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Application of protein ghccoaomt7 in regulation of plant heat tolerance

A technology of protein and heat resistance, applied in the fields of application, plant peptides, plant products, etc., can solve the problems of cotton bud shedding, boll development deformity, cotton production reduction, etc.

Active Publication Date: 2021-08-31
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Frequent occurrence of high temperature (≥35°C) adversity leads to severe cotton bud shedding and deformed boll development, resulting in reduced cotton yield and fiber quality

Method used

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  • Application of protein ghccoaomt7 in regulation of plant heat tolerance
  • Application of protein ghccoaomt7 in regulation of plant heat tolerance
  • Application of protein ghccoaomt7 in regulation of plant heat tolerance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Cloning of the coding gene (GhCCoAOMT7 gene) of embodiment 1, protein GhCCoAOMT7

[0061] 1. Using the Trizo1 method to extract the total RNA from the leaves of upland cotton TM-1 seedlings grown to 14 days, and then using reverse transcriptase AMV to reverse transcribe the first-strand cDNA to obtain the cDNA of upland cotton TM-1.

[0062] 2. Using the cDNA of Upland Cotton TM-1 obtained in step 1 as a template, PCR amplification was performed using a primer pair consisting of Forward Primer: 5'-ATGATTTGTTTCTCAAGCATATATTTG-3' and Reverse Primer: 5'-TCAACGAATTCGCCTGCA-3' to obtain approximately 750bp double-stranded DNA molecule.

[0063] The reaction system is 50 μL, consisting of 2 μL template (containing 1 μg of upland cotton TM-1 cDNA), 1 μL Phusion enzyme, 2.5 μL Forward Primer aqueous solution (concentration is 10 μM), 2.5 μL Reverse Primer aqueous solution (concentration is 10 μM), 10 μL 5×PhusionBuffer, 10 μL dNTP mixture (the concentrations of dATP, dTTP, dGT...

Embodiment 2

[0066] Example 2, the acquisition and identification of Arabidopsis transgenic GhCCoAOMT7

[0067] 1. Acquisition of recombinant plasmids 35S::GhCCoAOMT7-EYFP and GV3101 / 35S::GhCCoAOMT7-EYFP

[0068] 1. Insert the double-stranded DNA molecule shown in position 1 to 744 of Sequence 1 in the sequence table between the recognition sequences of restriction enzymes Sal I and BamH I of the vector pEZR-LNY to obtain a recombinant plasmid 35S::GhCCoAOMT7-EYFP.

[0069] The recombinant plasmid 35S::GhCCoAOMT7-EYFP expresses the protein GhCCoAOMT7 shown in Sequence 2 in the Sequence Listing.

[0070] 2. The recombinant plasmid 35S::GhCCoAOMT7-EYFP was introduced into Agrobacterium tumefaciens GV3101 to obtain the recombinant Agrobacterium, named GV3101 / 35S::GhCCoAOMT7-EYFP.

[0071] The vector pEZR-LNY was introduced into Agrobacterium tumefaciens GV3101 to obtain the recombinant Agrobacterium, named as GV3101 / pEZR-LNY.

[0072] 2. Acquisition of transgenic Arabidopsis thaliana

[0...

Embodiment 3

[0107] Example 3, Acquisition and Identification of Upland Cotton TM-1 Silent Strain

[0108] Infection solution: containing 10mM MES, 10mM MgCl 2 and 200 mM acetosyringone in water.

[0109] 1. Construction of recombinant plasmid PTRV2-GhCCoAOMT7 and acquisition of recombinant Agrobacterium

[0110] 1. Using the Trizo1 method to extract the total RNA from the leaves of upland cotton TM-1 seedlings grown to 14 days, and then using reverse transcriptase AMV to reverse transcribe the first-strand cDNA to obtain the cDNA of upland cotton TM-1.

[0111] 2. Using the cDNA of upland cotton TM-1 obtained in step 1 as a template, a primer pair consisting of primer F: 5'-CGACGACAAGACCCTAAAGATAATGAAGGGAGTTTTGATTTT-3' and primer R: 5'-GAGGAGAAGAGCCCTTCTATGATCGACCTCCTCGATT-3' was used for PCR amplification to obtain PCR amplified product.

[0112] The reaction system was 50 μL, consisting of 2 μL template (containing 1 μg of upland cotton TM-1 cDNA), 1 μL Phusion enzyme, 2.5 μL primer ...

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Abstract

The invention discloses the application of the protein GhCCoAOMT7 in regulating the heat resistance of plants, and its amino acid sequence is shown as sequence 2 in the sequence list. Experiments have shown that by increasing the expression and / or activity of the protein GhCCoAOMT7 in wild-type Arabidopsis, transgenic Arabidopsis is obtained; compared with wild-type Arabidopsis, the heat tolerance of transgenic Arabidopsis is improved, which is specifically expressed in the survival rate Increased pollen germination rate and increased pollen tube length. Inhibiting the expression and / or activity of the protein GhCCoAOMT7 in Upland cotton TM-1, and obtaining the upland cotton TM-1 silent strain; compared with the upland cotton TM-1, the heat resistance of the upland cotton TM-1 silent strain is reduced, and the specific performance It was due to the reduction of plant height and the shortening of the time for the shoot tip to reach half wilting after high temperature treatment. The protein GhCCoAOMT7 can regulate the heat tolerance of plants. The invention has important application value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to the application of protein GhCCoAOMT7 in regulating plant heat resistance. Background technique [0002] With the advancement of industrialization and the intensification of human activities, the trend of global warming is increasing. In 2018, the "China Climate Change Blue Book" issued by the China Meteorological Administration pointed out that 2017 was the second warmest year since the world had complete meteorological observation records. China is a sensitive area and a significant impact area of ​​global climate change. Extremely high temperature weather has occurred frequently in my country in recent years, and the regional average temperature has shown an upward trend, while the rate of temperature increase in the northwest region is more obvious than that in the south, especially the high temperature (≥35°C) adversity occurs frequently in the cotton region of northw...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/60A01H6/20
CPCC07K14/415C12N15/8218C12N15/8271
Inventor 朱生伟黄丹隆浩罗小敏蒙福宁曹务强陈霞毛玉伟
Owner INST OF BOTANY CHINESE ACAD OF SCI