New Application of Arabidopsis tt4 Gene in Plant Salt Tolerance

A technology of Arabidopsis and genes, which is applied in the field of genetic engineering applications and can solve problems such as variation

Active Publication Date: 2019-07-12
HENAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing studies have found that, TT4 The loss of flavonoid synthesis after gene mutation causes the variation of Arabidopsis seed coat color, and these polyketides play an important role in plant organ coloring, pest protection, and ultraviolet radiation resistance. However, other uses of this gene, especially There is no report on the research on plant resistance to salt

Method used

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  • New Application of Arabidopsis tt4 Gene in Plant Salt Tolerance
  • New Application of Arabidopsis tt4 Gene in Plant Salt Tolerance
  • New Application of Arabidopsis tt4 Gene in Plant Salt Tolerance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] This embodiment is mainly for TT4 The relationship between gene (At5g13930 gene) and plant seed vigor is briefly explained.

[0018] During the preliminary basic research process, the inventor believes that TT4 The gene (At5g13930 gene) is related to the vigor of plant seeds. Therefore, the inventors used Arabidopsis wild type (WT), tt4-2 and tt4-3 Mutants were taken as an example to verify this conclusion by comparative experiments. The related experimental process is introduced as follows.

[0019] Dried Arabidopsis wild-type (WT), tt4-2 and tt4-3 The mutant seeds were respectively placed in a solution containing triphenyltetrazolium chloride (TTC) at a temperature of 30°C, and dyed in the dark for 48 hours, and the coloring status of the seeds was observed and counted.

[0020] said tt4-2 and tt4-3 Mutants were obtained from the Arabidopsis Resource Center TT4 The gene (At5g13930 gene) was knocked out by gene knockout of three bases and one more base m...

Embodiment 2

[0024] On the basis of Example 1, the inventor thinks TT4 The vigor of the seeds can be improved after gene mutation, and its salt tolerance ability may also be improved, so the inventor has done further experimental verification. The related process is briefly introduced as follows.

[0025] Arabidopsis wild type (WT), tt4-2 and tt4-3 The mutant seeds were sown on MS medium containing 100 mM NaCl, 150 mM NaCl, and 200 mM NaCl respectively, and MS medium without adding other substances was set as a control, and the mass content of MS medium agar was 0.6%.

[0026] said tt4-2 and tt4-3 Mutant sources are the same as in Example 1.

[0027] After the seeds were sown, the cultivation environment was as follows: the light / dark cycle was 8 / 16h, the temperature was 22°C, the dark temperature was 18°C, the relative humidity was 70%, and the light intensity was 90 µmol·m -2 ·s -1 .

[0028] Specific observations were made on the germination process of Arabidopsis thaliana s...

Embodiment 3

[0032] Based on the research result of embodiment 2, the inventor thinks that it is necessary to TT4 The expression of genes in Arabidopsis tissues was further analyzed, and the relevant experimental procedures are briefly introduced as follows.

[0033] right TT4 When analyzing the expression of genes in Arabidopsis tissues, the technical scheme adopted is: TT4 The gene promoter was fused with the coding region of the GUS reporter gene on the pCAMBIA1391 vector to construct the pTT4-pCAMBIA1391 vector, and the transgenic plants transformed with wild-type Arabidopsis thaliana into TT4-GUS were obtained by using the method of Agrobacterium infecting Arabidopsis tidbits, and then right TT4 Gene expression in Arabidopsis tissues was analyzed in detail.

[0034] The specific experimental process is introduced as follows.

[0035] (1) Preparation of pTT4-pCAMBIA1391 vector

[0036] The At5g13930 gene promoter sequence was obtained from the Arabidopsis website (http: / / www.arabi...

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Abstract

The invention belongs to the technical field of genetic engineering application and particularly relates to application of an Arabidopsis thaliana TT4 gene (Arabidopsis thaliana genome code At5g13930) to culture of new varieties of salt-resistant plants. The TT4 gene with a CDS length being 1188bp encodes proteins of 395 amino acids and is related to seed activity, and the gene enables release of Arabidopsis thaliana seed activity after genetic mutation, namely the gene is capable of enhancing seed activity to accelerate seed germination and improve seed germinability after genetic mutation. The TT4 gene mainly serves as a first specific enzyme in a flavonoid synthesis route and participates in regulation of development of seed coats and seed coat pigments of Arabidopsis thaliana. According to researches of the TT4 gene in simulated dry or high-salinity adverse situations, plants under specific regulation of the gene has an osmotic stress reaction to salts and the like, and after silencing of the gene, mutant plants show favorable salt resistance at an initial germination stage.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering application, in particular to Arabidopsis thaliana TT4 The application of the gene (Arabidopsis thaliana genome coded as At5g13930) in breeding new varieties of salt-tolerant plants. Background technique [0002] At present, the shortage of global water resources is increasing day by day, and the extensive use of chemical fertilizers by humans has caused soil salinization to become increasingly serious, restricting the sustainable production and development of agriculture. With the deepening of molecular biology and genetic engineering research, many scientific and technological workers in the world are also trying to find genes related to plant salt resistance. They plan to improve the salt resistance of crops through genetic engineering technology, but there are very few achievements available at present. . [0003] Arabidopsis ( Arabidopsis thaliana ) belongs to the Brassicaceae...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/10C12N15/82A01H5/00A01H6/20
CPCC12N9/1037C12N15/8273C12Y203/01074
Inventor 张骁赵翔赵青平蔡欣媛王琳丹
Owner HENAN UNIVERSITY
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