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Application of photorespiratory branch protein in regulation and control of plant traits

A photorespiration and plant technology, applied in the fields of applications, plant peptides, plant products, etc., can solve the problems of the photorespiration metabolism of food crops and other reports

Pending Publication Date: 2021-08-06
SHANDONG SHUNFENG BIOTECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, there is no report on the

Method used

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  • Application of photorespiratory branch protein in regulation and control of plant traits
  • Application of photorespiratory branch protein in regulation and control of plant traits
  • Application of photorespiratory branch protein in regulation and control of plant traits

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1I

[0440] Example 1 Regulatory effect of II-GOC photorespiratory branch on rice traits

[0441] In the present invention, OsGLO, KatE and OsOXO are co-expressed to construct II-GOC photorespiratory branch in rice.

[0442] 1.1 Acquisition of TPC-OsGLO3, TPC-OsOXO3, and TPC-KatE fusion protein expression genes

[0443] (1) Acquisition of TPC-OsGLO3 fusion protein expression gene

[0444] Primers were designed according to the cDNA sequences of OsGLO3 and TPC provided by NCBI (http: / / www.ncbi.nlm.nih.gov / ).

[0445] Using cDNA of 2-week-old japonica rice seedling leaves as a template, under the guidance of primers OsGLO3-F and OsGLO3-R, and primers TPC-1F and TPC-1R, the OsGLO3 and TPC genes were amplified by conventional methods, respectively. After the reaction, the PCR amplified product was subjected to 1% agarose gel electrophoresis, and the DNA fragments of OsGLO3 (about 1100bp) and TPC (about 150bp) were recovered and purified; The ligase was used as a template for the sec...

Embodiment 12

[0452] Example 1.2 Obtaining of TPC-OsGLO3, TPC-OsOXO3, TPC-KatE Fusion Protein Expression Cassette Sequences

[0453] (1) Acquisition of TPC-OsGLO3 fusion protein expression cassette Pubi-TPC-OsGLO3-Tnos

[0454] Primers were designed according to the expression gene sequence of TPC-OsGLO3 fusion protein, using the pMD18-TPC-OsGLO3 vector described in Example 1.1 as a template, under the guidance of primers TPC-OsGLO3-F and TPC-OsGLO3, amplify TPC-OsGLO3 by conventional methods The gene was recovered by electrophoresis and cloned into the recombinant donor vector Pubi-DI (for the vector map, see figure 2 ) Between the PstI and BamHI restriction sites of the multiple cloning site, the vector Pubi-DI-TPC-OsGLO3 was obtained.

[0455] Design primers according to the Pubi promoter sequence and Tnos terminator sequence on the Pubi-DI vector, use the above-mentioned Pubi-DI-TPC-OsGLO3 vector as a template, and amplify using conventional methods under the guidance of primers Pubi-...

Embodiment 13

[0462] Example 1.3 Obtaining GOC-pYL1305 carrier for transformation of GOC photorespiratory metabolic branch

[0463] With Pubi-DI-TPC-OsGLO3, 2×P35s-DII-TPC-OsOXO3 and Pubi-DI-TPC-KatE as donor vectors, pYL1305 (for the vector map, see Figure 4 ) is the acceptor carrier, referring to the method of Lin et al. (10):5962-5967), through three rounds of recombination, the GOC photorespiratory metabolic transformation branch carrier GOC-pYL1305 was finally obtained, and the vector map is as follows Figure 5 shown. The specific reorganization method is as follows:

[0464] (1) Preparation of donor plasmid and recipient plasmid

[0465] Collect 10 mL of the overnight bacterial solution, extract the plasmid with a plasmid mini-extraction kit, detect and measure the plasmid concentration by electrophoresis, and control the plasmid concentration at 100-200 ng / μL.

[0466] (2) Co-transfection and recombination of donor plasmid and recipient plasmid

[0467] Mix 3-4 μL of plasmids ...

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Abstract

The invention relates to an application of a photorespiratory branch protein in regulation and control of plant traits. Specifically, the invention provides a reagent combination and a fusion protein, and the reagent combination or the fusion protein comprises glycollic acid oxidase or coding nucleic acid thereof and/or oxalate oxidase or coding nucleic acid thereof and/or catalase or coding nucleic acid thereof. The agronomic traits of plants can be remarkably regulated and controlled by introducing the reagent combination or the fusion protein disclosed by the invention into plant cells.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of photorespiratory shunt proteins in regulating plant traits, more specifically, to a combination of reagents and fusion proteins for regulating plant agronomic traits, and their application in regulating plant traits. Background technique [0002] Photorespiration, also known as the C2 cycle, refers to the use of light energy by plant green tissues to absorb O 2 and release CO 2 The process of photorespiration requires the joint participation of chloroplasts, mitochondria, peroxisomes and cytoplasm, and depends on light and O 2 , both are indispensable. Photorespiration is the second largest metabolic flow in C3 plants after photosynthesis. Under normal environmental conditions, C3 plants can consume 25-30% of their photosynthetic products. loss will be more severe. High photorespiration will not only reduce the photosynthetic efficiency of plants, but also the ...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/29C12N15/62C12N5/10A01H5/00C07K19/00
CPCC12N15/825C12N15/8269C12N9/0006C12N9/0008C12N9/0065C12Y101/03C12Y102/03004C12Y111/01006C07K2319/02A01H5/00C07K14/415C07K19/00C12N5/10C12N15/62C12N15/82
Inventor 不公告发明人
Owner SHANDONG SHUNFENG BIOTECH CO LTD
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