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Plant stress resistance related protein GsNAC019, encoding gene and application thereof

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

Active Publication Date: 2016-12-21
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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  • Plant stress resistance related protein GsNAC019, encoding gene and application thereof
  • Plant stress resistance related protein GsNAC019, encoding gene and application thereof
  • Plant stress resistance related protein GsNAC019, encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Embodiment 1, the cloning of soybean transcription factor GsNAC019 gene

[0085] 1. Processing of plant material

[0086] Pick plump wild soybean G07256 seeds in concentrated H 2 SO 4 Treat in medium for 10min to remove the waxy film, pour out the 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.

[0087] 2. RNA extraction

[0088] The total RNA of the roots of the 3-week-old wild soybean seedlings obtained in step 1 above was extracted by RNAprep pure kit (TRANSGEN BIOTECH).

[0089] 3. Acquisition of cDNA ...

Embodiment 2

[0099] Example 2, Analysis of the expression characteristics of wild soybean transcription factor GsNAC019 gene

[0100] 1. Analysis of the expression pattern of GsNAC019 gene in wild soybean roots and leaves under alkali stress treatment at different times

[0101] 1. Processing of plant material

[0102] 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.

[0103]When the wild soybean seedlings grow to 3 weeks old, respectively, in 50mM NaHCO 3 (pH 8.5) and 200mM NaCl conditions for 0h, 1h, 3h, 6h, 12h, 24h and then quickly cut young leaves and roots, and stored at -80°C.

[0104] 2. Extraction of total RNA and acquisition of cDNA

[0105] The ...

Embodiment 3

[0115] Example 3. Transient expression of GsNAC019 gene mediated by particle gun bombardment and subcellular localization of target protein

[0116] 1. Construction of subcellular localization vector

[0117] Using the total cDNA of wild soybean as a template, GsNAC019-YS and GsNAC019-YAS primers were used for PCR amplification to obtain the PCR amplification product, that is, the GsNAC019 gene. flanked by protective bases):

[0118] GsNAC019-YS: 5'-ACGCGTCGACATGGGAGTTCCCAGAGAAAGACCC-3';

[0119] GsNAC019-YAS: 5'-AAAACTGCAG ATTSCTGACCCGAACCCG-3';

[0120] 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).

[0121] PCR reaction conditions: 98°C for 8 min; 98°C for 10 s, 60°...

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Abstract

The invention discloses a plant stress resistance related protein GsNAC019, an encoding gene and an application thereof. The protein GsNAC019 is a protein a), a protein b) or a protein c), wherein the protein a) is a protein with the amino acid sequence shown in sequence 2; the protein b) is a fusion protein obtained by connecting the N-terminal and / or C-terminal of the protein as shown in the sequence 2 with a tag; the protein c) is a protein with the same function which is obtained by substituting and / or deleting and / or adding one or more amino acid residues for / from / in the amino acid sequence shown in the sequence 2. Experiments prove that the GsNAC019 gene is overexpressed in arabidopsis thaliana, the resistance of the obtained transgenic arabidopsis thaliana to carbonate stress is obviously higher than that of wild type arabidopsis thaliana, the GsNAC019 protein has the function of regulating and controlling the plant alkali resistance, and a foundation is laid for the research of cultivating transgenic plants with carbonate stress resistance.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a protein GsNAC019 related to plant stress resistance, its coding gene and application. Background technique [0002] Saline-alkali stress restricts agricultural production in Northeast China and even the whole country. Among them, alkali stress is an important environmental limiting factor affecting plant growth, development and geographical distribution, and seriously affects crop yield and quality. Developing and utilizing saline-alkali land and tapping the production potential of agro-ecological areas in adversity are major issues to be solved urgently to maintain the sustainable and efficient development of my country's agriculture and ensure my country's food security. So far, there are few reports on the molecular mechanism of plant alkali tolerance. [0003] Soybean is an important crop in my country, especially in Heilongjiang Province. It provides both protein a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00
CPCC12N15/8271C07K14/415
Inventor 朱延明丁晓东曹蕾于洋陈超段香波宋雪薇
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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