Aluminum-resistant related gene GmNAC069 and encoding protein and application thereof
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A protein and encoding technology, applied in the field of aluminum resistance related gene GmNAC069 and its encoded protein and application
Active Publication Date: 2021-06-04
SOUTH CHINA AGRI UNIV
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[0004] So far, no gene associated with aluminum tolerance has been screened and identified in soybean
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Embodiment 1
[0051] Example 1. Cloning and expression of soybean aluminum tolerance-related gene GmNAC069
[0052] 1. Cloning of Soybean Aluminum Tolerance Related Gene GmNAC069
[0053] The root total RNA of soybean material Huaxia No. 3 (aluminum-tolerant soybean variety) was extracted with a plant total RNA extraction kit (TR02, GeneMark), and its integrity was detected by 1% agarose electrophoresis.
[0054] The total RNA was reverse-transcribed to obtain cDNA, and the cDNA was synthesized according to the instructions of the PrimeScriptTMRT reagent Kit with gDNA Eraser kit.
[0055] Design the following primers:
[0056] Sequence 3: 5'-ATGAAGGGAGAATTAGAGTTG-3'
[0057] Sequence 4: 5'-TCACATCTTCTGTAGGTACATG-3'
[0058] 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.
[0059] The above PCR amplification system is a PCR reaction solution (50 μl system) configured in the follo...
Embodiment 2
[0083] Example 2, the acquisition of transgenic GmNAC069 soybean
[0084] 1. Transgenic GmNAC069 soybeans
[0085] 1. Obtaining the recombinant vector
[0086] Recombinant plant expression vector pTF101.1-GmNAC069 (such as figure 1Shown) is the plasmid obtained by replacing the fragment between the Xba I and Sac I restriction sites of the pTF101.1 vector with the GmNAC069 gene shown in sequence 1, and the full-length sequence of GmNAC069 is inserted forward into the CaMV35S promoter.
[0087] 2. Recombinant bacteria
[0088] The recombinant vector pTF101.1-GmNAC069 was transformed into Agrobacterium EHA105 (BioVector NTCC Inc.) with a Bio-rad electric shock transformation instrument to obtain recombinant Agrobacterium EHA105 / pTF101.1-GmNAC069 (using the primers shown in sequence 5 and sequence 6 For PCR amplification of bacterial solution, 180bp is positive).
[0089] 3. Transgenic GmNAC069 soybean
[0090] Soybean was transformed by Agrobacterium EHA105-mediated transfor...
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Abstract
The invention discloses an aluminum-resistant related gene GmNAC069 and an encoding protein and application thereof. The protein provided by the invention is named as protein GmNAC069 and is shown as follows: (1) a protein consisting of an amino acid sequence as 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 GmNAC069 disclosed 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 soybean can be enhanced and the accumulation of aluminum ions at the root tip can be reduced by over-expressing the gene in the soybean, which indicates that the protein can lay a foundation for the research of culturing transgenic plants with relatively high 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 GmNAC069 and its encoded protein and application. Background technique [0002] Aluminum toxicity on acidic soils is a major factor limiting plant growth. Under acidic conditions, aluminum ions in the soil are active and toxic, which seriously inhibits the growth of plant roots, affects the absorption of water and other nutrients by plants, and aggravates other stress problems on acidic soils. Soybean is an important food and oil crop, which is widely cultivated in the world and is the main source of human edible oil and vegetable protein. However, soybean production on acidic soils in tropical and subtropical regions is limited by Al stress. During the long-term evolution of plants, a series of aluminum tolerance mechanisms have been formed. [0003] Studies have found that NAC transcription factors have a conserved ...
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