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A protein related to plant stress resistance and its coding gene gsskp21 and application

A plant stress resistance, protein technology, applied in the direction of plant gene improvement, application, plant peptides, etc., can solve the problem of limited research on signal transduction pathways, and achieve the effect of improving plant stress resistance

Inactive Publication Date: 2017-10-27
NORTHEAST AGRICULTURAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the mechanism studies mainly focus on salt stress, and the research on the signal transduction pathway of alkali stress is still very limited.

Method used

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  • A protein related to plant stress resistance and its coding gene gsskp21 and application
  • A protein related to plant stress resistance and its coding gene gsskp21 and application
  • A protein related to plant stress resistance and its coding gene gsskp21 and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1. Acquisition of the gene GsSKP21 encoding the alkali-resistance-related protein in wild soybean

[0031] 1. Plant material processing

[0032] Pick plump wild soybean G07256 seeds in concentrated H 2 SO 4 Treat for 10 minutes to remove the mud film. Pour Net Concentration 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. When the bud grows to about 1 to 2 cm, it is transferred to a seedling pot filled with 30% peat soil and 70% common soil, and placed in an artificial climate box for cultivation. When the seedlings grow to 3 weeks old, quickly take fresh leaves and roots 3cm, and store them at -80°C.

[0033] 2. Extraction of plant total RNA

[0034] For the specific steps of RNA extraction, please refer to the instruction manual of the RNA prep pure kit from TIANGEN Company.

[0035] 3. Synthesis of the first strand of cDNA

[0036] Use OligodT as a primer to ...

Embodiment 2

[0044] Example 2, Analysis of subcellular localization of GsSKP21 in plant cells

[0045]1. Preparation of onion epidermal cells

[0046] Peel off 1-2 outer slices of the onion on the ultra-clean workbench, cut the slices near the heart into small pieces of about 1.5cm x l.5cm, carefully tear off the inner skin, and spread it on 1 / 2MS Incubate on solid medium in the dark at 25°C for 6 hours.

[0047] 2. Gene gun bombardment of subcellular localization vector

[0048] For washing of gold powder and embedding of plasmid DNA, please refer to the manual of Bio-Rad Helios gene gun system in the United States. Using SalI and BamHI restriction enzyme sites, the full-length CDS region of GsSKP21 was cloned into the subcellular localization vector pBSK II-eGFP to obtain the GsSKP21 subcellular localization vector pBSK II-GsSKP21-eGFP. The primer sequences are as follows:

[0049] S-GsSKP21sense:5'-GC GTC GAC ATGTCAGAAATTGACATGGCAG-3'

[0050] S-GsSKP21antisense:5'-CG GGATCC AG...

Embodiment 3

[0056] Example 3, Analysis of the expression characteristics of GsSKP21 under saline-alkali stress conditions

[0057] 1. Processing of plant material

[0058] Take 3-week-old wild soybean seedlings and place them in 200mM NaCl (high salt stress), 50mM NaHCO 3 (Simulated alkali stress) After treatment for 0h, 1h, 3h, 6h, 12h, 24h, 48h, quickly cut young leaves and roots and store them at -80°C; 12h, 24h, 48h young leaves and roots were used as controls.

[0059] 2. RNA Extraction and Genomic DNA Removal from Wild Soybean Materials

[0060] 1. Take about 100 mg of wild soybean materials treated by the above different methods, grind with liquid nitrogen, and extract RNA with the plantRNAprep Plant Kit (TIANGEN, cat no: DP402) and refer to the kit instructions.

[0061] 2. Use DNase I from TaKaRa Company to react at 37°C for 20-30 minutes to remove genomic DNA in RNA. The reaction system is as follows: RNA 40 μL, 10×DNase I Buffer 5 μL, DNase I 2 μL, RNAase Inhibitor 0.5 μL, ...

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Abstract

The invention discloses a protein related to plant resistance and an encoding gene GsSKP21 and application thereof. The protein related to the plant resistance is protein a) or b): a) protein formed by an amino acid sequence shown as a sequence 2 in a sequence table; b) protein obtained by substituting and / or deleting and / or adding one or more amino acid residues of the amino acid sequence shown as the sequence 2 in the sequence table, and related to the plant resistance. Experiments prove that the protein provided by the invention has a function of improving the plant resistance and has an important application value in plant breeding.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a protein related to plant stress resistance, its coding gene GsSKP21 and its application. Background technique [0002] Salinity-alkali stress is one of the main factors affecting crop production, which seriously affects the yield and quality of crops. It is an important problem that restricts agricultural production in my country and needs to be solved urgently. Compared with salt stress, alkali stress keeps the soil at a higher pH (>8.0) in addition to the osmotic stress caused by the decrease of soil water potential and the ion toxicity caused by the accumulation of high-concentration salt in cells. Due to the alkaline salt ion CO 3 2- and HCO 3 - The existence of , can directly destroy the structure and function of root cells, making Ca 2+ , Mg 2+ , H 2 PO 4 - Precipitation of ions occurs, thereby disrupting the dynamic ion balance of plant cells. Plants have comple...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/74C12N1/21C12N15/84A01H5/00
CPCC07K14/415C12N15/8205C12N15/8273
Inventor 朱延明孙晓丽刘艾林端木慧子肖佳雷于洋孙明哲段香波
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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