CN101831439A - Rice mutator gene associated with high chlorophyll content and recombinant plant overexpression vector thereof - Google Patents
Rice mutator gene associated with high chlorophyll content and recombinant plant overexpression vector thereof Download PDFInfo
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- CN101831439A CN101831439A CN 201010177039 CN201010177039A CN101831439A CN 101831439 A CN101831439 A CN 101831439A CN 201010177039 CN201010177039 CN 201010177039 CN 201010177039 A CN201010177039 A CN 201010177039A CN 101831439 A CN101831439 A CN 101831439A
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Abstract
The invention discloses a rice mutator gene associated with high chlorophyll content, which has a nucleotide sequence SEQ ID No.1. The invention also discloses a recombinant plant overexpression vector containing the mutator gene. Thus, a powerful tool for researching rice transgenosis is provided, and the researches on the breeding of high-yield and high-quality rice are promoted.
Description
Technical field
The present invention relates to a kind of gene, particularly paddy rice high chlorophyll content mutator gene DET1 also relates to the recombinant plant overexpression vector of this mutator gene.
Background technology
Paddy rice is C
3Plant, its photorespiration is stronger, and effectively photosynthetic rate is lower, and especially in the filling stage of high temperature dry season district paddy rice, because the temperature height, its photorespiration will be wasted a large amount of photosynthates, not only can influence output, also can have a strong impact on a meter matter.Therefore, improving effective photosynthetic rate is an effective way that improves rice yield and improve rice matter.The approach that improves effective photosynthetic rate is to reduce photorespiration or improve photosynthetic rate, and C
3The physiology of plant and its high photorespiration characteristic of having dissected structures shape, because chlorophyll is the basic substance of paddy rice photosynthesis indispensability, therefore, to improve photosynthetic rate be to improve an effective way of the effective photosynthetic rate of paddy rice by improving chlorophyll content.
The contriver has found a mutating strain series by the high chlorophyll proterties of simple inheritance in previous research work, and its genetic behavior and physiological function are studied, and the result shows that this proterties controlled by the dominance single-gene; The major physiological function of this gene is to realize the raising of total chlorophyll content by the content that significantly improves chlorophyll b, thereby improve photosynthetic rate by " seizure " ability that improves luminous energy, can improve biological yield and economic yield more than 16%, reduce the white grain of chalk rate about 17%; The molecule marker positioning result shows that this assignment of genes gene mapping is in paddy rice the 1st the short arm of a chromosome, and the genetic distance between microsatellite marker RM 6464, RM6340 and the RM462 be respectively 0cM, 0.588cM and 1.18cM (Li Xianyong etc. the discovery of a paddy rice high chlorophyll content gene. southwestern agriculture journal, 2002,15 (4): 122-123; Li Xian is brave etc. the biological characteristic research of a paddy rice high chlorophyll gene. and southwestern agriculture journal, 2004,17 (3): 292-294; Wang FH et al.Heredity, physiology and mapping of a chlorophyll content gene of rice (Oryza sativa L.) .Journal of Plant Physiology, 2008,165 (3): 324-330).Therefore, determine the nucleotide sequence of this gene, will help the breeding research of high yield and high quality paddy rice, promote new step on the Rice Production.
Summary of the invention
In view of this, one of purpose of the present invention is to provide the nucleotide sequence of paddy rice high chlorophyll content mutator gene; Two of purpose is to provide the recombinant plant overexpression vector of described paddy rice high chlorophyll content mutator gene, thereby for the paddy rice transgenic research provides strong instrument, promotes the breeding research of high yield and high quality paddy rice.
For achieving the above object, at first, the invention provides paddy rice high chlorophyll content mutator gene DET1, have the nucleotide sequence shown in SEQ ID No.1.
The present invention by predictive genes, homology search and gene order diversity ratio, has determined that tentatively paddy rice high chlorophyll content mutator gene is DET1 (DE-ETIOLATED 1) earlier on the basis of the assignment of genes gene mapping in early stage.Subsequently, the present invention is a material with paddy rice high chlorophyll mutant, has cloned the cDNA of mutator gene DET1.Sequencing result shows: mutator gene DET1 full-length cDNA is 1536bp, is made up of 11 exons, 512 amino acid of encoding.Compare with the wild-type Zhenshan 97B, mutator gene DET1 has the conversion of T-C on the 7th exon, an EcoR I restriction enzyme site has been introduced in this change, and causes the 328th leucine (L) to change into Serine (S), and the mutational site is in the high conservative zone.The amino acid sequence homology of other higher plants such as the aminoacid sequence of mutator gene DET1 and corn is up to more than 62%.
Secondly, the invention provides the recombinant plant overexpression vector that contains described paddy rice high chlorophyll content mutator gene DET1.
Further, described recombinant plant overexpression vector is to insert paddy rice high chlorophyll content mutator gene DET1 and obtain in the multiple clone site of pCUPHN carrier; Described pCUPHN carrier is transformed by the pCAMBIA-1300 carrier, promptly introduce corn ubiquitin promoter gene by restriction enzyme PstI at the multiple clone site place of pCAMBIA-1300 carrier, introduce HA label gene by Sac I and Spe I afterwards, introduce terminator NOS gene by Spe I and EcoR I again.
Beneficial effect of the present invention is: the invention provides one and improve relevant mutator gene DET1 with chlorophyll content of rice, and made up its recombinant plant overexpression vector, for the paddy rice transgenic research provides strong instrument, can promote the breeding research of high yield and high quality paddy rice.
Description of drawings
In order to make the purpose, technical solutions and advantages of the present invention clearer, the present invention is described in further detail below in conjunction with accompanying drawing, wherein:
Fig. 1 is that the agarose gel electrophoresis of the RT-PCR product of gene DET1 identifies that 1 swimming lane is DL2,000
DNA Marker, 2 swimming lanes are blank, and 3 swimming lanes are the RT-PCR product of DET1 (high chlorophyll mutant), and 4 swimming lanes are the RT-PCR product of DET1 (wild-type);
Fig. 2 cuts evaluation for the enzyme of recombinant vectors pMD19T-DET1,1 swimming lane is DNA Marker III, 2 swimming lanes are that Spe I enzyme is cut pMD19T-DET1 (high chlorophyll mutant), 3 swimming lanes are that Spe I/BamH I enzyme is cut pMD19T-DET1 (high chlorophyll mutant), 4 swimming lanes are that Spe I enzyme is cut pMD19T-DET1 (wild-type), and 5 swimming lanes are that Spe I/BamH I enzyme is cut pMD19T-DET1 (wild-type);
Fig. 3 comprises the partial nucleotide sequence comparing result in mutational site for gene DET1, the EcoR I restriction enzyme site that underscore is partly introduced for sudden change;
Fig. 4 is the amino acid identity comparing result of gene DET1 proteins encoded, and arrow indication place is the mutational site of gene DET1, and the zone is the high conservative zone of gene DET1 shown in the underscore;
Fig. 5 is the systematic evolution tree of gene DET1;
Fig. 6 is the wetting ability and the hydrophobicity analysis of gene DET1 proteins encoded;
Fig. 7 is the structure synoptic diagram of mutator gene DET1 recombinant plant overexpression vector pCUPHN-DET1;
Fig. 8 is that the PCR of mutator gene DET1 recombinant plant overexpression vector pCUPHN-DET1 identifies, 1 swimming lane is DNAMarker III, 2 swimming lanes are blank, 3 swimming lanes are the PCR product of pCUPHN-DET1,4 swimming lanes are the PCR product (negative control) of pCUPHN, and 5 swimming lanes are the PCR product (positive control) of pMD19T-DET1;
Fig. 9 cuts evaluation for the enzyme of mutator gene DET1 recombinant plant overexpression vector pCUPHN-DET1,1 swimming lane is λ-Hind III DNA Marker, 2 swimming lanes are that Spe I enzyme is cut pCUPHN-DET1,3 swimming lanes are that Spe I/BamH I enzyme is cut pCUPHN-DET1,4 swimming lanes are that Spe I enzyme is cut pCUPHN, 5 swimming lanes are that Spe I/BamH I enzyme is cut pCUPHN, and 6 swimming lanes are DNA Marker III.
Embodiment
Hereinafter with reference to accompanying drawing, the preferred embodiments of the present invention are described in detail.The experimental technique of unreceipted actual conditions in the preferred embodiment, usually according to normal condition, the molecular cloning experiment guide (third edition for example, J. work such as Sa nurse Brooker, Huang Peitang etc. translate, Science Press, 2002) described in condition, or the condition of advising according to manufacturer.
The material that uses in the preferred embodiment: wild-type rice material Zhenshan 97B (Wt.) and high chlorophyll mutant (Mut.) are provided by Chongqing City's research of agricultural science institute; M-MLV reversed transcriptive enzyme, high-fidelity DNA polymerase PFU, Taq archaeal dna polymerase, T4DNA ligase enzyme, restriction enzyme, pMD19-T carrier, Trizol test kit, dna gel reclaim test kit, plasmid extraction kit, λ-Hind III DNA Marker and DL2, and 000DNA Marker is available from TaKaRa company; DNA Marker III is available from TIANGEN Biotech (Beijing) Co., Ltd.; Penbritin (Ampicillin, Amp) and kalamycin (Kanamycin Kan) is Sigma company product; Primer is synthetic to be finished by the handsome Bioisystech Co., Ltd in Shanghai with dna sequencing; Other chemical reagent is available from Beijing ancient cooking vessel state biotechnology limited liability company; E. coli jm109, Agrobacterium GV3101 are preserved by this laboratory.
The present invention is on the basis of gene Fine Mapping in early stage, at first, determined that tentatively paddy rice high chlorophyll content mutator gene is DET1 by online predictive genes (http://mendel.cs.rhul.ac.uk) and the online comparison of BLAST (http://blast.ncbi.nlm.nih.gov/).Gene DET1 has report in tomato, Arabidopis thaliana, corn, Chinese sorghum.In tomato, gene DET1 improves total chlorophyll content, and whole plant all shows as deep green.In Arabidopis thaliana, gene DET1 regulates and control the startup ability of light regulation and control promotor, and when the sun plant Arabidopis thaliana was in the dark surrounds, gene DET1 regulated the startup ability of light regulation and control promotor by space-time; In addition, gene DET1 can also regulate and control the clock of discipline round the clock of Arabidopis thaliana by the degraded of regulating proteasome.
Subsequently, the present invention is a material with paddy rice high chlorophyll mutant, cloned the cDNA of mutator gene DET1, utilize the information biology means physico-chemical property of the nucleotide sequence of mutator gene DET1, evolutionary tree, proteins encoded and hydrophilic and hydrophobic etc. are analyzed, and made up the transgenic research that mutator gene DET1 recombinant plant overexpression vector is used for paddy rice.
One, the clone of mutator gene DET1 and order-checking
According to the fine gene DET1 sequence of GenBank listed paddy rice Japan, utilize Primer5.0 and Oligo6.0 software design special primer: upstream primer DET1F:5 '-cggatccggtgatggcgaccttcttc-3 ' (SEQID No.2), underscore partly are BamH I restriction enzyme site; Downstream primer DET1R:5 '-ggactagtccg-ctcacaactgttacaaatac-3 ' (SEQ ID No.3), underscore partly is a Spe I restriction enzyme site.
The spire 2g in the rice wild-type of fetching water respectively and two weeks of high chlorophyll mutant illumination cultivation puts into the liquid nitrogen grind into powder rapidly, extracts total RNA according to Trizol test kit specification sheets.The electrophoresis result of gained paddy rice wild-type and the total RNA of high chlorophyll mutant shows the master tape complete display, and the band luminance factor of 28S and 18S is about 2: 1, illustrates that the concentration of RNA and purity meet requirement of experiment, can be used for synthetic double chain cDNA.
Be template with gained paddy rice wild-type and the total RNA of high chlorophyll mutant respectively,, use Oligo (dT) primer to carry out reverse transcription and obtain cDNA according to M-MLV reversed transcriptive enzyme specification sheets; Be template with reverse transcription synthetic cDNA again, adopt special primer DET1F/DET1R and high-fidelity DNA polymerase PFU to carry out pcr amplification, the PCR reaction conditions is: 94 ℃ of pre-sex change 4 minutes; 94 ℃ of sex change are 30 seconds then, 60 ℃ of renaturation 30 seconds, and 72 ℃ were extended totally 32 circulations 2 minutes; Last 72 ℃ were extended 10 minutes.The RT-PCR product is carried out 1.0% (g/mL) agarose gel electrophoresis detect, reclaim the test kit specification sheets according to dna gel again and cut glue recovery purifying; The dna fragmentation of purifying and pMD19-T carrier are connected in 16 ℃ under the effect of T4DNA ligase enzyme and spend the night, connect product transformed into escherichia coli JM109 competent cell, with the LB plate screening positive colony that contains penbritin, extract plasmid, enzyme checks order after cutting and identifying clip size, gets recombinant vectors pMD19T-DET1.
The agarose gel electrophoresis of the RT-PCR product of gene DET1 is identified as shown in Figure 1, as seen the RT-PCR product of DET1 (wild-type) and DET1 (high chlorophyll mutant) all is the single specificity band at about 1600bp place, conforms to the gene DET1 size of GenBank report.
The enzyme of recombinant vectors pMD19T-DET1 is cut qualification result as shown in Figure 2, as seen pMD19T-DET1 (wild-type) and pMD19T-DET1 (high chlorophyll mutant) all can obtain the purpose fragment of about 1600bp with Spe I/BamH I double digestion, conform to expected results, confirm that the purpose fragment has been connected into the pMD19-T carrier and has been the forward connection.
Sequencing result shows that the DET1 full-length cDNA of paddy rice wild-type and high chlorophyll mutant is 1536bp, 512 amino acid of encoding.Wherein, the DET1 full length cDNA sequence of high chlorophyll mutant is shown in SEQ ID No.1.The partial nucleotide sequence comparing result that gene DET1 comprises the mutational site as shown in Figure 3, compare with wild-type, the gene DET1 of high chlorophyll mutant is made up of 11 exons, 1 base mutation has taken place on the 7th exon, promptly the 983rd T sports C, and an EcoR I restriction enzyme site has been introduced in this sudden change, and has caused the difference of proteins encoded the 328th amino acids, wild-type is leucine (Leu), and the high chlorophyll mutant is Serine (Ser).
Two, the bioinformatic analysis of mutator gene DET1
Utilize the ORF Finder (http://www.ncbi.nlm.nih.gov/gorf/gorf.html) among the NCBI to carry out open reading frame identification; Utilize CDD (conserved domain database) (http://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi) to carry out the domain analysis of protein conserved structure; Utilize Clustalx and DNA Man software to carry out the comparison of aminoacid sequence and the generation of evolutionary tree; Utilize ExPASy (http://cn.expasy.org/) on-line analysis protein structure and physicochemical property thereof.
ORF Finder software analysis show mutator gene DET1 by one complete and successive open reading frame form.
CDD analyzes and shows that gene DET1 has two high conservative zones, is respectively the from the 282nd to 336 amino acids and the 400th to 442 amino acids, and the mutational site of mutator gene DET1 occurs in first high conservative zone.
The aminoacid sequence comparison result of mutator gene DET1 proteins encoded as shown in Figure 4, the homology of mutator gene DET1 (being expressed as OsDET1 among the figure) and Arabidopis thaliana AtDET1 (the GenBank accession number is NP_192756), tomato S1DET1 (the GenBank accession number is Q9ZNU6), Chinese sorghum SbDET1 (the GenBank accession number is XP_002443935), corn ZmDET1 (the GenBank accession number is NP_001147932) and castor-oil plant RcDET1 (the GenBank accession number is XP_002519399) proteins encoded is up to more than 62%, and the mutational site occurs in the high conservative zone.
The systematic evolution tree of mutator gene DET1 as shown in Figure 5, the affinity of mutator gene DET1 (being expressed as OsDET1 among the figure) and Chinese sorghum SbDET1 is nearer, takes second place with the affinity of corn ZmDET1.
The composition and the analysis of physical and chemical property of mutator gene DET1 proteins encoded show: its proteins encoded is made up of 512 amino acid, and relative molecular mass is 59126.8, and molecular formula is C
2683H
4040N
716O
768S
16, iso-electric point (pI) is 7.65, serine content is up to 10.6%.
By the ProtParam instrument among the online software ExPASy mutator gene DET1 proteins encoded is carried out the hydrophilic and hydrophobic analysis, the result as shown in Figure 6, statistical result showed GRAVY (Grand average ofhydropathicity) value is for-0.098, illustrate that mutator gene DET1 encoded protein matter is hydrophilic protein matter, this may be relevant with the function of gene, because chlorophyllous synthesizing is a process that needs water to participate in, the wetting ability of this gene product increases, help the transportation of moisture, thereby accelerate chlorophyllous biosynthesizing.
Three, the structure of mutator gene DET1 recombinant plant overexpression vector
The pCUPHN carrier is transformed by the pCAMBIA-1300 carrier, promptly introduce corn ubiquitin promoter (Ubi Promter) gene by restriction enzyme Pst I at the multiple clone site place of pCAMBIA-1300 carrier, introduce HA label (tag) gene by Sac I and SpeI afterwards, introduced terminator NOS gene by Spe I and EcoR I again.The corn ubiquitin promoter has stronger startup ability, is particularly suitable for using in grass.Although HA tag is not necessary in the binary expression vector, its expression of gene of the separation and purification in the future product that is introduced as provides convenience.The pCUPHN carrier is applicable to the expression of all plant genes.
The recombinant vectors PMD19T-DET1 BamH I/Spe I double digestion that will contain mutator gene DET1, reclaim mutator gene DET1 fragment, spend the night with under the effect of T4DNA ligase enzyme, being connected equally again in 16 ℃ through the pCUPHN carrier of BamH I/Spe I double digestion, connect product transformed into escherichia coli JM109 competent cell, with the LB plate screening positive colony that contains kalamycin, extract plasmid, PCR and enzyme are cut evaluation, promptly get mutator gene DET1 recombinant plant overexpression vector pCUPHN-DET1, its building process as shown in Figure 7.Subsequently pCUPHN-DET1 is transformed Agrobacterium GV3103 ,-80 ℃ of preservations are standby.
The PCR of mutator gene DET1 recombinant plant overexpression vector pCUPHN-DET1 identifies and enzyme is cut qualification result respectively as Fig. 8 and shown in Figure 9, and purpose fragment mutator gene DET1 has been connected in the pCUPHN carrier.
Explanation is at last, above embodiment is only unrestricted in order to technical scheme of the present invention to be described, although by invention has been described with reference to the preferred embodiments of the present invention, but those of ordinary skill in the art is to be understood that, can make various changes to it in the form and details, and the spirit and scope of the present invention that do not depart from appended claims and limited.
Sequence table
＜110〉University Of Chongqing
＜120〉paddy rice high chlorophyll content mutator gene DET1 and recombinant plant overexpression vector thereof
<160>3
<210>1
<211>1536
<212>DNA
＜213〉paddy rice (Oryza Sativa L)
<220>
＜223〉high chlorophyll content mutator gene DET1
<400>1
atggcgacct?tcttccgcag?cgccaacctc?gcctccaggg?tcttcgaccg?ccagttcctc 60
tccccacgcc?ccggcgctac?tgttaacact?gttagacagt?tctatgagaa?cttggttccg 120
agttacacta?tatgtgacat?agattgccca?gattattcat?ttcgtaaatt?cactgatgat 180
ggcaactatc?ttgtggcttt?tagccggaac?caccaagatt?taatcgtgta?ccgtcccatc 240
tggccaacat?tttcatgcaa?tgagccatgc?gactctcatg?atcttccacc?caaagcaaag 300
aaattcgaca?gcttctttaa?gcagctctac?tcaatttcac?ttgcatctag?caatgaatac 360
atctgcaagg?actttttcct?ttacatggag?tgccatcagt?ttggcttatt?tgccacatca 420
actgcacaaa?gcaacgattc?aagtgccact?gagggtgcta?ttcatggggt?accatcaatt 480
gagaaaatta?ctttctatct?tgtcaggcta?gacgacggcg?caatactaga?tgagaaagct 540
ttccgcaatg?attttatcaa?tctggctcat?agtattggag?cttacttgta?tgaggacctg 600
ctatgcattg?tctctcttcg?atatcaaaca?attcatgtcc?tacagatacg?agattcaggc 660
aaccttgttg?aagtacgcaa?aattggtgca?ttttgccagg?aagatgatga?gctatttctc 720
cattcacatg?gtcaggctgc?acgaggtgtt?tcttttcttc?ctggcatcaa?gcaacggttg 780
ttgtcattta?tttttcgcaa?gacatggaat?gaggaatctg?atcagacctt?gagagtccag 840
catctgaaga?agaagttcta?ttttcacttt?caagactatg?ttgatcttat?catatggaag 900
gtgcagtttt?tggatcgtca?tcacctattt?atcaagtttg?gtagtgtgga?tggaggggtt 960
tctcgcagca?ctgaacagaa?ttcagcattc?tttgctgtgt?acaacatgga?gacgacagat 1020
attgtttcac?tctatcagaa?ttcttcggag?gagctctatt?cattgtttga?atatttttat 1080
gaccattttc?acacaaatcc?acaaaattca?tcgcatggaa?attttatctc?gtcacattcc 1140
aataatgttc?atgctctcga?tcaacttcgg?acaattaaaa?ataaagcaaa?cagcacctca 1200
cagtttgtga?aaaagatgat?ggcgtcacta?ccttacactt?gtcaatcaca?aagtccttcg 1260
ccatactttg?acctatcatt?gtttcggtat?gatgagaagc?tgatttcagc?gattgatcgg 1320
cacagacact?gtacagagca?tccaataaaa?tttatatcag?tgaaacagcc?aaatgttgtc 1380
aaattcaaga?taaaaccagg?ttcagattct?ggtgcttctg?acagcagggc?aaaaagaata 1440
tcttccttct?tgtttcaccc?atttttccct?ctcgcacttt?ctattcagca?gacatatatg 1500
cagccaactg?ttgtcaattt?acatttcagg?agatag 1536
<210>2
<211>26
<212>DNA
＜213〉artificial sequence
<220>
＜223〉the special primer DET1F of amplification gene DET1
<400>2
cggatccggt?gatggcgacc?ttcttc 26
<210>3
<211>31
<212>DNA
＜213〉artificial sequence
<220>
＜223〉the special primer DET1 R of amplification gene DET1
<400>3
ggactagtcc?gctcacaact?gttacaaata?c 31
Claims (3)
1. paddy rice high chlorophyll content mutator gene DET1 is characterized in that: have the nucleotide sequence shown in SEQ ID No.1.
2. the recombinant plant overexpression vector that contains the described paddy rice high chlorophyll content of claim 1 mutator gene DET1.
3. recombinant plant overexpression vector according to claim 2 is characterized in that: described recombinant plant overexpression vector is to insert paddy rice high chlorophyll content mutator gene DET1 and obtain in the multiple clone site of pCUPHN carrier; Described pCUPHN carrier is transformed by the pCAMBIA-1300 carrier, promptly introduce corn ubiquitin promoter gene by restriction enzyme Pst I at the multiple clone site place of pCAMBIA-1300 carrier, introduce HA label gene by Sac I and Spe I afterwards, introduce terminator NOS gene by Spe I and EcoR I again.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102993284A (en) * | 2011-09-14 | 2013-03-27 | 中国科学院植物研究所 | New use of synechocystis protein in paddy rice property improvement |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006121757A2 (en) * | 2005-05-05 | 2006-11-16 | Monsanto Technology, Llc | Plants containing a heterologous flavohemoglobin gene and methods of use thereof |
| CN101096676A (en) * | 2007-06-08 | 2008-01-02 | 南京农业大学 | Rice chlorophyll synthase mutant gene and its application in gene engineering |
-
2010
- 2010-05-19 CN CN 201010177039 patent/CN101831439A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006121757A2 (en) * | 2005-05-05 | 2006-11-16 | Monsanto Technology, Llc | Plants containing a heterologous flavohemoglobin gene and methods of use thereof |
| CN101096676A (en) * | 2007-06-08 | 2008-01-02 | 南京农业大学 | Rice chlorophyll synthase mutant gene and its application in gene engineering |
Non-Patent Citations (2)
| Title |
|---|
| 《GenBank》 20091007 Huang,J等 GQ898934.1 1-2 1-3 , 2 * |
| 《Journal of Plant Physiology》 20081231 Fenghua Wang等 Heredity, physiology and mapping of a chlorophyll content gene of rice (Oryza sativa L.) 324-330 1-3 , 2 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102993284A (en) * | 2011-09-14 | 2013-03-27 | 中国科学院植物研究所 | New use of synechocystis protein in paddy rice property improvement |
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Application publication date: 20100915 |