CN114807221A - Mutant, protein and expression vector of rice root elongation regulatory gene OsDRP1C and application thereof - Google Patents
Mutant, protein and expression vector of rice root elongation regulatory gene OsDRP1C and application thereof Download PDFInfo
- Publication number
- CN114807221A CN114807221A CN202210463032.XA CN202210463032A CN114807221A CN 114807221 A CN114807221 A CN 114807221A CN 202210463032 A CN202210463032 A CN 202210463032A CN 114807221 A CN114807221 A CN 114807221A
- Authority
- CN
- China
- Prior art keywords
- osdrp1c
- ala
- gene
- rice
- leu
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/146—Genetically Modified [GMO] plants, e.g. transgenic plants
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Physics & Mathematics (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to the technical field of rice breeding, and particularly discloses a mutant, a protein, an expression vector and application of a rice root elongation regulatory gene OsDRP 1C. The gene OsDRP1C is applied to the regulation of the growth of the rice root system, and the growth of the rice root system is regulated through missense mutation of one or more base pairs in a DNA sequence of the gene OsDRP 1C; the serial number of the gene OsDRP1C in TIGR is LOC _ Os03g50520, and the gene OsDRP1 8920 contains a nucleotide sequence shown as SEQ ID NO: 1. According to the invention, the gene OsDRP1C with the number of LOC _ Os03g50520 in TIGR is regulated and controlled, and the elongation of main roots, adventitious roots, lateral roots and root hair cells of rice is inhibited or promoted through missense mutation of one or more base pairs in a DNA sequence of the gene OsDRP1C, so that the regulation of the growth performance of the root system of the rice can be realized.
Description
Technical Field
The invention relates to the technical field of rice breeding, in particular to a rice root elongation regulatory gene OsDRP1C mutant, protein, an expression vector and application thereof.
Background
The rice root system is an important component of rice, is not only a physical support of the ground, but also an important organ for absorbing nutrient components and water from the ground. The rice root system is a fibrous root system, can continuously generate adventitious roots from cells in a meristematic region of a stem node closest to a stelar canal, and finally forms a root system structure by a plurality of adventitious roots and lateral roots thereof. The root type is determined by several parameters such as length, quantity and space range, researches find that the root length determines the genetic expression of other root system characters, the root system characters are the most important characters in all root system characters, and 7 overground part agronomic characters such as single plant yield and the like are in extremely obvious positive correlation with the root length. One of the major cytological bases of root elongation is cell division of the root tip meristem, and defects in root tip meristem function or structure can lead to short root development. Elongation of the root system another major cytological basis is the elongation of cells in the elongation zone of the root tip, which is associated with cell wall formation, cellulose synthesis and microtubular organization. Although the root length plays an important role in the yield and resistance of plants, the root growth increases the difficulty of research in soil, so that the genetic background and molecular mechanism of root elongation are poorly understood, and the root length factor is not fully utilized in rice breeding and cultivation. Therefore, the gene for controlling the rice root length is mainly excavated, a series of genes for controlling the rice root length are separated and cloned, and the rice root system character is convenient to directionally improve, for example, the invention patent with the granted publication number of CN104031928B discloses a gene OsKASI related to the rice root system growth, wherein the gene OsKASI and the coded protein have mutation of some basic groups and amino acids, and the rice root system is expressed as a short root system.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a rice root elongation regulation gene OsDRP1C, which can inhibit or promote the elongation of the rice root system through missense mutation of one or more base pairs in a DNA sequence, and can create conditions for rice root system improvement and cross breeding or transgenic plant development.
In order to solve the problems, the technical scheme adopted by the invention is as follows:
the gene OsDRP1C is applied to the regulation of the elongation of the rice root system, and the elongation of the rice root system is regulated through missense mutation of one or more base pairs in a DNA sequence of the gene OsDRP 1C;
the serial number of the gene OsDRP1C in TIGR is LOC _ Os03g50520, and the gene OsDRP1 8920 contains a nucleotide sequence shown as SEQ ID NO: 1.
Compared with the prior art, the invention can inhibit or promote the elongation of rice main root, adventitious root, lateral root and root hair cells by regulating the gene OsDRP1C which is numbered LOC _ Os03g50520 in TIGR and missense mutation of one or more base pairs in the DNA sequence, thereby realizing the adjustment of the elongation performance of the rice root system.
A mutant of a rice root elongation regulatory gene OsDRP1C has a nucleotide sequence shown in SEQ ID NO: 1, the base of the DNA sequence at 2858bp generates missense mutation, and thymine (T) is replaced by guanine (A). By inducing the replacement of thymine (T) at 2858 position of DNA sequence of gene OsDRP1C with guanine (A), the elongation of main root, adventitious root, lateral root and root hair cell of rice containing the mutant of gene OsDRP1C can be inhibited, and the root of rice can grow slowly.
A protein encoded by a rice root elongation regulatory gene OsDRP1C mutant, wherein the amino acid sequence of the protein is shown in SEQ ID NO: 2, the 233 th tyrosine (Tyr) of the amino acid sequence is mutated into a stop codon. The protein coded by the mutant of the rice root elongation regulatory gene OsDRP1C can inhibit the elongation of cells of the root system of rice containing the protein, and the root growth is slow.
An expression vector containing the rice root elongation regulatory gene OsDRP 1C. The expression vector containing the rice root elongation modulating gene OsDRP1C can promote or inhibit the elongation of the root system of rice containing the expression vector.
The expression vector of the rice root elongation regulatory gene OsDRP1C is applied to the improvement of rice root system characters. The expression vector is applied to the aspect of improving the rice root system character, and the rice root system character can be optimized.
The rice root elongation regulation gene OsDRP1C mutant is applied to regulation of rice root system characters. The mutant of the rice root elongation regulation gene OsDRP1C is applied to the regulation of rice root system characters, so that the rice root system can be expressed into a short root system.
Drawings
FIG. 1 is a comparison of the phenotype of Wild Type (WT) and Osdrp1c mutants in normal hydroponic culture for 7 days, a being the total illumination of 7-day-old Wild Type (WT) and Osdrp1c mutants at bar 2.0 cm; b and c are observations of the main root of WT and Osdrp1c under the stereoscope, respectively, bars 1.0 mm;
FIG. 2 is a comparison of resin sections from taproots of Wild Type (WT) and Osdrp1c mutant normally hydroponically cultured for 7 days;
FIG. 3 is a map-based cloning and gene structure diagram of OsDRP1C gene, a is the fine localization result of OsDRP1C gene, RM1350N is SSR marker used for localization, In1, In2, In3 and In4 are newly developed polymorphic markers, OSJNBa0014G15 represents BAC clone, Rec represents recon; b is the structure and mutation site of gene OsDRP1C, intron is indicated by solid line, untranslated region is indicated by open square box, exon is indicated by solid square box, and arrow indicates mutation site;
FIG. 4 shows Wild Type (WT), Osdrp1c mutant and two strains T 2 Generation transgenic reversion lines (OV) 1 And OV 2 ) A tabular diagram of (a);
FIG. 5 is a schematic structural diagram of a modified overexpression transformation vector pCAMBIA 1300.
Detailed Description
The present invention will be further illustrated with reference to the following examples, which are intended to illustrate the invention and are not intended to limit the scope of the invention.
Example 1
The embodiment provides the application of the gene OsDRP1C in the regulation of the elongation of the rice root system, and aims to regulate the elongation of the rice root system through missense mutation of one or more base pairs in a DNA sequence of the gene OsDRP 1C;
the serial number of the gene OsDRP1C in TIGR is LOC _ Os03g50520, and the gene OsDRP1 8920 contains a nucleotide sequence shown as SEQ ID NO: 1.
Compared with the prior art, the invention can inhibit or promote the elongation of main root, adventitious root, lateral root and root hair cell of rice by regulating the gene OsDRP1C with the number of LOC _ Os03g50520 in TIGR and leading to the defect of the function or structure of the meristem of the root tip through missense mutation of one or more base pairs in the DNA sequence, thereby realizing the regulation of the elongation performance of the root system of rice.
Example 2
The embodiment provides a mutant of a rice root elongation regulatory gene OsDRP1C and a protein coded by the mutant, wherein the nucleotide sequence of the rice root elongation regulatory gene OsDRP1C is shown in SEQ ID NO: 1, the base at 2858bp of the DNA sequence has missense mutation, and thymine (T) is replaced by guanine (A); the amino acid sequence of the protein is shown in SEQ ID NO: 2, the 233 th tyrosine (Tyr) of the amino acid sequence is mutated into a stop codon.
By inducing the replacement of thymine (T) at 2858 position of DNA sequence of gene OsDRP1C by guanine (A), and mutating tyrosine (Tyr) at 233 position of amino acid sequence of coded protein to stop codon, the elongation of rice main root, adventitious root, lateral root and root hair cell containing mutant of gene OsDRP1C or coded protein thereof can be inhibited, and the growth of rice root can be slowed down.
The embodiment also provides application of the mutant of the rice root elongation regulatory gene OsDRP1C in regulation of rice root system characters. The mutant of the rice root elongation regulation gene OsDRP1C is applied to the regulation of rice root system characters, so that the rice root system can be expressed into a short root system.
EXAMPLE 3 phenotypic and genetic analysis of mutants
A rice short root system mutant Osdrp1c is screened from a Kasalath mutant library of indica rice (Oryza Sativa L. ssp indica) mutagenized by ethylmethane sulfonate. The root elongation of the 7-day-old Osdrp1c mutant was severely inhibited, and the main root, adventitious root, lateral root and root hair were all shortened (fig. 1). Inhibition of elongation of mutant Osdrp1c cells resulted in a significantly shorter main root length than wild type, wider cells in the mature and elongation regions of the mutant but slightly narrower cells in the meristematic region, resulting in a thicker mutant root system but slightly smaller root tips (fig. 2). Therefore, the OsDRP1C gene can further regulate and control the growth and development of the root system by controlling the length and width of the rice root system cells. The mutant Osdrp1c is taken as a male parent and hybridized with wild type Kasalath to obtain a first filial generation F1 plant which is completely consistent with the wild type Kasalath, and the result shows that Osdrp1c is recessive mutation. In F2 obtained by F1 generation self-pollination, the segregation ratio of normal plants to short-rooted plants is 379/131-2.89, and the chi-square detection result is 0.13<χ 2 0.05,1 The ratio of normal to short root plants was consistent with a pair of gene-controlled 3: 1 segregation ratio, indicating that the mutant is a recessive monogenic mutant.
Example 4 mapping of OsDRP1C Gene
Short-root mutant individuals were isolated from the progeny F2 of the cross between mutant Osdrp1c and Nipponbare, and the mutant genes were mapped using Map-based Cloning. Initial localization (30F 2 mutant individuals) mapped the mutant gene between chromosome 3 SSR markers RM1350N and RM 5813N. Thereafter, the localization population was expanded and 4 pairs of new InDel molecular markers with polymorphisms, named In1, In2, In3 and In4, were developed within this interval, with the following sequences:
In1U-5’ACCTCCTCCTCCGCCAGCTCC 3’
In1L-5’AAATCTCCTCGCCCTTCTTCCC 3’
In2U-5’TGCCCACTGATTAGCCGTAG 3’
In2L-5’GGATAGAGATGCCCATAGATTTG 3’
In3U-5’GCGGTGTTTATCAGATTGGT 3’
In3L-5’TAAATAGATACTCCATCGGCTTA 3’
In4U-5’CGCCAATGGTGTGTAGGAGGA 3’
In4L-5’CCGTTATGCCGTCCACAGATG 3’
finally, the gene is locked between InDel markers In2 and In3 (the specific physical positions on chromosome 3 are 28821343bp and 28952920bp respectively), recombinants are 3/1112 and 3/1112 respectively, and the recombination numbers are different. This interval was 131.2kb in size (FIG. 3).
Example 5 Gene prediction and sequence analysis
According to the fine localization results, the OsDRP1C gene was located on BAC clone OSJNBa0014G15 (FIG. 3). According to the rice gene annotation information of TIGR (http:// www.tigr.org/tdb/e2k1/osa1/), gene prediction analysis is carried out on positioned chromosome segments, 19 predicted genes are shared in the region, the 19 candidate genes are amplified by Kasalath wild type and Osdrp1c mutant DNA templates, amplification products are respectively sequenced, the sequencing results are subjected to alignment analysis, 2858bp thymine (T) of the DNA sequence of the gene with the gene number of LOC _ Os03g50520(Dynamin related protein) is replaced by guanine (A), thus tyrosine (Tyr) at position 233 of the coded amino acid sequence is mutated into a stop codon, and the gene is named as OsDRP 1C.
Example 6OsDRP1C functional complementation assay
According to CDS sequence information of the OsDRP1C gene, primers for amplifying complete CDS are designed, and the primer sequences are as follows (underlined is a restriction enzyme cutting site):
OsDRP1C-CDSU：5’TGAGCCGGAGACTACAGCGAC 3’
OsDRP1C-CDSL：5’AACCGATCATTTCCACGCGAC 3’；
OsDRP1C-OVU：5’AAAGAGCTCCTTTGCCCGAGATGGCGACGA3’
OsDRP1C-OVL：5’AAATCTAGAAACCGATCATTTCCACGCGAC3’；
the total RNA of the rice kasalash leaves is extracted by adopting a Shanghai worker RNA extraction kit (SK1321), oligo (dt) -18 is used as a primer, and the extracted total RNA is used as a template to carry out reverse transcription to synthesize first strand cDNA. Using the cDNA as a template and OsDRP1C-CDSU and OsDRP1C-CDSL as primers, amplifying a full-length ORF by using PrimeSTAR HS DNA Polymerase, and carrying out PCR conditions: pre-denaturation at 98 ℃ for 10s, then entering a cycling reaction (98 ℃ for 10s, 58 ℃ for 10s, 72 ℃ for 2min), wherein the number of cycles is 30, and finally, extending for 10 min. The PCR product was recovered by agarose electrophoresis tapping, purified and ligated into pEASY-Blunt Simple Cloning Vector. Transforming Escherichia coli DH5 alpha competent cells by hot shock of the ligation products, culturing at 37 ℃ for 16h, selecting positive monoclonal for sequencing, taking the recombinant vector as a template and OsDRP1C-OVU and OsDRP1C-OVL as primers after sequencing identification, amplifying full-length ORF by using PrimeSTAR HS DNA Polymerase, and carrying out PCR conditions: pre-denaturation at 98 ℃ for 10s, then entering a cycling reaction (98 ℃ for 10s, 58 ℃ for 10s, 72 ℃ for 2min), wherein the number of cycles is 30, and finally, extending for 10 min. The PCR product was recovered by agarose electrophoresis tapping, double digested overnight at 37 ℃ with SacI and XbaI, and ligated into the same double digested pCAMBIA1300 modified (35S) vector (FIG. 5), designated 35S-OsDRP1 COVER. The ligation product is transformed into escherichia coli DH5 alpha competence by heat shock in the same method, a single clone is picked after being coated on an LB plate containing kan resistance and cultured for 16h, plasmid double digestion is extracted by shaking bacteria, and agrobacterium tumefaciens EHA105 is transformed by electric shock after being detected to be correct by agarose gel.
The plasmid with correct enzyme digestion detection is introduced into the mutant Osdrp1c through a rice genetic transformation system mediated by an agrobacterium strain EHA105, and the transgenic plant is obtained through infection, co-culture, selection of hygromycin-resistant callus, differentiation, rooting, training and transplanting. The agrobacterium (EHA105) mediated rice genetic transformation system is optimized mainly by applying the method reported by Hiei et al (1994). The root length of the Osdrp1c mutant isolated from the transgenic plant is restored to be wild type (FIG. 4), which shows that the phenotype of Osdrp1c is caused by OsDRP1C mutation.
Although the present disclosure has been described above, the scope of the present disclosure is not limited thereto. Those skilled in the art can make various changes and modifications without departing from the spirit and scope of the present disclosure, and such changes and modifications will fall within the scope of the present invention.
Sequence listing
<110> Ningbo university college of science and technology
<120> rice root elongation regulatory gene OsDRP1C mutant, protein, expression vector and application thereof
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 5948
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 1
ggtactaaaa aaaatttttc cttcgatttc tctcgcctcc gcttctccac tctccggtag 60
atcccaccct ccaccgccgc cgccgccgcc gtgctccggt gagccgccgg tgagccggag 120
actacagcga cgtcgcgcag agccgttggg cgggtaggcg gttgggggct ttgcccgaga 180
tggcgacgat ggggagcctg atcgggctgg tgaaccggat ccagcgggcg tgcaccgtcc 240
tcggcgacca cgggggcggc ggcgaggggg gttccctctg ggaggcgctg ccctccgtcg 300
ccgtcgtcgg aggccaggta cgcgtgctcg ccaccaccct gcgtacctac ctctcagatc 360
caaagccatg gcggccagat ctggatatgg cgctcacccc ccttcggttt gcgtttgtga 420
tgcatccaga gttccgggaa gtcgtcggtg ctcgagagca tagtggggag ggacttcctg 480
cctcgtggat ccggtaatgc cgacgctgga tcgctcgttt ttttttcttt tttcgcgcgc 540
catttcgtct ggatcggctt ggtttgcgag ggattaatgc gcattctctc ggtttgatta 600
ggaatcgtga cgaggaggcc tcttgtgctg cagctgcaca agacggaggg tgggcaggag 660
tacgccgagt tcctccacgc cccgcggaag cgtttcactg actttggtaa gggatcattg 720
ccatatttgt gctttgcagt ccagcaggtt gatttgtgta ttcaatttca tttattagag 780
ttagtgttgt gcaaacttca aagttgtctt aattccaacg agtagacttt tccacgagag 840
atcagcaagg aggatgatac cgctttccct ggttatagtt tacttcactg aagtttgcct 900
atcaacctaa aaacaagaac atatacaatt agcatccaca cattgctata tgcagtgtga 960
actaaggaac taatgacgtt taggagtaga tctcacaagt cacaatcact gtaggatact 1020
agtgagtatc tgagcaaagt tatttcccgg ttcattacta gtgtcctaac atagctattg 1080
caagagatgt tcagttgaaa tatttaaatt ctttgtataa ttgcagctgc tgttaggaaa 1140
gagattgctg atgaaaccga tcgcattact ggaaaaacga aagctatatc aaatattccc 1200
atccatttga gtatatattc tccacatggt aagtcttgat gcttgttttg aacttttggt 1260
atgtgatgtg tactatgaat gtttcattca aacagaacct gcttcgctga ctcgctctca 1320
gctcccccaa atgcctgtaa actttaaatt gtatgttgtg gatgaaattt gggatcaata 1380
ggatgtagac tgataatttc tactattact tttacatttt taaatttgga ttcctttcag 1440
gatttatccc ttataattct tcaaaacttg agagctggca tatgtttcct ttaaaaaaaa 1500
tgtatatttg ctctgctctc cacctgttct tagttctgat gttggtctaa acccctccat 1560
tgtggtcctt tcagactaat cacaccaaac tgtatgaggc atctctgcag atttatatta 1620
gcacttcaaa atttcatgtt tgatattaat aatcgaccct gctttatatg caacaaaatt 1680
aagcatcaaa atattgacta tagaaacaat tcaaaaggtg tacataaaag gaatcattct 1740
ggaaatctat ttcttaaccc tgaggaacaa ataatcatat tatgataatt tgaaagtagt 1800
tacattgtgc ttgtgctgca gcatatgacc tgttatatag agacgttttg tgctcagcag 1860
atcctagtcc ttgtcataaa ttttttcact aatgtaattt tgcacccatt ttcttgcagt 1920
tgtaaacctg acacttattg atcttcctgg attgacgaag gttgctgtag gtatgttttc 1980
ttggatgctt ctagttttct atgcgtactc tttttatgtg attgtttgtg acacgaactt 2040
tttattaaca gaggggcaac aagaatctat tgtccaagat attgaaaaca tggttcggtc 2100
ttatgttgat aaggtaaaag aagtctgttg catgccatta gtgccatgtt tattttagct 2160
ccatgtgcta cattcactta atgagtactg tccaccctgc catatcaatt ctaagcttta 2220
ctattgttct tgtccaatgt ttatttgtgg tgacagtttt tttttctggg gtgggtagtg 2280
ggtactacga gtcaagtcta gaaacattat tcttttgttc tctggcctgt cttatggtca 2340
tgctaatata acttaatgta tttctatatg ctgcaaattc tgcttggtca tgttctgatt 2400
aaaactgttc atttttgttc ctacatttgt tctttttgga aacttcagcc aaattctatc 2460
atactggcca tatctccagc taatcaagat atagcaacat cagatgctat caagcttgct 2520
agggatgttg atccttcagg tacattttat attttcttca ctgtggcatt tcaaatattt 2580
ctcacatatg acaataattt tatgcatatt ttattttatt gccatacagc agtattgcag 2640
tgatctgtct tgttccattc ttatctgttg acaatttatt tcataatatt tttaggtgac 2700
agaacctttg gagtgttgac aaagcttgat ttgatggata agggtaccaa tgctgttgat 2760
gtatgtgact tgtcattttt tcttaactct tgtacgctat ggacacagca ctcattttca 2820
acgccatgta atcctaggta cttgaaggga ggcagtatcg tttgcaacac ccctgggtgg 2880
gaattgtcaa ccggtcacaa gctgatatca acaggaatgt cgacatgcta gcagcaagac 2940
gcaaagaaaa ggagtacttt gaaagtagcc cagattatgg tcacttggca cataaaatgg 3000
gtgcagagta ccttgctaag cttctgtctc aggtcagggc attttgctat tgtggtgtac 3060
tttccctctc cccaaaaaat aactggattg aaaaggctga ctgctgatac acttctattt 3120
acacattcag cacttagagg ctgtgatcag agcaaaaatt ccaagtatta tagccatgat 3180
taacaaaaca attgatgaaa ttgaagctga gttggatcgc cttggtaggc caattggagg 3240
tgatgctggg gtaagaaaaa ccgtactttg cttgttgtaa ctttctgcaa gtgttcaggt 3300
tcatgttgtt cgtgccacaa cctgcaacat tcatttgaat ggcaccctag cacattatct 3360
attcccaaac catgtctaca tcatgctgtc tcataaactt atcataatct acaggcacaa 3420
ctgtacacaa tattggacat gtgtcgtgcg tttgaccgag tttttaaaga gcacctagat 3480
ggcgggtaag atattttatt aggccctttt cttgagtgct gttgcttttt atgccatata 3540
tattgataaa tagtatccca tagatgaatg gcaaagaatc tacaggcatg ttaataacac 3600
tcaattattt tttgttctct tagtcggcca ggtggagatc gtatttatgg tgtctttgac 3660
caccaattac cagcagcact gaaaaagctt ccatttgata agcatctttc attgcaaaat 3720
gttcggaaag tcatttcaga ggctgatggt tatcaacccc acttgattgc ccctgagcaa 3780
gggtacagaa ggcttataga tagttcgctc cactacttca ggggtccagc ggaagcttca 3840
gttgatgcgg tatacttctc atcaatagat cttctttagg cacttttaat ttaattatta 3900
agcaagtagg atcctgttcc aatgactgat atatagactt acataaacta gccttttctt 3960
atccaatata tacattttcc ccctgtgttg aacaactaaa acatcatatg agaagattgc 4020
agaaccatga cgagcttctt gtcaaagtag tgaatataag agtgcttaag ttcatcaagt 4080
gctcaattgc ggaaagagag ggatactata gatttaattg gaatgtacca tttgtttatt 4140
caacaaagga aggcatttga ttccagtgct tttgggggca ttttcatatg tcaagccata 4200
ttgcactacg tctgttttgt tcactacatc tggttgatta tgttctcgta tctgctaggt 4260
tcatttggtc ttgaaggaac ttgttcgtag atcgattgca gcaacagagg tttgtgttga 4320
ttttcaattc tcagtgaacc tcagtagata ttactgatca ctcaatgaca ctcctgcatt 4380
gactgtttac attggttatc ttggatgcca caggagttga agcgtttccc aacacttcag 4440
acagatatag ctgctgcagc aaatgaaagc ctggaaagat ttcgtgagga tggtcgaaag 4500
acagttattc gtcttgttga gatggaggcc agctacctaa ctgtagaatt tttcagaaaa 4560
ctccctacag aaccagataa gggggctaat aataacactc cagccaatga cagatatcag 4620
gacaaccatt taagaagaat cggtaacaat tcaacattct attttgtttc tttaatggca 4680
attgattgag tcaactgttc taagacctca acacagaggt cacacgaagt acttaacaaa 4740
ctgacttaat aagcagaatc taagatgtaa catagggtac tagtgccatt cacgttctgc 4800
tgagtgctga cactagcaaa catgtcaaga tatgaccata tgtgaatttc gatgacaaca 4860
attatgcgaa ctaatttttt aaatcgacaa agtatattca tgacatcatc attgttttat 4920
tcagggtcaa atgtatcatc ttacattaac atggtttgtg agacattgag gaacaccatt 4980
ccaaaagctg tggtgcactg tcaagtgaag gaggcaaaaa gaaacttgct taaccgtttc 5040
tatgctcatg tgggaagcaa ggaggtatgt tttatactac cttagttcaa aactaaaaaa 5100
agataacgtg tttcatagtg attagaaagt tggctcataa ttaatatcga gtaaattgca 5160
ctctacttca aaatattaac catccatttc aaaatatatg tacactgtat ttgtggctaa 5220
atgaaaattt tcttgtttta tccttgtgtc gcattgattt gcacatttat attaacttgg 5280
acaccatttg ctttttattt ccataacgca gaagaaacag ctcagcgcga tgttggatga 5340
ggatcccgct ttgatggaga agagggattc cttagtcaag aggctagagc tgtacaaatc 5400
tgcccggaac gagatcgact cagtcgcgtg gaaatgatcg gttagcgaag tcacagctgg 5460
aagagataga catacatctg cgacacacag atcattgccg tagaatgctc taattccatt 5520
tacgtctgga ttcacatgga gatccgtaca ctattctttc tttctttttt ctttcatgcg 5580
agtgtccttt ctaccttgtc agacaagtat atttccttgg ccatcttggc ctttttgtat 5640
gatgttgatg atgatgagct ggtcctccgg gatcacactg atatgaatgc agatatatat 5700
attctttgtt gatttagcac ctccactgat atatcatgag agattttaga tgccttgttg 5760
ccaactgaaa ggatccgcat tcttggtaga gctacatttt ttgctaatgg aggcggctct 5820
tgaagtcgtc ccatggaaga tccttctcca ggtactcgat gttggtggga ttgaacgggc 5880
ccctcaccac gtccaccgtc tgtatgatcg ccctctccgg cagcacgctc atcgctggat 5940
ccgccgcc 5948
<210> 2
<211> 611
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 2
Met Ala Thr Met Gly Ser Leu Ile Gly Leu Val Ala Ala Ile Gly Ala
1 5 10 15
Ala Cys Thr Val Leu Gly Ala His Gly Gly Gly Gly Gly Gly Gly Ser
20 25 30
Leu Thr Gly Ala Leu Pro Ser Val Ala Val Val Gly Gly Gly Ser Ser
35 40 45
Gly Leu Ser Ser Val Leu Gly Ser Ile Val Gly Ala Ala Pro Leu Pro
50 55 60
Ala Gly Ser Gly Ile Val Thr Ala Ala Pro Leu Val Leu Gly Leu His
65 70 75 80
Leu Thr Gly Gly Gly Gly Gly Thr Ala Gly Pro Leu His Ala Pro Ala
85 90 95
Leu Ala Pro Thr Ala Pro Ala Ala Val Ala Leu Gly Ile Ala Ala Gly
100 105 110
Thr Ala Ala Ile Thr Gly Leu Thr Leu Ala Ile Ser Ala Ile Pro Ile
115 120 125
His Leu Ser Ile Thr Ser Pro His Val Val Ala Leu Thr Leu Ile Ala
130 135 140
Leu Pro Gly Leu Thr Leu Val Ala Val Gly Gly Gly Gly Gly Ser Ile
145 150 155 160
Val Gly Ala Ile Gly Ala Met Val Ala Ser Thr Val Ala Leu Pro Ala
165 170 175
Ser Ile Ile Leu Ala Ile Ser Pro Ala Ala Gly Ala Ile Ala Thr Ser
180 185 190
Ala Ala Ile Leu Leu Ala Ala Ala Val Ala Pro Ser Gly Ala Ala Thr
195 200 205
Pro Gly Val Leu Thr Leu Leu Ala Leu Met Ala Leu Gly Thr Ala Ala
210 215 220
Val Ala Val Leu Gly Gly Ala Gly Thr Ala Leu Gly His Pro Thr Val
225 230 235 240
Gly Ile Val Ala Ala Ser Gly Ala Ala Ile Ala Ala Ala Val Ala Met
245 250 255
Leu Ala Ala Ala Ala Leu Gly Leu Gly Thr Pro Gly Ser Ser Pro Ala
260 265 270
Thr Gly His Leu Ala His Leu Met Gly Ala Gly Thr Leu Ala Leu Leu
275 280 285
Leu Ser Gly His Leu Gly Ala Val Ile Ala Ala Leu Ile Pro Ser Ile
290 295 300
Ile Ala Met Ile Ala Leu Thr Ile Ala Gly Ile Gly Ala Gly Leu Ala
305 310 315 320
Ala Leu Gly Ala Pro Ile Gly Gly Ala Ala Gly Ala Gly Leu Thr Thr
325 330 335
Ile Leu Ala Met Cys Ala Ala Pro Ala Ala Val Pro Leu Gly His Leu
340 345 350
Ala Gly Gly Ala Pro Gly Gly Ala Ala Ile Thr Gly Val Pro Ala His
355 360 365
Gly Leu Pro Ala Ala Leu Leu Leu Leu Pro Pro Ala Leu His Leu Ser
370 375 380
Leu Gly Ala Val Ala Leu Val Ile Ser Gly Ala Ala Gly Thr Gly Pro
385 390 395 400
His Leu Ile Ala Pro Gly Gly Gly Thr Ala Ala Leu Ile Ala Ser Ser
405 410 415
Leu His Thr Pro Ala Gly Pro Ala Gly Ala Ser Val Ala Ala Val His
420 425 430
Leu Val Leu Leu Gly Leu Val Ala Ala Ser Ile Ala Ala Thr Gly Gly
435 440 445
Leu Leu Ala Pro Pro Thr Leu Gly Thr Ala Ile Ala Ala Ala Ala Ala
450 455 460
Gly Ser Leu Gly Ala Pro Ala Gly Ala Gly Ala Leu Thr Val Ile Ala
465 470 475 480
Leu Val Gly Met Gly Ala Ser Thr Leu Thr Val Gly Pro Pro Ala Leu
485 490 495
Leu Pro Thr Gly Pro Ala Leu Gly Ala Ala Ala Ala Thr Pro Ala Ala
500 505 510
Ala Ala Thr Gly Ala Ala His Leu Ala Ala Ile Gly Ser Ala Val Ser
515 520 525
Ser Thr Ile Ala Met Val Cys Gly Thr Leu Ala Ala Thr Ile Pro Leu
530 535 540
Ala Val Val His Cys Gly Val Leu Gly Ala Leu Ala Ala Leu Leu Ala
545 550 555 560
Ala Pro Thr Ala His Val Gly Ser Leu Gly Leu Leu Gly Leu Ser Ala
565 570 575
Met Leu Ala Gly Ala Pro Ala Leu Met Gly Leu Ala Ala Ser Leu Val
580 585 590
Leu Ala Leu Gly Leu Thr Leu Ser Ala Ala Ala Gly Ile Ala Ser Val
595 600 605
Ala Thr Leu
610
Claims (6)
1. The application of the gene OsDRP1C in regulation and control of rice root elongation is characterized in that the application is to regulate and control the rice root elongation by missense mutation of one or more base pairs in a DNA sequence; the serial number of the gene OsDRP1C in TIGR is LOC _ Os03g50520, and the gene OsDRP1 8920 contains a nucleotide sequence shown as SEQ ID NO: 1.
2. A mutant of a rice root elongation regulatory gene OsDRP1C is characterized in that the nucleotide sequence of the rice root elongation regulatory gene OsDRP1C is shown in SEQ ID NO: 1, the base at 2858bp of the DNA sequence has missense mutation, and thymine (T) is replaced by guanine (A).
3. The protein encoded by the rice root elongation regulatory gene OsDRP1C mutant as claimed in claim 2, wherein the amino acid sequence of the protein is shown in SEQ ID NO: 2, the 233 th tyrosine (Tyr) of the amino acid sequence is mutated into a stop codon.
4. An expression vector comprising the gene OsDRP1C according to claim 1.
5. The application of the expression vector containing the rice root elongation regulatory gene OsDRP1C in the aspect of rice root system character improvement in claim 4.
6. The application of the mutant of the rice root elongation regulatory gene OsDRP1C in regulating rice root system traits as claimed in claim 2.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210463032.XA CN114807221B (en) | 2022-04-28 | 2022-04-28 | Mutant, protein and expression vector of rice root elongation regulatory gene OsDRP1C and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210463032.XA CN114807221B (en) | 2022-04-28 | 2022-04-28 | Mutant, protein and expression vector of rice root elongation regulatory gene OsDRP1C and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114807221A true CN114807221A (en) | 2022-07-29 |
| CN114807221B CN114807221B (en) | 2023-06-06 |
Family
ID=82509237
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210463032.XA Active CN114807221B (en) | 2022-04-28 | 2022-04-28 | Mutant, protein and expression vector of rice root elongation regulatory gene OsDRP1C and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114807221B (en) |
Citations (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110197316A1 (en) * | 2010-02-08 | 2011-08-11 | Clemson University | Methods and compositions for transgenic plants with enhanced abiotic stress resistance and biomass production |
| US20120284880A1 (en) * | 2009-12-23 | 2012-11-08 | Elich Tedd D | Regulatory polynucleotides and uses thereof |
| CN102858998A (en) * | 2009-11-25 | 2013-01-02 | 雀巢产品技术援助有限公司 | Novel genomic biomarkers for irritable bowel syndrome diagnosis |
| CN103214581A (en) * | 2013-03-29 | 2013-07-24 | 中国农业科学院作物科学研究所 | Application of synthetic transcription factor VP64-Os03g57670 |
| WO2014091255A1 (en) * | 2012-12-14 | 2014-06-19 | Isis Innovation Limited | Transgenic plants |
| CN103898115A (en) * | 2014-04-01 | 2014-07-02 | 复旦大学 | Rice high-expression promoter related to development stage and application thereof |
| US20140230093A1 (en) * | 2011-07-19 | 2014-08-14 | Monsanto Technology Llc | Regulatory polynucleotides and uses thereof |
| CN104031928A (en) * | 2014-05-19 | 2014-09-10 | 宁波大学 | Rice root elongation control gene OsKASI and encoded protein |
| CN104211809A (en) * | 2013-05-31 | 2014-12-17 | 中国农业科学院作物科学研究所 | Application of synthetic transcription factor VP64-linker-Os03g11370 to improve paddy rice grain properties |
| CN104341514A (en) * | 2013-07-31 | 2015-02-11 | 中国农业科学院作物科学研究所 | Application of oryza sativa transcription factor Os03g05590.1 gene CDS sequence |
| US20150082475A1 (en) * | 2013-04-26 | 2015-03-19 | International Rice Research Institute | Root Growth, Nutrient Uptake, and Tolerance of Phosphorus Deficiency in Plants and Related Materials and Methods |
| EP2957637A2 (en) * | 2009-04-29 | 2015-12-23 | BASF Plant Science Company GmbH | Plants having enhanced yield-related traits and a method for making the same |
| CN105505981A (en) * | 2015-12-30 | 2016-04-20 | 中国农业科学院作物科学研究所 | Glyphosate-resistant transgenic soybean as well as preparation method and application thereof |
| CN111826392A (en) * | 2020-07-28 | 2020-10-27 | 河南大学 | Application of rice gene LJS5-2 and homologous gene thereof in controlling growth of leaf pillow and leaf angle of rice |
| CN112280784A (en) * | 2020-10-30 | 2021-01-29 | 宁波大学科学技术学院 | Rice lateral root development control gene OsLRD2, encoding protein and application thereof |
| US11220694B1 (en) * | 2018-01-29 | 2022-01-11 | Inari Agriculture, Inc. | Rice cells and rice plants |
-
2022
- 2022-04-28 CN CN202210463032.XA patent/CN114807221B/en active Active
Patent Citations (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2957637A2 (en) * | 2009-04-29 | 2015-12-23 | BASF Plant Science Company GmbH | Plants having enhanced yield-related traits and a method for making the same |
| CN102858998A (en) * | 2009-11-25 | 2013-01-02 | 雀巢产品技术援助有限公司 | Novel genomic biomarkers for irritable bowel syndrome diagnosis |
| US20120284880A1 (en) * | 2009-12-23 | 2012-11-08 | Elich Tedd D | Regulatory polynucleotides and uses thereof |
| US20110197316A1 (en) * | 2010-02-08 | 2011-08-11 | Clemson University | Methods and compositions for transgenic plants with enhanced abiotic stress resistance and biomass production |
| US20140230093A1 (en) * | 2011-07-19 | 2014-08-14 | Monsanto Technology Llc | Regulatory polynucleotides and uses thereof |
| WO2014091255A1 (en) * | 2012-12-14 | 2014-06-19 | Isis Innovation Limited | Transgenic plants |
| CN103214581A (en) * | 2013-03-29 | 2013-07-24 | 中国农业科学院作物科学研究所 | Application of synthetic transcription factor VP64-Os03g57670 |
| US20150082475A1 (en) * | 2013-04-26 | 2015-03-19 | International Rice Research Institute | Root Growth, Nutrient Uptake, and Tolerance of Phosphorus Deficiency in Plants and Related Materials and Methods |
| CN104211809A (en) * | 2013-05-31 | 2014-12-17 | 中国农业科学院作物科学研究所 | Application of synthetic transcription factor VP64-linker-Os03g11370 to improve paddy rice grain properties |
| CN104341514A (en) * | 2013-07-31 | 2015-02-11 | 中国农业科学院作物科学研究所 | Application of oryza sativa transcription factor Os03g05590.1 gene CDS sequence |
| CN103898115A (en) * | 2014-04-01 | 2014-07-02 | 复旦大学 | Rice high-expression promoter related to development stage and application thereof |
| CN104031928A (en) * | 2014-05-19 | 2014-09-10 | 宁波大学 | Rice root elongation control gene OsKASI and encoded protein |
| CN105505981A (en) * | 2015-12-30 | 2016-04-20 | 中国农业科学院作物科学研究所 | Glyphosate-resistant transgenic soybean as well as preparation method and application thereof |
| US11220694B1 (en) * | 2018-01-29 | 2022-01-11 | Inari Agriculture, Inc. | Rice cells and rice plants |
| CN111826392A (en) * | 2020-07-28 | 2020-10-27 | 河南大学 | Application of rice gene LJS5-2 and homologous gene thereof in controlling growth of leaf pillow and leaf angle of rice |
| CN112280784A (en) * | 2020-10-30 | 2021-01-29 | 宁波大学科学技术学院 | Rice lateral root development control gene OsLRD2, encoding protein and application thereof |
Non-Patent Citations (5)
| Title |
|---|
| "Rice Brittleness Mutants: A Way to Open the ‘Black Box’ of Monocot Cell Wall Biosynthesis" * |
| "Rice Brittleness Mutants: A Way to Open the ‘Black Box’ of Monocot Cell Wall Biosynthesis", JOURNAL OF INTEGRATIVE PLANT BIOLOGY * |
| TSUBASA KAWAI 等: "Auxin Distribution in Lateral Root Primordium Development Affects the Size and Lateral Root Diameter of Rice" * |
| 范路生;薛轶群;王晓华;王钦丽;林金星;: "隐失波荧光显微镜及其在植物细胞生物学中的应用" * |
| 范路生;薛轶群;王晓华;王钦丽;林金星;: "隐失波荧光显微镜及其在植物细胞生物学中的应用", 电子显微学报 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114807221B (en) | 2023-06-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| ES2260770T3 (en) | GENETIC CONTROL OF FLOWERING. | |
| US9663794B2 (en) | Heat-resistance rice gene OsZFP, screening marker and separation method thereof | |
| JPH10508481A (en) | Genetic control of flowering | |
| CN111172173A (en) | Method for reducing plant height of corn or delaying flowering | |
| EP0803572A2 (en) | Cpc gene for regulating initiation of root hair formation for arabidopsis (thaliana), and transgenic (arabidopsis) plant overexpressing the cpc gene | |
| CN109456396B (en) | Rice leaf senescence and panicle type regulation gene HK73, and protein, molecular marker and application encoded by gene HK73 | |
| CN109295071B (en) | Rice flower organ development regulation gene PEH1, and encoded protein and application thereof | |
| CN115894646B (en) | OsJDG1 gene and application thereof in regulation of rice grain type and thousand grain weight | |
| US20030177535A1 (en) | Genes for altering mitochondrial function and for hybrid seed production | |
| CN112280784B (en) | Rice lateral root development control gene OsLRD2, encoding protein and application thereof | |
| CN108341857B (en) | Rice yield related protein and coding gene and application thereof | |
| CN114807221B (en) | Mutant, protein and expression vector of rice root elongation regulatory gene OsDRP1C and application thereof | |
| AU764930B2 (en) | Plant photoperiod sensitivity gene and use of the same | |
| CN110950944B (en) | OsHCRF1 functional protein and application of coding gene thereof in rice breeding | |
| KR20000015750A (en) | Method for reducing interval of inflorescence section according to the introduction of transcription factor, petspl2 obtained from petunia | |
| CN109913468B (en) | Wheat tillering character related gene TaTAC1, expression product thereof, expression vector thereof and application | |
| EP2128251A1 (en) | Genes having activity of promoting endoreduplication | |
| EP1164193A1 (en) | Plant-signalling ligand like proteins | |
| CN109852624B (en) | Rice root elongation control gene OsNDPK4 and encoded protein | |
| CN112226459A (en) | Common wild rice grain type related coding gene and application thereof | |
| CN113754746A (en) | Rice male fertility regulation gene, application thereof and method for regulating rice fertility by using CRISPR-Cas9 | |
| CN113929756A (en) | Application of GL11 protein and gene for encoding GL11 protein in regulation of rice grain shape and thousand kernel weight | |
| CN109628462B (en) | Rice root elongation control gene OsKSR7 and encoded protein | |
| WO2003100062A1 (en) | Ehd1 GENE PROMOTING PLANT FLOEWERING AND UTILIZATION THEREOF | |
| CN115216487B (en) | Gene NSP1 for regulating rice yield formation and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |