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Application of rice GT transcription factor family gene OsGT gamma-1 in controlling salt tolerance of rice

A technology of salt tolerance and genetics, applied in the fields of application, genetic engineering, plant genetic improvement, etc.

Inactive Publication Date: 2011-09-28
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Rice GT-2 and Arabidopsis GT-1 can activate transcription in vivo (Le et al., Transcriptional activation by Arabidopsis GT-1 may be through interaction with TF II A-TBP-TATA complex. Plant J.1999, 18:663-668 ; Ni et al., GT-2: in vivo transcriptional activation activity and definition of novel twin DNA binding domains with reciprocal target sequence selectivity. Plant Cell. 1996, 8: 1041-1059), Arabidopsis thaliana PTL is involved in regulating the development of floral organs (Brewer et al., PETAL LOSS, a trihelix transcription factor gene, regulates perianth architecture in the Arabidopsis flower. Development. 2004, 131: 4035-4045), but whether this family is involved in the stress response of plants has not yet been reported

Method used

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  • Application of rice GT transcription factor family gene OsGT gamma-1 in controlling salt tolerance of rice
  • Application of rice GT transcription factor family gene OsGT gamma-1 in controlling salt tolerance of rice
  • Application of rice GT transcription factor family gene OsGT gamma-1 in controlling salt tolerance of rice

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Experimental program
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Effect test

Embodiment 1

[0026] Example 1: Carrying out bioinformatics analysis on the rice GT family and isolating and cloning a DNA fragment containing the OSGTγ-1 gene segment

[0027] Based on the reported plant GT protein sequence in NCBI (National Center for Biotechnology Information, http: / / www.ncbi.nlm.nih.gov), TIGR (The Institute for Genomic Research, http: / / www.tigr.org) and TAIR (TheArabidopsis Information Research, http: / / www.arabidopsis.org) database to conduct BLAST search results, collect GT transcription factor family member sequences, manually remove repetitive sequences. Phylogenetic analysis of GT family members in rice and Arabidopsis and GT factor protein sequences reported in some other plant species was carried out using MrBayes version 3.0 software (open access software) (results in figure 1 ), it was found that members of the subfamily GTγ have not been reported yet, and OsGTγ-1, a member of this subfamily, was selected as the research object. The gene corresponds to the cDN...

Embodiment 2

[0029] Example 2: Analysis of tissue and stress expression profiles of rice endogenous gene OSGTγ-1

[0030] The rice variety "Zhenshan 97" (a rice variety that has been popularized and widely used in China) was used as the material, and the treatments of drought, cold injury, high-salt stress and injury were respectively carried out at the 3-leaf stage. Dry early treatment is to cut off water from the seedlings, high-salt stress is to soak the roots of the seedlings in 200mM / L NaCl solution, and low-temperature treatment is to place the seedlings in a 4°C growth box at four time points of 0, 1, 3, and 6 hours Sampling, injury stress sampling time points are 0, 1, 10, 30 minutes. The seven hormones used in hormone spraying treatment are: A, abscisic acid (ABA); I, indole acetic acid (IAA); G, gibberellin (GA3); K, cytokinin (KT); J, jasmine acid (JA); E, ethephon (Eth); S, salicylic acid (SA). Concentrations are: 26.43mg / L, 10mg / L, 40mg / L, 100mg / L, 21.03mg / L, 43.35mg / L, 138....

Embodiment 3

[0031] Example 3: Subcellular localization of OsGTγ-1 protein

[0032] Using the rice stable transformation system, we fused the cDNA fragment containing the predicted nuclear localization sequence of OsGTγ-1 with the reporter gene GFP, and expressed it in rice plants through the drive of a strong promoter Ubiquitin in maize. The results observed under the laser confocal microscope showed that the OsGTγ-1 protein was localized in the nucleus (such as image 3 shown). The fusion vector construction method is as follows: OsGTγ-1 partial cDNA sequence (corresponding to the coding region sequence except the 3′-terminal 5 amino acids and stop codon) is passed through the primer 5′-CTA GGTACC GAGCTTCTCTTTGGAATTTGTG-3' (sequence-specific primer plus linker KpnI site) and 5'-TAA CCCGGG CTTTGGTTTGATACCCATCTC-3' (sequence-specific primer plus linker SmnaI site) was amplified by PCR, and the product corresponded to 391-1850 bp of the sequence shown in SEQ ID NO: 1 in the present inve...

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Abstract

The invention relates to the technical field of rice genetic engineering, in particular to the application of a GT transcription factor family gene OsGT gamma-1 in controlling the salt tolerance of rice. By functional analysis and identification, the gene OsGT gamma-1 for controlling the salt tolerance of rice is obtained, and the application of the gene in adverse resistance genetic improvement of the salt tolerance of rice is verified. The gene OsGT gamma-1 is one of the following nucleotide sequences: 1) the nucleotide sequence shown from 475th to 1707th in a sequence table SEQ NO: 1; or the nucleotide sequence of protein with the same code as that of the protein encoded by 1). The nucleotide sequence containing the gene OsGT gamma-1 is transferred to rice after being connected with an exogenous promoter, and the salt tolerance of the transgenic rice is obviously enhanced.

Description

technical field [0001] The invention relates to the field of rice genetic engineering. It specifically involves isolating and cloning a rice GT transcription factor family gene OsGTγ-1 that can improve high-salt tolerance, and realizing its application in genetic improvement of rice stress resistance through functional verification. The present invention uses the method of screening T-DNA insertion rice mutant library to isolate and clone the gene OsGTγ-1 that controls the salt tolerance of rice. On the one hand, it is verified that the mutant is closely linked with the high-salt sensitive phenotype through co-segregation detection, and on the other hand On the one hand, the overexpression of OSGTγ-1 gene improves the tolerance of transgenic rice to high salt, confirming the function of the gene. Background technique [0002] The growth and development of plants not only depends on their genetic background, but also is affected by many environmental factors. The environmen...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415A01H5/00
Inventor 熊立仲方玉洁
Owner HUAZHONG AGRI UNIV