CN105462992B - Rice Os RhoGAP2 gene is improving the application in rice effective tillering - Google Patents
Rice Os RhoGAP2 gene is improving the application in rice effective tillering Download PDFInfo
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- CN105462992B CN105462992B CN201610002409.6A CN201610002409A CN105462992B CN 105462992 B CN105462992 B CN 105462992B CN 201610002409 A CN201610002409 A CN 201610002409A CN 105462992 B CN105462992 B CN 105462992B
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- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
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- 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
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Abstract
The invention discloses riceOsRhoGAP2Gene is improving the application in rice effective tillering, by rice Rho gtpase activating protein encoding geneOsRhoGAP2Building is overexpressed plant vector, by the method rice transformation callus of mediated by agriculture bacillus, obtains T2 generation overexpressionOsRhoGAP2The transgenic paddy rice of gene, compared with the control, the tiller number of transgenic paddy rice improve 32.35%-38.75%.
Description
Technical field
The invention belongs to agricultural and technical field of molecular biology, and in particular to rice Os RhoGAP2 gene is improving water
Application in rice effective tillering.
Background technique
Global nearly half population is all food with rice.Rice, scientific name: Oryza sativa L. is most important in the world
One of cereal crops.At this stage, the crisis of food shortage is got worse, and the yield of cereal crops yield especially rice compels to be essential
It improves.Rice genome project has been completed, more and more Main Agronomic Characters genes and the assignment of genes gene mapping (quantitative character
Site) it is identified to come successively by researcher.According to its progress, can be answered with a series of knowledge of molecular biology
For rice breeding scheme, the method for Lai Tigao rice yield.
The tiller and heading of rice are the important characters for determining its grain yield.First identified control rice come out
The gene of tiller is single tiller gene (MOC1) of rice, is achieved in terms of determining important gene relevant to rice yield great
Progress, to be achieved to the hereditary basis for illustrating rice yield correlated traits.Some of which important gene is can be with
Molecular breeding for high-yield rice.
As more and more evidences show that rice yield is controlled by quantity shape site.Rice tillering is one important
Agronomic character, influence whether the yield of rice, for example the tillering quantity of single plant determines that spike number, available tillering determine tiller
The quantity of the spike number and number of grain per ear that grow up to.The gene relevant to tiller number found later is also: TAD1, TE, LAX1,
LAX2, OsTB1, OsMADS57, FC1, D3, D17, D10, D14, D27, DLT etc., these genes all pass through directly or indirectly
To have adjusted the fringe branch of rice.
Summary of the invention
The object of the present invention is to provide rice Os RhoGAP2 genes to improve the application in rice effective tillering, i.e., will contain
There is the expression vector of OsRhoGAP2 gene to be transfected into rice, then cultivates rice and obtain T2 generation overexpression OsRhoGAP2 gene
Transgenic paddy rice.
The present invention adopts the following technical scheme that rice Os RhoGAP2 gene is effective in raising rice to achieve the above object
Application in tiller, it is characterised in that rice Os RhoGAP2 gene order is as shown in SEQ ID NO.1.
Rice Os RhoGAP2 gene of the present invention is improving the application in rice effective tillering, it is characterised in that tool
Body process are as follows: design primer P1:5 '-TATCTGCAGATGGAGGGCGAGGTGCC-3 ' contains PstI restriction enzyme site and primer
P2:5 '-GCGAAGCTTTACACAGGCTTGTCCCGC-3 ' is by PCR amplification containing III restriction enzyme site of Hind
PstI and III restriction enzyme site of Hind are added in the end of OsRhoGAP2 gene cDNA encoding area 5 ' and 3 ' ends respectively, are connected to plant expression
The corresponding site of carrier pCAMBIA1302, then table is crossed by the method rice transformation callus acquisition T2 generation of mediated by agriculture bacillus
Up to the transgenic paddy rice of OsRhoGAP2 gene.
Rice Rho protein regulation factor code gene OsRhoGAP2 is constructed and is overexpressed plant vector by the present invention, passes through agriculture
The method rice transformation callus that bacillus mediates obtains the transgenic paddy rice for being overexpressed OsRhoGAP2 gene in T2 generation, and compares
It compares, the tiller number of transgenic paddy rice improves 32.35%-38.75%.
Detailed description of the invention
Fig. 1 is to compare rice and OsRhoGAP2 gene overexpression transgenic paddy rice T2 for the statistics of different strain tiller numbers
Compare, and wherein WT is control rice OryzasativaLcv.Nipponbare, and OE-OsRhoGAP2 is transgenic paddy rice；
Fig. 2 is control rice and OsRhoGAP2 gene overexpression transgenic paddy rice T2 for different strain available tillerings
Statistics compares, and wherein WT is control rice OryzasativaLcv.Nipponbare, and OE-OsRhoGAP2 is transgenic paddy rice；
Fig. 3 is the building of OsRhoGAP2 plant over-express vector, and the code area cDNA that wherein A is OsRhoGAP2 expands glue
Recovery product agarose gel electrophoresis, B is pMD18-T-OsRhoGAP2 carrier Hind III and I double digestion of Pst is identified, figure C is
PCAMBIA1302-OsRhoGAP2-eGFP expression vector Hind III and the identification of I double digestion of Pst；
Fig. 4 is that the PCR in OsRhoGAP2 overexpression transgenic paddy rice T2 generation is detected, M:DNA Maker (DL2000) in figure,
1, wild type detects, 2, negative control, 3, positive control, 4-13, the PCR product for being overexpressed plant T2 generation, can by PCR detection
To screen positive plant.The correct and bright representative positive plant of band.
Fig. 5 is the expression quantity detection that OsRhoGAP2 is overexpressed transgenic paddy rice T2 generation, be can detecte by expression quantity detection
Expression of the gene in this plant, from figure we it can be concluded that there is the expression quantity of the gene in transgenic paddy rice
Different degrees of raising, least to have 14 times, most has 229 times.
Specific embodiment
Above content of the invention is described in further details by the following examples, but this should not be interpreted as to this
The range for inventing above-mentioned theme is only limitted to embodiment below, and all technologies realized based on above content of the present invention belong to this hair
Bright range.
Embodiment 1
1, the building of rice Os RhoGAP2 gene overexpression carrier
Design primer P1:(5 '-TATCTGCAGATGGAGGGCGAGGTGCC-3 ' contains PstI restriction enzyme site) and primer
P2:(5 '-GCGAAGCTTTACACAGGCTTGTCCCGC-3 ' contains III restriction enzyme site of Hind), be by PCR amplification
PstI and III restriction enzyme site of Hind are added in the end of OsRhoGAP2 gene cDNA encoding area 5 ' and 3 ' ends respectively, are connected to plant expression
The corresponding site of carrier pCAMBIA1302 (being purchased from CAMBIA company).Connection product converts bacillus coli DH 5 alpha, blocks that containing
Transformant is screened on the LB plate of mycin, after digestion and PCR identification, confirmation has been sequenced.
2, in the genetic transformation of rice, screening and T2 generation, turn the analysis of OsRhoGAP2 trans-genetic hybrid rice character
Using conventional rice transformation method, using During Agrobacterium method by OsRhoGAP2 gene overexpression carrier
Conversion to Rice Callus (will go the rice paddy seed program request of glume on calli induction media N6D2.About after a week, water
Rice can grow callus.After removing bud and root, callus is placed on new N6D2 culture medium and continues to induce, about 10 days
It is capable of forming harder callus, can be used for Agrobacterium and infect.) hygromycin selection positive callus is utilized, into one
One-step inducing cultivates regeneration positive transgenic plant, and carries out Molecular Identification.Transgenic paddy rice screen to T2 for when, to same batch
Control rice and T2 observed for transgenic paddy rice, measure tiller number (as Figure 2-3), carry out statistical analysis, unite
The mean tillering number of each 20 plants of sample of meter, the control rice of the non-transgenosis of statistical result showed is 13.90 ± 2.77, transgenosis water
The mean tillering number of rice is 18.93 ± 4.20, and compared with the control, the tiller number of transgenic paddy rice improves 32.35%-
38.75%.
Embodiment above describes basic principles and main features of the invention and advantage, the technical staff of the industry should
Understand, the present invention is not limited to the above embodiments, and the above embodiments and description only describe originals of the invention
Reason, under the range for not departing from the principle of the invention, various changes and improvements may be made to the invention, these changes and improvements are each fallen within
In the scope of protection of the invention.
Claims (2)
1. riceOsRhoGAP2Gene is improving the application in rice effective tillering, it is characterised in that the riceOsRhoGAP2Base
Because sequence is as shown in SEQ ID NO.1.
2. rice according to claim 1OsRhoGAP2Application of the gene in raising rice effective tillering, feature exist
In detailed process are as follows: design primer P1:5 '-TATCTGCAGATGGAGGGCGAGGTGCC-3 ' containsPstI restriction enzyme site and draw
Object P2:5 '-GCGAAGCTTTACACAGGCTTGTCCCGC-3 ', containsHindIII restriction enzyme site is by PCR amplificationOsRhoGAP2The end of gene cDNA encoding area 5 ' and 3 ' ends are added respectivelyPstI andHindIII restriction enzyme site is connected to plant expression
The corresponding site of carrier pCAMBIA1302, then table is crossed by the method rice transformation callus acquisition T2 generation of mediated by agriculture bacillus
It reachesOsRhoGAP2The transgenic paddy rice of gene.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610002409.6A CN105462992B (en) | 2016-01-06 | 2016-01-06 | Rice Os RhoGAP2 gene is improving the application in rice effective tillering |
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| CN201610002409.6A CN105462992B (en) | 2016-01-06 | 2016-01-06 | Rice Os RhoGAP2 gene is improving the application in rice effective tillering |
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| CN105462992A CN105462992A (en) | 2016-04-06 |
| CN105462992B true CN105462992B (en) | 2019-03-05 |
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106434690A (en) * | 2016-09-18 | 2017-02-22 | 河南师范大学 | Application of rice OsRhoGAP2 gene to changing rice grain form |
| CN107236026B (en) * | 2017-07-03 | 2020-12-29 | 河北师范大学 | GBP protein and application of coding gene thereof in regulating and controlling plant yield |
| CN110904118B (en) * | 2019-12-04 | 2023-04-25 | 河南师范大学 | Application of rice OsRopGAP2 gene in preparation of glume rice |
| CN115820721B (en) * | 2022-07-26 | 2024-03-22 | 贵州大学 | Method for improving tillering and yield of Guizhou high-quality special rice and large gingko glutinous rice, promoter core sequence and application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1477112A (en) * | 2002-08-20 | 2004-02-25 | 中国科学院遗传与发育生物学研究所 | Rice tiller control gene MOC1 and its application |
| WO2010076766A1 (en) * | 2008-12-30 | 2010-07-08 | Institute Of Genetics And Developmental Biology | Genes associated with plant tiller number and uses thereof |
| CN102690838A (en) * | 2012-06-05 | 2012-09-26 | 中国科学院植物研究所 | Application of OsMADS57 protein or coding gene thereof in promotion of rice tillering |
-
2016
- 2016-01-06 CN CN201610002409.6A patent/CN105462992B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1477112A (en) * | 2002-08-20 | 2004-02-25 | 中国科学院遗传与发育生物学研究所 | Rice tiller control gene MOC1 and its application |
| WO2010076766A1 (en) * | 2008-12-30 | 2010-07-08 | Institute Of Genetics And Developmental Biology | Genes associated with plant tiller number and uses thereof |
| CN102690838A (en) * | 2012-06-05 | 2012-09-26 | 中国科学院植物研究所 | Application of OsMADS57 protein or coding gene thereof in promotion of rice tillering |
Non-Patent Citations (3)
| Title |
|---|
| Control of tillering in rice;Li X，等;《Nature》;20030410;第422卷;第618-621页 |
| Oryza sativa (japonica cultivar-group) Rho GTPase activating protein 2 mRNA,complete cds.;Liang W等;《Genbank登录号：AY364311.1》;20030909;参见全文 |
| 水稻分蘖基因的研究概况;李万昌，等;《作物杂志》;20120630(第3期);第19-22页 |
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