenosis for improving the saline alkali tolerance of the plants. In order to solve the technical problem, the invention adopts the technical scheme that the application of the TT1 gene in improving the saline alkali tolerance of the plants is provided; and proved by experiments, the seed germination rate of the plants with the TT1 genes in saline alkali environment is obviously improved, and the content of proline in grown plants is also improved, which proves that the TT1 genes can effectively improve the saline alkali tolerance of the plants. A method for cultivating the plants with the saline alkali tolerance is also simple, convenient and effective and provides a new effective selection for improving the saline alkali tolerance of the plants.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of TT1 gene in improving the salt-alkaline tolerance of plants. Background technique [0002] With the rapid development of molecular biotechnology and the continuous improvement of gene cloning technology, the study of plant genetic engineering is developing in depth, and the research on resistance genes has changed from resistance to biological stress (such as disease resistance and insect resistance) to resistance to abiotic stress ( Such as cold resistance, drought resistance, salt resistance, etc.) transfer. [0003] At present, there are more than 100 countries in the world with different types of saline-alkali land and 1 billion hm 2 , about 10% of the world's arable land. The area of ​​saline-alkali land in my country reaches 99.13 million hm 2 , mainly distributed in arid and semi-arid regions such as North China, Northwest China and Northeast ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82
Inventor 杨毅
Owner SICHUAN BIODESIGN GENE ENG
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