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A protein gshdz4 related to plant stress resistance and its coding gene and application

A technology of plant stress resistance and stress resistance, applied in the direction of plant gene improvement, application, plant peptides, etc.

Inactive Publication Date: 2018-07-31
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there are few reports on the molecular mechanism of plant alkali tolerance

Method used

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  • A protein gshdz4 related to plant stress resistance and its coding gene and application
  • A protein gshdz4 related to plant stress resistance and its coding gene and application
  • A protein gshdz4 related to plant stress resistance and its coding gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Embodiment 1, the cloning of soybean transcription factor Gshdz4 gene

[0097] 1. Processing of plant material

[0098] Pick plump wild soybean G07256 seeds in concentrated H 2 SO 4 Medium treatment for 10min to remove the mud film, pour the net concentrated H 2 SO 4 , washed with sterile water 3 to 4 times, placed on wet filter paper, cultured in dark at 25°C for 3 days to accelerate germination, and when the buds grew to about 1 to 2 cm, they were transferred to a bowl filled with Hoagland's culture medium. Fix it with space cotton, soak the buds in the culture solution, and place them in an artificial climate box for cultivation. When the seedlings grow to 3 weeks old, take 3 cm of their roots and put them into EP tubes, and store them at -80°C.

[0099] 2. RNA extraction

[0100] Total RNA was extracted from the roots of the 3-week-old wild soybean seedlings using the RNAprep pure kit (TRANSGEN BIOTECH).

[0101] 3. Acquisition of cDNA

[0102] Using the abo...

Embodiment 2

[0111] Example 2, Analysis of the expression characteristics of the soybean transcription factor Gshdz4 gene

[0112] 1. Analysis of the expression pattern of Gshdz4 gene in wild soybean roots and leaves under alkali stress treatment at different times

[0113] 1. Processing of plant material

[0114] Pick plump wild soybean G07256 seeds in concentrated H 2 SO 4 Medium treatment for 10min to remove the mud film, pour the net concentrated H 2 SO 4 , rinsed with sterile water 3 to 4 times, placed on wet filter paper, cultured in dark at 25°C for 3 days to accelerate germination, and when the buds grew to about 1 to 2 cm, removed, and used Hoagland liquid medium for hydroponics.

[0115] When wild soybean seedlings grow to 3 weeks old, in 50mM NaHCO 3 (pH 8.5) After treatment for 0h, 1h, 3h, 6h, 12h, 24h, the young leaves and roots were quickly clipped and stored at -80°C.

[0116] 2. Extraction of total RNA and acquisition of cDNA

[0117] The RNAprep pure kit (TRANSGEN B...

Embodiment 3

[0138] Example 3, Transient expression of Gshdz4 gene mediated by particle gun bombardment and analysis of subcellular localization of target protein

[0139] 1. Construction of subcellular localization vector

[0140] Using the total cDNA of wild soybean as a template, Gshdz4-YS and Gshdz4-YAS primers were used for PCR amplification to obtain the PCR amplification product, that is, the Gshdz4 gene. flanked by protective bases):

[0141] Gshdz4-YS: 5'-TAATAT GGATCC CATGAATCATCGACCACCTTTTC-3';

[0142] Gshdz4-YAS: 5'-AATTAT ACTAGT GCATATACAGATTAATCCATTCCATG-3';

[0143] PCR reaction system: 20μL 5×PrimeSTAR TM HS PCR buffer, 8 μL dNTP mix (A, G, T, C, each 2.5 mM), 2 μL upstream and downstream primers (10 μM), 1 μL universal template diluted 100 times (containing the plasmid of the target gene), 1 μL high-fidelity enzyme[ PrimeSTAR DNA Polymerase (TaKaRa)], sterile ddH 2 O to make up the volume (total volume 100 μL).

[0144] PCR reaction conditions: 98°C for 8min; 9...

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Abstract

The invention discloses a plant adverse resistance associated protein Gshdz4 and coding gene and application thereof The protein is the following a), b) or c): a) a protein with the amino acid sequence SEQ ID No. 2; b) a protein obtained by connecting label to the N terminal and / or C terminal of the protein shown in SEQ ID No.2; and c) a plant adverse resistance associated protein obtained by substitution and / or deletion and / or addition of one or more amino acid residues on the amino acid sequence shown in SEQ ID No.2. Experiments prove that the protein provided by the invention has the function of improving plant stress resistance, and has important applications in plant breeding.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a protein Gshdz4 related to plant stress resistance, its coding gene and application. Background technique [0002] Saline-alkali adversity severely restricts the agricultural production in Northeast China and even the whole country, and is a major problem facing the agricultural development in adversity ecological zones in our country. According to the incomplete statistics of UNESCO and FAO, the area of ​​saline-alkali land in my country is 99.13 million hm 2 , there is 3.73 million hm of saline-alkali land in the west of Songnen Plain in Northeast China 2 , of which the area of ​​saline-alkali land in Heilongjiang Province is as high as 1.88 million hm 2 . In order to ensure my country's food security, the arable land area of ​​the country must be guaranteed to be more than 1.8 billion mu, but the arable land area has been decreasing year by year, and it is now appro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/84C12N15/11A01H5/00A01H6/20
CPCC07K14/415C12N15/11C12N15/8205C12N15/8273
Inventor 朱延明曹蕾肖佳雷于洋高杰孙皓修
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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