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Aluminum-resistant related gene GsERF1 and encoding protein and application thereof

A protein and coding technology, which is applied to the aluminum resistance-related gene GsERF1 and its encoded protein and application fields, can solve problems such as differences in the ability to resist aluminum toxicity, and achieve the effect of increasing relative elongation.

Pending Publication Date: 2021-06-11
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, within a species, plant species and cultivars vary widely in resistance to Al toxicity

Method used

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  • Aluminum-resistant related gene GsERF1 and encoding protein and application thereof
  • Aluminum-resistant related gene GsERF1 and encoding protein and application thereof
  • Aluminum-resistant related gene GsERF1 and encoding protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1. Cloning and expression of soybean aluminum tolerance-related gene GsERF1

[0045] 1. Cloning of soybean aluminum tolerance-related gene GsERF1

[0046] The root total RNA of wild soybean material BW69 (aluminum-resistant wild soybean resource) was extracted with a plant total RNA extraction kit (TR02, GeneMark), and its integrity was detected by 1% agarose electrophoresis.

[0047] The total RNA was reverse transcribed to obtain cDNA, and the cDNA was synthesized according to PrimeScript TM RT reagent Kit with gDNA Eraser kit manual operation.

[0048] Design the following primers:

[0049] Sequence 3: 5'-ATGGAGAAAGAGAGAGGAGAG-3'

[0050] Sequence 4: 5'-TTAGTCTTCGTTATTCCTTC-3'

[0051] Using the above cDNA as a template, PCR amplification is performed with the primers shown in sequence 3 and sequence 4 to obtain a PCR amplification product.

[0052] The above PCR amplification system is a PCR reaction solution (50 μl system) configured in the following o...

Embodiment 2

[0075] Embodiment 2, the acquisition of transgenic GsERF1 Arabidopsis

[0076] 1. Transgenic GsERF1 Arabidopsis

[0077] 1. Obtaining the recombinant vector

[0078] The recombinant plant expression vector pTF101.1-GsERF1 is a plasmid obtained by replacing the fragment between the Xba I and Sac I restriction sites of the pTF101.1 vector with the GsERF1 gene shown in Sequence 1, and the full-length sequence of GsERF1 is inserted forward to the CaMV35S promoter.

[0079] 2. Recombinant bacteria

[0080] The recombinant vector pTF101.1-GsERF1 was transformed into Agrobacterium EHA105 (BioVector NTCC Inc.) with a Bio-rad electric shock transformation instrument to obtain recombinant Agrobacterium EHA105 / pTF101.1-GsERF1, and PCR was performed using sequence 5 and sequence 6 primer pairs Amplified, 126bp is positive recombinant bacteria.

[0081] 3. Transgenic GsERF1 Arabidopsis

[0082]Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method[J]...

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Abstract

The invention discloses an aluminum-resistant related gene GsERF1 and an encoding protein and application thereof. The protein provided by the invention is named as protein GsERF1 and is as shown in the following (1) or (2): (1) a protein consisting of an amino acid sequence shown in a sequence 2 in a sequence table; and (2) a protein which is derived from (1), has the same function and is obtained by substitution and / or deletion and / or addition of one or more amino acid residues on the amino acid sequence as shown in the sequence 2 in the sequence table. The gene GsERF1 provided by the invention has a positive effect on aluminum stress resistance of soybeans. Experiments prove that the relative elongation of the main root of the transgenic arabidopsis thaliana can be improved and the proline content can be increased when the gene is over-expressed in the arabidopsis thaliana, which indicates that the protein can lay a foundation for the research on the cultivation of transgenic plants with stronger aluminum resistance.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to an aluminum resistance-related gene GsERF1 and its encoded protein and application. Background technique [0002] Aluminum toxicity is a major factor limiting crop production in acidic soils, which account for approximately 40% of the world's arable land and 70% of potentially arable land. When the soil pH value is below 5.0, aluminum will exist in the form of ions, Al 3+ Dissolved into the soil, strongly inhibits root growth and function, thereby reducing crop yields. However, within a species, resistance to Al toxicity varies considerably between plant species and cultivars. To survive in acidic soils, some plant species or varieties have evolved higher levels of tolerance mechanisms. In southern my country, wild soybeans grow in acidic soil for a long time, which is likely to form a strong aluminum tolerance. Therefore, wild soybean is an important ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/54
CPCC07K14/415C12N15/8271
Inventor 马启彬李璐年海程艳波蔡占东李新岗
Owner SOUTH CHINA AGRI UNIV
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