Photorespiration metabolism modified branch and application of photorespiration metabolism modified branch in C3 plant

A technology of metabolic transformation and photorespiration, applied in the field of plant genetic engineering, can solve metabolic problems and achieve the effects of reducing CO2 emissions, increasing biomass, and increasing photosynthetic rate

Active Publication Date: 2018-08-24
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the photorespiration metabolism of food crops. At the same time, new and more efficient photorespiratory metabolic transformation branches need to be further explored.

Method used

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  • Photorespiration metabolism modified branch and application of photorespiration metabolism modified branch in C3 plant
  • Photorespiration metabolism modified branch and application of photorespiration metabolism modified branch in C3 plant
  • Photorespiration metabolism modified branch and application of photorespiration metabolism modified branch in C3 plant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Obtaining of TPC-OsGLO3, TPC-OsOXO3, and TPC-OsCATC fusion protein expression genes

[0050] (1) Acquisition of TPC-OsGLO3 fusion protein expression gene

[0051] Primers were designed according to the cDNA sequences of OsGLO3 and TPC provided by NCBI (http: / / www.ncbi.nlm.nih.gov / ), and the primer sequences were as follows:

[0052] OsGLO3-F: 5′-catg agatct atggagctaatcacaaac-3' (the underline is the recognition site of the restriction endonuclease BglII).

[0053] OsGLO3-R: 5′-tta atggtgatggtgatgatg cctgtcgctgtcggtgat-3' (the underlined part is the HIS tag sequence).

[0054] TPC-1F: 5'-ATGGCCCCCTCCGTGATG-3';

[0055] TPC-1R: 5′-ctac agatct catgcacctgatcctgcc-3' (the underline is the recognition site of the restriction endonuclease BglII).

[0056] Using the cDNA of 2-week-old seedling leaves of the japonica rice variety Zhonghua 11 as a template, under the guidance of primers OsGLO3-F and OsGLO3-R, and primers TPC-1F and TPC-1R, the OsGLO3 and TPC g...

Embodiment 2

[0068] Example 2 Obtaining of TPC-OsGLO3, TPC-OsOXO3, and TPC-OsCATC Fusion Protein Expression Cassette Sequences

[0069] (1) Acquisition of TPC-OsGLO3 fusion protein expression cassette Pubi-TPC-OsGLO3-Tnos

[0070] Primers were designed according to the TPC-OsGLO3 fusion protein expression gene sequence (SEQ ID NO: 1), and the primer sequences were as follows:

[0071] TPC-OsGLO3-F: 5′-gact ctgcag atggccccctccgtgatg-3' (PstI recognition site is underlined).

[0072] TPC-OsGLO3-R:5′-cagt ggatcc ctaatggtgatggtgatgatg-3' (the BamHI recognition site is underlined).

[0073] Using the pMD18-TPC-OsGLO3 vector described in Example 1 as a template, under the guidance of primers TPC-OsGLO3-F and TPC-OsGLO3, the TPC-OsGLO3 gene was amplified by conventional methods, and the gene was cloned into recombinant Donor vector Pubi-DI (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.

[0...

Embodiment 3

[0092] Example 3 Obtaining GOC-pYL1305 carrier for transformation of GOC photorespiratory metabolic branch

[0093] With Pubi-DI-TPC-OsGLO3, 2×P35s-DII-TPC-OsOXO3 and Pubi-DI-TPC-OsCATC 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:

[0094] (1) Preparation of donor plasmid and recipient plasmid

[0095] 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.

[0096] (2) Co-transfection and recombination of donor plasmid and recipient plasmid

[0097] Mix 3-4 μL of plasmids ...

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Abstract

The invention discloses a photorespiration metabolism modified branch and an application of the photorespiration metabolism modified branch in C3 plant, which relate to the technical field of a plantgenetic engineering. The branch comprises three proteins of OsGLO3, OsCAT2 and OsOXO3. The branch can modify the C3 plant photorespiration, so that photorespiration is reduced, the photosynthetic efficiency is increased, and the plant biomass is increased. The experiment result shows that GOC photorespiration metabolism modified branch transferred to the plant appears that photosynthetic rate is increased, the biomass is increased, and chlorophyll is increased. The photorespiration metabolism modified branch has deep meaning for explicating high-light effect mechanism, producing crops and renewable energy sources, and reducing CO2 discharge, and during the practical application, the GOC photorespiration metabolism modified branch is transferred to different C3 plants, so that high-yield variety can be cultured.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to a branch of photorespiratory metabolism transformation, related genes and the application of the branch in C3 plants. 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 (Bauwe H, Hagemann M, Fernie A R. Photorespiration: players, partners and origin [J]. Trends in Plant Science, 2010, 15(6): 330-336). 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 serious (Leegood R C, Lea P J, Adcock M D, et al..The regulation and control of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N9/08C12N9/04C12N15/62C12N15/82A01H5/00A01H6/46
CPCC07K14/415C07K2319/02C12N9/0006C12N9/0065C12N15/8269C12Y101/03013C12Y111/01006Y02P60/20
Inventor 彭新湘沈博然张建军
Owner SOUTH CHINA AGRI UNIV
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