OsFLP protein related to plant salt tolerance as well as related biological materials and application thereof

A biomaterial and protein technology, applied in angiosperms/flowering plants, applications, plant peptides, etc., can solve problems such as the intricate regulation network of plant adversity

Active Publication Date: 2021-06-15
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The plant stress regulation network is intricate, and its research is full of challenges and significance. The exploration and application of rice salt-tolerant genes provides a new idea to solve the current problems in agricultural production, and is of great significance to molecular breeding and the development of salt-tolerant varieties.

Method used

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  • OsFLP protein related to plant salt tolerance as well as related biological materials and application thereof
  • OsFLP protein related to plant salt tolerance as well as related biological materials and application thereof
  • OsFLP protein related to plant salt tolerance as well as related biological materials and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Example 1. Acquisition of plant salt-tolerance-related gene OsFLP

[0074] 1. Sequence analysis of rice OsFLP protein

[0075]Using the amino acid sequence of Arabidopsis AtFLP in NCBI (http: / / www.ncbi.nlm.nih.gov / ) and the rice genome database (http: / / rice.plantbiology.msu.edu / ), to find Homologous proteins of Arabidopsis AtFLP. As a result, a protein having 33.89% similarity to AtFLP's amino acid sequence was obtained and named as OsFLP, and its amino acid sequence is shown in Sequence 1 in the Sequence Listing.

[0076] Through comparative analysis with several known R2R3-MYB family protein sequences, including Arabidopsis AtFLP, AtMYB88, AtMYB121, ATMYB0 (ATGL1) and rice OsMPS protein sequences, the results are shown in figure 1 , found that there are two MYB domains, R2 and R3, at the N-terminal of OsFLP protein. Among them, R2 contains three conserved motifs of H1, H2 and H3, and R3 contains three conserved motifs of H1, H2 and H3, which proves that OsFLP is a ...

Embodiment 2

[0093] Embodiment 2, the acquisition of gene OsFLP mutant

[0094] 1. Using TILLING technology to obtain osflp-1 mutants

[0095] TILLING (Targeting Induced Local Lesions IN Genomes) technology is a new reverse genetics research method, using the chemical mutagen ethyl methanesulfonate to mutate the seeds, and then using high-throughput Sequencing methods quickly and efficiently identify point mutations from mutagenized populations. In order to better analyze the function of OsFLP in rice, we used TILLING technology to obtain osflp mutant lines at different mutation sites, see Table 1 for details. We observed and compared the obtained strains at different mutation sites, and found a strain A2349-2 with a different stomatal phenotype from the wild type, and named the mutant osflp-1 mutant.

[0096] Table 1

[0097]

[0098]

[0099] 2. Mutation site analysis of osflp-1 mutant

[0100] We designed corresponding primers to amplify OsFLP genomic DNA in ZH11 and osflp-1 m...

Embodiment 3

[0101] Embodiment 3, the acquisition of transgenic OsFLP rice

[0102] 1. Construction of OsFLP gene overexpression vector and acquisition of rice transgenic materials

[0103] OsFLP gene overexpression vector is using pH7WG2D.1 (plasmid map see Figure 5 ) as a starting carrier, adopting the gateway system method to construct, the specific implementation is as follows: amplify the cDNA fragment of OsFLP (the nucleotide sequence is sequence 2), after gel recovery, connect the cDNA fragment of OsFLP to the entry vector TOPOvector, and name the obtained plasmid is OsFLP-TOPO. Digest the plasmid OsFLP-TOPO with PVUⅠ restriction endonuclease (recognition site: CGATCG, the cut fragment is sticky, it belongs to restriction endonuclease type II) at 37°C for 3 hours, and put The enzyme products were separated by agarose gel electrophoresis and recovered by cutting the gel. Mix 1.5 μL of recovered product, 0.5 μL of pH7WG2D.1 plasmid, and 0.5 μL of Gateway LR ClonaseⅡenzyme mix, and...

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Abstract

The invention discloses an OsFLP protein related to plant salt tolerance as well as a related biological materials and application thereof. The OsFLP protein can be a protein of A1), A2) or A3) as follows: A1) a protein with an amino acid sequence shown as a sequence 1 in a sequence table; A2) a protein which is obtained by substitution and / or deletion and / or addition of one or more amino acid residues of the protein of A1), has 90% or more of identity with the protein of A1) and has the activity of improving salt tolerance; and A3) a fusion protein obtained by connecting a protein tag to the N terminal or / and C terminal of A1) or A2). The OsFLP protein and related biological materials thereof can be used for improving the salt tolerance of plants.

Description

technical field [0001] The invention relates to an OsFLP protein related to plant salt tolerance and related biological materials and applications in the field of biotechnology. Background technique [0002] With global climate change, the impact of abiotic stress on plant growth has become one of the major challenges faced by human beings, seriously restricting the growth and development of plants. Abiotic stress refers to the adverse effects of any abiotic factors on plants in a specific environment, including drought, low temperature, salinity and other factors. Drought and salinity are the two most important stress factors restricting plant growth. Especially as the degree of salinization of the global land is deepening and the area of ​​arable land is decreasing, the research on the response genes and their mechanisms of action to plant responses to abiotic stress has become one of the hottest topics today. It is very important to screen and breed genetic varieties re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8273
Inventor 乐捷张洁张春霞
Owner INST OF BOTANY CHINESE ACAD OF SCI
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