CN107987139A - A kind of Dof transcription factors and its application in terms of plant salt tolerance is improved - Google Patents
A kind of Dof transcription factors and its application in terms of plant salt tolerance is improved Download PDFInfo
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- C12N15/8286—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance for insect resistance
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Abstract
The invention discloses a kind of Dof transcription factors and its application in terms of plant salt tolerance is improved.The amino acid sequence of the transcription factor is as shown in sequence table SEQ ID NO.1.Research of the albumen and its encoding gene of the present invention for the resistance to inverse mechanism of plant, and the improvement for improving the resistance of reverse and correlated traits of plant have important theory and practical significance, will play a significant role, and have a extensive future in the resistance to inverse genetic engineering improvement of plant.
Description
Technical field
The invention belongs to biological gene technical field, and in particular to a kind of Dof transcription factors and its improve plant salt tolerance
The application of aspect.
Background technology
According to UNESCO and FAO's incomplete statistics, the whole world there are about 1,000,000,000 hectares it is saline and alkaline
Ground, China there are about 100,000,000 hectares of salt-soda soils, these salinized soil are a parts for the important land resource in China.In addition, China is existing
In arable land, at least also 8,000,000 hectares of soil causes salt accumulation in soil due to irrigation and improper fertilization.Due to soil
Salinization of soil harm weight, the growth and development to crop has a negative impact, beach saline-alkali area Per Unit Area Grain Yield be only in the high place of production
25%-40%.These land resources of rational exploitation and utilization will be related to whether China can keep 1,800,000,000 mu of farmland red lines, steady
Determine agricultural sustainable development, ensure the major action of grain and Environmental security.
Although the key technologies such as the quick desalinization of soil by flooding or leaching, control salt can effectively reduce saline and alkaline harm, increase crop yield, utilize
Salt-tolerant trait, the New Crop Varieties of cultivation saline-alkali tolerant of crop in itself can not only improve crop yield, but also can make salt
Utilized alkali, brought more land under cultivation.Though resistance of reverse improvement of traditional breeding technique for crop has certain effect, by
In its selection and breeding cycle, long and trait phenotypes foresight is poor；And can be according to advance by biotechnology cultivation resistance of reverse rice varieties
Design specific gene is transformed and shifted, breeding process can be made more accurate, more proactive, more efficient, thus
More development potentiality, becomes the important channel of rearing new variety.Therefore, excavate the degeneration-resistant begetting power of rice, make full use of it is resistance to inverse
Genetic resources cultivates resistance to inverse rice varieties, it has also become the important measure of China's grain-production sustainable development.
The content of the invention
It is an object of the invention to provide a kind of Dof transcription factors and its application in terms of plant salt tolerance is improved.
A kind of Dof transcription factors, the amino acid sequence of the transcription factor is as shown in sequence table SEQ ID NO1.
, can be in the amino terminal or carboxyl terminal of the amino acid sequence of aforementioned polypeptides in order to make above-mentioned protein easy to purifying
One or several labels as shown in Table 1 in connection, as shown in table 1.
The sequence of 1 label of table
| Label | Residue | Sequence |
| Poly-Arg | 5-6 (being usually 5) | RRRRR |
| Poly-His | 2-10 (being usually 6) | HHHHHH |
| FLAG | 8 | DYKDDDDK |
| Strep-tagⅡ | 8 | WSHPQFEK |
| c-myc | 10 | EQKLISEEDL |
Aforementioned polypeptides can be artificial synthesized, also can first synthesize its encoding gene, then carry out biological expression and obtain.It is above-mentioned
Polypeptide also can be by will lack the codon of one or several amino acid residues, and/or progress in sequence shown in SEQ ID NO.2
The missense mutation of one or several base-pairs, and/or hold the coded sequence for connecting the label shown in table 1 to obtain at its 5 ' end or 3 '
Arrive.
OsDof15 provided by the invention contains typical zinc finger-Dof domains, is the 78 to 132nd amino acid residue；EAR
Transcription repression domain, is the 253 to 258th amino acid residue.
The gene order of the Dof transcription factors is as shown in sequence table SEQ ID NO.2.
Genophore containing above-mentioned Dof transcription factors.
The carrier is GFP fusion expression vectors, specially passes through the gene (sequence shown in SEQ ID NO.2)
XbaI and BamHI restriction enzyme sites are connected to pAN580 carriers and obtain.
The carrier for BD carriers be specially by the gene (sequence shown in SEQ ID NO.2) by EcoRI and
BamHI restriction enzyme sites are connected to pGBKT7 carriers and obtain.
The recombinant bacterium or cell line of genophore containing above-mentioned Dof transcription factors.
Expand the primer of any fragment in above-mentioned rice transcription factor.
A kind of Dof transcription factors mutant, the Dof transcription factors mutant are amino acid sequence shown in SEQ ID NO.1
Row are by substitution, missing and/or the addition of one or several amino acid residues and relevant by SEQ ID with Salt Resistance of Rice
NO:Polypeptide derived from 1 sequence.
The gene (sequence shown in SEQ ID NO.2) is passed through BamHI and XhoI enzymes by the recombinant plant expression vector
Enzyme site is connected to pCAMBIA1307 (6 × Myc) carrier, obtains pCAMBIA1307 (6 × Myc)-OsDof15 carriers；It is described
DNA fragmentation shown in the SEQ ID NO.4 of engineer and its reverse complementary sequence are passed through pNW55 intermediate carriers by interference carrier
Connect, be then connected to by multiple cloning sites in plant expression vector pCAMBIA5300, obtain pCAMBIA5300-
RNAi-OsDof15 carriers.The recombinant expression carrier can be by using Ti-plasmids, Ri plasmids, plant viral vector, direct DNA
The conventional biology methods such as conversion, microinjection, conductance, agriculture bacillus mediated convert plant cell or tissue, and by the plant of conversion
Thing tissue cultivating is into plant.
The plant can be monocotyledon, or dicotyledon, such as rice crops.At one of the present invention
In embodiment, the plant is monocotyledon rice, is specially rice varieties Nipponbare.
An example in above-mentioned Dof transcription factors mutant, the amino acid sequence of the Dof transcription factors mutant is such as
Shown in sequence table SEQ ID NO.3.
The application of above-mentioned Dof transcription factors or above-mentioned Dof transcription factors mutant in terms of plant salt tolerance is improved.
Application of the above-mentioned Dof transcription factors mutant in pest-resistant.
Beneficial effects of the present invention：The present invention turns OsDof15 trans-genetic hybrid rice strain by acquisition and interference strain obtains
Laboratory phenotype：Patience of the rice seedling to salt stress can significantly be improved by being overexpressed OsDof15, plant is being run into the non-life such as salt
Energy normal growth when thing is coerced, is overexpressed transgenic line and wilts appearance evening relative to wild-type leaves, through coercing after a while
After compeling processing, survival rate is higher；Interference OsDof15 genes make rice seedling drop the tolerance of salt stress after reducing its expression
Low, interference of transgene strain is more early wilted relative to wild-type leaves, and survival rate is decreased obviously.The albumen and its volume of the present invention
Research of the code gene for the resistance to inverse mechanism of plant, and the improvement for improving the resistance of reverse and correlated traits of plant have important reason
Practical significance is referred to, will play a significant role, and have a extensive future in the resistance to inverse genetic engineering improvement of plant.
Brief description of the drawings
Fig. 1 show OsDof15 gene expression detection results in OsDof15 genes overexpression and interference plant.
Fig. 2 show the Salt Tolerance Analysis of OsDof15 transfer-gen plants.
Fig. 3 show the Subcellular Localization of OsDof15 albumen.
Fig. 4 show the transcripting activating characteristic testing result of OsDof15 albumen.
Embodiment
The present invention will be further described with specific embodiment below in conjunction with the accompanying drawings.
Experimental method in following embodiments, is conventional method unless otherwise specified.Examination used in following embodiments
Material is tested, is to be commercially available from routine biochemistry reagent shop unless otherwise specified.
Embodiment 1：The acquisition of OsDof15 overexpression vectors and artificial microRNA interference carriers
First, the structure of OsDof15 overexpression vectors pCAMBIA1307 (6 × Myc)-OsDof15
From OsDof15 genes (Gene ID:4334219) coding initiation site ATG starts design 5 ' and holds primer, in end
3 ' end primers are only designed before codon：
Primer 1：5’-ggatccatgatccaagaactccttggagg-3’；
Primer 2：5'-ctcgagtgggaacgcgccgttgggagtgg-3'；
The restriction enzyme site that GGATCC sequences in primer 1 are restriction enzyme BamHI, the sequence of underscore mark are
The coded sequence of OsDof15 genes；In primer 2 CTCGAG sequences be restriction enzyme XhoI restriction enzyme site, underscore mark
The sequence of knowledge is the coded sequence of OsDof15 genes.
Using the cDNA of japonica rice variety Nipponbare as template, PCR amplification is carried out with above-mentioned specific primer, recycles PCR amplification
Product, carries out double digestion with restriction enzyme BamHI and XhoI, recycles digestion products；With restriction enzyme BamHI and
SalI double digestions pCAMBIA1307 (6 × Myc), recycles skeleton carrier；The digestion products and the skeleton carrier are connected
Connect, be then sequenced, sequencing result shows, has obtained pCAMBIA1307 (6 × Myc)-OsDof15 carriers.
2nd, the structure of the artificial microRNA interference carriers of OsDof15
Using the transcript sequence of OsDof15 genes, in WMD3 websites (http://wmd3.weigelworld.org/
Cgi-bin/webapp.cgi the microRNA target sites of the gene) are analyzed, are then picked out from the amiRNA of candidate special
For the preferable sequence construct of property on intermediate carrier pNW55, process is exactly to pass through three pairs of primers：Primer I and primer IV, primer G-
4368 and primer II, primer III and primer G-4369, expand to obtain three fragments respectively using pNW55 as template, then with this three
A fragment expands to obtain big containing two microRNA that can be complementary for template by primer G-4368 and primer G-4369
Fragment, is then connected in plant expression vector pCAMBIA5300 by multiple cloning sites, has obtained pCAMBIA5300-
RNAi-OsDof15 carriers.
Primer I：5’-agtctgattaaccgaaatcgcaccaggagattcagtttga-3’；
Primer II：5’-tggtgcgatttcggttaatcagactgctgctgctacagcc-3’；
Primer III：5’-ctgtgcgttttgggttaatcagattcctgctgctaggctg-3’；
Primer IV：5’-aatctgattaacccaaaacgcacagagaggcaaaagtgaa-3’
Primer G-4368：5’-ctgcaaggcgattaagttgggtaac-3’；
Primer G-4369：5’-gcggataacaatttcacacaggaaacag-3’.
Embodiment 2：The acquisition of OsDof15 genetically modified plants
Using electric shocking method by recombinant expression carrier pCAMBIA1307 (6 × Myc)-OsDof15 and pCAMBIA5300-
RNAi-OsDof15 imports Agrobacterium AGL0, obtains recombinational agrobacterium.
The Agrobacterium of pCAMBIA1307 (6 × Myc)-OsDof15 and pCAMBIA5300-RNAi-OsDof15 will be contained
AGL0 disseminates Nipponbare Rice Callus respectively；The callus of dip dyeing is blotted on sterile filter paper, returns again to co-cultivation base
Upper 24 DEG C of light cultures 2-4 days；The callus of cleaning, which is gone to, carries out resistance screening on the Selective agar medium containing hygromycin；It is chosen
Kanamycin-resistant callus tissue afterwards goes to pre- differential medium and differential medium is returned again to after 7-10 days because carrying out illumination cultivation；Treat seedling grow to
Go in the test tube equipped with root media and grow 2-3 weeks during 2-4cm, well-grown seedling is after the hardening of 2-3 days
Transplanting is into greenhouse (T0 generations).Seedling is passed on respectively, obtains T1 generations.
The wild rice (Nipponbare) and T1 for taking 2 weeks respectively carry out reverse transcription, obtain for rice seedling, extraction mRNA
Pass through the expression (sense primer of OsDof15 in fluorescent quantitative PCR technique analyzing rice after cDNA：5’-
ccgtggccggacttgcccactc-3’；Anti-sense primer：5’-tcacactgattaaccgaaaccac-3’).By wild type water
The expression quantity of OsDof15 genes is as 1 in rice, overexpress strain OX-1, OX-5, OX-6, OX-13 and interference strain RI-1,
The expression of OsDof15 genes is as shown in Figure 1 in RI-2, RI-3, RI-4.The result shows that：In four overexpression strains
The expression quantity of OsDof15 genes reaches more than 2.5 times of wild rice, in four interference strains in addition to RI-3 OsDof15
The expression quantity of gene reaches 40% of wild rice or so.
Embodiment 3：The Salt Tolerance Analysis of OsDof15 transfer-gen plants
The T1 of OsDof15 is harvested for transgenic paddy rice seed, choose respectively two overexpression strains (OX-1, OX-13) and
Two interference strains (RI-1, RI-2) carry out NaCl analog salt Stress treatments.With its sprouting of 3 angel of water logging kind culture, Ran Houyong
50mg/L hygromycin carries out resistance screening, and seedling is moved to soil after sprouting 3 days carries out earth culture, the wild type day grown with the same period
This is fine as compareing, and experiment is repeated 3 times.
Salt treatment (150mM NaCl) is carried out to the earth culture seedling for growing to 2 weeks, after handling 7 days, observation blade treats its appearance
After wilting, brine is changed through repeatedly displacement, observes blade.Strain is overexpressed after salt treatment and shows stronger salt tolerant than wild type
Property, and disturb strain to show salt tolerance reduction than wild type, illustrate that OsDof15 can improve tolerance of the rice to salt stress
Property.The results are shown in Figure 2.
Embodiment 4：OsDof15 protein subcellular positioning analysises
First, OsDof15-GFP fusion vectors are built
The OsDof15 sequences with XbaI and BamHI restriction enzyme sites are obtained by method primer 3 in embodiment 1 and primer 4
Fragment, then with restriction enzyme XbaI and BamHI digestion pAN580, recycle skeleton, be connected, obtain with above-mentioned fragment
OsDof15-pAN580 carriers.
Primer 3：5'-tctagaatgatccaagaactccttggagg-3'
Primer 4：5'-ggatcctgggaacgcgccgttgggagtgg-3'
The restriction enzyme site that TCTAGA sequences in primer 3 are restriction enzyme XbaI, the sequence of underscore mark are
The coded sequence of OsDof15 genes；The restriction enzyme site that GGATCC sequences in primer 4 are restriction enzyme BamHI, underscore
The sequence of mark is the coded sequence of OsDof15 genes.
2nd, the Subcellular Localization of OsDof15-GFP fusion proteins
By OsDof15-pAN580 fusion vectors by PEG mediated method rice transformation protoplasts after, fluorescence microscope see
Examine GFP signals.As shown in figure 3, it was observed that after empty carrier pAN580 conversion protoplasts, GFP has in cytoplasm and nucleus
Expression；And GFP shows that OsDof15 is positioned at carefully with after OsDof15 amalgamation and expressions, only seeing GFP fluorescence signals in nucleus
In karyon.
Embodiment 5：OsDof15 protein transcriptions activate Characteristics Detection
First, OsDof15-BD vector constructions
The OsDof15 sequences with EcoRI and BamHI restriction enzyme sites are obtained by method primer 5 in embodiment 1 and primer 4
The fragment of row, then with restriction enzyme EcoRI and BamHI digestion pGBKT7, recycles skeleton, is connected, obtains with above-mentioned fragment
To OsDof15-BD carriers.
Primer 5：5'-gaattcatgatccaagaactccttggagg-3'
The restriction enzyme site that GAATTC sequences in primer 5 are restriction enzyme EcoRI, the sequence of underscore mark are
The coded sequence of OsDof15 genes.
2nd, OsDof15-BD transformed yeasts detection activation characteristic
The OsDof15-BD expression vectors built are transformed into yeast gold109, it is to be identified it is correct after by transformant
Bacterium solution be applied on SD/-Trp/X- α-gal tablets and the SD/-Trp/X- α-gal/AbA containing 0.1% AbA resistances it is flat
On plate.Cultivated 2-3 days in 30 DEG C of incubators.As shown in figure 4, positive control is in SD/-Trp/X- α-gal tablets and SD/-Trp/X-
Normal growth and blue reaction can be shown on α-gal/AbA defect culture medium flat plates, and negative control is in SD/-Trp/X-
Normal growth but blue reaction can not be shown on α-gal plates, while cannot be in SD/-Trp/X- α-gal/AbA defect culture mediums
Upper growth.The yeast strain for turning OsDof15-BD shows blue reaction on SD/-Trp/X- α-gal plates, and can be
Normal growth on SD/-Trp/X- α-gal/AbA Selective agar mediums, illustrates that OsDof15 albumen has activation LacZ bases in yeast
Because of the transcriptional activation activity of expression.
The pest-resistant performance of embodiment 5Dof transcription factor mutant
PCR clones OsDof15 genes, and builds plant expression vector p2300-35S-OsDof15, utilizes p2300-35S-
OsDof15 plant expression vectors are masterplate, and mutant is obtained by the method for Overlap extension PCR, and mutational site is the 42nd and the
43 amino acids residue SS sport QQ.Plant expression vector p2300-35S-OsDof15 ' is rebuild, and passes through Agrobacterium
Mediated method rice transformation Nipponbare, obtains the transgenic rice plant of the OsDof15 ' genes of 35S promoter driving.
Plantation turns the rice and rice Nipponbare of OsDof15 ' genes, grows into 40 days, takes and grows in the same size, the number of blade
The consistent rice plant of mesh, turns 30 plants of the rice plant of OsDof15 ' genes, 30 plants of rice Nipponbare, and 100 are connect on every plant of rice
The ripe aptery aphid of head, 30 it is small when after, count aphid inhibiting rate：Aphid inhibiting rate %=(remaining aphid on 100- blades)/
100。
Experimental result：The aphid inhibiting rate for turning the rice of OsDof15 ' genes is 78%, and the aphid of rice Nipponbare suppresses
Rate is 1%.
Plantation turns the rice and rice Nipponbare of OsDof15 ' genes, grows into 40 days, takes and grows in the same size, the number of blade
The consistent rice plant of mesh, turns 30 plants of the rice plant of OsDof15 ' genes, 30 plants of rice Nipponbare, and 100 are connect on every plant of rice
Head brown paddy plant hopper, 30 it is small when after, count brown paddy plant hopper inhibiting rate：Brown paddy plant hopper inhibiting rate %=(remaining brown paddy plant hopper on 100- blades)/
100。
Experimental result：The brown paddy plant hopper inhibiting rate for turning the rice of OsDof15 ' genes is 69%, the brown paddy plant hopper of rice Nipponbare
Inhibiting rate is 0%.
Plantation turns the rice and rice Nipponbare of OsDof15 ' genes, grows into 40 days, takes and grows in the same size, the number of blade
The consistent rice plant of mesh, turns 30 plants of the rice plant of OsDof15 ' genes, 30 plants of rice Nipponbare, and 100 are connect on every plant of rice
Head rice leaf roller, 30 it is small when after, count rice leaf roller inhibiting rate：Rice leaf roller inhibiting rate %=(is remained on 100- blades
Remaining rice leaf roller)/100.
Experimental result：The rice leaf roller inhibiting rate for turning the rice of OsDof15 ' genes is 3%, and the rice of rice Nipponbare is indulged
Leaf roll snout moth's larva inhibiting rate is 3%.
Sequence table
<110>Biological Technology institute, Chinese Academy of Agricultural Sciences
<120>A kind of Dof transcription factors and its application in terms of plant salt tolerance is improved
<160> 15
<170> SIPOSequenceListing 1.0
<210> 1
<211> 359
<212> PRT
<213>Rice (Oryza sative L)
<400> 1
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1 5 10 15
Ala Ser Ala Leu Asn His Ala Ser Leu Pro Val Val Leu Gln Pro Ile
20 25 30
Val Ser Asn Pro Ser Pro Thr Ser Ser Ser Ser Thr Ser Ser Arg Ser
35 40 45
Ser Ala Gln Ala Thr Gln Gln Arg Ser Ser Ser Ala Thr Ser Ser Pro
50 55 60
His Gly Gln Gly Gln Gly Gly Gly Ala Ala Glu Gln Ala Pro Leu Arg
65 70 75 80
Cys Pro Arg Cys Asn Ser Ser Asn Thr Lys Phe Cys Tyr Tyr Asn Asn
85 90 95
Tyr Asn Leu Thr Gln Pro Arg His Phe Cys Lys Thr Cys Arg Arg Tyr
100 105 110
Trp Thr Lys Gly Gly Ala Leu Arg Asn Val Pro Ile Gly Gly Gly Cys
115 120 125
Arg Lys Pro Arg Pro Met Pro Ala Pro Val Ala Lys Pro Pro Met Ser
130 135 140
Cys Lys Ala Ala Pro Pro Leu Gly Leu Gly Gly Gly Pro Val Ser Trp
145 150 155 160
Ala Ser Gly Gln Gln Ala Ala Thr Ala His Leu Met Ala Leu Leu Asn
165 170 175
Ser Ala Arg Gly Val Gln Gly His Gly Gly Ser Asn Val His Arg Leu
180 185 190
Leu Gly Leu Asp Thr Met Gly His Leu Gln Ile Leu Pro Gly Ala Pro
195 200 205
Asn Gly Ala Gly Ala Gly Thr Ala Ala Ser Leu Trp Pro Gln Ser Ala
210 215 220
Pro Arg Pro Val Thr Pro Pro Pro Pro His Met Asp Ser Gln Leu Gly
225 230 235 240
Met Gly Thr Leu Gly His His Asp Val Leu Ser Ser Leu Gly Leu Lys
245 250 255
Leu Pro Ser Ser Ala Ser Ser Ser Pro Ala Ala Ser Tyr Tyr Ser Asp
260 265 270
Gln Leu His Ala Val Val Ser Asn Ala Gly Arg Pro Gln Ala Pro Tyr
275 280 285
Asp Val Ala Thr Ala Ser Leu Pro Cys Thr Thr Ala Val Thr Ser Leu
290 295 300
Pro Ser Ala Leu Ser Ser Val Ser Ala Ala Ala Pro Thr Ser Asn Thr
305 310 315 320
Val Gly Met Asp Leu Pro Pro Val Ser Leu Ala Ala Pro Glu Met Gln
325 330 335
Tyr Trp Asn Gly Pro Ala Ala Met Ser Val Pro Trp Pro Asp Leu Pro
340 345 350
Thr Pro Asn Gly Ala Phe Pro
355
<210> 2
<211> 1080
<212> DNA
<213>Rice (Oryza sative L)
<400> 2
atgatccaag aactccttgg agggacaacc atggaccagc tcaagggcgc cagcgctctg 60
aaccacgcct ccctgccggt ggtgctgcag cctatcgtgt ccaacccgtc gcccacgtcg 120
tcgtcgtcga cgtcgtcgcg ctcgtcggcg caggcgacgc agcagaggtc gtcgtcggcg 180
acctcgtcgc cgcacgggca ggggcagggt ggcggcgcgg cggagcaggc gccgctgcgg 240
tgcccgcggt gcaactcgtc gaacaccaag ttctgctact acaacaacta caacctcacc 300
cagccgcgcc acttctgcaa gacgtgccgc cggtactgga ccaagggcgg cgcgctccgc 360
aacgtcccca tcggcggcgg gtgccgcaag ccgcgcccca tgccggcgcc ggtcgccaag 420
ccgcccatgt cttgcaaggc cgcgccgccg ctcggcctcg gcggcgggcc agtgtcctgg 480
gcctccgggc agcaggccgc caccgcgcac ctcatggcgc tgctcaacag cgccagggga 540
gtgcagggcc acggcggcag caatgtccac cggcttcttg ggctggacac catgggtcac 600
ctccagatcc tgccaggcgc tcccaatggc gccggcgccg gcacggcggc gtcgctctgg 660
ccacagtccg cgccgcggcc ggtcactcca ccgccgccgc acatggactc ccagctcggc 720
atggggacgc tgggccacca cgacgtgctg tcgagcctcg gcctcaagct gccctcgtcg 780
gcgtcgtcct cgccggcggc gagctactac agcgaccagc tgcacgcggt ggtgagcaac 840
gcggggcgcc cccaggcgcc gtacgacgtc gccaccgcgt ccctcccttg caccaccgcg 900
gtgacctcac tcccgtcggc gctgtcgagc gtctccgccg ccgcgccgac cagcaacacg 960
gtcgggatgg acctgccacc cgtgtcgctc gccgcgccgg agatgcagta ctggaatggc 1020
ccggcggcga tgtcggtgcc gtggccggac ttgcccactc ccaacggcgc gttcccatga 1080
<210> 3
<211> 359
<212> PRT
<213>Rice (Oryza sative L)
<400> 3
Met Ile Gln Glu Leu Leu Gly Gly Thr Thr Met Asp Gln Leu Lys Gly
1 5 10 15
Ala Ser Ala Leu Asn His Ala Ser Leu Pro Val Val Leu Gln Pro Ile
20 25 30
Val Ser Asn Pro Ser Pro Thr Ser Ser Gln Gln Thr Ser Ser Arg Ser
35 40 45
Ser Ala Gln Ala Thr Gln Gln Arg Ser Ser Ser Ala Thr Ser Ser Pro
50 55 60
His Gly Gln Gly Gln Gly Gly Gly Ala Ala Glu Gln Ala Pro Leu Arg
65 70 75 80
Cys Pro Arg Cys Asn Ser Ser Asn Thr Lys Phe Cys Tyr Tyr Asn Asn
85 90 95
Tyr Asn Leu Thr Gln Pro Arg His Phe Cys Lys Thr Cys Arg Arg Tyr
100 105 110
Trp Thr Lys Gly Gly Ala Leu Arg Asn Val Pro Ile Gly Gly Gly Cys
115 120 125
Arg Lys Pro Arg Pro Met Pro Ala Pro Val Ala Lys Pro Pro Met Ser
130 135 140
Cys Lys Ala Ala Pro Pro Leu Gly Leu Gly Gly Gly Pro Val Ser Trp
145 150 155 160
Ala Ser Gly Gln Gln Ala Ala Thr Ala His Leu Met Ala Leu Leu Asn
165 170 175
Ser Ala Arg Gly Val Gln Gly His Gly Gly Ser Asn Val His Arg Leu
180 185 190
Leu Gly Leu Asp Thr Met Gly His Leu Gln Ile Leu Pro Gly Ala Pro
195 200 205
Asn Gly Ala Gly Ala Gly Thr Ala Ala Ser Leu Trp Pro Gln Ser Ala
210 215 220
Pro Arg Pro Val Thr Pro Pro Pro Pro His Met Asp Ser Gln Leu Gly
225 230 235 240
Met Gly Thr Leu Gly His His Asp Val Leu Ser Ser Leu Gly Leu Lys
245 250 255
Leu Pro Ser Ser Ala Ser Ser Ser Pro Ala Ala Ser Tyr Tyr Ser Asp
260 265 270
Gln Leu His Ala Val Val Ser Asn Ala Gly Arg Pro Gln Ala Pro Tyr
275 280 285
Asp Val Ala Thr Ala Ser Leu Pro Cys Thr Thr Ala Val Thr Ser Leu
290 295 300
Pro Ser Ala Leu Ser Ser Val Ser Ala Ala Ala Pro Thr Ser Asn Thr
305 310 315 320
Val Gly Met Asp Leu Pro Pro Val Ser Leu Ala Ala Pro Glu Met Gln
325 330 335
Tyr Trp Asn Gly Pro Ala Ala Met Ser Val Pro Trp Pro Asp Leu Pro
340 345 350
Thr Pro Asn Gly Ala Phe Pro
355
<210> 4
<211> 29
<212> DNA
<213> Artificial
<400> 4
ggatccatga tccaagaact ccttggagg 29
<210> 5
<211> 29
<212> DNA
<213> Artificial
<400> 5
ctcgagtggg aacgcgccgt tgggagtgg 29
<210> 6
<211> 40
<212> DNA
<213> Artificial
<400> 6
agtctgatta accgaaatcg caccaggaga ttcagtttga 40
<210> 7
<211> 40
<212> DNA
<213> Artificial
<400> 7
tggtgcgatt tcggttaatc agactgctgc tgctacagcc 40
<210> 8
<211> 40
<212> DNA
<213> Artificial
<400> 8
ctgtgcgttt tgggttaatc agattcctgc tgctaggctg 40
<210> 9
<211> 40
<212> DNA
<213> Artificial
<400> 9
aatctgatta acccaaaacg cacagagagg caaaagtgaa 40
<210> 10
<211> 25
<212> DNA
<213> Artificial
<400> 10
ctgcaaggcg attaagttgg gtaac 25
<210> 11
<211> 22
<212> DNA
<213> Artificial
<400> 11
ccgtggccgg acttgcccac tc 22
<210> 12
<211> 23
<212> DNA
<213> Artificial
<400> 12
tcacactgat taaccgaaac cac 23
<210> 13
<211> 29
<212> DNA
<213> Artificial
<400> 13
tctagaatga tccaagaact ccttggagg 29
<210> 14
<211> 29
<212> DNA
<213> Artificial
<400> 14
ggatcctggg aacgcgccgt tgggagtgg 29
<210> 15
<211> 29
<212> DNA
<213> Artificial
<400> 15
gaattcatga tccaagaact ccttggagg 29
Claims (9)
- A kind of 1. Dof transcription factors, it is characterised in that the amino acid sequence of the transcription factor such as sequence table SEQ ID NO.1 It is shown.
- 2. rice transcription factor according to claim 1, it is characterised in that the gene order such as sequence of the Dof transcription factors Shown in list SEQ ID NO.2.
- A kind of 3. genophore containing Dof transcription factors described in claim 1.
- 4. recombinant bacterium or cell line containing the genophore of Dof transcription factors described in claim 3.
- 5. expand the primer of any fragment in the rice transcription factor described in claim 1.
- 6. a kind of Dof transcription factors mutant, it is characterised in that the Dof transcription factors mutant is shown in SEQ ID NO.1 Amino acid sequence by one or several amino acid residues substitution, missing and/or addition and with Salt Resistance of Rice it is relevant by Polypeptide derived from SEQ ID NO.1 sequences.
- 7. Dof transcription factors mutant according to claim 6, it is characterised in that the ammonia of the Dof transcription factors mutant Base acid sequence is as shown in sequence table SEQ ID NO.3.
- 8. Dof transcription factors mutant described in Dof transcription factors described in claim 1 or claim 6 is improving plant salt tolerance The application of aspect.
- 9. application of the Dof transcription factors mutant in pest-resistant described in claim 7.
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| CN109355270A (en) * | 2018-11-16 | 2019-02-19 | 中国农业科学院生物技术研究所 | A kind of rice kinases OSK1 and its application |
| CN111518826A (en) * | 2019-02-01 | 2020-08-11 | 中国科学院植物研究所 | Method for improving stress tolerance of rice |
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| CN102174085A (en) * | 2005-11-10 | 2011-09-07 | 先锋高级育种国际公司 | Dof (dna binding with one finger) sequences and methods of use |
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| CN109355270A (en) * | 2018-11-16 | 2019-02-19 | 中国农业科学院生物技术研究所 | A kind of rice kinases OSK1 and its application |
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| CN111518826A (en) * | 2019-02-01 | 2020-08-11 | 中国科学院植物研究所 | Method for improving stress tolerance of rice |
| CN111518826B (en) * | 2019-02-01 | 2021-12-28 | 中国科学院植物研究所 | Method for improving stress tolerance of rice |
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