CN103709237B - Photosynthesis of plant associated protein OsPSF1 and encoding gene thereof and application - Google Patents

Abstract

The invention discloses a plant photosynthesis-related protein and its encoding gene and application. The amino acid sequence of the plant photosynthesis-related protein is shown in SEQ ID NO: 1, and the nucleotide sequence of the gene encoding the protein is shown in SEQ ID NO: 2 or shown in SEQ ID NO: 3, or its degenerate sequence. Introducing the gene of the present invention into rice for overexpression can complement the yellow leaf phenotype of the rice <i>psf1</i> mutant, and increase the photosynthetic rate of the transgenic plant leaves after oxidative stress treatment. The genome gene and the cDNA gene of the plant photosynthesis-related protein OsPSF1 provided by the invention provide a basis for cultivating high-light-efficiency crops with economic value. <b><u /></b>

Description

Plant photosynthesis related protein OsPSF1 and its coding gene and application

technical field

The invention belongs to the field of biotechnology, and relates to a plant photosynthesis-related protein OsPSF1 and its coding gene and application.

Background technique

The process by which plants use light energy to convert carbon dioxide and water into carbohydrates and release oxygen is called photosynthesis. Photosynthesis is the basis for the survival of the biological world and an important medium for the earth's carbon-oxygen cycle. Leaves are the main place for higher plants to carry out photosynthesis. In this process, the chlorophyll located in the chloroplast captures light energy and transmits it to the photosynthetic reaction center, thus starting the energy conversion process. Photosynthesis is considered to be the most important chemical reaction on the earth. Photosynthesis is the material basis for the formation of crop yields. 90-95% of the dry weight of plants comes from photosynthetic products. At present, the light energy utilization rate of rice in my country is very low, generally only about 0.5-1.0%, and the highest is only 2%. Theoretically, the light energy utilization rate of plants can reach 13~14%, and the ideal light energy utilization rate of rice should reach 3~5%, so it has great potential to improve the high light efficiency of rice.

How to make full use of the solar radiation that irradiates the earth's surface for photosynthesis is a fundamental problem in agricultural production. The use of light energy by crops is a comprehensive process. There are many reasons that affect the light energy utilization rate of crops, which can be roughly divided into the characteristics of the crop itself and the changes in the external environment such as light, temperature, carbon dioxide, water, drought and flood, salinity, and pests and diseases. Improving crop light efficiency requires improving light energy utilization efficiency under optimal conditions and crop adaptation to non-optimal external environments, which can be divided into strategies at the chloroplast level, single leaf level, plant level, and group level. The invention identifies a new related factor affecting plant photosynthesis, which can be used as a target for designing and cultivating germplasm with high light efficiency.

Contents of the invention

The object of the present invention is to provide a plant photosynthesis-related protein and its coding gene and application, the coding gene can be introduced into rice for overexpression, which can improve the photosynthesis efficiency of transgenic rice.

The plant photosynthesis-related protein (OsPSF1, Oryza sativa photosynthesisfactor 1) provided by the present invention is derived from the japonica rice variety "Nipponbare" (Oryza sativa subsp. japonicacv. Nipponbare), and its amino acid sequence is shown in SEQ ID NO:1.

The sequence of SEQ ID NO: 1 consists of 172 amino acid residues.

The present invention also provides a protein derived from the sequence of SEQ ID NO:1, which is related to plant stress tolerance through the substitution and/or deletion and/or addition of one or several amino acid residues to the amino acid sequence of SEQ ID NO:1.

In order to facilitate the purification of the plant photosynthesis-related protein (OsPSF1), the amino acid sequence shown in SEQ ID NO:1 can be connected to the amino-terminal or carboxyl-terminal of the protein, as shown in Table 1.

Table 1 Sequence of tags

Label Residues sequence Poly-Arg 5-6 (usually 5) RRRRR Poly-His 2-10 (usually 6) HHHHHH FLAG 8 DYKDDDDK Strep-tag 8 WSHPQFEK c-myc 10 EQKLISEEDL

The above-mentioned protein derived from the sequence of SEQ ID NO:1 can be artificially synthesized, or its coding gene can be synthesized first, and then obtained by biological expression, and its coding gene can be obtained by deleting one or more of the DNA sequences shown in SEQ ID NO:2. amino acid residues, and/or carry out a missense mutation of one or several base pairs, and/or connect the coding sequence of the tag shown in Table 1 at its 5' end and/or 3' end to obtain .

The present invention also provides a gene encoding the plant photosynthesis-related protein (OsPSF1), the nucleotide sequence of which is shown in SEQ ID NO: 2 or SEQ ID NO: 3, or its degenerate sequence.

At the same time, the present invention also provides a DNA molecule having more than 90% homology to the DNA sequence shown in SEQ ID NO: 2 or SEQ ID NO: 3 and encoding a stress tolerance-related protein.

The present invention also provides a DNA molecule that hybridizes to the DNA sequence shown in SEQ ID NO:2 or SEQ ID NO:3 under stringent conditions and encodes the protein; it also provides a DNA molecule that has more than 90% identity with the DNA molecule. DNA molecules that are native and encode stress tolerance-related proteins; the stringent conditions can be hybridized at 65 ^o C in a solution of 6×SSC, 0.5% SDS, and then mixed with 2×SSC, 0.1% SDS and 1 ×SSC, 0.1% SDS wash the membrane once.

The DNA molecule shown in SEQ ID NO: 3 is composed of 3734 nucleotides, from the 5' end 1-1495 is a promoter, 1496-1839 is the first exon, 2179-2299 is an intron, 2583- 3191 is the second exon.

The invention also provides the recombinant expression vector of the gene.

An existing plant expression vector can be used to construct a recombinant expression vector containing the gene.

The plant expression vectors include binary Agrobacterium vectors and vectors that can be used for plant microprojectile bombardment and the like. The plant expression vector can also include the 3' untranslated region of the foreign gene, that is, the polyadenylation signal and any other DNA fragments involved in mRNA processing or gene expression. The polyA signal can direct polyA to be added to the 3' end of the mRNA precursor, such as Agrobacterium crown gall tumor induction (Ti) plasmid gene (such as nopain synthase Nos gene), plant gene (such as soybean storage The untranslated region transcribed at the 3' end of protein gene) has similar functions.

When using the gene to construct a recombinant plant expression vector, any enhanced promoter or constitutive promoter can be added before its transcription start nucleotide, such as the cauliflower mosaic virus (CAMV) 35S promoter, maize Ubiquitin promoters (Ubiquitin), which can be used alone or in combination with other plant promoters; in addition, when using the gene of the present invention to construct a plant expression vector, enhancers, including translation enhancers or transcription enhancers, can also be used, These enhancer regions can be ATG initiation codons or adjacent region initiation codons, etc., but must be in the same reading frame as the coding sequence to ensure correct translation of the entire sequence. The sources of the translation control signals and initiation codons are extensive and can be natural or synthetic. The translation initiation region can be from a transcription initiation region or a structural gene.

In order to facilitate the identification and screening of transgenic plant cells or plants, the plant expression vectors used can be processed, such as adding genes (GUS gene, luciferase gene, etc.) Genes, etc.), antibiotic resistance markers (gentamicin markers, kanamycin markers, etc.), or chemical resistance marker genes (such as herbicide resistance genes), etc. Considering the safety of the transgenic plants, the transformed plants can be screened directly by adversity without adding any selectable marker gene.

The recombinant expression vector is a recombinant plasmid obtained by inserting the gene into the multiple cloning sites of pCAMBIA1300 and pCAMBIA2300.

Expression cassettes, transgenic cell lines and recombinant bacteria containing any of the above genes (OsPSF1) all belong to the protection scope of the present invention.

A pair of primers for amplifying the full length or any fragment of the gene (OsPSF1) also falls within the protection scope of the present invention.

The invention also provides the application of the gene in cultivating transgenic plants with improved photosynthetic efficiency.

The above application is to introduce the gene encoding the plant photosynthesis-related protein into the target plant (such as plant cells or tissues) to obtain a transgenic plant with higher photosynthetic efficiency than the target plant. Specifically, the recombinant expression vector is introduced into the target plant, and the photosynthetic efficiency of the transgenic plant is higher than that of the recipient plant.

By using any carrier capable of directing the expression of exogenous genes in plants, the gene encoding the protein is introduced into plant cells to obtain transgenic cell lines and transgenic plants with enhanced drought tolerance. The expression vector carrying the gene can transform plant cells or tissues by conventional biological methods such as Ti plasmid, Ri plasmid, plant virus vector, direct DNA transformation, microinjection, electrical conduction, Agrobacterium-mediated, and transform the transformed The plant tissue is grown into a plant. The transformed plant host can be either a monocotyledon or a dicotyledonous plant, such as: tobacco, lotus root, Arabidopsis, rice, wheat, corn, cucumber, tomato, poplar, lawn grass, alfalfa, etc. .

Experiments show that introducing the DNA sequence encoding and regulating plant photosynthesis-related factor OsPSF1 of the present invention into rice for overexpression can improve the photosynthesis efficiency of transgenic rice. The genome gene and cDNA gene of the invention encoding and regulating plant photosynthesis-related factor OsPSF1 provide a basis for cultivating other high-light-efficiency plants with economic value.

Description of drawings

Figure 1 shows the psf1 mutants screened from the mutant library, where A is the 2-week-old wild-type and psf1 mutant seedlings, B is the 2-week-old wild-type and psf1 mutant leaves.

Figure 2 shows the chlorophyll content of psf1 mutant leaves, where A is the total amount of chlorophyll a and chlorophyll b in wild type and psf1 mutant leaves, and B is the ratio of chlorophyll a to chlorophyll b in wild type and psf1 mutant leaves.

Figure 3 is the sequencing spectrum of the psf1 mutation site.

Figure 4 shows the chlorophyll content of the leaves of the complementary transgenic plants.

Figure 5 is the PCR identification of complementary transgenic plants.

Fig. 6 is the sequence map of complementary transgenic plants.

Figure 7 is the PCR identification of overexpression transgenic plants.

Figure 8 is the RT-PCR identification of overexpression transgenic plants.

Figure 9 is the determination of photosynthetic rate of leaves of overexpressed transgenic plants.

detailed description

The present invention will be further clarified through the detailed description of specific embodiments below, but it is not intended to limit the present invention, but only for illustration.

The experimental methods in the following examples, unless otherwise specified, are conventional methods, and the test materials used, unless otherwise specified, are purchased from conventional biochemical reagent stores. The experiments are all set to repeat three times, and the results are averaged .

Example 1: Screening of psf1 mutants and map-based cloning of mutant genes

1. Screening of rice mutant library

Take 2000g seeds of japonica rice variety "Nipponbare" (Oryza sativa subsp. japonicacv. Nipponbare) (Institute of Crop Science, Chinese Academy of Agricultural Sciences, No. WD-10576; original source is unclear), soak the seeds in water for 16 hours, and then use 0.5% methyl Ethyl sulfonate (EMS) was treated at 28°C for 12 hours, and then washed 10 times with a large amount of tap water to obtain M1 seeds, which were sown and harvested from individual plants to obtain the M2 mutant library.

2000 copies of M2 mutant seeds were germinated, and the leaf color was observed at the seedling stage of 2 weeks, and a light yellow mutant was found, named psf1, as shown in Figure 1 . 80% acetone was used to extract the leaf pigments of wild type and psf1 mutants, and the chlorophyll content was measured by colorimetry. It was 5 times that of the wild type, indicating that chlorophyll b decreased more than chlorophyll a (Fig. 2AB).

2. Map-based cloning of mutant genes of psf1 mutants

The psf1 mutant was backcrossed with Nippon Haru wild type, and the ratio of light yellow leaf plants to green leaf plants in the seedlings of the F2 backcross segregation population was counted. The results are shown in Table 2. The ratio of light yellow leaf to normal leaf color plants in 3 independent F2 backcross populations was analyzed by chi-square test and binomial test and found to be in line with the segregation ratio of 1:3 controlled by a single gene (Table 2).

Table 2 The segregation ratio of light yellow leaves and green leaves normal plants of backcross F2 generation plants of psf1 mutant

The psf1 mutant was crossed with Dular to obtain F1, and F1 was self-crossed to obtain F2 population. Linkage analysis of 352 F2 light yellow leaves isolated by SSR molecular markers found that the mutation site was closely linked with molecular markers Chr7-20.1 and Chr7-20.2 , all 8 ORFs in this segment were sequenced and found that the 284th position T of psf1 mutant ORF7 was mutated to G, and the encoded amino acid changed from neutral leucine to basic arginine, and the position of Dular was the same as that of wild type (image 3). ORF7 encodes a protein with ankyrin domain that is presumed to be located in the plastid, which we named OsPSF1 protein (LOC_Os07g33660).

Example 2: Cloning of OsPSF1 cDNA and Genomic Gene

1. Cloning of OsPSF1 cDNA

Take 0.2g leaves of japonica rice variety "Nipponbare" (Oryza sativa subsp. japonicacv. Nipponbare), grind with liquid nitrogen, and extract total RNA by TRIzol method. Take 2 μg of total RNA for reverse transcription with M-MLV reverse transcriptase to synthesize the first strand of cDNA, which is used as a template to carry out PCR reaction with specific primers. After the PCR product was separated and recovered by electrophoresis, it was cloned into pEASY-T (Beijing Quanshijin Biotechnology Co., Ltd.), named pEASY-T-OsPSF1ORF, and sequenced.

The specific primer pair A is as follows:

Upstream primer: 5'-ATGGCATCCATCCCGTGCAC-3';

Downstream primer: 5'-TCAGGCGGCCAAGGTGGCGG-3'.

Sequencing results show that the nucleotide sequence of the fragment is shown in SEQ ID NO:2, encoding the protein shown in SEQ ID NO:1.

2. Acquisition of OsDRPSF1 genome gene

Genomic DNA of Nipponbare leaves was extracted by CTAB method, and the genomic DNA was used as a template to react with specific primers. After the PCR products were separated and recovered by electrophoresis, they were cloned into pEASY-T (Beijing Quanshijin Biotechnology Co., Ltd.) for sequencing.

The specific primer pair B is as follows:

Upstream primer: 5'-GTGTGCTGTTGTCCCAGTATG-3';

Downstream primer: 5'-AACCTGTTAGCCCTACGAGTG-3'.

Sequencing results showed that the nucleotide sequence of the fragment is shown in SEQ ID NO:3.

The protein shown in SEQ ID NO: 1 was named OsPSF1, and the gene encoding OsPSF1 was named OsPSF1. The recombinant plasmid obtained by linking the nucleotide shown in SEQ ID NO: 2 with pEASY-T was named pEASY-T-OsPSF1.

Example 3: Functional complementation of psf1 mutants

1. Construction of pCAMBIA1300-OsPSF1 expression vector

1. Digest the recombinant plasmid pEASY-T-OsPSF1 in Implementation 1 with restriction endonucleases XbaI and KpnI, and reclaim the 3.7kb fragment comprising the OsPSF1 sequence.

2. Digest pCAMBIA1300 (Cambia, GPO Box 3200, Canberra, ACT 2601, Australia) with restriction enzymes XbaI and KpnI, and recover the backbone.

3. Ligate the fragment obtained in step 1 with the fragment obtained in step 2 to obtain pCAMBIA1300-OsPSF1.

The acquisition of transgenic plants

1. Introduce the recombinant expression vector pCAMBIA1300-OsPSF1 into Agrobacterium AGL0 (ATCC®BAA-100™, www.atcc.org) by electroporation.

5. Agrobacterium AGL0 containing pCAMBIA1300-OsPSF1 was used to infect the embryo-type callus induced by the psf1 mutant, and then the resistant callus was selected in MS medium (containing 30 mg/L hygromycin), 15 times per generation. 3 generations in total, and then the resistant callus was induced into complete plants, transplanted in the field, and the T1 generation seeds of the transgenic plants were harvested.

The T1 generation seeds of the psf1 mutant transformed with OsPSF1 were germinated, and the leaf color of the seedlings returned to normal after 2 weeks. The chlorophyll content was determined to be the same as that of the wild-type control, but significantly higher than that of the psf1 mutant (Figure 4).

Leaf DNA was extracted by CTAB method, and the hygromycin phosphotransferase gene (hpt) specific primers 5'-CATCGAAATTGCCGTCAACC-3' and 5'-AGCCTGACCTATTGCATCTC-3'PCR detection, the results showed that the psf1 mutant complementary transgenic plants (com) There are specific amplification bands, while the wild-type and psf1 mutant DNA have no specific amplification bands (Figure 5). This indicated that the screening marker hpt gene on the vector had been introduced into the psf1 mutant complementary transgenic plants.

Because the wild-type OsPSF1 DNA fragment and the hpt gene on the pCAMBIA1300-OsPSF1 plasmid vector are located in the T-DNA region in tandem, it can be speculated that the wild-type OsPSF1 DNA fragment and the hpt gene fragment are co-introduced into psf1 mutant complementation transgenic plants. Because the psf1 mutant is a point mutation, the PCR method cannot separate the introduced wild-type OsPSF1 DNA from the mutant fragment in the psf1 mutant genome. We used OsPSF1 gene-specific primers 5'-ATGGCATCCATCCCGTGCAC-3' and 5'-TCAGGCGGCCAAGGTGGCGG-3 'Respectively PCR amplified from the wild type, psf1 mutant and complementary transgenic plant genomic DNA, and then sequenced the PCR products, the results are shown in Figure 6. The complementation transgenic plant (com) was T/G heterozygous at position 284 of OsPSF1 ORF, which clearly proved that the wild-type OsPSF1 fragment had been introduced into the psf1 mutant.

The above results indicated that the introduction of OsPSF1 gene sequence into the psf1 mutant could transform the light yellow phenotype into normal leaf color, which further proved that the OsPSF1 gene mutation was the gene that caused the light yellow phenotype.

Example 4: Overexpression of OsPSF1 in rice

1. Construction of pCAMBIA2300-Act-OsPSF1 expression vector

1. Using the recombinant plasmid pEASY-T-OsPSF1ORF in Implementation 1 as a template, amplify the OsPSF1 sequence with the following specific primers.

Upstream primer: 5'-TCCCCCGGGATGGCATCCATCCCGTGC-3';

Downstream primer: 5'-AACTGCAGTCAGGCGGCCAAGGTGGCGG-3'.

After the PCR products were separated by electrophoresis, a 520bp DNA fragment was recovered by cutting the gel, and then digested with restriction enzymes XmaI and PstI.

2. Digest pCAMBIA2300-ACT (Actin promoter) (Cambia, GPO Box 3200, Canberra, ACT 2601, Australia) with restriction enzymes XmaI and PstI, and recover the backbone.

3. Ligate the fragment obtained in step 1 with the fragment obtained in step 2 to obtain pCAMBIA2300-Act-OsPSF1.

The acquisition of transgenic plants

1. The recombinant expression vector pCAMBIA2300-Act-OsPSF1 was introduced into Agrobacterium AGL0 by electric shock method.

5. Agrobacterium AGL0 containing pCAMBIA2300-Act-OsPSF1 was used to infect the embryo-type callus produced by Nipponbare wild-type induction, and then the resistant callus was screened in MS medium (containing 100mg/L G418), 15 times per generation. 3 generations in total, and then the resistant callus was induced into complete plants, transplanted in the field, and the T1 generation seeds of the transgenic plants were harvested.

3. Molecular detection of transgenic plants

Leaf DNA was extracted by CTAB method, using Act-OsPSF1 specific primers 5'-TCTGCGATCCGCCGTTGTTG-3' and 5'-GCGCCTCCATCTTGTCGTAG

-3'PCR detection, the results showed that there were specific amplification bands in the transgenic plants, while the wild-type DNA had no specific amplification bands (Fig. 7), which indicated that Act-OsPSF1 had been introduced into the transgenic plants.

Total RNA was extracted from leaves of 2-week-old wild-type and Act-OsPSF1 transgenic plants (ox1, 2, 3) by TRIzol method, and 2 μg of total RNA was treated with 2 u RQ1 RNase-free DNase (Promega) at 37 °C for 30 min. After terminating the reaction solution, use polyA as a primer, use M-MLV reverse transcriptase at 42 °C for 1 hour to synthesize the first strand of cDNA, then use cDNA as a template, and use Actin (primers 5'-GCCAATCGTGAGAAGATGAC-3' and 5'- CTATGAAGGAAGGCTGGAAG-3') was used as an internal reference, and the expression of PSF1 gene (primers 5'-ATGGCATCCATCCCGTGCAC-3' and 5'-TCAGGCGGCCAAGGTGGCGG-3') was detected by PCR method, and the results are shown in Figure 8. The expression of Actin gene was basically the same among the plants, while the expression level of PSF1 gene in Act-OsPSF1 transgenic plants (ox1, 2, 3) was about 5-10 times that of the control plants.

4. Determination of photosynthetic rate

The leaves of 3-week-old wild-type and Act-OsPSF1 transgenic plants (ox1, 2, 3) were sprayed with 50 μmol L ^-1 methyl viologen (MV), with water as the control, and 3 days later with LI6400XT (LI-COR, Inc ) portable photosynthetic instrument to measure the net photosynthetic rate of 3-week-old wild-type and Act-OsPSF1 transgenic plants (ox1, 2, 3) leaves, the parameters were set as CO2 400 μmol CO2mol ^-1 , light intensity 800 μmol m ^-2 s ^-1 , temperature 30 °C, the results showed that the net photosynthetic rate between the transgenic plants (ox1, 2, 3) and the wild-type leaves had no significant difference under the control condition, while the transgenic plants (ox1, 2, 3) after treatment with methyl viologen The net photosynthetic rate of leaves was higher than that of the wild type (Fig. 9). This indicates that overexpression of OsPSF1 gene can increase the photosynthetic rate of transgenic plants after oxidative stress.

<110> Institute of Biotechnology, Chinese Academy of Agricultural Sciences

<120>Plant photosynthesis related protein OsPSF1 and its coding gene and application

<160> 3

<210> 1

<211>172

<212> amino acid

<400> 1

Met Ala Ser Ile Pro Cys Thr Phe Gln Leu Ser Ala Arg Ala Ser Ser Ala SerAla Ala 20

Ala Ala Ala Arg Arg Ser Pro Arg Ala Ala Ala Arg Leu Gly Trp Leu Arg Pro Ser Arg 40

Leu Ser Ala Val Val Pro Ala Ser Glu Ser Gly Arg Val Gly Pro Thr Cys Phe Phe Lys 60

Phe Gly Asn Lys Asp Ala Glu Gly Ala Gly Ile Tyr Gly Ser Gln Gly Arg Asp Asp Phe 80

Asp Arg Asp Asp Val Glu Gln Tyr Phe Asn Tyr Met Gly Met Leu Ala Val Glu Gly Thr 100

Tyr Asp Lys Met Glu Ala Leu Leu Asn Gln Asp Ile His Pro Val Asp Ile Leu Leu Met 120

Leu Ala Ala Ser Glu Gly Asp Lys Pro Lys Leu Glu Glu Leu Leu Arg Ala Gly Ala Lys 140

Tyr Asp Val Lys Asp Val Asp Gly Arg Thr Ala Leu Asp Arg Ala Ala Asp Asp Thr Arg 160

Glu Phe Ile Leu Gly Phe Ala Ala Thr Leu Ala Ala 172

<210> 2

<211>519

<212>DNA

<400> 2

ATGGCATCCATCCCGTGCACCTTCCAGCTGAGCGCGAGGGCGTCGTCGGC 50

GTCGGCGGCGGCGGCGGCGAGGAGGTCGCCGCGGGCGGCGGCGAGGCTGG 100

GGTGGCTGCGGCCGTCGCGGCTGAGCGCGGTGGTGCCGGCGAGCGAGAGC 150

GGGAGGGTGGGGCCGACGTGCTTCTTCAAGTTCGGGAACAAGGACGCCGA 200

GGGCGCCGGCATCTACGGCAGCCAGGGCAGGGACGACTTCGACCGCGACG 250

ACGTCGAGCAGTACTTCAACTACATGGGGATGCTGGCGGTGGAGGGCACC 300

TACGACAAGATGGAGGCGCTGCTGAACCAGGACATCCACCCGGTGGACAT 350

CCTCCTCATGCTCGCCGCCTCCGAGGGCGACAAGCCCAAGCTCGAGGAGC 400

TCCTCCGCGCCGGCGCCAAGTACGACGTCAAGGACGTCGACGGCCGGACG 450

GCGCTCGACCGCGCCGCCGACGACACCAGGGAGTTCATCCTCGGCTTCGC 500

CGCCACCTTGGCCGCCTGA 519

<210> 3

<211>3734

<212>DNA

<400> 3

GTGTGCTGTTGTCCCAGTATGGCACGGATCAAATTTATCCATCTTTTTTT 50

TCACCTTTACTTTTTACTGAAATTTATACACATACCCTTTTGAAGCAGAG 100

TTGCATACCCATTTTCTTTTTTTGGTTGCTATACAACGGAATCTCCTTCA 150

GACGTGATGATACCGATTAGTTATCAAGTCCTAATACCTAATCGGTATCG 200

GGTGATACATATCAAGTATCATACGATTCTACCACGTATTAGGTGATACT 250

TGGTTATTTCCTATTAGGTATACGATTCTTTACCCATAGAAGGTGATACTT 300

ACGAGATATTAGGTGATACCTATTAGGTATCATGCGATTCTTATCACGTA 350

GAAGGCTATACTTGCGAGATATCAGGTGATACCTATGAAGTATCAGGCGA 400

TACCTACTAGATAATAGGTGATTTCTACTGGGTATCAGAAGCTGGACATA 450

CCTGACGCCGACGCCGACGACCACTACCGCCCTTCCTCCCCTTGGCTCTA 500

GCCACCACCGCCGCTCCCTTTCCTGGCTCCGGCTACCACCGCTGCCCCTC 550

CCTCCTCTAGCTCTTGCCACCGTCACCACTGCGTTCCTTAGCTTCGGCCA 600

CCGACGCCCCTCGTCTCTTCGTTGATGTATCCCCCGCCGCCGGCTTGGCC 650

GCGACGCGCCGCTGCCTCGACGCGACACAGATGGGGTCCTCGACACCACG 700

GCAGCTCACCACAAGGGGCTCGTCGCCGGCGGCCTCGTCCTCTTCTCGCC 750

GAGCAGGTCCTCTTCCCCAGCCCGCGCCGAAGGTCCTCGCCAATAAGCGG 800

CAACCACCTCCTCCTCATGGCTCAGGCGTTCAGCTCTCGCTTCTCTTTGG 850

ATAGGCAAGGAGGACAAAGGATCCCGTCCCAACTGGATCCCGGTGCGTAG 900

CATTTCCGTTTCTTTTTTTCACATTTTTTTATGGGTGTCTATTTCACATGG 950

TAATTAGACAGCTAATTCCAAGCTTAAAACCTGGCTTAATGCTACTGCTT 1000

TTTATCCTAGGAATCTATTTTGATTTCTCCAAGAGGATGTCTTCTCGTAAG 1050

CTTAAAAACCATTCAAATGGTTATAAAAAAAATTCTAAAAAAAATTAACAA 1100

CACTCATACAATTTACTTCCTCCGTTCAAAAAAAACCAACGGATGTGACC 1150

CCCTCCTAATACAGTAAATCTAAAACAGCACTTAATCCAAATTTGTTATAT 1200

TAGGAGATGTCACATCCTTTTAGGTTGTTTTTTTTGGTGACGGATTTTTTT 1250

TTTACGGAGGGATGGAGTATATCTCCACACAGCTCTAGCACGTACTCCTAT 1300

CTCACACAGATTTATCACAAATCATACACTTTTTTTTAATATCTTCTACA 1350

CTGCACTTTATGAATATCTCATGTCCTGTCTTCCACAGGAAAAAAGTGA 1400

AATAGTGAAAAGATTCAGAATTAGGAAGCATCTAATCACCTAACTTGACA 1450

GCAATCAGTTTCAAGTGGCCAAGTGCCAAACAGTTTCAAGATTACTTACA 1500

ACTTATCCGTTCCTCTCATATATCTCTCTCCCCTCTTCTCCACCTCTCGC 1550

TACTCCACCATCCAACGTGCGAGAAGCCATGGCATCCATCCCCGTGCACCT 1600

TCCAGCTGAGCGCGAGGGCGTCGTCGGCGTCGGCGGCGGCGGCGGCGAGG 1650

AGGTCGCCGCGGGCGGCGGCGAGGCTGGGGTGGCTGCGGCCGTCGCGGCT 1700

GAGCGCGGTGGTGCCGGCGAGCGAGAGCGGGAGGGTGGGGCCGACGTGCT 1750

TCTTCAAGTTCGGGAACAAGGACGCCGAGGGCGCCGGCATCTACGGCAGC 1800

CAGGGCAGGGACGACTTCGACCGCGACGACGTCGAGCAGGTGAGCCAGCT 1850

TCTCGTTTCTTCCTCCTCCCCATGTTGCCTTAGGTGTTCCTTGATTGCTCC 1900

AGTCCATGGGGAATGTGCTGACGAATGACTGAGATGCTACCTTACCACTG 1950

CTATATTACTTGGTGGATTGGTAATTCTGATCCTAATGTGAGACATACGA 2000

AGTAGTTGATTGCTATACTTGCTAGTTTGGTTTGAGGACAAATGATGCAG 2050

CATTTGATTTGAGTTGAATTAAGGCATGTCTTTTTTCCTTATGCTTATACT 2100

TATCAACCAACATTTGAATTTTAAATTTGGAGTTGATTTAGAGGTTTTTT 2150

CATTGAAGTATATTTTTCAGCATTTGCTTTTAAATCTCTAAGAATATACA 2200

TATAAAAGTTTTATTTACAAATTACTTTTTATTTGCAAATATTCCGTTTC 2250

GCTTGCTTCGCTTATTCCGCGAATAAGCAAAACGATGGCTCGTATCATAG 2300

CCAGCAGTAGCAAAGAGGAAGAGAAGAAAAGAAAAAAACAACAATCACCT 2350

CGCAATTTAGTCTTTGTAGTTTCATGGACAAAATGCAAATTGCAGCTGCA 2400

GGGTATCTGAAATTCTGAATATGCTAATCAAGAATTCAAGATAGCTGTAG 2450

ACTAGCACATTCAGATTGAAGAACAGCACCTGAACATGGAAAGCTCTAGT 2500

GACAAGAAAGATTCAGAATAGTAACTTTGATTCCTGAGATGAGATTCTGA 2550

ATGTATCTGAATCTTTCTGTCTGCGCTTGTAGTACTTCAACTACATGGGG 2600

ATGCTGGCGGTGGAGGGCACCTACGACAAGATGGAGGCGCTGCTGAACCA 2650

GGACATCCACCCGGTGGACATCCTCCTCATGCTCGCCGCCTCCGAGGGCG 2700

ACAAGCCCAAGCTCGAGGAGCTCCTCCGCGCCGGCGCCAAGTACGACGTC 2750

AAGGACGTCGACGGCCGGACGGCGCTCGACCGCGCCGCCGACGACACCAG 2800

GGAGTTCATCCTCGGCTTCGCCGCCACCTTGGCCGCCTGATCGATCGCTT 2850

CGCCGACGACGACGACCTTGATGCCTGGATATGTTTCGGCTATACTAGTA 2900

TACTACGTATTTATCTTTTTCTTATTTTTTTTCTTTTCAAGTTTCTCCAAG 2950

TTTTTCTTGTGTTTGGTAGAGTTTAGCCGGGAAATTCACAAGTTGTTTAC 3000

AGTTCAAGAGGAAGTAGCTTCATCTGTATCTTTCACTTTCTTCGCTGTGT 3050

GAGGCTATGCTTTGTAGAAATTGTTGTTACAAGTAGTATCTTCCACTGTT 3100

TCAATATGAAAGTAAGAAGAAGCTTTGGATGCATGATACTATTGGAATTG 3150

GAAGCTTTGATGTTCACATAATAACATGTTAAGATGCCAAATCCTAACAG 3200

CAAGCAATAGCATGAACTTCAAGCAAAGCATTACTGCAACAGCAAGCAAA 3250

ACTTCATCCTTTATTGTCTCAGATTAACTGCTATGTCCATGTTGCCGTCA 3300

GTACATGTTCTATGTACTAGCCTAGAATTACAAAACAGCATTTCGCCGAAA 3350

GATTCACTTGGGGGAAACAAACAGTCTGCGACCACAGCCTCAACAACGAA 3400

ATTACGAGTATAAACTCTATAAACCCCCTCATGAATCCGCCAGATCATAT 3450

TATTCTTTTCTTTCTTATATTCATCTTGGTATATATTTCTTATAAGGACT 3500

GTGAATGTACTCCTACCTACAAAAACATTGGGTAAATGGCAGTTGTACTTC 3550

AACTTTGGATGTGACCGGAGCGCTTGCTGGAGCGCCGTGTCCACCGTCAC 3600

GCCCTCGCGGAACAGCTTGTCATACAGGAGGCAGATGAACTGAGCCATGT 3650

ACTCCTCATCATCGAAATCTTTGCTGCGATCGCCATTTTTCTCCATGTCG 3700

CTGTCTTCCCAGTCACTCGTAGGGCTAACAGGTT 3734

Claims (4)

1. The application of a plant photosynthesis-related protein coding gene in cultivating transgenic plants with improved photosynthetic efficiency, wherein the amino acid sequence of the related protein is as shown in SEQ ID NO:1. 2. The application according to claim 1, wherein the nucleotide sequence of the gene is shown in SEQ ID NO:2 or SEQ ID NO:3. 3. The application according to claim 1 or 2, characterized in that: the expression vector containing the gene is introduced into the plant to obtain a transgenic plant whose photosynthetic efficiency is higher than that of the recipient plant. 4. The application according to claim 3, characterized in that: the transgenic plant is transgenic rice.