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Protein related with heat resistance of plants as well as encoding gene and application of protein

A heat-tolerance, coding technology, applied in the direction of plant gene improvement, application, plant peptides, etc., can solve problems such as poor adaptability

Active Publication Date: 2014-03-26
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, wheat is a cool-loving C3 crop, and its adaptability to high temperature is relatively poor.

Method used

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  • Protein related with heat resistance of plants as well as encoding gene and application of protein
  • Protein related with heat resistance of plants as well as encoding gene and application of protein
  • Protein related with heat resistance of plants as well as encoding gene and application of protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Example 1. Obtaining of black rice bean ferritin gene (Vefer)

[0072] 1. Extraction of total RNA from black rice bean

[0073] The roots of the black rice bean plants in hydroponic culture for 10 days were placed in 100mM Fe 2+ -Treated in EDTA aqueous solution for 24 hours, and the young roots were taken and immediately frozen in liquid nitrogen for future use. Total RNA was extracted from 100 mg of black rice bean radicle thoroughly ground by liquid nitrogen with a Trizol kit. For details, refer to the kit instruction manual.

[0074] 2. Obtain cDNA by reverse transcription

[0075] Add the following system to an RNase-free 0.2mL PCR tube: 2 μg of total RNA, Primer T 15 (10 μM) 1 μL with DEPC ddH 2 O to make up to 10 μL.

[0076] Briefly centrifuge at low speed to concentrate all components of the system to the bottom of the PCR tube. Denature on a PCR instrument at 70°C for 5 minutes, place on ice immediately, and prepare the following reverse transcription re...

Embodiment 2

[0089] Example 2, construction and identification of recombinant expression vector pBAC47P-ubi-fer

[0090] 1. Construction of recombinant expression vector pBAC47P-ubi-fer

[0091] The recombinant plasmid pTE-fer obtained in Example 1 was digested with BamHI and KpnI, and the target band with a size of about 880bp was recovered, which was combined with the large backbone fragment of the pBAC47P-ubi vector (see below for the construction method) that had undergone the same double digestion connected to obtain a recombinant plasmid, which was named pBAC47P-ubi-fer (plasmid map as shown in figure 1 shown in A).

[0092] Among them, the enzyme digestion system (20 μL) is as follows: plasmid DNA 3 μL, 10×L buffer 2 μL, BamH Ⅰ 1 μL, Kpn Ⅰ 1 μL, H 2 O13 μL. Enzyme digestion at 37°C for 2h.

[0093] The construction method of the pBAC47P-ubi vector and the subsequent screening and identification process can be found in the literature "Sambrook J, Fritsch E F, Maniatis T. Molecul...

Embodiment 3

[0099] Embodiment 3, the acquisition and identification of transgenic Vefer wheat

[0100] 1. Acquisition of Vefer genetically modified wheat

[0101] Callus induction medium SD 2 : MS basic medium (phyto Technology Laboratories TM company, product number M524) + VB 1 1mg / L+asparagine 150mg / L+2,4-D2mg / L+sucrose 30g / L+plant gel 2.8g / L, pH5.8. Each concentration is the final concentration of the corresponding component in the medium.

[0102] Hyperosmotic medium: callus induction medium SD 2 +Sorbitol 200mM+Mannitol 200mM. Each concentration is the final concentration of the corresponding component in the medium.

[0103] recovery medium SD 0 : MS basic medium (phyto Technology Laboratories TM company, product number M524) + VB11mg / L+ asparagine 150mg / L+ sucrose 30g / L+ plant gel 2.8g / L, pH5.8. Each concentration is the final concentration of the corresponding component in the medium.

[0104] Screening differentiation medium: 1 / 2MS medium + 0.5mg / L VB1 + 0.25mg / L VB6 +...

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Abstract

The invention discloses protein related with the heat resistance of plants as well as an encoding gene and an application of the protein. The protein related with the heat resistance of the plants is the following (a) or (b): (a) protein consisting of amino acid sequences represented in sequence 1 in a sequence table; (b) protein formed by performing substitution and / or absence and / or addition of one or more amino acid residues on the amino acid sequences represented by the sequence 1, derived from the sequence 1 and related to the heat resistance of the plants. Experiments prove that after heat stress on transgenic plants, the content of malondialdehyde in plant blades and the relative electrical conductivity are obviously lower than those of receptor plants, which proves that the heat stability and the heat resistance of cell membranes of transgenic lines are enhanced. Thus it can be seen that the protein and the encoding gene of the protein have important theoretical and practical significance in cultivating wheat with enhanced heat resistance and other new varieties of plants and have wide application prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a protein related to plant heat resistance and its coding gene and application, in particular to a protein derived from black rice bean and related to plant heat resistance, its coding gene and its application. Background technique [0002] Wheat is the third largest grain crop in my country, and its output and quality are directly related to people's daily life. At the same time, wheat is a cool C3 crop and has poor adaptability to high temperature. In the main wheat-producing areas of my country, high temperatures above 30°C are often encountered during the grain-filling stage of wheat grains, resulting in a substantial reduction in wheat production. In my country's Yellow River, Huaihe River, Haihe River Basin and Xinjiang wheat area, high temperature is often accompanied by drought, forming dry hot wind, which can damage 2 / 3 of the area's wheat planting area and reduce wheat produ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N15/11A01H5/00
CPCC07K14/415C12N15/8271
Inventor 孙其信梁荣奇倪中福李保云尤明山彭惠茹解超杰秦丽燕王树斌姚颖垠杜金昆刘志勇
Owner CHINA AGRI UNIV
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