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Application of LHAP1 protein and its coding gene in regulation of plant photosynthesis

A technology for photosynthesis and transgenic plants, which is applied in the fields of application, plant peptides, and plant products, and can solve the problems of in-depth research on interrelationships and influencing factors

Inactive Publication Date: 2018-04-20
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although in higher plants, LHCB protein is currently the most studied protein family with a relatively clear structure, the research on the functions of each member of the family and their interrelationships and influencing factors is still not in-depth

Method used

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  • Application of LHAP1 protein and its coding gene in regulation of plant photosynthesis
  • Application of LHAP1 protein and its coding gene in regulation of plant photosynthesis
  • Application of LHAP1 protein and its coding gene in regulation of plant photosynthesis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Embodiment 1, the acquisition of LHAP1 gene

[0074] 1. Mutant acquisition and phenotype

[0075] 1. Obtaining mutants

[0076] The seeds of wild-type Arabidopsis and mutants (T-DNA insertion mutants ordered from the NASC germplasm resource bank, the seed number is SALK_031802C) were treated with low-temperature vernalization (4°C light-proof treatment after soaking in water), and after 72 hours of treatment Sterilize with 10% sodium hypochlorite for 10 minutes, wash with sterile water 4 times, sow on MS medium containing 2% sucrose and 0.8% agar, place at 22°C, light intensity 100umolm -2 the s -1 and a photoperiod of 12h / 12h (light / dark) were cultivated to obtain wild-type Arabidopsis seedlings (WT) and mutant plants (lhap1).

[0077] The results showed that: compared with the wild-type plants, the mutant plants (lhap1) germinated later than the wild-type plants, and grew slowly. The leaf yellowing phenotype was aggravated during drought ( figure 1 ).

[0078] ...

Embodiment 2

[0102] Embodiment 2, acquisition of complementary plants and analysis of photosynthetic function

[0103] 1. Acquisition of Complementary Plants

[0104] 1. Construction of complementary vectors

[0105] Insert the DNA molecule shown in Sequence 1 in the sequence listing between the BamHI and KpnI restriction sites of the pSN1301 vector (purchased from BioVector Plasmid Vector Strain Cell Gene Collection Center, the product number is Biovector105802), and keep other sequences of the pSN1301 vector unchanged , to obtain the recombinant vector.

[0106] 2. Acquisition of recombinant bacteria

[0107] The recombinant vector obtained in step 1 was transformed into EHA105 Agrobacterium (purchased from Beijing Zhuangmeng International Biogene Technology Co., Ltd., product catalog number is ZC142) to obtain recombinant bacteria.

[0108] 3. Conversion

[0109] The homozygous mutant plants obtained in Step 1 of Example 1 were infected with the recombinant bacteria in Step 2 by inf...

Embodiment 3

[0135] Example 3. Yeast two-hybrid verification of the interaction between LHAP1 and LHCII transporter LTD

[0136] In this example, through the soluble yeast two-hybrid and other protein interaction experiments, it was found that LHAP1 had a strong interaction with the LHCII transporter LTD.

[0137] 1. Soluble yeast two-hybrid

[0138] Yeast two-hybrid detection system Matchmaker Gold Yeast Two-Hybrid was purchased from Clontech, and the transformation method was completely in accordance with the operation manual. The specific steps are as follows: According to ChlorolP prediction, the first 54 amino acids of the N-terminal of LHAP1 are transit peptides, and the amino acids (Aa55-382) after removing the transit peptide of LHAP1 are divided into the following six segments: LHPA1-1C-terminal (amino acids 55-110), LHPA1-2C-terminal (amino acid 134-147), LHPA1-3C-terminal (amino acid 171-190), LHPA1-4C-terminal (amino acid 214-253), LHPA1-5C-terminal (amino acid 277-313) and LH...

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Abstract

The invention discloses an application of an LHAP1 protein and its coding gene in the regulation of plant photosynthesis. The transmembrane protein LHAP1 localized in chloroplast is found, and it is found that the LHAP1 deletion mutant plant (lhap1) of the LHAP1 gene has a yellow-green leaf color and a reduced photosynthetic efficiency. The regulation effect of the LHAP1 in the transport process of the plant light-capturing chromoprotein LHCII is determined through the separation of the LHAP1 and the identification and analysis of gene functions, so the content of the LHCII protein can be regulated by using the gene actual production through a transgenic means.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to the application of LHAP1 protein and its coding gene in regulating plant photosynthesis. Background technique [0002] In higher plants, the light-harvesting pigment protein complex (LHC) is a kind of pigment protein complex that captures light energy and quickly transfers the energy to the reaction center to cause photochemical reactions. Although both photosystem I (LHCI) and photosystem II (LHCII) in higher plants contain their own light-harvesting pigment protein complexes, since LHCII binds nearly 50% of the pigments and about 1 / 3 of the proteins in the photosynthetic membrane, their physiological functions It is complex and easy to be extracted in large quantities, so it has attracted the attention of researchers. The components of LHCII are a family of pigment-protein complexes (LHCB) formed by proteins encoded by nuclear genes (Lhcb) and pigments, which are simila...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00
CPCC07K14/415C12N15/8269
Inventor 张立新刘旖旎马今方
Owner INST OF BOTANY CHINESE ACAD OF SCI
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