CN116676284A - Gene for regulating crop dwarfing, lodging resistance and yield and application thereof - Google Patents
Gene for regulating crop dwarfing, lodging resistance and yield and application thereof Download PDFInfo
- Publication number
- CN116676284A CN116676284A CN202310795542.1A CN202310795542A CN116676284A CN 116676284 A CN116676284 A CN 116676284A CN 202310795542 A CN202310795542 A CN 202310795542A CN 116676284 A CN116676284 A CN 116676284A
- Authority
- CN
- China
- Prior art keywords
- gene
- seq
- rice
- sequence
- dwarf
- 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
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 138
- 230000001105 regulatory effect Effects 0.000 title claims abstract description 25
- 241000209094 Oryza Species 0.000 claims abstract description 91
- 235000007164 Oryza sativa Nutrition 0.000 claims abstract description 84
- 235000009566 rice Nutrition 0.000 claims abstract description 84
- 241000196324 Embryophyta Species 0.000 claims abstract description 44
- 230000001276 controlling effect Effects 0.000 claims abstract description 11
- 238000012217 deletion Methods 0.000 claims abstract description 11
- 230000037430 deletion Effects 0.000 claims abstract description 11
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 32
- 239000012620 biological material Substances 0.000 claims description 29
- 102000004169 proteins and genes Human genes 0.000 claims description 25
- 239000002773 nucleotide Substances 0.000 claims description 19
- 125000003729 nucleotide group Chemical group 0.000 claims description 19
- 238000000034 method Methods 0.000 claims description 16
- 238000009395 breeding Methods 0.000 claims description 15
- 239000013598 vector Substances 0.000 claims description 10
- 241000894006 Bacteria Species 0.000 claims description 8
- 241000702632 Rice dwarf virus Species 0.000 claims description 8
- 230000002829 reductive effect Effects 0.000 claims description 8
- 238000009396 hybridization Methods 0.000 claims description 7
- 230000006872 improvement Effects 0.000 claims description 7
- 229920001184 polypeptide Polymers 0.000 claims description 7
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 7
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 7
- 239000002299 complementary DNA Substances 0.000 claims description 5
- 230000000694 effects Effects 0.000 claims description 5
- 108700026220 vif Genes Proteins 0.000 claims description 5
- 125000000539 amino acid group Chemical group 0.000 claims description 4
- 230000002401 inhibitory effect Effects 0.000 claims description 3
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 2
- 230000000295 complement effect Effects 0.000 claims description 2
- 238000012546 transfer Methods 0.000 claims description 2
- 230000001488 breeding effect Effects 0.000 abstract description 16
- 238000010353 genetic engineering Methods 0.000 abstract description 4
- 238000012214 genetic breeding Methods 0.000 abstract description 3
- 108010021466 Mutant Proteins Proteins 0.000 abstract description 2
- 230000035772 mutation Effects 0.000 description 7
- 108020004414 DNA Proteins 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 235000013339 cereals Nutrition 0.000 description 4
- 238000003556 assay Methods 0.000 description 3
- 238000002703 mutagenesis Methods 0.000 description 3
- 231100000350 mutagenesis Toxicity 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 101001066230 Gibberella fujikuroi (strain CBS 195.34 / IMI 58289 / NRRL A-6831) Cytochrome P450 monooygenase 2 Proteins 0.000 description 2
- 230000002411 adverse Effects 0.000 description 2
- 230000009418 agronomic effect Effects 0.000 description 2
- 238000012364 cultivation method Methods 0.000 description 2
- 239000000796 flavoring agent Substances 0.000 description 2
- 230000004807 localization Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 235000013599 spices Nutrition 0.000 description 2
- 101150028074 2 gene Proteins 0.000 description 1
- 108700028369 Alleles Proteins 0.000 description 1
- 241000893384 Ceratorhiza oryzae-sativae Species 0.000 description 1
- 101710201890 Gibberellin 20 oxidase 2 Proteins 0.000 description 1
- 102000008300 Mutant Proteins Human genes 0.000 description 1
- 208000031481 Pathologic Constriction Diseases 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 238000009402 cross-breeding Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000003828 downregulation Effects 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000003209 gene knockout Methods 0.000 description 1
- 238000010363 gene targeting Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000005065 mining Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000005204 segregation Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000036262 stenosis Effects 0.000 description 1
- 208000037804 stenosis Diseases 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
-
- 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
- C12Y—ENZYMES
- C12Y114/00—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14)
- C12Y114/11—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14) with 2-oxoglutarate as one donor, and incorporation of one atom each of oxygen into both donors (1.14.11)
- C12Y114/11012—Gibberellin-44 dioxygenase (1.14.11.12)
-
- 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)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Medicinal Chemistry (AREA)
- Virology (AREA)
- Botany (AREA)
- Developmental Biology & Embryology (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention relates to the technical field of plant genetic engineering and breeding, in particular to a gene for regulating and controlling dwarfing, lodging resistance and yield of crops and application thereof. The gene is shown as a gene sequence shown as a sequence A, and the application comprises the following steps: regulating and controlling the plant height, the lodging resistance capability, the maturing rate and the yield of the rice. The invention also discloses a rice stalk dwarf control mutant gene, which is a gene with 14 base deletions from 4492 th to 4505 th positions in a gene sequence shown in a sequence A. The stalk dwarf control gene, protein, mutant gene and mutant thereof can be used for reducing the plant height of conventional rice and hybrid rice, improving the lodging resistance of the rice, reducing the yield loss caused by lodging and having important application value.
Description
Technical Field
The invention relates to the technical field of plant genetic engineering and breeding, in particular to a gene for regulating and controlling dwarfing, lodging resistance and yield of crops and application thereof.
Background
Rice is one of the most important grain crops in China, and more than 60% of people in China take rice as main food. Cultivation of lodging-resistant rice is a powerful guarantee for realizing high yield and stable yield of rice, cultivation of short-stalk rice is an effective means for improving lodging-resistant capability of rice, and dwarf rice breeding has proved to be an ideal way for solving contradiction between high yield and lodging of rice. At present, the rice production mainly depends on green gene sd1 and each allele thereof to reduce the plant height, and the dwarf source is single. The single low source tends to pose a risk of potential genetic background stenosis, and further improvement of variety yield is limited. In addition, rice breeding practices show that even though the sd1 gene is contained, the plant height of some rice varieties is too high, and the weight of the spike part is heavy, so that lodging and yield reduction of rice occur throughout the year. Therefore, the mining of new dwarf gene resources remains an important item in the study of genetic breeding of rice.
Disclosure of Invention
The invention provides a gene for regulating and controlling dwarfing, lodging resistance and yield of crops and application thereof, which are used for solving the defect of single dwarf source in the prior art.
The invention provides sdlr5 muteins, genes encoding them or biological materials containing genes encoding them; the biological material is an expression cassette, a vector, a host cell, recombinant bacteria or hybrid dwarf hybrid rice.
The sdlr5 mutein has any one of the following amino acid sequences:
(1) An amino acid sequence as shown in SEQ ID NO. 5;
(2) An amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity to the amino acid sequence shown in SEQ ID NO.5, said protein having a function of regulating plant dwarfing;
(3) An amino acid sequence obtained by substituting and/or deleting and/or adding one or more amino acid residues in the amino acid sequence shown in SEQ ID NO.5, wherein the protein has the function of regulating plant dwarfing.
The mutant proteins of the present invention are non-natural proteins.
The invention provides a sdlr5 mutant gene, a protein encoded by the same or a biological material containing the sdlr5 mutant gene, wherein the sdlr5 mutant gene is a gene deleted at 14 bases from 4492 th to 4505 th in a gene sequence shown in a sequence A; or the cDNA sequence of the sdlr5 gene is a sequence of 14 base deletion at 914-927 bits in the cDNA sequence shown in SEQ ID NO. 3; the biological material is an expression cassette, a vector, a host cell, recombinant bacteria or hybrid dwarf hybrid rice.
The sequence A is formed by sequentially connecting SEQ ID NO.1 and SEQ ID NO.2, wherein the 1 st to 6660bp is SEQ ID NO.1, and the 6661 st to 10389bp is SEQ ID NO.2.
The invention provides sdlr5 proteins, genes encoding the same, or biological materials containing the genes encoding the same; the biological material is an expression cassette, a vector, a host cell or a recombinant bacterium;
the sdlr5 protein has any one of the following amino acid sequences:
(1) An amino acid sequence as shown in SEQ ID NO. 4;
(2) An amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity to the amino acid sequence shown in SEQ ID NO.4, said protein having a function of regulating plant dwarfing;
(3) An amino acid sequence obtained by substituting and/or deleting and/or adding one or more amino acid residues in the amino acid sequence shown in SEQ ID NO.4, wherein the protein has the function of regulating plant dwarfing. Compared with sd1, the sdlr5 can further reduce plant height and improve lodging resistance without adverse effect on yield.
The present invention provides an sdlr5 gene, a protein encoded thereby, or a biological material containing the sdlr5 gene, the sdlr5 gene having any one of the following nucleotide sequences:
(1) A nucleotide sequence shown as a sequence A or SEQ ID NO.3 or a complementary sequence thereof;
(2) A sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity with the nucleotide sequence shown in sequence a or SEQ ID No.3, which encodes a polypeptide having a function of regulating plant dwarfing;
(3) A sequence which hybridizes under stringent conditions to sequence A or SEQ ID NO. 3;
(4) The nucleotide sequence shown in the sequence A or SEQ ID NO.3 is obtained by substituting and/or deleting and/or adding one or more nucleotides, and the coded polypeptide has the function of regulating plant dwarfing.
The biological material is an expression cassette, a vector, a host cell or a recombinant bacterium.
The vector of the invention is a plant expression vector.
The vector of the present invention may be a plasmid.
The present invention provides the use of a protein, gene or biological material in any one, two, three or four of the following:
(1) Application in regulating and controlling rice plant height;
(2) Application in rice lodging resistance;
(3) Application in regulating and controlling the maturing rate of rice;
(4) The application in regulating and controlling the rice yield.
Preferably, the application is any one, two, three or four of the following:
(1) Application in reducing rice plant height;
(2) The application of the anti-lodging agent in improving the lodging resistance of rice;
(3) The application of the method in not reducing (preferably improving) the rice setting rate.
(4) The application in improving the rice yield.
The protein, gene or biological material is selected from any one of the following:
(1) The sdlr5 mutein, its encoding gene or biological material containing its encoding gene;
(2) The sdlr5 mutant gene, the protein encoded by the same, or a biological material containing the sdlr5 mutant gene;
(3) The sdlr5 protein, its coding gene, or a biological material containing its coding gene;
(4) The sdlr5 gene, a protein encoded by the same, or a biomaterial containing the sdlr5 gene.
According to the application, the regulation is negative.
The invention provides a method for creating a rice dwarf lodging-resistant strain, which comprises any one of the following steps:
(1) Knocking out or changing the nucleotide sequence shown as sequence A or SEQ ID NO.3 to make the sequence deleted or mutated, so that the activity level of the gene is reduced or the function is lost.
(2) Inhibiting or reducing expression of a nucleotide sequence encoding an amino acid sequence as set forth in SEQ ID NO.4, and reducing the level of expression of a polypeptide corresponding to said amino acid sequence.
Preferably, selecting a conventional rice variety, treating and cultivating to obtain the rice dwarf line, wherein the treatment is that,
(a) The nucleotide sequence shown as the sequence A or SEQ ID NO.3 is knocked out or changed by adopting natural mutation, mutagenesis or conventional genetic engineering methods, so that the sequence is deleted or mutated, and the activity level of the gene is reduced or the function is lost.
Or (b) inhibiting or reducing expression of a nucleotide sequence encoding an amino acid sequence as set forth in SEQ ID NO.4 by conventional genetic engineering methods, and reducing expression levels of polypeptides corresponding to the amino acid sequence.
According to the method for creating the rice dwarf lodging-resistant strain provided by the invention, the nucleotide sequence is shown as SEQ ID NO. 3.
The invention provides a method for creating a rice dwarf lodging-resistant strain, which comprises the step of deleting 14 bases at 914-927 in a sequence shown in SEQ ID NO. 3.
The invention provides a method for creating a rice dwarf lodging-resistant strain, which comprises the step of deleting 14 bases from 4492 th to 4505 th in a sequence shown in a sequence A.
The invention provides an application of a rice stalk dwarf control mutant in conventional rice improvement, wherein the mutant is introduced into a conventional rice variety by means of cross breeding, so that the plant height is reduced, and the lodging resistance is improved.
Preferably, the mutant transfers the mutant gene to a conventional rice variety through a hybridization breeding means, so that the rice plant height is reduced to a proper range, and the lodging resistance is improved.
The rice stalk dwarf control is prepared by a method for preparing the rice stalk lodging-resistant strain.
The invention provides a method for cultivating hybrid dwarf hybrid rice, which comprises (1) enabling a sterile line to carry dwarf control mutant genes, enabling a restorer line to carry dwarf control genes, and combining the sterile line and the restorer line to produce hybrid rice.
Or (2) the restorer line carries the dwarf control mutant gene, the sterile line carries the dwarf control gene, and the sterile line and the restorer line are matched to produce hybrid rice.
Or (3) the sterile line carries dwarf control mutant gene, the restorer line carries dwarf control mutant gene, and the sterile line and the restorer line are matched to produce homozygous dwarf hybrid rice.
The cultivation method can reduce the plant height of the rice, improve the lodging resistance of the rice, not reduce (preferably improve) the seed setting rate of the rice and improve the yield of the rice.
The dwarf control mutant gene is sdlr5 mutant gene.
The dwarf control gene is sdlr5 gene.
The dwarf control gene is a nucleotide sequence which codes for an amino acid sequence shown as SEQ ID NO. 4.
According to the cultivation method of the hybrid dwarf hybrid rice, the nucleotide sequence of the dwarf control gene is shown as SEQ ID NO. 3.
The invention has the beneficial effects that:
the invention clones a new regulated rice stem dwarf gene LOC_Os05g34854 (a gene sequence shown by SDLR 5-sequence A), which codes gibberellin 20oxidase (the cDNA sequence is shown as SEQ ID NO.3, and the protein sequence is shown as SEQ ID NO. 4). The mutant gene sdlr5 is obtained by mutating the gene sequence shown in the sequence A and deleting 14 bases. The stalk dwarf control gene, the protein, the mutant gene and the mutant thereof can be used for reducing the plant height of rice, realizing dwarf breeding of the rice, improving the lodging resistance of the rice and improving the yield of the rice. The gene can be directly applied to conventional rice and hybrid rice, reduces the plant height of the rice to a proper range, improves the lodging resistance, has no obvious influence on the yield and the yield, and has important application value in rice breeding. Provides a new idea and method for rice lodging-resistant breeding.
Drawings
In order to more clearly illustrate the invention or the technical solutions of the prior art, the following description will briefly explain the drawings used in the embodiments or the description of the prior art, and it is obvious that the drawings in the following description are some embodiments of the invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 is a graph comparing the phenotype of the gene 9311B with the short stem mutant short 11B; a is short 11B and B is a group 9311B.
FIG. 2 is a graph showing the gene targeting results of the Mutmap method.
FIG. 3 shows the mutation sites within the localization intervals of the group 9311B and the short 11B; a is the position of the reference genome (http:// www.elabcaas.cn/price/index. Html) on chr5, B is the resequencing result of short 11B, the deletion of 14 bases, C is the resequencing result of the gene 9311B, and D is the sequence of the reference genome 9311.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings, and it is apparent that the described embodiments are some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
EXAMPLE 1 obtaining of Rice dwarf mutant
The group 9311A is an excellent three-line sterile line, and the hybrid rice matched with the group has the characteristics of high yield, excellent rice quality and the like, and more than 140 combinations are approved and applied in large-area production. However, the partial hybrid combination using the girl 9311A as the female parent is poor in lodging resistance, and lodging often occurs under severe weather conditions. Dwarf 11B is a dwarf mutant obtained by screening mutation of the maintainer line of the human 9311A through EMS mutagenesis. The plant height of the short 11B is 77.02 +/-3.02 cm, the plant height of the girl 9311B is 98.42+/-4.12 cm, the plant height of the hybrid F1 generation (hereinafter referred to as F1) of the short 11B and the girl 9311B is 89.03 +/-4.55 cm, and the difference among the three reaches a very significant level, which indicates that the short stalk character of the short 11B is semi-dominant character.
Further measuring the ear length and the length from one inverted node to six inverted nodes, finding that the ear length of short 11B is 20.11 cm, the difference between the ear length of short 11B and the difference between the ear length of short 11B (22.44 cm) and the difference between the ear length of short 11B and the length of short 11F 1 (22.72 cm) reach a significant level, and the difference between the ear lengths of short 11B and the ear length of short 11F 1 are not significant; the difference of the lengths of the three parts between the two sections does not reach a significant level, and the difference of the lengths between the two sections and the three sections is a significant difference; short 11B is not significantly different from F1 in the length of the four, five and six internodes, but is significantly lower than the length of the 9311B. Meanwhile, the investigation shows that the three have no obvious differences in tillering number, grain per spike number and setting rate.
The single tillering fresh weight, basal fracture resistance and lodging index of the short 11B and the girl 9311B were detected, and it was found that the difference between the single tillering fresh weight and basal fracture resistance of the short 11B and the girl 9311B was not significant, but the lodging index (6.11) of the short 11B was extremely significantly smaller than the girl 9311B (9.61) (the lodging index was smaller, the lodging resistance was stronger), mainly due to the fact that the short 11B was significantly shorter than the girl 9311B.
Example 2 Gene preliminary targeting
In order to locate genes that control the dwarf 11B phenotype, hybridization with short 11B to the group 9311B resulted in the F2 population. F2, planting a group F2, selecting 30 plants of single plants of short and high stems, mixing the leaves, extracting DNA, and respectively extracting DNA of a female parent sample and DNA of a male parent sample. Genome resequencing was performed according to the BSA method (isolate-group mixed assay or mixed-group assay, also known as group separation assay, bulked Segregation Analysis), and the short stalk gene was located by the Mutmap method, and the results are shown in fig. 2. As can be seen from FIG. 2, SNP-Index shows a distinct peak only at the 21Mb position of the fifth chromosome and breaks through the 95% and 99% confidence lines. This suggests that the dwarf trait of dwarf 11B is controlled by a single major gene, and that the mutant gene was designated sdlr5 #Semi-DwarfandLodgingResistance5)。
Example 3 further mapping, sequencing and cloning of genes
Short 11B is a mutant of the gene 9311B, and the number of different loci on the genome is small. Comparing the resequenced data of the group 9311B and the group 9311B revealed that the group 9311B had a deletion mutation of 14bp relative to the group 9311B in the initial localization region (fig. 3), and the deletion of 14bp (TCCAGGACGGCGAC) was found to occur on the second exon of loc_os05g34854 by the annotation genome (Rice Genome Annotation Project (uga.edu)) of MSU 7.0. In combination with the first generation sequencing, it was verified that the gene sequence shown in the sequence A of the short 11B lacks 14 bases from 4492 th to 4505 th. LOC_Os05g34854 encodes a putative gibberellin 20oxidase (gibberellin 20oxidase 2,putative). Further, F3 family verification of 1388 short 11B/quan 9311B shows that the family or the single plant lacking 14 bases is short.
EXAMPLE 4 mutant Gene and application of mutant in conventional Rice improvement
The mutant gene is used for improving 19-flavor and agricultural flavor 32, so that the plant height is reduced and the lodging resistance is improved. The dwarf 11B is hybridized with the 19-spice plants and the agricultural spice plants 32 respectively, F2 groups are planted, the dwarf plants are selected, DNA is extracted, and the detection finds that the selected dwarf plants all contain deletion mutation of 14bp, can pick up the plants with better agronomic characters, and can reduce the height of the dwarf plants by 5-30 cm than the height of the 19-spice plants or the agricultural spice plants 32. The sdlr5 can be applied to rice plant height improvement of different backgrounds, particularly rice varieties with higher plant heights, and has no obvious adverse effect on agronomic characters.
Example 5 mutant Gene and use of mutant in improving hybrid Rice
The short 11A is used as female parent and the short 11B is used as male parent, and the short-stem excellent three-line sterile line short 11A is bred through continuous backcrossing. 10 hybridization combinations are respectively configured by taking short 11A and the girl 9311A as female parent, taking R4-3 and 27 as well as tsukuba 110, 19117R and Yu He as male parent. The hybridization combinations are planted, and the hybridization combinations taking short 11A as female parent are 8-12 cm shorter than the hybridization combinations taking short 9311A as female parent, and the average plant height is reduced by 9.89 cm. And the length between the two, three and four internodes of the matched offspring of the short 11A is extremely obviously shorter than that of the matched offspring of the same male parent 9311A. The lodging index measurement finds that the lodging index of the offspring of the low 11A matched set is smaller than that of the offspring of the old 9311A (the smaller the lodging index, the better the lodging resistance). Meanwhile, the tillering number, the grain number per spike, the fruiting rate and the average single-plant grain weight are measured, and no obvious difference exists between the low 11A matched set offspring and the matched set offspring of the girl 9311A. In the later period of maturity, the combined part 9311A/27 is inclined and is in a lodging trend, while the part 11A/27 is still kept in an upright posture, so that the lodging resistance is obviously improved.
As can be seen from the application, the invention can generate plant dwarf phenotype by utilizing the deletion mutation of 14 bases on the sdlr5 and introducing the deletion mutation into other conventional rice or hybrid rice materials, and can be directly utilized in breeding.
Example 6 other applications
As the sdlr5 gene is cloned, the gene has the function of controlling the dwarf of rice stalks and has no obvious linkage encumbrance, so the gene can be directly applied to dwarf rice breeding, lodging-resistant breeding and high-yield breeding. The cultivation of short-stalk rice can improve lodging resistance and avoid or reduce a series of problems of yield reduction, spike germination, rice quality reduction and the like caused by lodging. The gene is a negative regulation gene, and the breeding purpose can be realized by means of mutagenesis, gene knockout and the like, or by means of gene engineering to down regulate the expression of the sdlr5 gene or the translation level of the sdlr5 protein. The above genes and various biological materials containing the genes can also be directly utilized to carry out rice breeding improvement by a transgenic mode. In addition, the mutant containing the mutant gene can be directly adopted to hybridize with other rice materials to carry out rice breeding improvement, especially the conventional rice or hybrid rice with higher plant height and poorer lodging resistance can be used for reducing the plant height of the rice to a proper range, and the mutant has important application value in rice breeding.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and are not limiting; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention.
Claims (10)
- An sdlr5 mutein, a gene encoding the same or a biological material comprising the gene encoding the same; the biological material is an expression cassette, a vector, a host cell or a recombinant bacterium;the sdlr5 mutein has any one of the following amino acid sequences:(1) An amino acid sequence as shown in SEQ ID NO. 5;(2) An amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity to the amino acid sequence shown in SEQ ID NO.5, said protein having a function of regulating plant dwarfing;(3) An amino acid sequence obtained by substituting and/or deleting and/or adding one or more amino acid residues in the amino acid sequence shown in SEQ ID NO.5, wherein the protein has the function of regulating plant dwarfing.
- An sdlr5 mutant gene, a protein encoded by the same, or a biological material containing the sdlr5 mutant gene, characterized in that the sdlr5 mutant gene is a gene deleted from 14 bases from 4492 th to 4505 th in a gene sequence shown in sequence a; or the cDNA sequence of the sdlr5 gene is a sequence of 14 base deletion at 914-927 bits in the cDNA sequence shown in SEQ ID NO. 3;the sequence A is formed by sequentially connecting SEQ ID NO.1 and SEQ ID NO.2, wherein the 1 st to 6660bp is SEQ ID NO.1, and the 6661 st to 10389bp is SEQ ID NO.2;the biological material is an expression cassette, a vector, a host cell or a recombinant bacterium.
- An sdlr5 protein, a gene encoding the same, or a biological material containing the gene encoding the same; the biological material is an expression cassette, a vector, a host cell or a recombinant bacterium;the sdlr5 protein has any one of the following amino acid sequences:(1) An amino acid sequence as shown in SEQ ID NO. 4;(2) An amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity to the amino acid sequence shown in SEQ ID NO.4, said protein having a function of regulating plant dwarfing;(3) An amino acid sequence obtained by substituting and/or deleting and/or adding one or more amino acid residues in the amino acid sequence shown in SEQ ID NO.4, wherein the protein has the function of regulating plant dwarfing.
- An sdlr5 gene, a protein encoded thereby, or a biological material comprising an sdlr5 gene, wherein the sdlr5 gene has any one of the following nucleotide sequences:(1) A nucleotide sequence shown as a sequence A or SEQ ID NO.3 or a complementary sequence thereof;(2) A sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity with the nucleotide sequence shown in sequence a or SEQ ID No.3, which encodes a polypeptide having a function of regulating plant dwarfing;(3) A sequence which hybridizes under stringent conditions to sequence A or SEQ ID NO. 3;(4) A sequence obtained by substituting and/or deleting and/or adding one or more nucleotides from the nucleotide sequence shown in the sequence A or SEQ ID NO.3, and the coded polypeptide has the function of regulating plant dwarfing;the sequence A is formed by sequentially connecting SEQ ID NO.1 and SEQ ID NO.2, wherein the 1 st to 6660bp is SEQ ID NO.1, and the 6661 st to 10389bp is SEQ ID NO.2;the biological material is an expression cassette, a vector, a host cell or a recombinant bacterium.
- 5. Use of a protein, gene or biological material in any one, two, three or four of the following:(1) Application in regulating and controlling rice plant height;(2) Application in rice lodging resistance;(3) Application in regulating and controlling the maturing rate of rice;(4) Application in regulating and controlling rice yield;the protein, gene or biological material is selected from any one of the following:(1) The sdlr5 mutein of claim 1, a gene encoding thereof, or a biological material containing a gene encoding thereof;(2) The sdlr5 mutant gene, the protein encoded by the same, or the biological material containing the sdlr5 mutant gene of claim 2;(3) The sdlr5 protein of claim 3, the coding gene thereof, or a biological material containing the coding gene thereof;(4) The sdlr5 gene of claim 4, a protein encoded thereby, or a biomaterial containing the sdlr5 gene;preferably, the application is any one, two, three or four of the following:(1) Application in reducing hybrid rice plant height;(2) Application in improving lodging resistance of hybrid rice;(3) The application in not reducing the maturing rate of hybrid rice;(4) Application in improving the yield of hybrid rice;preferably, the modulation is negative.
- 6. The method for creating the rice dwarf lodging-resistant strain is characterized by comprising any one of the following steps:(1) Knocking out or changing the nucleotide sequence shown as the sequence A or SEQ ID NO.3 to make the sequence deleted or mutated, so that the activity level of the gene is reduced or lost;(2) Inhibiting or reducing expression of a nucleotide sequence encoding an amino acid sequence as set forth in SEQ ID NO.4, and reducing expression levels of polypeptides corresponding to said amino acid sequence;the sequence A is formed by sequentially connecting SEQ ID NO.1 and SEQ ID NO.2, wherein the 1 st to 6660bp is SEQ ID NO.1, and the 6661 st to 10389bp is SEQ ID NO.2.
- 7. The method for creating a rice dwarf-resistant strain according to claim 6, which is characterized by comprising the deletion of 14 bases from 4492 th to 4505 th in the sequence represented by the sequence A.
- 8. The method for creating a rice dwarf-stump-resistant strain according to claim 6, comprising deletion of 14 bases from 914 to 927 in the sequence represented by SEQ ID NO. 3.
- 9. The application of the rice stalk dwarf control mutant in conventional rice improvement is characterized in that the mutant transfers mutant genes to conventional rice varieties through a hybridization breeding method, so that the rice plant height is reduced to a proper range, and the lodging resistance is improved;the rice stalk dwarf control is created by the method of any one of claims 6-8.
- 10. A method for cultivating hybrid short-stalk hybrid rice is characterized in that: comprises (1) making sterile line carry dwarf control mutant gene, restoring line carry dwarf control gene, and combining sterile line and restoring line to produce hybrid rice;or (2) the restorer line carries the dwarf control mutant gene, the sterile line carries the dwarf control gene, and the sterile line and the restorer line are matched to generate hybrid rice;or (3) the sterile line carries dwarf control mutant gene, the restorer line carries dwarf control mutant gene, and the sterile line and the restorer line are matched to produce homozygous dwarf hybrid rice:the dwarf control mutant gene is a gene according to any one of claims 1 to 2;the dwarf control gene is a gene according to any one of claims 3 to 4;preferably, the dwarf control gene is a nucleotide sequence encoding an amino acid sequence shown as SEQ ID NO. 4;preferably, the nucleotide sequence of the dwarf control gene is shown as SEQ ID NO. 3.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310795542.1A CN116676284B (en) | 2023-06-30 | 2023-06-30 | Gene for regulating crop dwarfing, lodging resistance and yield and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310795542.1A CN116676284B (en) | 2023-06-30 | 2023-06-30 | Gene for regulating crop dwarfing, lodging resistance and yield and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN116676284A true CN116676284A (en) | 2023-09-01 |
| CN116676284B CN116676284B (en) | 2024-05-07 |
Family
ID=87787320
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310795542.1A Active CN116676284B (en) | 2023-06-30 | 2023-06-30 | Gene for regulating crop dwarfing, lodging resistance and yield and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116676284B (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007046403A1 (en) * | 2005-10-19 | 2007-04-26 | National University Corporation Nagoya University | Plant belonging to genus oryza having dominant dwarf trait, and use thereof |
| CN102703497A (en) * | 2012-03-28 | 2012-10-03 | 上海交通大学 | Establishing method and application of paddy short-stalk strain, and method for recovering short stalk character |
| CN103936843A (en) * | 2013-03-25 | 2014-07-23 | 袁隆平农业高科技股份有限公司 | Rice Os05g26890.1 protein, gene coding protein, and application of gene |
| CN103993018A (en) * | 2014-03-13 | 2014-08-20 | 中国科学院遗传与发育生物学研究所 | Gene for controlling Oryza sativa plant height, enhancing lodging resistance, increasing effective tiller number and yield and its application |
| CN105543242A (en) * | 2015-11-27 | 2016-05-04 | 中国农业科学院作物科学研究所 | Rice STD1 gene, encoding protein and application thereof |
| WO2023029587A1 (en) * | 2021-08-31 | 2023-03-09 | 中国科学院合肥物质科学研究院 | Mutation site of ideal brittle stalk mutant ibc in rice, control gene ibc, and application |
-
2023
- 2023-06-30 CN CN202310795542.1A patent/CN116676284B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007046403A1 (en) * | 2005-10-19 | 2007-04-26 | National University Corporation Nagoya University | Plant belonging to genus oryza having dominant dwarf trait, and use thereof |
| CN102703497A (en) * | 2012-03-28 | 2012-10-03 | 上海交通大学 | Establishing method and application of paddy short-stalk strain, and method for recovering short stalk character |
| CN103936843A (en) * | 2013-03-25 | 2014-07-23 | 袁隆平农业高科技股份有限公司 | Rice Os05g26890.1 protein, gene coding protein, and application of gene |
| CN103993018A (en) * | 2014-03-13 | 2014-08-20 | 中国科学院遗传与发育生物学研究所 | Gene for controlling Oryza sativa plant height, enhancing lodging resistance, increasing effective tiller number and yield and its application |
| CN105543242A (en) * | 2015-11-27 | 2016-05-04 | 中国农业科学院作物科学研究所 | Rice STD1 gene, encoding protein and application thereof |
| WO2023029587A1 (en) * | 2021-08-31 | 2023-03-09 | 中国科学院合肥物质科学研究院 | Mutation site of ideal brittle stalk mutant ibc in rice, control gene ibc, and application |
Also Published As
| Publication number | Publication date |
|---|---|
| CN116676284B (en) | 2024-05-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112390865B (en) | Application of Zm5008 gene in regulating and controlling plant height and internode distance of corn | |
| CN105646684B (en) | A kind of rice grain shape GAP-associated protein GAP GLW2 and its encoding gene and application | |
| CN104774836A (en) | Polygene pyramiding early-breeding method for raising litter size of sheep | |
| CN114990139A (en) | Application of CsHLS1 gene or protein coded by same in regulation and control of organ size of cucumber plant | |
| CN114292868A (en) | Application of rice plant height gene LOC _ Os03g64415 in rice plant type improvement | |
| CN112522282B (en) | Gene for regulating and controlling soluble solid content of tomato and application thereof | |
| CN116676284B (en) | Gene for regulating crop dwarfing, lodging resistance and yield and application thereof | |
| CN103130884B (en) | Protein relevant to plant spike shape and encoding gene and appliance thereof | |
| CN107338255A (en) | Application of the PINNATE PENTAFOLIATA2 genes in legume leaflet quantity is regulated and controled | |
| CN114395580B (en) | Gene for controlling maize plant height | |
| CN114561407B (en) | Gene for regulating corn root system included angle and lodging resistance and application thereof | |
| CN114230648B (en) | Application of rice gene PANDA in improving plant yield | |
| CN113563439B (en) | Fruit shape development related protein, and coding gene and application thereof | |
| CN114957420A (en) | Rice false smut related panicle gene spr9, protein and application | |
| KR101983902B1 (en) | A Novel mutant tomato plants with modified determinate shoot architecture | |
| CN111172171A (en) | Gene for controlling plant height and flowering phase of corn and application thereof | |
| CN112625099A (en) | Rape dwarf gene BND2 and application thereof in rape crossbreeding | |
| CN114921471B (en) | Gene and molecular marker for controlling rape specific inflorescence characteristics and application | |
| CN113429468B (en) | Barley male sterility gene msg3002 and application thereof | |
| CN113248585B (en) | Wheat TaARF25-B gene, mutant thereof and application thereof in regulating plant height and grain traits | |
| KR102010726B1 (en) | A Novel mutant tomato plants with modified determinate shoot architecture | |
| KR102010727B1 (en) | A Novel mutant tomato plants with modified determinate shoot architecture | |
| CN109354615B (en) | LEPTO1 and application of encoding protein thereof in rice fertility control | |
| US20240180095A1 (en) | Stay green cucurbitaceae plant | |
| CN115838736A (en) | A pair of genes ClSLB/Clslb for controlling length of lateral branches of watermelons 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 | ||
| CB02 | Change of applicant information | ||
| CB02 | Change of applicant information |
Address after: 231201 Shenggu Agricultural Inspection Building in the south of Forest Avenue, Zipeng Town, Feixi County, Hefei City, Anhui Province Applicant after: Anhui Shenggu Agricultural Technology Co.,Ltd. Applicant after: Guizhou Zhunong High tech Seed Industry Co.,Ltd. Address before: 231201 Shenggu Agricultural Inspection Building in the south of Forest Avenue, Zipeng Town, Feixi County, Hefei City, Anhui Province Applicant before: Anhui Shenggu Agricultural Technology Co.,Ltd. Applicant before: Guizhou zhunongke Seed Industry Co.,Ltd. |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |