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The application of hmt1 gene of wheatgrass

A technology of HMT1, Jiejiemai, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problem that gene function research has not been reported yet.

Active Publication Date: 2021-01-19
CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The sequence of the HMT1 gene has been published, but the functional study of the gene has not yet been reported

Method used

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  • The application of hmt1 gene of wheatgrass
  • The application of hmt1 gene of wheatgrass
  • The application of hmt1 gene of wheatgrass

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Materials: The diploid Ae.tauschii ssp.strangulataAS2407 was provided by Sichuan Agricultural University. Three liters of wild-type tobacco was used for the experiment. The above materials were propagated and preserved by the Key Laboratory of Crop Molecular Breeding in Qinghai Province.

[0037] Select AS2407 seeds with full grains and put them into a petri dish, add distilled water, and spring flowers in a low-temperature room in a refrigerator at 4°C for 2 days. After germination, transplant them into flower pots and cultivate them in a light cultivation room. After one week, randomly pick leaves. RNA was extracted after the samples were snap-frozen in liquid nitrogen.

[0038] Using Agrobacterium tumefaciens LBA4404 to mediated genetic transformation of HMT1 gene into tobacco, the physiological characteristics of transgenic tobacco were analyzed. Specific implementations such as figure 1 .

[0039] Access to genetically modified tobacco

[0040] 1 Specific enzyme...

Embodiment 2

[0052] Establishment of Genetic Transformation System of Tobacco with HMT1 Gene in Transjoint Wheat

[0053]1 Transformation of Agrobacterium tumefaciens

[0054] Transform the constructed eukaryotic expression vector (named pBI-AetHMT1) into LBA4404 Agrobacterium, the detailed steps are as follows:

[0055] (1) Take LBA4404 Agrobacterium competent cells stored at -80°C and melt them in an ice-water bath.

[0056] (2) Under sterile conditions, add 1 μg of recombinant plasmid pBI-AetHMT1 to the competent cell suspension just thawed. Mix gently, and let stand in an ice-water bath for 10 min.

[0057] (3) Place the centrifuge tube in liquid nitrogen for quick freezing for 5 minutes.

[0058] (4) Then quickly place the centrifuge tube in a 37°C water bath for 5 minutes.

[0059] (5) Put the centrifuge tube back into the ice bath for 5 minutes.

[0060] (6) Add 800 μL of antibiotic-free LB liquid medium under sterile conditions, and shake at 28° C. for 3 hours.

[0061] (7) C...

Embodiment 3

[0082] A Preliminary Study on Stress Resistance of Tobacco with HMT1 Gene in Transjoint Wheat

[0083] Planting of Aseptic Tissue Cultured Seedlings of Tobacco and Determination of Moisture Content

[0084] On the ultra-clean workbench, a proper amount of wild-type tobacco (three liters) and transgenic tobacco seeds with plump seeds were randomly selected, placed in a 1.5mL EP (eppendorf) tube, and sterilized with 75% ethanol in a sterile ultra-clean workbench. sterilized for 1 min, discarded the waste liquid, washed twice with sterile water, sterilized with 20% sodium hypochlorite solution for 10 min, discarded the waste liquid, washed with sterile water for 6 times, blotted the water on the seed surface with sterile filter paper, and placed in MS A number of wild-type tobacco (three liters) and AetHMT1 gene transgenic tobacco were cultivated on solid medium (without other elements). At the three-leaf one-heart stage, select the tobacco with uniform growth, transfer it to a ...

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Abstract

The present invention relates to use of Aegilops tauschii HMT (AetHMT1) gene, and belongs to the technical field of functional identification of genes. The present invention provides the use of AetHMT1 gene in improving plant stress resistance, total antioxidant capacity, selenium tolerance, and plant selenium absorptive and accumulating capacities. The AetHMT1 gene improves plant stress resistance, total antioxidant capacity, and selenium tolerance, and enhances plant selenium absorptive and accumulating capacities.

Description

technical field [0001] The invention relates to the technical field of gene function identification, in particular to the application of the HMT1 gene of barley barley. Background technique [0002] Selenocysteine ​​methyltransferase (SMT) is considered to be a methylselenocysteine ​​methyltransferase that can specifically catalyze the production of non-protein selenocysteine ​​(SeCys) with methanol methylation, Thereby reducing the concentration of selenocysteine ​​and selenomethionine in the cells. The SMT protein of Astragalus bisulcatus has more than 40% homology with Escherichia coli yagD, and the latter protein has the ability to methyl-catalyze homocysteine, selenhomocysteine ​​and selenocysteine. [0003] Comparing the amino acid sequence of Astragalus SMT in various databases, it was found that the similarity between this sequence and homocysteine ​​methyltransferase (Homocysteine ​​S-Methyltransferase, HMT, EC 2.1.1.10) of Arabidopsis thaliana and rice reached 70%...

Claims

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Application Information

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IPC IPC(8): C12N15/54C12N15/84A01H5/00A01H5/10A01H6/82
CPCC12N9/1007C12N15/8243C12N15/8271C12Y201/0101
Inventor 张波张怀刚吴丽军刘宝龙刘韬周让让陈红
Owner CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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