CN109295071B - Rice flower organ development regulation gene PEH1, and encoded protein and application thereof - Google Patents
Rice flower organ development regulation gene PEH1, and encoded protein and application thereof Download PDFInfo
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Abstract
The invention provides a rice floral organ development regulation gene PEH1, and a protein coded by the gene PEH1 and application thereof, wherein the gene PEH1 has a nucleotide sequence shown as SEQ ID No. 1; 1 nucleotide sequence, allele or derivative generated by adding, substituting, inserting or deleting one or more nucleotides in the SEQ ID No; the protein has an amino acid sequence shown as SEQ ID No. 2, and the amino acid sequence or the derivative generated by adding, substituting, inserting or deleting one or more amino acids in the amino acid sequence of the SEQ ID No. 2; the gene PEH1 and the protein coded by the gene are applied to the growth regulation of rice floral organs. The gene is used for regulating and controlling the development of rice floral organs, and has important theoretical and practical significance for deeply researching the molecular mechanism of the development of rice reproductive organs.
Description
[ technical field ] A method for producing a semiconductor device
The invention relates to the field of plant genetic engineering, in particular to a rice floral organ development regulating gene PEH1, and a protein coded by the gene and application of the gene.
[ background of the invention ]
Rice is not only a model plant for monocot genome research, but also an important food crop. Rice is the staple food of one third of the world's population, and its flowers are not only reproductive organs, but also the basis for the formation of seed particles. The normal development of rice flowers directly affects the rice yield and rice quality, so the research on the rice flower development is always the focus of attention of rice genetic breeding workers. The research on the development of rice flowers not only has important theoretical significance, but also has important guiding significance on the genetic breeding of the rice yield and quality.
In the 90 s of the 20 th century, plant developmental biologists discovered that floral organ formation of dicotyledonous plants was controlled by multiple genes, which were related to each other, through studies on homeotic mutants of Arabidopsis and grass flowers, and proposed an ABC model with milestone significance (Coen ES and Meyerowitz EM.the war of the world: genetic interactions controlling flower depth. Nature,1991,353(5): 31-37.). With the progress of the study and the increasing number of floral allotypes cloned, the ABC model has been extended to the ABCDE model (Theissen G., Development of floral organization: store from the MADS house. curr Opin Plant biol.,2001,4(1): 75-85.).
The research of rice molecular genetics finds that the part of the ABCDE flower organ regulation model obtained from the research of species such as dicotyledonous model plants Arabidopsis thaliana and snapdragon is suitable for explaining the molecular regulation of the development of the flower organ of monocotyledonous plants such as rice (Yoshida H and Nagato Y. flower definition in rice.2011, J Exp Bot.,62(14): 4719-. At present, a plurality of MADS-box genes (in New, Wangjiajun, Wangchailin, molecular mechanism of rice floral organ development, molecular plant breeding, 2013,11(4): 617-624), which have similar functions and homologous products with dicotyledonous plants Arabidopsis and antirrhinum, are cloned, however, the rice has a far difference in evolution with Arabidopsis and the like, the morphological structure difference of flowers is large, rice flowers have stamen and pistil reproductive organs similar to those of the dicotyledonous plants, but have no obvious calyx and petal structures of the dicotyledonous plants, and periandral and pistil reproductive floral organs are husk, hull and serous which are obviously different from calyx and petals of the dicotyledonous plants and other monocotyledonous plants.
With the intensive research and the isolation of new mutant genes related to rice floral development, more and more researches show that the major difference between the organ morphologies of monocotyledonous rice and dicotyledonous arabidopsis thaliana flowers still needs to be solved (Hitoshi Y and Yasuo N.flower definition in rice.J Exp Bot,2011,62(14): 4719-. Firstly, whether the palea and palea are homologues of the calyx or not is not supported by sufficient experimental evidence; secondly, the inner and outer lemma are the same organ or different organs without obtaining sufficient evidence; third, it is still unclear whether the rice glumes are organic components of a degenerated flower or an entire flower. Therefore, the genetic regulation mechanism of the rice floral development process cannot form an overall outline like that of arabidopsis thaliana, and new gene cloning and research are needed to clarify some basic problems of rice floral development.
[ summary of the invention ]
The invention aims to solve one of the technical problems and provides a rice floral organ development regulating gene PEH1 and application thereof, wherein the gene is used for regulating and controlling the rice floral organ development, and has important theoretical and practical significance for deeply researching the molecular mechanism of rice reproductive organ development.
The present invention achieves one of the above technical problems:
a rice floral organ development regulation gene PEH1, wherein the gene PEH1 has a nucleotide sequence shown as SEQ ID No. 1.
Further, the nucleotide sequence, allele or derivative generated by adding, substituting, inserting or deleting one or more nucleotides in the nucleotide sequence of SEQ ID No. 1.
Further, the rice floral organ development regulation gene PEH 1-containing plasmid, a plant expression vector and a host cell.
Furthermore, the gene PEH1 and the nucleotide sequence, the allele or the derivative generated by adding, substituting, inserting or deleting one or more nucleotides in the nucleotide sequence thereof are applied to the regulation of the development of the floral organs of rice.
Furthermore, the plasmid, the plant expression vector and the host cell containing the rice floral organ development regulation gene PEH1 are applied to the rice floral organ development regulation.
The second technical problem to be solved by the invention is to provide a protein encoded by a rice floral organ development regulating gene PEH1 and application thereof, wherein the protein is used for regulating and controlling rice floral organ development, and has important theoretical and practical significance for deeply researching a molecular mechanism of rice reproductive organ development.
The invention realizes the second technical problem in the following way:
a protein coded by a rice floral organ development regulation gene PEH1, which has an amino acid sequence shown in SEQ ID No. 2.
Furthermore, the amino acid sequence or the derivative generated by adding, substituting, inserting or deleting one or more amino acids in the amino acid sequence of SEQ ID No. 2.
Furthermore, the protein and the amino acid sequence thereof are added, substituted, inserted or deleted with one or more amino acids to generate the amino acid sequence or the derivative, and are applied to the regulation of the development of the floral organs of rice.
The invention has the following advantages:
the gene or protein is vital to the normal development of rice floral organs (palea, male and female organs, and the like), the mutation of the gene or protein causes the abnormal formation of rice male and female embryos, the morphological deformity of the palea and the palea to form pepper-like glume flowers, and the gene or protein can be used for modifying the rice floral organs, so that the gene or protein has important theoretical and practical significance for deeply researching the molecular mechanism of rice reproductive organ development.
[ description of the drawings ]
The invention will be further described with reference to the following examples with reference to the accompanying drawings.
FIG. 1 is a structural diagram of a mutation of the PEH1 gene of the present invention.
FIG. 2 is a sequence diagram of the PEH1 gene of the present invention, wherein a is a wild type and b is a mutant type.
FIG. 3 is a bar graph showing the expression levels of the PEH1 gene of the present invention in mutant fro1(t) and Minghui 86.
[ detailed description ] embodiments
The invention relates to a rice floral organ development regulation gene PEH1, wherein the gene PEH1 has a nucleotide sequence shown as SEQ ID No. 1.
The nucleotide sequence, allele or derivative generated by adding, substituting, inserting or deleting one or more nucleotides in the nucleotide sequence of SEQ ID No. 1.
The gene PEH1 and the nucleotide sequence, the allele or the derivative generated by adding, substituting, inserting or deleting one or more nucleotides in the nucleotide sequence thereof are applied to the regulation of the development of the floral organs of rice.
The invention also relates to a plasmid, a plant expression vector and a host cell containing the rice floral organ development regulating gene PEH 1.
The plasmid, the plant expression vector and the host cell containing the rice floral organ development regulation gene PEH1 are applied to the rice floral organ development regulation.
The invention also relates to a protein coded by the rice floral organ development regulatory gene PEH1, and the protein has an amino acid sequence shown as SEQ ID No. 2.
The amino acid sequence or the derivative generated by adding, substituting, inserting or deleting one or more amino acids in the amino acid sequence of SEQ ID No. 2.
The protein encoded by the rice floral organ development regulatory gene PEH1 is an amino acid sequence or a derivative generated by adding, substituting, inserting or deleting one or more amino acids in an amino acid sequence of the protein and the amino acid sequence, and is applied to the rice floral organ development regulation.
The gene obtaining and function verification steps are as follows:
PEH1 gene mapping: in order to separate genes, the invention firstly constructs a positioning population, normal plants (the genotype is wild type or heterozygote type) in the population are separated by the mutant fro1(t) and hybridized with the indica rice variety 9311 to construct the positioning population, and candidate genes are quickly positioned by adopting a segregant mixed analysis (BSA-seq) method based on high-throughput sequencing.
And (3) verifying candidate mutation sites: the information of the candidate gene is found by using the Rice Genome mutation Project, a pair of specific sequencing primers are designed at two sides of the candidate locus, the DNA of the homozygous wild type and the DNA of the mutant are respectively subjected to PCR amplification, and the products are sent to sequencing and are compared to determine the candidate gene. And analyzing the expression quantity of the wild type and the mutant on RNA by Real-time PCR to further determine the candidate gene.
Functional analysis of the PEH1 gene: transgenic rice with normal phenotype restored by genetic transformation was obtained by transferring a fragment containing the entire PEH1 gene into mutant fro1(t) for functional complementation test.
The invention is based on early discovery of a rice floral organ development mutant fro1(t) (Maobigang, Liuhuaqing, Chenjian, Chengjie, Pengyonghong, Wanfeng, two rice reproductive organ mutants with morphological characteristics and genetic analysis. molecular plant breeding, 2008,6(2): 233-.
Compared with the wild type rice Minghui 86, the mutant fro1(t) has the mutation phenotype mainly shown in that: the plant type is short and small, and the growth vigor is weak; the inner and outer palea slices are twisted, deformed and cohered, the appearance of the spikelet is exactly like that of hot pepper, but the spikelet is always closed and does not bloom, and finally withers; the majority of the minor ingredients are degenerated; transformation of the serosa into a thin leaf organ; the stamen filament is expanded and twisted, the surface of the anther is twisted and deformed, no pollen exists, the number of the stamens is 2-6, part of the stamens are converted into a bottom leaf shape, and the top of the stamen is provided with a pistil-shaped organ similar to a stigma; the ovary develops normally, but most of the style fuses, and 3-4 styles cluster. The mutant fro1(t) cannot fruit and can only be stored in the form of hybrid strains, and is a mutant controlled by a single recessive gene.
The present invention will be further described with reference to the following specific examples.
Example 1: acquisition of rice floral organ development regulation gene PEH1
Rice material: the rice mutant fro1(t) is from tissue-cultured progeny of indica rice variety Minghui 86, and the parent is located as indica rice variety 9311.
Constructing a positioning group: wild phenotype (genotype is wild homozygous or heterozygous) in the segregating population of the mutant fro1(t) is selected to be hybridized with indica rice variety 9311 to construct a locating population F2.
Gene localization: based on the F2 population, performing phenotype selection in the segregation progeny, selecting individuals with 'mutant phenotype' (namely the phenotype is consistent with that of the mutant parent), respectively shearing leaves, mixing the leaves in equal amount, extracting DNA, and constructing a recessive homozygous pool; randomly selecting 50 individuals with a wild phenotype (namely, the phenotype of the individuals is consistent with that of a wild parent), respectively shearing leaves, mixing the leaves in equal amount, extracting DNA, and constructing a dominant heterozygous pool; and (4) selecting a single mutant parent strain to extract DNA for sequencing. Sending the parent individual and the offspring mixed pool to a Nordhea immunogenic bioinformatics technology limited company for whole genome sequencing, and adopting an IlluminaPE (paired end) sequencing method, wherein the read length is 150bp, and the sequencing depth is about 40 multiplied. And (3) taking a Nipponbare genome sequence as a reference, and screening the variation sites based on the two-pool reading data. The mutation sites that match the desired genotype in both pools (homozygous normal genotype in the wild-type pool, homozygous mutant genotype in the mutant pool) are identified as candidate mutation sites.
The total genome was screened to 4 candidate mutant spots (shown in table 1 below), 3 of which were intergenic and therefore excluded. Another candidate mutation site occurs in the LOC _ Os08g39420 gene coding region (+800bp) with 1 base A deletion (TAAAAAT-TAAAAT), which results in a frame shift of the coding sequence, thereby causing premature termination of protein translation of the gene, specifically referring to FIG. 1 (black box indicates exon region, left white box indicates 5-UTR, right white box indicates 3-UTR, black arrow indicates mutation position, number indicates deletion site, number indicates protein translation termination). Therefore, the LOC _ Os08g39420 gene was identified as a candidate gene for the fro1(t) mutant and was designated PEH1 (i.e., abbreviation for PEPER HULL 1).
TABLE 1 annotation of candidate mutation sites and genes involved
Example 2: determination of mutation site and target gene
To verify the results of the above screening, a pair of primers (5'-TACAGAAAGCCCTCAAGGAAGC-3' (shown as SEQ ID No: 3); 5'-AACAACAAGAGGGGGAAATCAG-3' (shown as SEQ ID No: 4)) was designed on both sides of the mutation site of PEH1 gene, and the genomic DNA of Minghui 86 and the mutant were PCR-amplified and the amplified products were sequenced, specifically referring to FIG. 2, wherein the sequence a in FIG. 2 is the wild type and the sequence b in FIG. 2 is the mutant, and the sequencing results revealed that the mutation in the mutant was indeed present.
In order to further verify the relevance of the deletion mutation and the gene thereof to the development of rice floral organs, the total RNA of Minghui 86 and mutant strains (three plants respectively) in some glumes of the heading is extracted and then is reversely transcribed into cDNA. Primers (5'-CAGAAGTCAGCGGATGTAAGGT-3' (shown as SEQ ID No: 5) and 5'-GGAATCGGCACAGCAATCAA-3' (shown as SEQ ID No: 6)) specific to the PEH1 gene sequence are designed to carry out qRT-PCR related experiments. The qRT-PCR reaction condition is 95 ℃ and 30 s; 95 ℃ for 10 s; 20s at 55 ℃; 72 ℃ for 10 s; 40 Cycles. All reactions were performed in 3 biological and 3 technical replicates, with the reference gene for qRT-PCR being OsActin1(LOC _ Os03g50885), and the specific results are shown in fig. 3. A, B, C in FIG. 3 represents the difference of the expression level of the PEH1 gene in three different plants of the mutant fro1(t) and Minghui 86 respectively, and the result shows that the transcription level of the PEH1 in the mutant is obviously reduced compared with that of the wild type.
Example 3: complementary experiment verification of PEH1 gene function
According to the sequence of LOC _ Os08g39420 gene, finding the upstream and downstream sequences of the gene in Nipponbare BAC clone AP014964.1 containing the gene on a website (https:// blast. ncbi. nlm. nih. gov/blast. cgi), designing a whole gene amplification primer, wherein the primer pair respectively contains URS and DRS sequences of CE Entry Vector (Vazyme) and recognition sequences of two endonucleases, and one primer comprises a URS sequence and a recognition sequence of endonucleases EcoRI [5'-ggatcttccagagatgaattcGTTGAAGAACTCGCTGTCCGTG-3' (shown as SEQ ID No: 7) ]; the other primer comprises a DRS sequence and a recognition sequence of endonuclease HindIII (5'-ctgccgttcgacgataagcttGTCGTCAGTCCTCGCATTATTCA-3' (shown as SEQ ID No: 8)), a fragment 6589bp comprises a coding region of the whole gene, a promoter sequence 1320bp upstream of an initiation codon and a regulatory sequence 419bp downstream of a termination codon.
Extracting DNA of indica rice variety Minghui 86 byHS DNA Polymerase with GC Buffer (TAKALA), PCR conditions of 94 ℃ for 5 sec; 98 ℃ for 10 sec; 60 ℃ for 30 sec; 72 ℃ for 7 min; 32 Cycles; 72 ℃ for 5 min. Using 1% agarose gel electrophoresis, the corresponding DNA fragments were recovered by scooping. This fragment was directly cloned into CE Entry Vector using the Clonexpress Entry One Step Cloning Kit (Vazyme), and the recombinant product was transformed and then subjected to monoclonal sequencing. And (3) completely digesting the clone shake bacteria quality-improving particles with correct sequencing by using endonucleases EcoRI and HindIII, and after electrophoretic separation, cutting a DNA fragment and connecting the DNA fragment to pCAMBIA1300 to construct a complementary vector pCAMBIA1300+ promoter + PEH1+ nos. The plasmid was transformed into rice by electric shock into Agrobacterium tumefaciens (Agrobacterium tumefaciens) strain LBA 4404. Transformation of mutant fro1(t) mature embryo-induced callus by Agrobacterium-mediated genetic transformation (Sujun, Huchangquan, Dian Zhai, Yan Jing Wang, Chen Jie, Wangfeng, 2003), establishment of Agrobacterium-mediated high-efficiency stable transformation system for indica Minghui 86, Fujian agricultural science, 18(4): 209-213).
The invention obtains 23 rice plants which transfer PEH1 independently, and carries out PCR identification on the transgenic plants to find 21 positive plants and 2 negative plants. Comparing the positive PEH1 transgenic rice plant with the contemporary mutant, the spikelet shape is found to be recovered to normal, and the rice plant has 1 complete spike-axis, 2 auxiliary glumes, 1 lemma, 1 palea, 1 pair of serous discs, 6 stamens and 1 pistil. The fertility of the male and female pistils is recovered, and the fruit is normal. Thus, it is further proved by complementation experiments that the gene PEH1 is a floral organ development regulatory gene.
In conclusion, the invention utilizes early screened rice mutant to construct a mixed pool, performs whole genome sequencing, rapidly locates and determines a target gene PEH1, and the gene controls the development of floral organs. Through the function reading of the PEH1 and the relationship of genes at the upstream and the downstream thereof, the method lays a foundation for further exploring the molecular regulation mechanism of the monocotyledon floral development.
Although specific embodiments of the invention have been described above, it will be understood by those skilled in the art that the specific embodiments described are illustrative only and are not limiting upon the scope of the invention, and that equivalent modifications and variations can be made by those skilled in the art without departing from the spirit of the invention, which is to be limited only by the appended claims.
Sequence listing
<110> institute of biotechnology of academy of agricultural sciences of Fujian province
<120> rice floral organ development regulation gene PEH1, and protein coded by same and application thereof
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gaagcgtact catcagagaa gtcagaaagg gaggagagcc gtcgccgccg tgggatgtcg 60
ccgtcgtcgt cgtcgtcgag cacggggccg atgggggtgg cggtgtcgcc ggaggtggaa 120
gcggcgctgg cgcgcggcgg cgcggtcgtc gccctcgagt ccaccatcat ctgccacggt 180
aagctcgcgc ctcgccgcct cctccccacc ccgtctccct cccggattac atggctcact 240
ccggcgatgt gcgcccgcgg ctgggggacg cgtttggctt tcttcccatc ccaggtatgc 300
cctacccgaa gaatctccag accgccatgg aggtggaggc cgtcgtgagg gagaacgggg 360
cggttcctgc caccatagcc attctgaacg gcgtgccaca tgttggtgag tcctgtgatt 420
tttcctgtga ctcggagatc agatacatgt tcaactgttc aagtgctgga ggcgtgatcc 480
gtagttgcta gctactatga attattctcc aattaacacc ggataatttt cgtaatgtga 540
taggccttag cggcgagcaa ttgaagagct tggctgtaag tggaagacag tttcagaaga 600
cggctagaag ggatattgca catgttgtga gtaccttgca cactagaaat tggtcgttca 660
gaatgttttc caatgtgttc agttatatct gcactaaaac tttacagttg atatatgttt 720
ctcgtcttat tgcttcctgt aacgagtttt actgattgaa aggtggcctc tggtggtaat 780
ggtgcaacaa cagtttctgc cactatgttt ttcgctcata aggtcagagc caaaaacatc 840
ttactgctct tgttactgtt aatttatctt tccttgattt gatagccttt ttcttgttca 900
tctttgttac atgtagttgg tgccaatcat tcatctctat atcaaactaa agttgctctc 960
aaattcaggt tggcatacca attttcgtaa ctggagggat tggaggtgtt catagaaatg 1020
gtgaacagag taagtacctt cctccttcca atcgtgtaag gagtcccttg ctatctgcac 1080
attttacagt tgatgcttca atcagttttt ctgagttatg aatggatgct gtagccttcc 1140
attttaatat cgtttactat taccatgggt ctaaatgtgg attatagcag ctcatgcctg 1200
atagaggtca tgtggcttaa tagctctaaa agagggcaag tgaaatgaca tgtttattat 1260
atctagacga tgtaattttt tgttgattta taccatttcg atcagtttgt gtttcttcat 1320
gtgttgaaaa cttagcagaa atttaatatg cctcttgatt ttttgtaatc tagtttgtct 1380
atgcctcttg attttttgta atctagtttg ttttctttag cttttctgca tcctgttgtc 1440
attctctagt taatttaaga gtaaagtgca cgggcggtcc ttaaacttgc aggggtatgt 1500
catctaggtc tctaaactct caaaatacat atccaagtcc aagaacttgt catagtgtgt 1560
catttaggtc ccaaatcgcc ccagccccta taggatccta cgtggcactg atgtggcatg 1620
ccacacggac aagacgtggc atcattttga ctaacaaatg aaacccattt atctttttct 1680
tcttttctta tcttttcctc ctcctctttt ttttctttct tctccttttt ttttcttttc 1740
ccgagtagaa gaaaggaaaa aaaaagaaga atgagaagaa aacgaagaaa aaagaaggga 1800
aaaaggaaga aaaggacata tcacgtccat atggcatgcc atatcagcgc cacataggat 1860
tctatagggg ctatgtcgat ttgggaccta aatgatacac tttgacaagt tctggaacct 1920
agatgtgcat tttgagagtt tggggaccta gatgacacaa ccttacaagt ttaaggaccg 1980
cctatgcact ttactcttaa tttaagatgc tgatggtgtt ataccatcta ctcagtttga 2040
accctgaagc taaatattta tttatatcca ttatccacac gttctttgaa atacattacc 2100
gacagtttgt tgtatatact tgattaagac atcaattgat taagcacacg tatgtcctat 2160
taaacagttt gcactgtttg atgcagccat ggacatctcc tcagacttaa ctgaacttgg 2220
aaagactcct gtcactgtta tttcagctgg tgtgaaatct attttggaca taccacggac 2280
acttgagtac ttggtattac ctatcactcc cactcttcaa aagaatggct ctttgatctg 2340
ttaaaagtga acttccgtca gttcatgtag attgttttct tgttagaaca ttggatactt 2400
cacacactgt gtatcagtat tttctggatt ctgttcatga agtatgagtg gagcagctgt 2460
tttcaggggc tctccaacaa aattctgtag tcatatctgt tagaccactc tgatatagga 2520
atactaatgt agcattataa tttaaatatg aagtattaca aaacaaggtt tacctagtag 2580
caaagcaaaa aatgctgtgg aattggtagc ttatttttac tcctagtgat gtgctgccat 2640
aagtatgctg ttagctgcac cgttactatg ataggatgat acactgtgca aattacagag 2700
ccaggatctg atacagtaag aaaccaggaa ccagaaatta gttaccacga aaattagctc 2760
acttcctatg tattttttgt tgttatattt ccctcaagaa atagaatgct gttgcatcag 2820
aacctgaaag ttttctcatg tccatgataa ctccatccag acaagtaaaa ctgctagatc 2880
tacaactgag ggtttatagt tttccagaat catacgtttg ttcattttat tctccataat 2940
ctacgaccat ccttttcaag taagtcatga caattttctg gtgtttcttg gtatagacaa 3000
gttatttctt taggttttac tcgttaaatg tgctatattt gaaatttggt gtatttctgg 3060
ctaggcattt caatgaatgc aatgttgttt ggtgctcaat cgtataggtt aactgattgc 3120
attggcagga aactcaagga gtaacggttg ctgcttacaa aaccaatgaa tttcctgctt 3180
tctttacaga agtcagcgga tgtaaggtag aataatttgc ttatgtaaaa tgtttataag 3240
ctatatttaa tttttgtttt gtttgtttaa ggagattttc ttttctgtaa tctggtgtga 3300
atctgtataa aaaagaagag ctggaaatga gtatgggtgt tgatttttca atgttgtgta 3360
ttgtgttgat ctgaaggtcc cagaatcttt cctttatata ttttgccatg agcccttggt 3420
acagtatctt ttgtctcact gtactgcagt gttaggatgt ttgtactaac tggctcctgt 3480
ctgcttaatt taggtgccat gtcgcgttga ttctcccgaa gagtgtgcca agataatatg 3540
taagtattaa agttcgcatt taccccatct gtattcatat ctgtatcatt tgtgcgatat 3600
tggtttgaca tgcactgttg gcacatgatc cattaacccg aactgattga actccagcag 3660
acatccaata ccttttgacc tttttactct gttggttaca gaaagctact ggccaatgtt 3720
aattcagaat cacgtaaatt caacaccaat atcctattag aatctacaat tgagtcagca 3780
taagattcag aactgaaagc agcacataac acgaatcggg agttcactat tgtatagagt 3840
ttgattttat cctatgaatt actataactc aactggaaaa agaatttatt gtaacacaga 3900
gtttatcagc attgctgatt tatatctatt ttttacagat gcaaacaaga atttacatct 3960
aggatctggg attttgattg ctgtgccgat tcccaaggaa catgcagctt caggaaatgc 4020
tatagagtct gcaatacaga aagccctcaa ggaagcagag tattgtcctt actcccaacc 4080
tcctgctctc tcactttatc ttttctgtgt atttgtgata ctaacagaaa tgatatctaa 4140
tgactttcta tgttgtttca gggataaaaa tataataggc aacgcgatca ctcccttcat 4200
gcttgacaga gtgaaagtac taactggacg atcttcgcta gaagccagta tcctacaaat 4260
ttaatttcaa actttgctgt attttgaacc acgatgattt tcgtatttcc ctgtagtttt 4320
gagcattgat tagctattta tttctatttc aacatgcaat ttggcatttg gatcttttca 4380
agttggactt tatgagagaa aaatcaaaag gttcaaacgt ttagatgaat aacatctgat 4440
ttccccctct tgttgtttcc tgtgtcaata gaaacttctc cttagcatat tcagatattg 4500
cgcttgtaaa gaacaatgct cttgttggtg ctaaaattgc tgtggccctt tctgatcttc 4560
accagagagt aacaaacagt aaggttctct catcaataat aatagtaaaa tcctaattct 4620
gtaattatct ctacaactaa atacttaggc atcctggttc ataaagattt gcaaggttat 4680
ctgttcttct tgcatgtttt tagtttttac tggatttagt ctactgtctg tgtttattgt 4740
tggagctaga ccctagactt gtgatcacac tcacgtctgc tctgggccag tgatttgctt 4800
ttttatgtga gatatcacac tagcatctcc tctgcctaca tgttcaattg agacctaaca 4860
attttgaact tgctgtaaca ggatttcgga ggtctgcctt atagagtgca tgtacaaagg 4920
caagcgaggt ctgaaacatg gggcatccta ttcatcctca caatgtatgc ttgtacaact 4980
agttggaaac ttgtagcttc atccttttct ggcactatat aaatgaaacc tgaggtgcga 5040
gtgcatgatt tgtagcgaga tttgctactg tagacttaaa ttagtacatt ccactccagc 5100
ataacgtgtt cttcactttt tcgggcgtgt tcttcacttt tttgttaatc tgatcttctg 5160
ctttcacctt gaaaacttga aattaatttg aactgggcat tcagtcctcc tatttggagt 5220
gcatgaactg ttgtcaaact aatgtggaat ttgtgacttg tc 5262
<210> 2
<211> 327
<212> PRT
<213> (Oryza sative)
<400> 2
Met Ser Pro Ser Ser Ser Ser Ser Ser Thr Gly Pro Met Gly Val Ala
1 5 10 15
Val Ser Pro Glu Val Glu Ala Ala Leu Ala Arg Gly Gly Ala Val Val
20 25 30
Ala Leu Glu Ser Thr Ile Ile Cys His Gly Met Pro Tyr Pro Lys Asn
35 40 45
Leu Gln Thr Ala Met Glu Val Glu Ala Val Val Arg Glu Asn Gly Ala
50 55 60
Val Pro Ala Thr Ile Ala Ile Leu Asn Gly Val Pro His Val Gly Leu
65 70 75 80
Ser Gly Glu Gln Leu Lys Ser Leu Ala Val Ser Gly Arg Gln Phe Gln
85 90 95
Lys Thr Ala Arg Arg Asp Ile Ala His Val Val Ala Ser Gly Gly Asn
100 105 110
Gly Ala Thr Thr Val Ser Ala Thr Met Phe Phe Ala His Lys Val Gly
115 120 125
Ile Pro Ile Phe Val Thr Gly Gly Ile Gly Gly Val His Arg Asn Gly
130 135 140
Glu Gln Thr Met Asp Ile Ser Ser Asp Leu Thr Glu Leu Gly Lys Thr
145 150 155 160
Pro Val Thr Val Ile Ser Ala Gly Val Lys Ser Ile Leu Asp Ile Pro
165 170 175
Arg Thr Leu Glu Tyr Leu Glu Thr Gln Gly Val Thr Val Ala Ala Tyr
180 185 190
Lys Thr Asn Glu Phe Pro Ala Phe Phe Thr Glu Val Ser Gly Cys Lys
195 200 205
Val Pro Cys Arg Val Asp Ser Pro Glu Glu Cys Ala Lys Ile Ile Tyr
210 215 220
Ala Asn Lys Asn Leu His Leu Gly Ser Gly Ile Leu Ile Ala Val Pro
225 230 235 240
Ile Pro Lys Glu His Ala Ala Ser Gly Asn Ala Ile Glu Ser Ala Ile
245 250 255
Gln Lys Ala Leu Lys Glu Ala Glu Asp Lys Asn Ile Ile Gly Asn Ala
260 265 270
Ile Thr Pro Phe Met Leu Asp Arg Val Lys Val Leu Thr Gly Arg Ser
275 280 285
Ser Leu Glu Ala Asn Ile Ala Leu Val Lys Asn Asn Ala Leu Val Gly
290 295 300
Ala Lys Ile Ala Val Ala Leu Ser Asp Leu His Gln Arg Val Thr Asn
305 310 315 320
Arg Phe Arg Arg Ser Ala Leu
325
<210> 3
<211> 22
<212> DNA
<213> (Artificial sequence)
<400> 3
tacagaaagc cctcaaggaa gc 22
<210> 4
<211> 22
<212> DNA
<213> (Artificial sequence)
<400> 4
aacaacaaga gggggaaatc ag 22
<210> 5
<211> 22
<212> DNA
<213> (Artificial sequence)
<400> 5
cagaagtcag cggatgtaag gt 22
<210> 6
<211> 20
<212> DNA
<213> (Artificial sequence)
<400> 6
ggaatcggca cagcaatcaa 20
<210> 7
<211> 43
<212> DNA
<213> (Artificial sequence)
<400> 7
ggatcttcca gagatgaatt cgttgaagaa ctcgctgtcc gtg 43
<210> 8
<211> 44
<212> DNA
<213> (Artificial sequence)
<400> 8
ctgccgttcg acgataagct tgtcgtcagt cctcgcatta ttca 44
Claims (3)
1. The application of a rice floral organ development regulation gene PEH1 is characterized in that: the gene PEH1 consists of a nucleotide sequence shown as SEQ ID No. 1, and the gene PEH1 is applied to the regulation of the development of rice floral organs.
2. A plasmid, a plant expression vector and a host cell of a rice floral organ development regulation gene PEH1 are applied to the rice floral organ development regulation, and the gene PEH1 consists of a nucleotide sequence shown as SEQ ID No. 1.
3. The application of the protein coded by the rice floral organ development regulatory gene PEH1 is characterized in that: the protein consists of an amino acid sequence shown as SEQ ID No. 2, and is applied to the growth regulation of rice floral organs.
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CN201811187792.2A CN109295071B (en) | 2018-10-12 | 2018-10-12 | Rice flower organ development regulation gene PEH1, and encoded protein and application thereof |
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CN116790623B (en) * | 2023-07-10 | 2024-01-26 | 贵州省水稻研究所 | Rice flower organ development regulation gene OsROXY2, protein coded by same and application thereof |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102321633A (en) * | 2011-09-28 | 2012-01-18 | 福建农林大学 | Pleiotropic gene for controlling vegetative growth and development of floral organs of rice and application thereof |
CN108570474A (en) * | 2018-04-17 | 2018-09-25 | 浙江师范大学 | Rice flower development gene EH1 and its application |
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2018
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Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102321633A (en) * | 2011-09-28 | 2012-01-18 | 福建农林大学 | Pleiotropic gene for controlling vegetative growth and development of floral organs of rice and application thereof |
CN108570474A (en) * | 2018-04-17 | 2018-09-25 | 浙江师范大学 | Rice flower development gene EH1 and its application |
Non-Patent Citations (3)
Title |
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GenBank: AP014964.1,Oryza sativa Japonica Group DNA, chromosome 8, cultivar: Nipponbare, complete sequence;Kawahara,Y.等;《Genbank》;20151010;第1-3页 * |
水稻花器官发育的分子机理;于新等;《分子植物育种》;20130318;第11卷(第4期);第617-624页 * |
水稻花器官突变体lfl(leafy lodicules)的基因定位;袁小珍等;《西南农业学报》;20121028;第25卷(第5期);第898-901页 * |
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