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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 plants and pathogens, but it is not clear whether it regulates plant resistance to Phytophthora

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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