CN116463359A - Application of rice gene OSK21 - Google Patents
Application of rice gene OSK21 Download PDFInfo
- Publication number
- CN116463359A CN116463359A CN202310629865.3A CN202310629865A CN116463359A CN 116463359 A CN116463359 A CN 116463359A CN 202310629865 A CN202310629865 A CN 202310629865A CN 116463359 A CN116463359 A CN 116463359A
- Authority
- CN
- China
- Prior art keywords
- rice
- gene
- osk21
- fertility
- plant
- 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.)
- Withdrawn
Links
- 235000007164 Oryza sativa Nutrition 0.000 title claims abstract description 114
- 235000009566 rice Nutrition 0.000 title claims abstract description 114
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 84
- 240000007594 Oryza sativa Species 0.000 title 1
- 241000209094 Oryza Species 0.000 claims abstract description 121
- 208000035199 Tetraploidy Diseases 0.000 claims abstract description 57
- 230000035558 fertility Effects 0.000 claims abstract description 51
- 241000196324 Embryophyta Species 0.000 claims abstract description 30
- 230000001105 regulatory effect Effects 0.000 claims abstract description 14
- 238000010353 genetic engineering Methods 0.000 claims abstract description 6
- 238000000034 method Methods 0.000 claims description 24
- 238000011161 development Methods 0.000 claims description 23
- 108091033409 CRISPR Proteins 0.000 claims description 20
- 238000012217 deletion Methods 0.000 claims description 18
- 230000037430 deletion Effects 0.000 claims description 18
- 210000001161 mammalian embryo Anatomy 0.000 claims description 14
- 230000009466 transformation Effects 0.000 claims description 12
- 230000009261 transgenic effect Effects 0.000 claims description 11
- 230000035899 viability Effects 0.000 claims description 11
- 230000002159 abnormal effect Effects 0.000 claims description 10
- 230000014509 gene expression Effects 0.000 claims description 10
- 238000010354 CRISPR gene editing Methods 0.000 claims description 9
- 230000035772 mutation Effects 0.000 claims description 9
- 239000013598 vector Substances 0.000 claims description 9
- 231100000502 fertility decrease Toxicity 0.000 claims description 8
- 210000000349 chromosome Anatomy 0.000 claims description 6
- 239000000463 material Substances 0.000 claims description 6
- 108020004414 DNA Proteins 0.000 claims description 5
- 102000004169 proteins and genes Human genes 0.000 claims description 5
- 230000033228 biological regulation Effects 0.000 claims description 4
- 239000013612 plasmid Substances 0.000 claims description 4
- 230000001131 transforming effect Effects 0.000 claims description 4
- 241000209504 Poaceae Species 0.000 claims description 3
- 239000012620 biological material Substances 0.000 claims description 3
- 230000003247 decreasing effect Effects 0.000 claims description 3
- 239000012634 fragment Substances 0.000 claims description 3
- 241000894006 Bacteria Species 0.000 claims description 2
- 210000004027 cell Anatomy 0.000 claims description 2
- 238000004520 electroporation Methods 0.000 claims description 2
- 239000013604 expression vector Substances 0.000 claims description 2
- 238000000520 microinjection Methods 0.000 claims description 2
- 108700026220 vif Genes Proteins 0.000 claims description 2
- 239000013603 viral vector Substances 0.000 claims description 2
- 108091027544 Subgenomic mRNA Proteins 0.000 claims 3
- 230000006870 function Effects 0.000 abstract description 11
- 230000001276 controlling effect Effects 0.000 abstract description 9
- 230000009418 agronomic effect Effects 0.000 abstract description 5
- 238000009395 breeding Methods 0.000 abstract description 5
- 230000001488 breeding effect Effects 0.000 abstract description 4
- 238000003976 plant breeding Methods 0.000 abstract description 4
- 230000008827 biological function Effects 0.000 abstract description 2
- 238000012214 genetic breeding Methods 0.000 abstract description 2
- 230000009467 reduction Effects 0.000 abstract description 2
- 230000018109 developmental process Effects 0.000 description 19
- 210000001672 ovary Anatomy 0.000 description 15
- 210000001519 tissue Anatomy 0.000 description 13
- 238000004458 analytical method Methods 0.000 description 7
- 230000001850 reproductive effect Effects 0.000 description 7
- 238000011529 RT qPCR Methods 0.000 description 6
- 238000003209 gene knockout Methods 0.000 description 6
- 230000021121 meiosis Effects 0.000 description 6
- 210000000056 organ Anatomy 0.000 description 6
- 238000011160 research Methods 0.000 description 5
- 230000005856 abnormality Effects 0.000 description 4
- 238000004043 dyeing Methods 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 238000010186 staining Methods 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 230000004069 differentiation Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 238000011144 upstream manufacturing Methods 0.000 description 3
- 230000009105 vegetative growth Effects 0.000 description 3
- 238000012795 verification Methods 0.000 description 3
- 241000589158 Agrobacterium Species 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 238000009412 basement excavation Methods 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 238000001218 confocal laser scanning microscopy Methods 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 230000007850 degeneration Effects 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 238000010362 genome editing Methods 0.000 description 2
- 101150054900 gus gene Proteins 0.000 description 2
- 238000003780 insertion Methods 0.000 description 2
- 230000037431 insertion Effects 0.000 description 2
- 230000017346 meiosis I Effects 0.000 description 2
- 230000023439 meiosis II Effects 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 230000008119 pollen development Effects 0.000 description 2
- 238000012257 pre-denaturation Methods 0.000 description 2
- 238000003762 quantitative reverse transcription PCR Methods 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 238000007480 sanger sequencing Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 238000010453 CRISPR/Cas method Methods 0.000 description 1
- 208000020584 Polyploidy Diseases 0.000 description 1
- 238000002123 RNA extraction Methods 0.000 description 1
- 102000000341 S-Phase Kinase-Associated Proteins Human genes 0.000 description 1
- 108010055623 S-Phase Kinase-Associated Proteins Proteins 0.000 description 1
- 101150063416 add gene Proteins 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 150000001413 amino acids Chemical group 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000002559 cytogenic effect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000009274 differential gene expression Effects 0.000 description 1
- 210000005069 ears Anatomy 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- ZBQZBWKNGDEDOA-UHFFFAOYSA-N eosin B Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC([N+]([O-])=O)=C(O)C(Br)=C1OC1=C2C=C([N+]([O-])=O)C(O)=C1Br ZBQZBWKNGDEDOA-UHFFFAOYSA-N 0.000 description 1
- 238000007667 floating Methods 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 230000007614 genetic variation Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 230000002028 premature Effects 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 101150075675 tatC gene Proteins 0.000 description 1
- 230000002123 temporal effect Effects 0.000 description 1
- 210000002105 tongue Anatomy 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
Classifications
-
- 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
-
- 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
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
-
- 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)
- C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
- C12N15/8218—Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]
-
- 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
- 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
- C12N15/8287—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for fertility modification, e.g. apomixis
-
- 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
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/20—Type of nucleic acid involving clustered regularly interspaced short palindromic repeats [CRISPRs]
-
- 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)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Virology (AREA)
- Botany (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention belongs to the technical fields of genetic engineering and plant genetic breeding, and particularly relates to application of a rice gene OSK21. In order to develop a novel gene capable of regulating and controlling the fertility of novel tetraploid rice, 1 novel gene OSK21 for regulating and controlling the fertility of novel tetraploid rice is discovered and cloned, and the gene can lead to about 20 percent reduction of the fertility of novel tetraploid rice after the gene function is deleted. The gene only affects fertility of novel tetraploid rice, has no influence on fertility of diploid rice, does not affect other agronomic characters related to yield after function loss, has high application value, and can be used in a subsequent tetraploid plant breeding system. The invention reveals the biological function of the rice gene OSK21 for the first time, provides a new gene resource for tetraploid rice breeding, and has important application prospect.
Description
Technical Field
The invention belongs to the technical fields of genetic engineering and plant genetic breeding, and particularly relates to application of a rice gene OSK21.
Background
Rice is one of the important food crops and is also the main food source for more than half of the population worldwide. With the rapid increase in world population and the varied climate, future food safety may be severely challenged. To address the future potential challenges, the excavation and utilization of new germplasm is critical. The autotetraploid rice is polyploid rice obtained by doubling diploid rice by chromosome. Compared with diploid rice, the autotetraploid rice has the characteristics of strong stress resistance, high biological yield, obvious heterosis and the like. And the autotetraploid rice has richer genetic variation, increases the recombination probability of excellent genes, and has great yield-increasing potential. However, the autotetraploid cannot be directly used for production due to its low seed setting rate.
In order to solve the problem of low fertility of the autotetraploid rice, the subject group obtains novel tetraploid rice by utilizing the cross directional breeding of different types of autotetraploids, and the novel tetraploid rice has huge production prospects in the aspect of rice production due to the advantages of high fertility, strong hybrid vigor and stability (Guo Haibin, mendrikahy JeanNestor, xie Lei, deng Junfeng, lu Zijun, wu Jinwen, li Xiang, shahid Muhammad Qasim, liu Xiangdong. Transcriptime analysis of neo-tetraploid rice reveals specific differential gene expressions associatedwith fertility andheterosis, scientific Reports,2017,7 (40139): 1-11; liu Xiangdong, wu Jinwen, shahidMummad Qasaim. Novel tetraploid rice is developed by using the research and development of hybrid vigor thereof, biotechnology, 2021, 37 (8): 1-6.
At present, the research on the reason of the generation of the novel tetraploid rice fertility is less, and the excavation of the functional genes for regulating and controlling the novel tetraploid rice fertility is helpful for analyzing the molecular mechanism for regulating and controlling the novel tetraploid rice reproductive development. Therefore, the development of a novel gene capable of regulating and controlling the fertility of novel tetraploid rice has important application prospect.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention discovers and clones 1 new gene OSK21 (LOC_Os07 g22680/Os07g 0409500) for regulating the fertility of novel tetraploid rice, and the gene can be used for regulating the fruiting rate and/or fertility of tetraploid gramineae plants and can be used in a subsequent tetraploid plant breeding system.
In order to achieve the above purpose, the present invention is realized by the following technical scheme:
in a first aspect, the present invention provides the use of gene OSK21 for regulating the setting rate and/or fertility of a tetraploid gramineous plant.
The protein encoded by the gene OSK21 and the derivative protein having the same function and having one or more amino acids substituted, deleted or added in the sequence are included.
Preferably, the tetraploid gramineous plant is tetraploid rice.
Preferably, the purpose of regulating and controlling the fruiting rate and/or fertility of the tetraploid gramineae plant is achieved by regulating and controlling the pollen viability, and/or embryo sac fertility, and/or female gamete development.
According to the invention, the function of the gene OSK21 is verified in rice through a gene knockout experiment and an over-expression experiment, and after the novel tetraploid rice Huaduo No.1 and diploid rice stage 65 are respectively transformed through the constructed CRISPR/Cas9 gene knockout vector, the OSK21 is found to not influence the fertility of the diploid rice, but influence the fertility of the tetraploid rice, and the functional deficiency of the OSK21 in the tetraploid rice can lead to the reduced fertility of plants, the reduced pollen vigor and abnormal female gamete development. The invention also discovers that after the function of the OSK21 gene is lost, only the fertility of the novel tetraploid rice is reduced, other main agronomic traits are not affected, male organs and the main agronomic traits of the novel tetraploid rice are normal, and the OSK21 gene is a novel gene which only affects the fertility of the tetraploid rice.
Preferably, the OSK21 gene further comprises a biological material related thereto, including a protein encoding the OSK21 gene, a recombinant vector, an expression cassette, a transgenic cell line or a recombinant bacterium containing the OSK21 gene.
In a second aspect, the invention provides a method for reducing fertility or setting rate of rice or abnormal development of female gametes of rice, namely, knocking out or reducing gene OSK21 in rice by using genetic engineering means.
In one embodiment, the method for reducing fertility or setting rate of rice or causing abnormal development of female gametes of rice specifically comprises the following steps: and (3) using a gene engineering means based on CRISPR/Cas9, taking a gene OSK21 as a target, designing a sgRNA sequence based on CRISPR/Cas9, connecting a DNA fragment containing the coding the sgRNA sequence to a carrier carrying the CRISPR/Cas9, and transforming rice, thereby obtaining the transgenic rice material with the gene function deletion.
In one embodiment, the above method for reducing fertility or setting rate of rice or abnormal development of female gametes of rice, wherein the sgRNA sequence is shown in SEQ ID No.1 and SEQ ID No. 2.
In a third aspect, the present invention provides a method for restoring fertility or seed setting rate in a rice plant, which comprises introducing the gene OSK21 into a rice plant having reduced fertility or seed setting rate by a plasmid or integrating the gene OSK21 into a rice chromosome by genetic engineering means, wherein the plant has reduced fertility or seed setting rate is caused by mutation of the gene OSK21 in the rice.
In one embodiment, the plant fertility or setting rate recovery method for the decrease of the fertility or setting rate of rice caused by the mutation of the rice gene OSK21 specifically comprises the following steps: constructing the gene OSK21 on a plant binary expression vector (such as pCUBI 1390), and then transforming rice to obtain transgenic rice with over-expressed gene.
In one embodiment, the plant fertility or seed setting rate recovery method for the decrease in fertility or seed setting rate of rice caused by the mutation of the rice gene OSK21 comprises transformation of rice using Ti plasmid, transformation using plant viral vector, direct DNA transformation, microinjection transformation, electroporation transformation.
In a fourth aspect, the present invention provides the use of a transgenic rice obtained in the second or third aspect in plant breeding.
Preferably, the breeding methods include, but are not limited to, transgenesis, crosses, backcrosses, selfing, or asexual propagation.
Compared with the prior art, the invention has the beneficial effects that:
the invention discovers and clones 1 new gene OSK21 for regulating and controlling the fertility of novel tetraploid rice, and the gene can lead to about 20 percent of the reduction of the fertility of the novel tetraploid rice after the gene is in a loss of function. It is worth mentioning that the gene only affects the fertility of novel tetraploid rice, has no influence on the fertility of diploid rice, does not affect other agronomic characters related to yield after the function is deleted, has stronger application value, and can be used in a subsequent tetraploid plant breeding system. The invention reveals the biological function of the rice gene OSK21 for the first time, provides a new gene resource for tetraploid rice breeding, and has important application prospect.
Drawings
FIG. 1 is a screen of OSK21 gene and analysis of gene expression pattern;
in FIG. 1, the structure of the OSK21 gene and the domain encoding the protein thereof are shown in FIG. A; FIG. B.GUS staining results of OSK21 on young ears, anthers, leaves, leaf tongues, leaf sheaths, etc. of rice of different lengths; FIG. C. RT-qPCR analysis results of OSK21 in anther tissue of Huaduo No. 1; wherein a-PMC is an anther at the meiotic interval; a-PMA is an anther in the pre-meiotic phase; a-MAI is an anther at meiosis I; a-MAII is an anther in meiosis II; a-SCPE is an anther in the early phase of the microspore monocaryon; A-SCP is anther in the microspore stage; a-BCP is anther in the two-cell pollen stage; A-MP is anther in mature pollen stage; FIG. D. RT-qPCR analysis of OSK21 in ovary tissue of Huaduo No. 1; O-PMC is the ovary in the meiotic interval of the corresponding anther; O-PMA is the ovary in the pre-meiosis phase of the corresponding anther; O-MAI is the ovary at the corresponding anther meiosis I; O-MAII is the ovary in the corresponding anther meiosis II; O-SCPE is the ovary in the early stage of the corresponding anther monocaryon microspore; O-SCP is an ovary in the corresponding anther monocaryon microspore stage; O-BCP is the ovary in the corresponding anther two-cell pollen stage.
FIG. 2 is a comparative study of the construction and phenotype of OSK21 loss-of-function mutants in tetraploid rice;
in FIG. 2, FIG. A. OSK21 shows the gene knockout target and the sequence mutation site information in tetraploid rice; panel B-D.functional deletion mutant of OSK21 in tetraploid rice compared to wild type differences in plant phenotype; FIG. E-G.OSK21 functional deletion mutant in tetraploid rice compared to wild type differences in ear traits; FIG. H-P.functional deletion mutant of OSK21 in tetraploid rice was different from wild type in floral organ traits.
FIG. 3 is a comparative study of the construction and major agronomic traits of OSK21 functional deletion mutants in diploid rice;
in FIG. 3, FIG. A. OSK21 gene knockout target and sequence mutation site information; panel B-D.functional deletion mutant of OSK21 in diploid rice compared to wild type differences in plant phenotype; FIG. E-G.OSK21 functional deletion mutant in diploid rice compared to wild type differences in ear pattern; FIG. H-P.functional deletion mutant of OSK21 in diploid rice and wild type were different in floral organ traits.
FIG. 4 is a fertility trait study of OSK21 loss-of-function mutants in tetraploid rice;
FIG. 4 shows a comparison of the OSK21 loss-of-function mutant and its wild type in mature pollen fertility and pollen viability in tetraploid rice, wherein the arrows indicate abnormal embryo sacs, embryo sac degeneration and extreme nuclear differentiation, respectively; FIG. B. difference in pollen viability of OSK21 functional deletion mutant and wild type thereof in tetraploid rice, wherein No represents pollen-free viability, TTC cannot be dyed, low represents Low pollen viability, TTC dyeing degree is shallower, high represents pollen viability High, and TTC can be deeply dyed; panel C. comparison of the differences in embryo sac fertility between the functional deletion mutant of OSK21 and its wild type in tetraploid rice.
Detailed Description
The following describes the invention in more detail. The description of these embodiments is provided to assist understanding of the present invention, but is not intended to limit the present invention. In addition, the technical features of the embodiments of the present invention described below may be combined with each other as long as they do not collide with each other.
The experimental methods in the following examples, unless otherwise specified, are conventional, and the experimental materials used in the following examples, unless otherwise specified, are commercially available.
Example 1 screening of OSK21 Gene and analysis of spatial and temporal expression Pattern
In this example, OSK21 (LOC_Os07 g22680/Os07g 0409500) was used as a target gene containing the SKP1 domain in the bioinformatics database (FIG. 1A). First, the expression control pattern of the OSK21 gene was verified (fig. 1). The gene expression condition of OSK21 in each tissue and each development period is detected by using the wild material Huaduo No.1 root, stem, leaf sheath, young spike, anther and ovary cDNA in each period as templates and using qRT-PCR technology. The specific primers of the OSK21 gene are respectively as follows:
OSK21-F:5’-ACATGCTCAAGGGCAAGACCTC-3’(SEQ ID No.3)；
OSK21-R:5’-GTTCTGCTCCTTGATTGCCTTGTC-3’(SEQ ID No.4)；
wherein qRT-PCR amplification is performed according to a two-step PCR. The PCR reaction procedure was: pre-denaturation at 95℃for 30s, denaturation at 95℃for 10s, extension at 58℃for 20s,45 cycles; melting curve from 65 ℃ to 95 ℃. Negative controls were established for each qRT-PCR. qRT-PCR for each gene had 3 technical replicates and three biological replicates.
The detection result shows that: OSK21 was expressed in both anthers and ovary tissues at early reproductive development of the novel tetraploid rice huado No.1 (fig. 1C and 1D). OSK21 was expressed higher in anther tissue than in ovary tissue, and was expressed highest in meiosis intervals, gradually decreasing from meiosis to maturity (fig. 1C). In the ovary tissue, OSK21 was expressed at various times with the highest meiosis interval expression (fig. 1D).
Next, the expression site of OSK21 gene was further clarified by GUS staining. Amplifying a 2000bp sequence (SEQ ID No. 5) of an upstream promoter of an OSK21 gene by using a Primer STAR high-fidelity enzyme of Takara company by using a wild type template, and connecting the amplified target sequence to a pCAMBIA1305.1 linearization vector (purchased from adedge) which is subjected to enzyme digestion and carries a GUS gene after sequencing and verification to obtain a genetic transformation vector containing the fusion of the OSK21 gene promoter and the GUS gene; after genetic transformation is finished, screening to obtain a positive plant carrying an OSK21 gene upstream promoter sequence; selecting a proper amount of glumes of rice in different development periods for GUS dyeing, and putting the glumes into a 1.5mL centrifuge tube for GUS dyeing to completely immerse the materials into the solution; dyeing at 37deg.C for 2-8 hr, decolorizing with 75% ethanol, storing in fixing solution, observing, and photographing. For specific methods, reference is made to the methods of Lu et al, the laboratory (Lu Z, guo X, huang Z, et al Transcriptame and Gene Editing Analyses Reveal MOF1a Defect Alters the Expression of Genes Associated with Tapetum Development and Chromosome Behavior at Meiosis Stage Resulting in Low Pollen Fertility ofTetraploid Rice [ J ]. International Journal of Molecular Sciences,2020 (20): 7489.).
The sequence of the upstream promoter 2000bp (SEQ ID No. 5):
5’-CCATTCACTCATTTCCATTAACCTAAAATTGGTACTCTCTCTTTCACAATGTA AGTCATTCTAGTATTTCCTACATTCATATTGATGTTAATAAATCTAGACATATATATCTATCTAAATTCATTAATATCGATATGAATGTGGGAAATGCTAAAATGACTTACATTATGAAACGGAGGAAGTAAGAACAAAGAAAAAACCCTTAACAAAATATCCAATATGTTGATACCAGTTTAAAGAAATGTAGCACTCAGATTAGTTTTAAATCAATTAAAGCATCACTCAGGACAAGTTGCAAAAACCTAGTAAAAATGAAAAAAAAAGGACATTATTTTTTTTAAGATAATGGAAGATTTATTGAACTTGGCAAAATTTTTTAAGGACACTCCTAGCCTCTACACTACGAAAGTGCACACAACCAACACACCTCACACACAAAGAAAATAAAAACCAAACTAAGGTTCAAGAACCTTGCACCTGAAAACTATGTTGCCACCCATGCACCAAGGTGAAAAAAAAACTCATTGGCCACATGCTCCAAATGAATACAAGCCG
AAGTGACAAAACTATGTGAACCGGGCTTCTGGAGAATAGCCCATGCTCGAAGCTA
ATGTGTAACCAAGAAAATAACTAGCAAAGGAGAAGAAACATATTTGCTTTCAAAA
CCCATAATCATTCCTCGAAAGCCAAGTGACCCAATAAGTTGTTGCTGCCCCTACAA
GAAAAAGATGTCTGAGCTATTTGGTTTGTCGCAAAGTCCATTGCCCAAACATATGT
GACACAATCCTAGGCGAACAAATGTTCGAAGAAATCTGGATTAGAGACCAAACCG
ACCGAGCTAGCCGACAACAAGTCTGAGCTCCTCTCCTCTCCTGCCACCGCATTGCA
CGCCACTCCTCTCCCCCGCCGCTCGTGCACTCCATGGCCGACAATAGCCTTGTTACG
GTCGCCGCTGCAGGATCCGGCGGAGAGGGAGGAGGCGGGGAGGCCGGCGAGCGG
GTGGGGGAAGAAGGAGGCCAGCGAGCGGGCGGGGGGAGGAAGGAGGCCAGTGAG
GAGGCCGTCACAGAGCCTCAGCCGTCACCTTCGCGAGCCACACCTACCGTTGCTGA
GTCGCCACAACGCTGGCATGCCAACAACCGTAGCCGACTGACAAGGAAGCTCCAG
GAGGGAGGAAGAATGGCCAAGAGGGAGAGGGGAGAAGGAGGCGCCGACGTGCCG
CTGTCGAGGGCTTCTTCACCGACTCCGGCAACTTCTTCGGCCGTATCCACGCCCGCC
TCGTCGACACCTCCTCCTCTGCTCTCTCTTATCTGTCTCGCTGGTGTCGATTGGTTAA
GGAGCCACTCGCAGCTAACATGTGGGACCTATGTAGGGCCCACGCTGACTCAACCG
GAGGACATGTGAGCCCCATATTTTTTTAATTTTTAATTTGTATATTTTTCTTCTAGTG
CCATCATCACCACGTAGGACGATATTTTTCTTCTAGTGTCATCATCACCACGTAGGA
CGATGACTAAGTCAAACCAGCCACGTAGACGCCATGTCAACTTAAGCCACCGTTCA
AACCGTATTAGACCTTATTTTCACCGGTTTTGACAGTTGAGGGACCAGTGTTTCAGA
GATGAAATCGGTGTCAAGTGAAGGGACCTCAAATGAACTTATTCCCAAATAAATCA
ACACAGTAATTCTGGGCCCATTCTTCCCCTCTCTCGCAGGCCGCAGAACGGCCCAG
TAAGTTTACTATTCTTTCGTGCAAATCAAGAAACGGACGCGAAACCTGAGAGAAAA
CTGCAGTGGGAGACGGAAGCAAATTCCGGCACGCCTCCGCTCCAAGCGGGCTCAG
ACCCGAACCCGGCAAGCGCGGCGTCGTCGTCGTGTATAAAACGGCGGCCACGCCG
ATCATTCTTCGTTCCACCTCGCCTCGCATCCACCGTGAGCGGAGCGCCAACTCGCTAGCAAATCAGAAGAGGAGGTTAGAGTTTCGTCGTGAGCGAGCGAGAGTC-3’。
GUS tissue staining results showed that: no signal could be detected in the young root, stem and leaf of rice, indicating that OSK21 gene was not expressed in vegetative growth tissue. In the reproductive development stage of rice, the reproductive tissues such as floret, anther and ovary are detected, and the internal and external floating sheets, anther and ovary tissues of the floret are dyed. The anther detected a blue GUS signal at 2.0-5.0mm, with the color reaching the darkest at 4.5mm, and then slowly turning lighter (FIG. 1B). At 2.0-6.0mm in florets, the ovary tissue also detected a blue GUS signal (FIG. 1B). The research results show that the expression level of the OSK21 gene in the reproductive organ is at a higher level, and the function of the target gene is further verified.
Example 2 functional verification of OSK21 by Gene knockout
In order to determine the function of OSK21 in novel tetraploid rice, RNA extraction is performed on young ear tissue of the rice in meiosis stage by taking novel tetraploid rice Huaduo No. 1-4x (4 x represents 4 sets of chromosome groups and is tetraploid) as a research material. The cDNA obtained by RNA reverse transcription is used as a template, and the Primer STAR high-fidelity enzyme of Takara company is used for amplifying the CDS sequence of the OSK21 gene, and the complete CDS sequence of the OSK21 gene is obtained after sequencing verification. Specific primers for amplifying the CDS sequence of the OSK21 gene are respectively as follows:
OSK21-CDS-F:5’-CGCCAACTCGCTAGCAAATC-3’(SEQ ID No.6)；
OSK21-CDS-R:5’-GCATGCAGGAACAGATCAGC-3’(SEQ ID No.7)。
wherein, the amplification of CDS sequence of OSK21 gene adopts 50 mu L system (prepared according to the specification of Primer STAR high-fidelity enzyme of Takara company), and the PCR reaction procedure is as follows: pre-denaturation at 94℃for 3min; denaturation at 94℃for 30s, extension at 55℃for 45s, extension at 72℃for 1min,30 cycles; extending at 72℃for 5min.
CDS sequence of OSK21 gene (SEQ ID No. 8):
5’-ATGGAGGCGGACAAGAGCGGGGAGGGGGAGAAGGCCGGCGGCAAGACGA TCTCCTTCAGGTGCT CggacggacaggccttccacatgccggtggccgcggcgatgctaagcacggccatccgcaaaatgttcgacaagtaccccagcatcgaccacggcggcgtgatcgagctcccccaccaaatctcctccggtatcttccccaaggtcaaggaatactgcaccaagcacgccaaggtcgacgacaagggcaaccccaccgtctccaccaacaccggcgcggcggcggcggcgtcaagcagcagcaccgacgacgaggaggaggacctcaagaactgggacaaggagtttgtcaacatggaggtgaagcccctccacgacctcctcctcgtcgcccacctactcgacatcaaaggcctcttccacatcacctgccgcaaggtcgccgacatgctcaagggcaagacctccgaggagatgcgccagatcctcaacatccgcaacgacttcaccgaggaagaagacaaggcaatcaaggagcagaacccctgggtcttcccggaTCCGGAGTAG-3' (31 bp sequence at the underlined position corresponding to the deletion of the mutant, 498bp sequence at the lowercase position corresponding to the deletion of the mutant).
Second, OSK21 is edited using CRISPR/Cas9 technology. With the OSK21 gene (loc_os07 g 22680) as a target, a CRISPR/Cas 9-based sgRNA sequence [ target 1:5'-ATCTCCTTCAGGTGCTCGGA-3' (SEQ ID No. 1); target 2:5'-GGATCCGGAGTAGAAGCTA-3' (SEQ ID No. 2) the DNA fragment containing the sequence encoding the sgRNA (SEQ ID No.1 and SEQ ID No. 2) was ligated into a vector carrying CRISPR/Cas9 (pYLCRISPR/Cas 9 Pubi-H binary vector system available from the university of agricultural university of North, liu Yaoguang, laboratories, available from addgene). Then, the agrobacterium mediating method is used for transforming the novel tetraploid rice variety Huaduo No.1, and the Sanger sequencing technology is used for identifying positive transgenic plants, so that 2 independent transgenic knockout positive plants are obtained. The two homozygous mutant types obtained by the screening are respectively named osk21-1 -4x And osk21-2 -4x . Wherein mutant osk21-1 -4x A 498bp substitution between the 18 th base of target 1 and the 2 nd base of target 2 is TATC; wherein the 498bp sequence is as follows: 5' -GGACGGACAGGCCTTCCACATGCCGGTGGCCGCGGCGATGCTAAGCACGGCCAT CCGCAAAATGTTCGACAAGTACCCCAGCATCGACCACGGCGGCGTGATCGAGCTCCCCCACCAAATCTCCTCCGGTATCTTCCCCAAGGTCAAGGAATACTGCACCAAGCACGCCAAGGTCGACGACAAGGGCAACCCCACCGTCTCCACCAACACCGGCGCGGCGGCGGCGGCGTCAAGCAGCAGCACCGACGACGAGGAGGAGGACCTCAAGAACTGGGACAAGGAGTTTGTCAACATGGAGGTGAAGCCCCTCCACGACCTCCTCCTCGTCGCCCACCTACTCGACATCAAAGGCCTCTTCCACATCACCTGCCGCAAGGTCGCCGACATGCTCAAGGGCAAGACCTCCGAGGAGATGCGCCAGATCCTCAACATCCGCAACGACTTCACCGAGGAAGAAGACAAGGCAATCAAGGAGCAGAACCCCTGGGTCTTCCCGGA-3’。osk21-2 -4x 31bp of target 1 of (a), the deleted 31bp sequence is: CTCGGACGGACAGGCCTTCCACATGCCGGTG. Target 2 is a 1a base insertion. The above mutation types resulted in premature termination of OSK21 protein translation (fig. 2A). After comparative study of the phenotype of the OSK21 knockout homozygotic mutant and its wild type, it was found that mutant OSK was in vegetative growth phase -4x There was no significant difference in phenotype such as plant type, plant height, tillering, etc., compared to the wild type (FIGS. 2B-2G). During reproductive development, mutant osk21 -4x Normal flower organ phenotype (fig. 2H-2P), normal male gamete development; the functional deletion mutant osk-1 is found by field phenotype observation in the mature period, compared with the plant setting rate 68.91 +/-0.57% of the wild type Huaduo No.1 -4x And osk21-2 -4x The plant setting percentage is obviously reduced to 13.65 plus or minus 0.42 percent and 12.60 plus or minus 0.46 percent respectively.
In addition, the same gene editing vector (the target sequences contained in the vector are SEQ ID No.1 and SEQ ID No. 2) in novel tetraploid rice was used to transform 65-2x (2 x represents 2 sets of chromosome sets, is diploid) in diploid rice plants by using an agrobacterium-mediated method, and positive transgenic plants were identified by using Sanger sequencing technology, so that 2 independent transgenic knockout positive plants were obtained (FIG. 3A). The two homozygous mutant types obtained by the screening are respectively named osk21-1 -2x And osk21-2 -2x . Wherein mutant osk21-1 -2x 1bp of T base insertion exists at the target point 1, and 1bp of T base deletion exists at the target point 2; osk21-2 -2x 31bp base deletion exists at the target point 1, and the deletion sequence is as follows: CTCGGACGGACAGGCCTTCCACATGCCGGTG; at target 2 is a deletion of 1bp base.
By comparing and researching the homozygous mutant of OSK21 gene in the gene knockout of diploid rice and the phenotype of wild type, the mutant OSK is found in the vegetative growth phase -2x There was no significant difference in phenotype such as plant type, plant height and tillering compared with the wild type (FIGS. 3B to 3G). During reproductive development, mutant osk21 -4x Normal flower organ phenotype (FIGS. 3H-3P); observations by field phenotypes during maturityIt was found that osk21-1 -2x And osk21-2 -2x There was no significant difference in average seed setting rate between the two seasons of the late 2021 and early 2022 seasons compared to the wild type. Therein osk-21-1 -2x Average setting rates in two seasons of 2021 late season and 2022 early season decrease by 2.28% and 4.11%, respectively, osk21-2 -2x The average seed setting rates of the two seasons of the late and early 2021 and 2022 of the mutant were reduced by 4.25% and 2.14%, respectively. The above results indicate that the loss of OSK21 function does not result in a decrease in the seed setting rate of diploids. The result more confirms that the OSK21 gene only participates in the regulation and control of the fertility of the novel tetraploid rice, and the seed setting rate of the novel tetraploid rice is reduced.
Example 3 analysis of the observation of the osk21-4x mutant during the development of pollen and embryo sacs in Rice
To verify the mutant osk21 -4x And the difference of wild type Huaduo No. 1-4x in fertility, the mutant osk21 was initially observed by utilizing WE-CLSM (global eosin B staining transparent laser scanning confocal microscope) technique -4x And the development process of wild pollen and embryo sac (specific observation method is referred to "Li Xiang. Cytogenetic study of the hypofertility of autotetraploid rice [ D)]University of agricultural, south china, 2018 "), and summarised the development process (fig. 4).
The research results show that: in terms of pollen development, mutant osk21 -4x Consistent with the pollen development process of novel tetraploid rice Huaduo No. 1-4x, no obvious abnormality exists, but embryo sac abnormality appears, which respectively comprises embryo sac degeneration and extremely nuclear differentiation abnormality types, indicating that the female gamete is abnormal in development (figure 4A). Mutant osk21 -4x The abnormality of (2) is mainly expressed in mature pollen viability, compared with wild-type mature pollen viability (88.33%), osk-1- 4x Pollen with normal activity accounts for 81.39%, and pollen activity is reduced by 6.94%; osk21-2- 4x Pollen with normal viability accounted for 78.96% and pollen viability decreased by 9.37% (fig. 4B), which was insufficient to cause a significant decrease in seed setting rate. At the same time, for mutant osk21 -4x The embryo sac fertility of (2) is observed by WE-CLSM, and the research result shows that: mutant osk- 4x The average decrease in fertility of the mature embryo sac was 16.60% (FIG. 4C), the major abnormal embryo sac was extremeAbnormal nuclear differentiation, the earliest abortive stage of embryo sac begins with the eighth nuclear embryo sac stage.
The results of the studies in comprehensive examples 1-3 show that OSK21 is indeed involved in the regulation of fertility of novel tetraploid rice, but the gene does not affect the regulation of fertility of diploid rice. The OSK21 gene is indicated to be used for regulating and controlling the setting percentage and/or fertility of tetraploid rice, is used in a subsequent tetraploid rice breeding system, and has wide application prospect.
The embodiments of the present invention have been described in detail above, but the present invention is not limited to the described embodiments. It will be apparent to those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, and yet fall within the scope of the invention.
Claims (10)
1. The application of the gene OSK21 in regulating the setting percentage and/or fertility of tetraploid gramineous plants.
2. The use according to claim 1, wherein the tetraploid gramineous plant is tetraploid rice.
3. The use according to claim 1, wherein the regulation of the seed setting rate and/or fertility is achieved by regulating the pollen viability, and/or embryo sac fertility, and/or female gamete development of the tetraploid gramineae.
4. The use according to claim 1, wherein the OSK21 gene further comprises a biological material associated therewith, the associated biological material comprising a protein encoding an OSK21 gene, a recombinant vector, an expression cassette, a transgenic cell line or a recombinant bacterium comprising an OSK21 gene.
5. A method for reducing fertility or setting rate of rice or abnormal development of female gametes of rice, characterized in that gene OSK21 in rice is knocked out or reduced by genetic engineering means.
6. The method for reducing fertility or setting percentage of rice or abnormal development of female gametes of rice according to claim 5, wherein gene engineering means based on CRISPR/Cas9 is utilized, gene OSK21 is used as a target, a sgRNA sequence based on CRISPR/Cas9 is designed, a DNA fragment containing the encoding sgRNA sequence is connected to a carrier carrying CRISPR/Cas9, rice is transformed, and transgenic rice material with the gene function deletion is obtained.
7. The method for reducing fertility or setting rate of rice or abnormal development of female gametes of rice according to claim 6, wherein the sgRNA sequence is shown in SEQ ID No.1 and SEQ ID No. 2.
8. A method for restoring fertility or seed setting rate of a plant having reduced fertility or seed setting rate of rice by mutation of OSK21 gene of rice, characterized in that OSK21 gene is introduced into a rice plant having reduced fertility or seed setting rate by a plasmid or OSK21 gene is integrated into a rice chromosome by genetic engineering means.
9. The method for restoring fertility or setting rate of a plant having a decreased fertility or setting rate of a rice plant due to mutation of an OSK21 gene of a rice plant according to claim 8, wherein the OSK21 gene is constructed on a binary expression vector of the plant, and the rice plant is transformed to obtain a transgenic rice plant over-expressed by the gene.
10. The method for restoring fertility or seed setting rate in a plant having reduced fertility or seed setting rate in rice resulting from mutation of OSK21 in rice according to claim 9, wherein the method for transforming rice comprises transformation with Ti plasmid, transformation with plant viral vector, direct DNA transformation, microinjection transformation, electroporation transformation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310629865.3A CN116463359A (en) | 2023-05-31 | 2023-05-31 | Application of rice gene OSK21 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310629865.3A CN116463359A (en) | 2023-05-31 | 2023-05-31 | Application of rice gene OSK21 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116463359A true CN116463359A (en) | 2023-07-21 |
Family
ID=87177339
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310629865.3A Withdrawn CN116463359A (en) | 2023-05-31 | 2023-05-31 | Application of rice gene OSK21 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116463359A (en) |
-
2023
- 2023-05-31 CN CN202310629865.3A patent/CN116463359A/en not_active Withdrawn
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107988229B (en) | Method for obtaining tillering-changed rice by modifying OsTAC1 gene through CRISPR-Cas | |
| WO2017121411A1 (en) | Plant male fertility-related protein, and coding gene and application thereof | |
| CA3047163A1 (en) | Genome editing-based crop engineering and production of brachytic plants | |
| CN109207505B (en) | Method for creating tomato male sterile line through genome editing and application thereof | |
| WO2015035951A1 (en) | Use of genic male sterility gene and mutation thereof in hybridization | |
| WO2021244007A1 (en) | Rice male fertility regulatory gene, mutant of rice male fertility regulatory gene, use thereof and method for regulating rice fertility | |
| CN117209577B (en) | Plant meiosis related protein GmPRD1, and coding gene and application thereof | |
| CN116103304B (en) | Temperature-sensitive male sterile gene of rice and application thereof | |
| CN113862278B (en) | Upland cotton GhMS20 gene and application thereof in creation of cotton single dominant male sterile line | |
| CN110241126B (en) | Application of OsDGD2 beta gene in cultivation of male sterile rice variety | |
| CN112391395A (en) | Soybean sterile gene mutant, application and construction method of soybean sterile line | |
| US20220333125A1 (en) | GENE ZmPLD3 FOR INDUCING MAIZE MATERNAL HAPLOID PRODUCTION AND ITS APPLICATION THEREOF | |
| WO2022042619A1 (en) | Plant pollen tube growth gene and application | |
| CN116463359A (en) | Application of rice gene OSK21 | |
| CN110878314B (en) | Hl-2 gene for regulating and controlling tomato trichome and application thereof | |
| CN109504703B (en) | Method for creating maize dominant nuclear male sterile line by using p5126-ZmMs1D construct and breeding application method thereof | |
| CN112608937A (en) | Method for cultivating self-compatible cabbage based on CRISPR/Cas9 gene editing method and application thereof | |
| WO2023016097A1 (en) | Oryza sativa cytoplasmic male sterility restorer gene osrf19 and application thereof | |
| CN114410653B (en) | Soybean sterile gene mutant, construction method and application thereof in photosensitive fertility regulation and control | |
| CN117402887B (en) | Maize male fertility regulation gene ZmMS, mutant thereof and application | |
| CN113186199B (en) | Thermo-sensitive male sterile gene and application thereof | |
| CN112322638B (en) | Cabbage type rape hau CMS fertility restorer gene Rfh and application thereof | |
| CN108728479B (en) | Method for obtaining rice mutant with loss of genetic interference and application thereof | |
| CN116179565A (en) | Creation method of thermosensitive female sterile rice material | |
| CN118240864A (en) | Application of STR of rice OsFD gene promoter region or targeted deletion method of STR in rice molecular breeding |
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 | ||
| WW01 | Invention patent application withdrawn after publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20230721 |