CN103709237A

CN103709237A - Plant photosynthesis associated protein OsPSF1, and encoding gene and application thereof

Info

Publication number: CN103709237A
Application number: CN201310591977.0A
Authority: CN
Inventors: 权瑞党; 张海文; 张执金; 王娟; 黄荣峰
Original assignee: Biotechnology Research Institute of CAAS
Current assignee: Biotechnology Research Institute of CAAS
Priority date: 2013-11-22
Filing date: 2013-11-22
Publication date: 2014-04-09
Anticipated expiration: 2033-11-22
Also published as: CN103709237B

Abstract

The invention discloses plant photosynthesis associated protein OsPSF1, and encoding gene and application thereof. The amino acid sequence of the plant photosynthesis associated protein is shown as SEQ ID NO: 1, the nucleotide sequence of a gene encoding the protein is shown as SEQ ID NO: 2 or SEQ ID NO: 3 or degenerate sequences thereof. By introducing the gene provided by the invention into paddy rice for overexpression, the yellow-leaf phenotype of paddy rice psf1 mutant is complemented, and the photosynthetic rate of transgenetic plant leaves subjected to oxidation stress processing is raised. The genome gene and the cDNA gene of the plant photosynthesis associated protein OsPSF1 provide a base for cultivating high-photosynthetic-efficiency crops possessing economic value.

Description

Photosynthesis of plant associated protein OsPSF1 and encoding gene thereof and application

Technical field

The invention belongs to biological technical field, relate to a kind of photosynthesis of plant associated protein OsPSF1 and encoding gene and application.

Background technology

The process that plant utilization luminous energy is transformed into carbohydrate releasing oxygen by carbonic acid gas and water is called photosynthesis.Photosynthesis is the basis that organic sphere is depended on for existence, is also the important medium of earth carbon oxygen cycle.Blade is that higher plant carries out photosynthetic main place, in this process, first by the chlorophyll that is positioned at chloroplast(id), is caught luminous energy and is delivered to photosynthetic reaction center, thereby started the conversion process of energy.Photosynthesis is considered to most important chemical reaction on the earth, and photosynthesis is the basic substance that crop yield forms, and 90 ~ 95% plant dry weight is from photosynthate.At present, China's light energy of rice utilization ratio is very low, generally only has 0.5 ~ 1.0% left and right, the highest by only 2%.In theory, plant luminous energy utilization ratio can reach 13 ~ 14%, and the desirable efficiency of light energy utilization of paddy rice should reach 3 ~ 5%, so improve the high light efficiency of paddy rice, has very large potentiality.

How making full use of the solar radiant energy that is irradiated to earth surface and carry out photosynthesis, is a fundamental issue in agriculture production.Crop is a combined process to the utilization of luminous energy, and the reason that affects Efficiency of energy conversion photosynthesis potency is a lot, substantially can be divided into the external environments such as the feature of crop own and illumination, temperature, carbonic acid gas, moisture and drought and waterlogging, saline and alkaline, disease and pest and change.Improve crop light efficiency and need to improve the efficiency of light energy utilization and the adaptation of crop to non-best external environment under optimum condition, can be divided into the strategy of chloroplast(id) level, single leaf level, plant level and population level different levels.The present invention identifies a new correlation factor that affects photosynthesis of plant, can be used as the target spot that high light efficiency germplasm is cultivated in design.

Summary of the invention

The object of this invention is to provide a kind of photosynthesis of plant associated protein and encoding gene and application, by crossing and express in this encoding gene Introduced into Rice, can improve transgenic paddy rice photosynthetic efficiency.

Photosynthesis of plant associated protein provided by the invention (OsPSF1, oryza sativaphotosynthesis factor 1), derive from japonica rice variety " Japan fine " ( oryza sativasubsp. japonicacv. Nipponbare), its aminoacid sequence is as shown in SEQ ID NO:1.

The sequence of SEQ ID NO:1 is comprised of 172 amino-acid residues.

The present invention also provides the aminoacid sequence of SEQ ID NO:1 through the replacement of one or several amino-acid residue and/or disappearance and/or interpolation and relevant to plant stress tolerance by the derivative protein of SEQ ID NO:1 sequence.

In order to make described photosynthesis of plant associated protein (OsPSF1) be convenient to purifying, can connect label as shown in table 1 at N-terminal or the C-terminal of the protein being formed by the aminoacid sequence shown in SEQ ID NO:1.

The sequence of table 1 label

Label	Residue	Sequence
			Poly-Arg	5-6(is generally 5)	RRRRR
Poly-His	2-10(is generally 6)	HHHHHH
			FLAG	8	DYKDDDDK
Strep-tag	8	WSHPQFEK
			c-myc	10	EQKLISEEDL

Above-mentioned can synthetic by the derivative protein of SEQ ID NO:1 sequence, also can first synthesize its encoding gene, carrying out biological expression obtains again, its encoding gene can be by lacking the codon of one or several amino-acid residue in the DNA sequence dna shown in SEQ ID NO:2, and/or carry out the missense mutation of one or several base pair, and/or the encoding sequence that connects the label shown in table 1 at its 5 ' end and/or 3 ' end obtains.

The present invention also provides the gene of the described photosynthesis of plant associated protein of a kind of coding (OsPSF1), and its nucleotide sequence is as shown in SEQ ID NO:2 or shown in SEQ ID NO:3, or its degenerate sequence.

Meanwhile, the present invention also provides as shown in SEQ ID NO:2 or the DNA sequence dna shown in SEQ ID NO:3 has 90% above homology, and the DNA molecular of coding stress tolerance correlative protein.

The present invention also provide under stringent condition with DNA sequence dna hybridization shown in SEQ ID NO:2 or shown in SEQ ID NO:3 and the DNA molecular of encoding said proteins; Also provide with this DNA molecular simultaneously and there is 90% above homology, and the DNA molecular of coding stress tolerance correlative protein; Described stringent condition can be at 6 * SSC, in the solution of 0.5% SDS, 65 ^ounder C, hybridize, then use 2 * SSC, 0.1% SDS and 1 * SSC, 0.1% SDS respectively washes film once.

DNA molecular shown in SEQ ID NO:3 is comprised of 3734 Nucleotide, from 5 ' end 1-1495 position, is promotor, and 1496-1839 is First Exon, and 2179-2299 is intron, and 2583-3191 is Second Exon.

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

The recombinant expression vector that available existing plant expression vector construction contains described gene.

Described plant expression vector comprises double base agrobacterium vector and can be used for the carrier etc. of plant micropellet bombardment.Described plant expression vector also can comprise 3 ' end untranslated region of foreign gene, comprises the DNA fragmentation of polyadenylic acid signal and any other participation mRNA processing or genetic expression.The bootable polyadenylic acid of described polyadenylic acid signal joins 3 ' end of mRNA precursor, and the non-translational region of transcribing as Agrobacterium crown-gall nodule induction (Ti) plasmid gene (as kermes synthetic enzyme Nos gene), plant gene (as soybean stores protein gene) 3 ' end all has similar functions.

While using described gene constructed recombinant plant expression vector, before its transcription initiation Nucleotide, can add any enhancement type promotor or constitutive promoter, as the ubiquitin promoter (Ubiquitin) of cauliflower mosaic virus (CAMV) 35S promoter, corn, they can be used alone or are combined with other plant promoter; In addition, while using gene constructed plant expression vector of the present invention, also can use enhanser, comprise translational enhancer or transcriptional enhancer, these enhanser regions can be ATG initiator codon or neighboring region initiator codon etc., but must be identical with the reading frame of encoding sequence, to guarantee the correct translation of whole sequence.The source of described translation control signal and initiator codon is widely, can be natural, also can synthesize.Translation initiation region can be from transcription initiation region or structure gene.

For the ease of transgenic plant cells or plant are identified and are screened, can process plant expression vector used, the coding that can express in plant as added can produce the enzyme of colour-change or the gene of luminophor (gus gene, luciferase genes etc.), have the antibiotic marker thing (gentamicin marker, kantlex marker etc.) of resistance or anti-chemical reagent marker gene (as anti-weedkiller gene) etc.From the security consideration of transgenic plant, can not add any selected marker, directly with adverse circumstance screening transformed plant.

Described recombinant expression vector is the recombinant plasmid that the multiple clone site of described gene insertion pCAMBIA1300 and pCAMBIA2300 is obtained.

Contain above arbitrary described gene ( osPSF1) expression cassette, transgenic cell line and recombinant bacterium all belong to protection scope of the present invention.

Increase described gene ( osPSF1) primer pair of total length or arbitrary fragment also belongs to protection scope of the present invention.

The application of gene described in the present invention also provides in the transgenic plant of cultivating photosynthetic efficiency raising.

Above-mentioned application is that the gene of the described photosynthesis of plant associated protein of coding is imported in object plant (as vegetable cell or tissue), obtains photosynthetic efficiency higher than the transgenic plant of described object plant.Particularly, described recombinant expression vector is imported in object plant, obtain the photosynthetic efficiency of transgenic plant higher than described recipient plant.

Utilize any carrier that can guide foreign gene to express in plant, by the gene transfered plant cell of encoding said proteins, can obtain transgenic cell line and transfer-gen plant that drought-resistance ability strengthens.Carry described gene expression vector can by using, Ti-plasmids, Ri plasmid, plant viral vector, directly delivered DNA, microinjection, electricity be led, conventional biological method transformed plant cells or the tissue such as agriculture bacillus mediated, and the plant tissue of conversion is cultivated into plant.The plant host being converted can be both monocotyledons, can be also dicotyledons, as: tobacco, Root or stem of Littleleaf Indianmulberry, Arabidopis thaliana, paddy rice, wheat, corn, cucumber, tomato, willow, turfgrass, lucerne place etc.

Experiment shows, by crossing and express in the DNA sequence dna Introduced into Rice of coding regulating plant photosynthesis correlation factor OsPSF1 of the present invention, can improve transgenic paddy rice photosynthetic efficiency.The genomic gene of coding regulating plant photosynthesis correlation factor OsPSF1 of the present invention and cDNA gene thereof provide the foundation for cultivating other high light efficiency plants with economic worth.

Accompanying drawing explanation

Fig. 1 is the psf1 mutant that filters out from mutant library, and wherein A is wild-type and psf1 mutant seedling in 2 week age, and B is wild-type and psf1 mutant blade in 2 week age.

Fig. 2 is psf1 mutant chlorophyll content in leaf blades, and wherein A is wild-type and psf1 mutant leaf chlorophyll a and chlorophyll b total amount, and B is wild-type and psf1 mutant leaf chlorophyll a and chlorophyll b ratio.

Fig. 3 is psf1 mutational site order-checking spectrogram.

Fig. 4 is complementary rotaring gene plant blade chlorophyll content.

Fig. 5 is that complementary transfer-gen plant PCR identifies.

Fig. 6 is complementary transfer-gen plant order-checking collection of illustrative plates.

Fig. 7 was that express transgenic plant PCR identifies.

Fig. 8 was that express transgenic plant RT-PCR identifies.

Fig. 9 was the mensuration of express transgenic plant leaf photosynthetic rate.

Embodiment

Detailed description below by embodiment is further illustrated the present invention, but is not limitation of the present invention, only does example explanation.

Experimental technique in following embodiment, if no special instructions, is ordinary method, and test materials used if no special instructions, is and from routine biochemistry reagent shop, purchases availablely, and experiment all arranges three repetitions, results averaged.

embodiment 1: psf1the screening of mutant and the map based cloning of mutator gene

One, the screening of Rice mutant pool

Get 2000g japonica rice variety " Japan fine " ( oryza sativasubsp. japonicacv. Nipponbare) (Institute of Crop Science, Chinese Academy of Agricultural Science, numbering WD-10576; Primary source is unclear) seed, first water seed soaking 16 hours, then use 0.5% methylsulphonic acid second vinegar (EMS) at 28 ℃, to process 12 hours, then with a large amount of tap water, rinse 10 times, obtain M1 seed, by M1 planting seed, individual plant results, obtain M2 mutant library.

By 2000 parts of M2 mutant seed germinations, at 2 weeks Seedling Stages, observe leaf color, find a light yellow mutant, called after psf1, as shown in Figure 1.80% acetone extraction wild-type and psf1mutant blade pigment, colorimetric method for determining chlorophyll content, result as shown in Figure 2, psf1chlorophyll a and b all reduce, and total amount is only 50% of wild-type plant, and chlorophyll a/b is 5 times of wild-type, show amplitude that chlorophyll b reduces than chlorophyll a larger (Fig. 2 AB).

Two, the map based cloning of psf1 mutant mutator gene

Will psf1mutant and Japanese fine wild-type backcross, the backcross ratio of pale yellow leaf plant and greenery plant in segregating population seedling of statistics F2, and result is as shown in table 2.To 3, independently the pale yellow leaf of F2 backcross population is separated than (table 2) with the 1:3 that the ratio of normal leaf look plant meets Dominant gene by two kinds of methods analysts discoveries of chi square test and binomial test.

Table 2 psf1mutant backcrosses F2 for the pale yellow leaf of plant ratio separated with greenery normal plant

Will psf1the hybridization of mutant and Dular obtains F1, F1 selfing obtains F2 colony, utilize SSR molecule marker to find this mutational site and molecule marker Chr7-20.1 and Chr7-20.2 close linkage to 352 separated individual plant linkage analysises of the pale yellow leaf of F2, to this section, whole 8 ORF order-checkings are found psf1the 284th T of mutant ORF7 sports G, and coded amino acid becomes alkaline arginine from neutral leucine, and this site of Dular identical with wild-type (Fig. 3).Supposition of ORF7 coding is positioned the albumen with ankyrin structural domain of plastid, our called after OsPSF1 albumen (LOC_Os07g33660).

the clone of embodiment 2:OsPSF1 cDNA and genomic gene

One, the clone of OsPSF1 cDNA

Get 0.2g japonica rice variety " Japan fine " ( oryza sativasubsp. japonicacv. blade Nipponbare), liquid nitrogen grinding, TRIzol method is extracted total RNA.Get the total RNA of 2 μ g and carry out reverse transcription by M-MLV ThermoScript II, synthetic cDNA the first chain, as template, carries out PCR reaction with special primer to first.PCR product, after electrophoretic separation reclaims, is cloned into pEASY-T(Beijing Quanshijin Biotechnology Co., Ltd), called after pEASY-T-OsPSF1ORF, checks order.

Special primer is as follows to first:

Upstream primer: 5 '-ATGGCATCCATCCCGTGCAC-3 ';

Downstream primer: 5 '-TCAGGCGGCCAAGGTGGCGG-3 '.

Sequencing result shows, the nucleotide sequence of this fragment as shown in SEQ ID NO:2, the protein shown in coding SEQ ID NO:1.

Two, the acquisition of OsDRPSF1 genomic gene

CTAB method is extracted the genomic dna of Japanese fine blade, take this genomic dna as template, with special primer, second is reacted.PCR product, after electrophoretic separation reclaims, is cloned into pEASY-T(Beijing Quanshijin Biotechnology Co., Ltd), check order.

Special primer is as follows to second:

Upstream primer: 5 '-GTGTGCTGTTGTCCCAGTATG-3 ';

Downstream primer: 5 '-AACCTGTTAGCCCTACGAGTG-3 '.

Sequencing result shows, the nucleotide sequence of this fragment is as shown in SEQ ID NO:3.

By the protein called after OsPSF1 shown in SEQ ID NO:1, by the encoding gene called after of OsPSF1 osPSF1.Nucleotide shown in SEQ ID NO:2 is connected to the recombinant plasmid called after pEASY-T-obtaining with pEASY-T osPSF1.

embodiment 3: psf1having complementary functions of mutant

One, the structure of pCAMBIA1300-OsPSF1 expression vector

1, with restriction enzyme XbaI and KpnI enzyme, conscientiously execute recombinant plasmid pEASY-T-in 1 osPSF1,the 3.7kb fragment that recovery comprises OsPSF1 sequence.

2, with restriction enzyme XbaI and KpnI enzyme, cut pCAMBIA1300(Cambia, GPO Box 3200, Canberra, ACT 2601, Australia), reclaim skeleton.

3, the fragment that fragment step 1 being obtained obtains with step 2 is connected, and obtains pCAMBIA1300-OsPSF1.

Two, the acquisition of transgenic plant

1, utilize electric shocking method that recombinant expression vector pCAMBIA1300-OsPSF1 is imported to Agrobacterium AGL0(ATCC BAA-100www.atcc.org).

5, the Agrobacterium AGL0 that contains pCAMBIA1300-OsPSF1 is infected psf1the embryonic type callus that mutation induction produces then screens resistant calli, 15 days per generations in MS substratum (containing 30mg/L Totomycin), amounted to for 3 generations, then resistant calli is induced into whole plant, rice transplanting is in land for growing field crops, and the T1 of results transgenic plant is for seed.

OsPSF1 is transformed psf1mutant T1, for seed germination, finds after 2 weeks that seedling leaf look recovers normal, and it is identical with wild-type contrast that mensuration chlorophyll content is found, is significantly higher than psf1mutant chlorophyll content (Fig. 4).

CTAB method is extracted leaf DNA, with hygromycin phosphotransferase gene ( hpt) Auele Specific Primer 5 '-CATCGAAATTGCCGTCAACC-3 ' and 5 '-AGCCTGACCTATTGCATCTC-3 ' PCR detection, result shows psf1in the complementary transfer-gen plant of mutant (com), have specific amplification band, and wild-type and psf1mutant DNA is without specific amplified band (Fig. 5).This shows the selection markers on carrier hptgene imports psf1the complementary transfer-gen plant of mutant.

Because on pCAMBIA1300-OsPSF1 plasmid vector wild-type OsPSF1 DNA fragmentation with hptgene series connection is co-located at T-DNA district, can infer wild-type OsPSF1 DNA fragmentation with hptgene fragment imports jointly psf1in the complementary transfer-gen plant of mutant.Cause psf1mutant is point mutation, PCR method cannot by the wild-type OsPSF1 DNA being imported into psf1the fragment of suddenling change in mutant gene group separately, we with OsPSF1 gene-specific primer 5 '-ATGGCATCCATCCCGTGCAC-3 ' and 5 '-TCAGGCGGCCAAGGTGGCGG-3 ' respectively from wild-type, psf1pcr amplification in mutant and complementary transfer-gen plant genomic dna, then, by the order-checking of PCR product, result as shown in Figure 6.284 of complementary transfer-gen plant (com) OsPSF1 ORF are T/G heterozygosis, and this clearly proves that wild-type OsPSF1 fragment imports psf1mutant.

Above result shows, to psf1in mutant, importing OsPSF1 gene order, can to make light yellow Phenotype Transition be normal leaf look, further proved that OsPSF1 transgenation is to cause it to be the gene of light yellow phenotype.

the overexpression of embodiment 4:OsPSF1 in paddy rice

One, the structure of pCAMBIA2300-Act-OsPSF1 expression vector

1, take that to implement recombinant plasmid pEASY-T-OsPSF1ORF in 1 be masterplate, with following primer amplified, obtain OsPSF1 sequence.

Upstream primer: 5 '-TCCCCCGGGATGGCATCCATCCCGTGC-3 ';

Downstream primer: 5 '-AACTGCAGTCAGGCGGCCAAGGTGGCGG-3 '.

After PCR product electrophoretic separation, cut glue and reclaim 520bp DNA fragmentation, then with restriction enzyme XmaI and PstI enzyme, cut.

2, with restriction enzyme XmaI and PstI enzyme cut pCAMBIA2300-ACT (Actin promotor) (Cambia, GPO Box 3200, Canberra, ACT 2601, Australia), reclaim skeleton.

3, the fragment that fragment step 1 being obtained obtains with step 2 is connected, and obtains pCAMBIA2300-Act-OsPSF1.

Two, the acquisition of transgenic plant

1, utilize electric shocking method that recombinant expression vector pCAMBIA2300-Act-OsPSF1 is imported to Agrobacterium AGL0.

5, the Agrobacterium AGL0 that contains pCAMBIA2300-Act-OsPSF1 is infected to the embryonic type callus that Japanese fine wild-type induction produces, then in MS substratum (containing 100mg/L G418), screen resistant calli, 15 days per generations, amounted to for 3 generations, then resistant calli is induced into whole plant, rice transplanting is in land for growing field crops, and the T1 of results transgenic plant is for seed.

Three, the Molecular Detection of transgenic plant

CTAB method is extracted leaf DNA, with Act-OsPSF1 Auele Specific Primer 5 '-TCTGCGATCCGCCGTTGTTG-3 ' and 5 '-GCGCCTCCATCTTGTCGTAG

-3 ' PCR detects, and result shows in transfer-gen plant, there is specific amplification band, and wild-type DNA is without specific amplified band (Fig. 7), and this shows that Act-OsPSF1 has imported transfer-gen plant.

TRIzol method extraction wild-type in 2 week age and Act-OsPSF1 transfer-gen plant (ox1, 2, 3) the total RNA of blade, get the total RNA of 2 μ g, with 37 ° of C of 2 u RQ1 RNase-free DNase (Promega), process 30 min, add after termination reaction liquid, take polyA as primer, utilize 1 hour synthetic cDNA the first chain of 42 ° of C of M-MLV ThermoScript II, then take cDNA as template, the Actin (primer 5 '-GCCAATCGTGAGAAGATGAC-3 ' and 5 '-CTATGAAGGAAGGCTGGAAG-3 ') of take is internal reference, PCR method detection PSF1 gene (primer 5 '-ATGGCATCCATCCCGTGCAC-3 ' and 5 '-TCAGGCGGCCAAGGTGGCGG-3 ') express, result as shown in Figure 8.Between each plant, Actin genetic expression is basic identical, and the 5-10 that Act-OsPSF1 transfer-gen plant (ox1,2,3) PSF1 gene expression dose is about adjoining tree doubly.

Four, the mensuration of photosynthetic rate

3 week age wild-type and Act-OsPSF1 transfer-gen plant (ox1,2,3) blade spraying, 50 μ mol L ^-1methyl viologen (MV) is processed, and take water as contrast, after 3 days with LI6400XT (LI-COR, Inc) portable photosynthetic instrument mensuration wild-type in 3 week age and Act-OsPSF1 transfer-gen plant (ox1,2,3) Net Photosynthetic Rate of blade, parameter setting is CO2 400 μ mol CO2 mol ^-1, light intensity 800 μ mol m ^-2s ^-1, 30 ° of C of temperature, result shows transfer-gen plant (ox1 under collating condition, 2,3) Net Photosynthetic Rate and between wild-type blade does not have significant difference, and methyl viologen is processed rear transfer-gen plant (ox1,2,3) net photosynthetic rate is higher than wild-type (Fig. 9).This showed to express OsPSF1 gene can improve the photosynthetic rate after transfer-gen plant oxidative stress.

<110> Biological Technology institute, Chinese Academy of Agricultural Sciences

<120> photosynthesis of plant associated protein OsPSF1 and encoding 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 Ser Ala 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

TGGTTATTCCTATTAGGTATACGATTCTTACCACATAGAAGGTGATACTT 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

CATTTCCGTTTCTTTTTTTCACATTTTTTATGGGTGTCTATTTCACATGG 950

TAATTAGACAGCTAATTCCAAGCTTAAAACCTGGCTTAATGCTACTGCTT 1000

TTTATCCTAGGAATCTATTTTGATTTCTCAAGAGGATGTCTTCTCGTAAG 1050

CTTAAAAACCATTCAAATGGTTATAAAAAAAATTCTAAAAAAATTAACAA 1100

CACTCATACAATTTACTTCCTCCGTTCAAAAAAAACCAACGGATGTGACC 1150

CCCTCCTAATACAGTAAATCTAAACAGCACTTAATCCAAATTTGTTATAT 1200

TAGGAGATGTCACATCCTTTTAGGTTGTTTTTTTGGTGACGGATTTTTTT 1250

TTTACGGAGGGATGGAGTATATCTCACACAGCTCTAGCACGTACTCCTAT 1300

CTCACACAGATTTATCACAAATCATACACTTTTTTTTAATATCTTCTACA 1350

CTGCACTTTATGAATATCTCATGTCCTGTCTTCCACAGGAAAAAAAGTGA 1400

AATAGTGAAAAGATTCAGAATTAGGAAGCATCTAATCACCTAACTTGACA 1450

GCAATCAGTTTCAAGTGGCCAAGTGCCAAACAGTTTCAAGATTACTTACA 1500

ACTTATCCGTTCCCTCCATATATCTCTCTCCCCTCTTCTCCACCTCTCGC 1550

TACTCCACCATCCAACGTGCGAGAAGCCATGGCATCCATCCCGTGCACCT 1600

TCCAGCTGAGCGCGAGGGCGTCGTCGGCGTCGGCGGCGGCGGCGGCGAGG 1650

AGGTCGCCGCGGGCGGCGGCGAGGCTGGGGTGGCTGCGGCCGTCGCGGCT 1700

GAGCGCGGTGGTGCCGGCGAGCGAGAGCGGGAGGGTGGGGCCGACGTGCT 1750

TCTTCAAGTTCGGGAACAAGGACGCCGAGGGCGCCGGCATCTACGGCAGC 1800

CAGGGCAGGGACGACTTCGACCGCGACGACGTCGAGCAGGTGAGCCAGCT 1850

TCTCGTTCTTCCTCCTCCCCATGTTGCCTTAGGTGTTCCTTGATTGCTCC 1900

AGTCCATGGGGAATGTGCTGACGAATGACTGAGATGCTACCTTACCACTG 1950

CTATATTACTTGGTGGATTGGTAATTCTGATCCTAATGTGAGACATACGA 2000

AGTAGTTGATTGCTATACTTGCTAGTTTGGTTTGAGGACAAATGATGCAG 2050

CATTTGATTTGAGTTGAATTAAGGCATGTCTTTTTCCTTATGCTTATACT 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

TACTACGTATTTATCTTTTCTTATTTTTTTCTTTTTCAAGTTTCTCCAAG 2950

TTTTTCTTGTGTTTGGTAGAGTTTAGCCGGGAAATTCACAAGTTGTTTAC 3000

AGTTCAAGAGGAAGTAGCTTCATCTGTATCTTTCACTTTCTTCGCTGTGT 3050

GAGGCTATGCTTTGTAGAAATTGTTGTTACAAGTAGTATCTTCCACTGTT 3100

TCAATATGAAAGTAAGAAGAAGCTTTGGATGCATGATACTATTGGAATTG 3150

GAAGCTTTGATGTTCACATAATAACATGTTAAGATGCCAAATCCTAACAG 3200

CAAGCAATAGCATGAACTTCAAGCAAAGCATTACTGCAACAGCAAGCAAA 3250

ACTTCATCCTTTATTGTCTCAGATTAACTGCTATGTCCATGTTGCCGTCA 3300

GTACATGTTCTATGTACTAGCCTAGAATTACAAACAGCATTTCGCCGAAA 3350

GATTCACTTGGGGGAAACAAACAGTCTGCGACCACAGCCTCAACAACGAA 3400

ATTACGAGTATAAACTCTATAAACCCCCTCATGAATCCGCCAGATCATAT 3450

TATTCTTTTCTTCTTCTATATTCATCTTGGTATATATTTCTATAAGGACT 3500

GTGAATGTACTCCTACCTACAAAACATTGGGTAAATGGCAGTTGTACTTC 3550

AACTTTGGATGTGACCGGAGCGCTTGCTGGAGCGCCGTGTCCACCGTCAC 3600

GCCCTCGCGGAACAGCTTGTCATACAGGAGGCAGATGAACTGAGCCATGT 3650

ACTCCTCATCATCGAAATCTTTGCTGCGATCGCCATTTTTCTCCATGTCG 3700

CTGTCTTCCCAGTCACTCGTAGGGCTAACAGGTT 3734

Claims

1. a photosynthesis of plant associated protein, its aminoacid sequence is as shown in SEQ ID NO:1.

2. a gene for photosynthesis of plant associated protein described in the claim 1 of encoding, its nucleotide sequence is as shown in SEQ ID NO:2 or shown in SEQ ID NO:3, or its degenerate sequence.

3. the recombinant expression vector that comprises gene described in claim 2.

4. recombinant expression vector as claimed in claim 3, is characterized in that, it is restructuring plant expression vector.

5. recombinant expression vector as claimed in claim 3, is characterized in that: it is recombinant plasmid.

6. recombinant expression vector as claimed in claim 3, is characterized in that: described recombinant plasmid is that the multiple clone site of gene insertion pCAMBIA1300 or pCAMBIA2300 described in claim 2 is obtained.

7. the recombinant host bacterium that contains recombinant expression vector as claimed in claim 3.

8. the application of gene claimed in claim 2 in the transgenic plant of cultivating photosynthetic efficiency raising.

9. application as claimed in claim 8, is characterized in that: expression vector as claimed in claim 3 is imported in plant, obtain photosynthetic efficiency higher than the transgenic plant of described recipient plant.

10. application as claimed in claim 9, is characterized in that: described transgenic plant are transgenic paddy rices.