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Brachypodium distachyon Drought Tolerance Tolerance Gene And Its Encoded Protein And Its Application

A gene and drought technology, which is applied to Brachypodium erinaceus drought resistance and salt resistance gene and expression vector and its encoded protein and application fields, can solve problems such as deterioration, quality decline, crop yield reduction, etc., and achieves reduction of oxidative damage and improvement of tolerance. the effect of enhancing stability

Active Publication Date: 2020-10-30
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Unfavorable environments such as drought and high salinity affect the normal growth and development of plants by affecting photosynthesis, lipid metabolism and protein synthesis and metabolism of plants, and cause crop yield and quality decline
Ecological imbalance and environmental pollution make the growing environment of plants more complex and deteriorating, and the normal growth of plants is seriously threatened

Method used

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  • Brachypodium distachyon Drought Tolerance Tolerance Gene And Its Encoded Protein And Its Application
  • Brachypodium distachyon Drought Tolerance Tolerance Gene And Its Encoded Protein And Its Application
  • Brachypodium distachyon Drought Tolerance Tolerance Gene And Its Encoded Protein And Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Isolation of Brachypodium distachyon BdGF14a gene

[0037] 1. Cloning of BdGF14a gene

[0038] The 14-3-3 gene sequence (BRADI1G11290.1, GenBank: KU933262.1) obtained by Phytozome v12.1 (https: / / phytozome.jgi.doe.gov / pz / portal.html) Blast was submitted to NCBI (https: / / www.ncbi.nlm.nih.gov) for BLAST comparison analysis. Use the Primer6 primer design software to specifically amplify the primers for Brachypodium distachyon BdGF14a. The primer sequences are shown in SEQ ID NO.3 and SEQ ID NO.4. The PCR reaction system is shown in Table 1. The PCR program is as follows: pre-denaturation at 98°C for 3 minutes; Denaturation at 98°C for 15s; annealing at 65°C for 5s; extension at 72°C for 30s; final extension at 72°C for 10min; storage at 4°C. Among them, the three consecutive steps of denaturation, annealing and extension set up 35 cyclic reactions.

[0039] Table 1 PCR reaction system

[0040]

[0041] 2. Gel recovery of target fragments

[0042] Spot th...

Embodiment 2

[0065] Example 2: Construction of pBI121-BdGF14a-GFP eukaryotic expression vector

[0066] 1. PCR amplification of BdGF14a gene

[0067] Using the cloning vector pMD18-T-BdGF14a containing the correctly sequenced Brachypodium distachyon BdGF14a gene as a template, use the Oligo7 primer design software to design the 5' end with pBI121 vector multi-cloning site region XbaI restriction site and BamHI enzyme The gene-specific primers at the cleavage site were used for PCR amplification. The primer sequences are shown in Tables SEQ ID NO.5 and SEQ ID NO.6. The PCR reaction system is shown in Table 3 / same as Table 1. The PCR program is as follows: pre-denaturation at 98°C for 2 minutes; 98°C Denaturation at ℃ for 10s; annealing at 65℃ for 5s; extension at 72℃ for 30s; final extension at 72℃ for 10min; storage at 4℃. Among them, the three consecutive steps of denaturation, annealing and extension set up 35 cyclic reactions.

[0068] Table 3 PCR reaction system

[0069]

[0070]...

Embodiment 3

[0116] Example 3: Subcellular localization of Brachypodium distachyon BdGF14a protein

[0117] 1. Preparation and transformation of Agrobacterium competent

[0118] All the operation steps of the preparation and transformation experiment of Agrobacterium competent cells were carried out aseptically in the ultra-clean workbench. The specific process is as follows:

[0119] 1) Take the preserved Agrobacterium strain EHA105 from a -80°C ultra-low temperature refrigerator and inoculate it on the YEB solid medium containing 50 mg / L Str (streak), and culture it in the dark at 28°C for 2-3 days.

[0120] 2) Pick large and round monoclonal colonies, inoculate them into 1 mL of YEB liquid medium containing 50 mg / L Str, and incubate in the dark at 200 rpm at 28°C for 12 to 16 hours.

[0121] 3) Take 1 mL of the above-mentioned bacterial solution and transfer it to 100 mL of YEB (containing 50 mg / L streptomycin), expand the culture on a shaker at 28 ° C at 200 rpm, and measure its OD va...

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Abstract

The invention provides an application of coded proteins of drought resisting and salt resisting gene of brachypodium distachyon. The nucleotide sequence of the gene is a sequence as shown in SEQ ID NO.1 or a nucleotide sequence complementary as shown in SEQ ID NO.1. The protein is obtained by coding the gene according to the claim 1, and the amino acid sequence of the protein is as shown in SEQ IDNO.2. The gene is applied to improving tolerance of plants to drought and salt resistance. The gene adjusted plant keeps cell moisture by accumulating through matters. ROS accumulation is reduced andthe cytomembrane oxidizing damage caused by active oxygen is reduced by improving the activity of an antioxidant enzyme system. By accelerating closing of pores under drought, the drought and salt resistance of transgenic tobacco is improved by way of reducing water damaged by means of transpiration, improving the expression level of stress related genes and the like. Drought and salt resisncve of the transgenic tobacco with the gene is dependent on adjustment of ABA synthesis and an ABA signal channel.

Description

technical field [0001] The invention belongs to the technical field of biogenetic engineering, and in particular relates to a drought-resistant and salt-resistant gene of Brachypodium distachyon, its expression vector, its encoded protein and its application. Background technique [0002] Unfavorable environments such as drought and high salinity affect the normal growth and development of plants by affecting photosynthesis, lipid metabolism and protein synthesis and metabolism of plants, and cause crop yield and quality decline. Ecological imbalance and environmental pollution make the growing environment of plants more complex and deteriorating, and the normal growth of plants is seriously threatened. Because they cannot avoid environmental stresses by moving themselves to favorable positions like animals, plants have formed a complete system of sensing, transmitting and responding to various stresses at the molecular, cellular and physiological levels during evolution. C...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/70C12N1/21C12N15/82A01H5/00A01H6/82C12N1/19
CPCC07K14/415C12N15/70C12N15/8273
Inventor 何光源杨广笑常俊丽张扬赵洪艳何圆
Owner HUAZHONG UNIV OF SCI & TECH
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