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Novel application of Arabidopsis thaliana TT4 gene to salt resistance of plants

An Arabidopsis thaliana, new application technology, applied in the field of genetic engineering applications, can solve problems such as variation

Active Publication Date: 2016-06-08
HENAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing studies have found that the loss of flavonoid synthesis after TT4 gene mutation causes the variation of Arabidopsis seed coat color, and these polyketide compounds play an important role in plant organ coloring, pest protection, and ultraviolet radiation resistance. Other uses of the gene, especially studies on plant salt resistance have not been reported

Method used

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  • Novel application of Arabidopsis thaliana TT4 gene to salt resistance of plants
  • Novel application of Arabidopsis thaliana TT4 gene to salt resistance of plants
  • Novel application of Arabidopsis thaliana TT4 gene to salt resistance of plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] This example mainly briefly describes the relationship between the TT4 gene (At5g13930 gene) and the vigor of plant seeds.

[0018] In the previous basic research process, the inventor believed that the TT4 gene (At5g13930 gene) was related to the vigor of plant seeds. For this reason, the inventor took Arabidopsis wild type (WT), tt4-2 and tt4-3 mutants as examples for this conclusion A comparative experiment was carried out to verify. The related experimental process is introduced as follows.

[0019] The dried Arabidopsis wild-type (WT), tt4-2 and tt4-3 mutant seeds were respectively placed in a solution containing triphenyltetrazolium chloride (TTC) at a temperature of 30°C and darkened for 48 hours. Observe and count the coloring situation of the seeds.

[0020] The tt4-2 and tt4-3 mutants are mutants of the TT4 gene (At5g13930 gene) obtained from the Arabidopsis Resource Center by knocking out three bases and one more base respectively (the specific mutation con...

Embodiment 2

[0024] On the basis of Example 1, the inventor believes that the mutated TT4 gene can improve the vigor of the seed, and may also improve its salt tolerance, 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 mutant seeds were sown on MS medium containing 100mMNaCl, 150mMNaCl, and 200mMNaCl, respectively, and MS medium without adding other substances was set as a control, MS culture The mass content of base agar is 0.6%.

[0026] The sources of the tt4-2 and tt4-3 mutants are the same as in Example 1.

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

[0028] Specific observations were made on the germination process of Arabidopsis thaliana seeds and the germination rate of the seeds was co...

Embodiment 3

[0032] Based on the research results of Example 2, the inventor believes that it is necessary to further analyze the expression of the TT4 gene in Arabidopsis tissues, and the relevant experimental procedures are briefly introduced as follows.

[0033] When analyzing the expression of the TT4 gene in Arabidopsis tissues, the technical scheme adopted is: fuse the TT4 gene promoter with the coding region of the GUS reporter gene on the pCAMBIA1391 vector to construct the pTT4-pCAMBIA1391 vector, and use agricultural The method of Bacillus infecting Arabidopsis thaliana tidbits obtained the transgenic plants transformed from wild-type Arabidopsis thaliana into TT4-GUS, and then specifically analyzed the expression of TT4 gene in Arabidopsis tissues.

[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.arabidopsis....

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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, and specifically relates to the application of Arabidopsis thaliana TT4 gene (the Arabidopsis thaliana genome code is At5g13930) in cultivating 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 thaliana (Arabidopsisthaliana) belongs to ...

Claims

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Application Information

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IPC IPC(8): C12N9/10C12N15/82A01H5/00
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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