Plant stress resistance-related GmSIZ1a/b protein and coding gene and application thereof

A technology for plant stress resistance and coding genes, which is applied in the field of GmSIZ1a/b protein and its coding genes and applications, and can solve unclear problems

Active Publication Date: 2016-04-20
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Current studies have found that SUMOylation is closely related to the interaction between...

Method used

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  • Plant stress resistance-related GmSIZ1a/b protein and coding gene and application thereof
  • Plant stress resistance-related GmSIZ1a/b protein and coding gene and application thereof
  • Plant stress resistance-related GmSIZ1a/b protein and coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1, the acquisition of GmSIZ1a / b gene

[0045] 1. Primers: F1 (forward primer): 5'-CGGGATCCATGGATTTGGTACCGAGCG-3'; R1 (reverse primer): 5'-CGGGATCCTCTCTGAATCTGAATCAATAGAA-3'.

[0046] 2. Extract RNA from soybean leaves and reverse transcribe to obtain cDNA.

[0047] 3. Using the above cDNA as a template, using the primer pair designed in step 1, PCR amplifies the GmSIZ1a / b gene.

[0048] 4. The amplified product was sequenced, and the sequencing results showed that: the GmSIZ1a protein is shown in sequence 2 of the sequence listing, and its coding gene sequence is shown in sequence 1 of the sequence listing; the GmSIZ1b protein is shown in sequence 4 of the sequence listing, and its coding The gene sequence is shown in sequence 3 of the sequence listing.

Embodiment 2

[0049] Embodiment 2, the acquisition of transgenic plants

[0050] 1. Construction of the carrier

[0051] The GmSIZ1a / b full-length cDNA obtained in Example 1 and the pCambia1302 vector were double digested with restriction enzymes HindIII and NcoI respectively, and ligated to obtain a recombinant vector. Then, the AtSIZ1 promoter region was connected into the recombinant vector respectively to obtain the pCambia1302-ProAtSIZ1:GmSIZ1a:GFP vector and the pCambia1302-ProAtSIZ1:GmSIZ1b:GFP vector, which were verified by sequencing.

[0052] Sequencing results show that: the GmSIZ1a gene sequence inserted between the HindIII and NcoI restriction sites of the pCambia1302-ProAtSIZ1:GmSIZ1a:GFP vector is shown in the 1st-2643rd nucleotide in sequence 1, indicating that the vector is correct; in pCambia1302-ProAtSIZ1 : GmSIZ1b: The GmSIZ1b gene sequence inserted between the HindIII and NcoI restriction sites of the GFP vector is shown in nucleotides 1-2640 in Sequence 3, indicating ...

Embodiment 3

[0056] Example 3, GmSIZ1a / b Complementary Transgenic Plants Detecting Heat Stress-Induced SUMOylation Levels

[0057] After placing the transgenic Arabidopsis sample obtained in Example 2 at 37° C. for 30 minutes, the total protein was extracted. The protein extraction solution included: NaCl, 5% (w / v) SDS, 0.5% (v / v) NP40, 6mMEDTA, 3mMDTT, 1mMPMSF and 30% (v / v) glycerol.

[0058] The protein concentration of the transgenic Arabidopsis sample obtained in Example 2 was determined to be 20 g / L by Bradford protein assay reagent (Cat: CW0013, CWBIO, Beijing, China). The extracted protein was separated by polyacrylamide gel electrophoresis, and after transfer to the membrane, it was detected by western blot with anti-AtSUMO1 antibody.

[0059] The results showed that compared with Arabidopsis siz1-2 mutants and plants transformed only with empty vectors, the SUMOconjugates content of plants transformed with GmSIZ1a / b increased and returned to the level of wild-type Col-0, indicati...

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Abstract

The invention discloses a plant stress resistance-related GmSIZ1a/b protein and its coding gene and application. The protein is a protein selected from the following a) or b): a) a protein composed of an amino acid sequence as shown in the sequence 2/4 in a sequence table; and b) a protein which is obtained after one or more amino acid residues substitution and/or deletion and/or addition of an amino acid sequence as shown in the sequence 2/4 in the sequence table. Experimental results prove that a transgenic line for inhibiting expression of GmSIZ1a/b in soybean obviously raises resistance to phytophthora sojae.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a GmSIZ1a / b protein related to plant stress resistance and its coding gene and application. Background technique [0002] Soybean is one of the most important commercial crops in the world, yet it is seriously infected by various pathogens. Phytophthorasojae, a plant pathogen of the oomycete class that causes soybean root rot, is one of the most destructive soybean diseases, causing losses of $1-2 billion annually. Soybean plants resist Phytophthora sojae through at least two different mechanisms: effector-induced immune response (ETI) mediated by a single gene (Rpsgene) and partial resistance regulated by multiple genes, which may be related to basal defense. ) are associated with weak ETI. Currently, about twenty Rps genes have been mapped to four chromosomes. Only Rps1k, RpsYD29, Rps10 and RpsJS were fine-mapped among these Rps genes. However, the molecular mechanism by which ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N1/15C12N1/21C12N1/19C12N15/82C12N15/113
Inventor 金京波蔡斌
Owner INST OF BOTANY CHINESE ACAD OF SCI
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