CN1313486C - Plant DREB transcription factor and its coding gene and use - Google Patents
Plant DREB transcription factor and its coding gene and use Download PDFInfo
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Abstract
The present invention discloses a plant DREB transcription factor, a coding gene thereof and the application thereof. The plant DREB transcription factor is protein having one of the following amino acid residue sequences: (1) SEQ ID No: 1 in a sequence table; (2) protein which replaces and/or deletes and/or adds an amino acid residue sequence of SEQ ID No: 1 in the sequence table by one or a plurality of amino acid residue bases and is relevant to the stress resistance of plants. The protein and the coding gene thereof can enhance the stress resistance of the plants and particularly can enhance the resistance of low temperature stress.
Description
Technical field
The present invention relates to a kind of plant DREB transcription factor and encoding gene and application in the plant biotechnology field, particularly the DREB transcription factor relevant and an encoding gene and an application that derives from rice grass (Spartina anglica) with resistance of reverse.
Background technology
Under environment stress, can produce a series of responsing reactions in the plant materials, the variation that is accompanied by many Physiology and biochemistries and grows.Illustrate the reaction mechanism of plant, will provide scientific foundation for the molecular breeding of plant stress-resistance to adverse circumstance.
Many genes of plant have been proved to be inducing of being subjected to that abiotic stress coerces.With coerce relevant gene product and can be divided into two big classes: the product of first kind genes encoding comprises that ionophorous protein, aquaporin, the osmoregulation factor (sucrose, proline(Pro) and trimethyl-glycine etc.) synthetic enzyme etc. participate in the gene product that plant stress is replied directly; The product of second genoid coding comprises the protein factor that participates in coercing relevant signal transmission and genetic expression adjusting, as protein kinase, transcription factor etc.Wherein, transcription factor plays an important role in the gene expression regulation that plant stress is replied.
Transcription factor is also referred to as trans-acting factor, is can be conjugated protein with the DNA of cis-acting elements generation specific effect in the eukaryotic gene promoter region, by between them and and other associated protein between interaction, activate or suppress and transcribe.The DNA land of transcription factor has determined it and cis-acting elements bonded specificity, and transcription regulatory region has determined it that genetic expression is risen to activate or restraining effect.In addition, himself activity also is subjected to appraising and deciding the influence of effects such as position and oligomerization.
Known transcription factor relevant with abiotic stress in plant mainly contains at present: the DREB class transcription factor family with EREBP (element responsive to ethylene is conjugated protein, ethylene responsive element binding protein)/AP2 (APETALA2) structural domain, bZIP (basic region/leucine zipper motif transcription factors) the class transcription factor that contains alkalescence zone and leucine zipper, contain the MYC family of alkaline helix-loop-helix (bHLH) and leucine zipper and have MYB family of tryptophane bunch (Trp cluster) or the like.
Yamaguchi-Shinozaki and Shinozaki are in the research to the promoter region of adverse circumstance response gene rd29A gene, found that an environment stress replys the functional element of taking advantage of a situation, be dehydration response element (DRE, dehydration-responsive element).After this, find that many adverse circumstance response gene promoter regions contain the DRE element.People such as Liu utilize yeast-one-hybrid system first, from the cDNA expression library of Arabidopis thaliana, be cloned into the transcription factor dehydration response element conjugated protein (DREB of two kinds of codings and DRE combination of elements, dehydration-responsive element binding protein) cDNA, difference called after DREB1A, DREB2A.They do not have significant homogeny on aminoacid sequence, but all contain one section very conservative DNA calmodulin binding domain CaM (EREBP/AP2 structural domain) of being made up of 58 amino acid.This class contains the transcription factor of EREBP/AP2 structural domain, transmits relevant with the adverse circumstance signal.Liu Qiang etc. think that a dreb gene can be regulated and control a plurality of and plant arid, high salt and low temperature patience function associated expression of gene.Kasuga etc. studies confirm that, the DREB1A gene that imports to Arabidopis thaliana can promote the expression of gene rd29, rd17, kin1, cor6.6, cor15a and the erd10 relevant with environment stress patience simultaneously, and the resistance of transfer-gen plant strengthens greatly.Equally, the low temperature tolerance ability of the transfer-gen plant of low temperature patience transcription factor CBF1 (dreb1b) is significantly increased.Because the stress tolerance of plant is the complex character by the polygene regulation and control, rely on to import the comprehensive raising that the individual feature protein gene is difficult to realize stress resistance of plant.Therefore, utilize a key transcription factor to promote the expression of a plurality of functional genes, thereby strengthen the comprehensive resistance of plant, become the engineered research focus of plant stress-resistance.
Summary of the invention
The purpose of this invention is to provide a kind of plant DREB transcription factor and an encoding gene thereof.
Plant DREB transcription factor provided by the present invention, name is called DMY-DREB, and it derives from the rice grass, is the protein with one of following amino acid residue sequences:
1) the SEQ ID № in the sequence table: 1;
2) with SEQ ID № in the sequence table: 1 amino acid residue sequence is through replacement and/or disappearance and/or the interpolation and the protein relevant with plant stress tolerance of one or several amino-acid residue.
Wherein, the sequence in the sequence table 1 is made up of 223 amino-acid residues.From the 53rd of the aminoterminal of sequence 1 to the 111st amino acids residue is the AP2/EREBP conserved domain.
The replacement of described one or several amino-acid residue and/or disappearance and/or interpolation are meant replacement and/or the disappearance and/or the interpolation of no more than ten amino-acid residues.
Above-mentioned plant DREB transcription factor encoding gene (DMY-DREB) also belongs to protection scope of the present invention.
The cDNA gene of above-mentioned plant DREB transcription factor can have one of following nucleotide sequence:
1) SEQ ID № in the sequence table: 2 dna sequence dna;
2) SEQ ID № in the code sequence tabulation: the polynucleotide of 1 protein sequence;
3) under the rigorous condition of height can with SEQ ID № in the sequence table: the nucleotide sequence of the 2 dna sequence dnas hybridization that limit.
The rigorous condition of above-mentioned height can be 0.1 * SSPE (or 0.1 * SSC), in the solution of 0.1%SDS, under 65 ℃, hybridize and wash film.
Wherein, the SEQ ID № in the sequence table: 1 is made up of 669 deoxynucleotides, is encoding sequence from the 5 ' 1st to 669 deoxynucleotides.
Contain expression carrier of the present invention, clone, host bacterium and expression cassette and all belong to protection scope of the present invention.
Utilize existing molecular biological method can obtain containing the expression vector of DMY-DREB, as pC2300:Prd29A-dmydrebA (physical map as shown in Figure 1).
Described expression cassette comprises plant promoter, DMY-DREB gene and terminator.
Described plant promoter can be composing type, induction type or tissue-specific promoter.
DMY-DREB gene of the present invention can add any constitutive promoter, tissue-specific promoter or inducible promoter in being building up to plant expression vector the time before its transcription initiation Nucleotide.For the ease of transgenic plant cells or plant being identified and screening, can process employed carrier, as the antibiotic marker thing (gentamicin, kantlex etc.) that adds the alternative mark (gus gene, luciferase genes etc.) of plant or have resistance.By the plant transformed host both can be monocotyledons, it also can be dicotyledons, wherein, monocotyledons can be turfgrass, wheat, barley, oat, paddy rice or corn etc., and dicotyledons can be potato, tobacco, cotton, lettuce, tomato, muskmelon, cucumber, pea, oil grain, beet or Sunflower Receptacle etc.
Carry that the DMY-DREB expression carrier can Ti-plasmids, Ri plasmid, plant viral vector, directly DNA conversion, microinjection, electricity be led, conventional biological method transformed plant cells or tissue such as agriculture bacillus mediated by using, and plant transformed is become plant through tissue cultivating.
Experiment shows that DMY-DREB albumen of the present invention and encoding gene thereof can strengthen plant to adverse circumstance, particularly cold patience of coercing.This gene will be widely used in cultivates the gene plant of anti-the reverse, has far-reaching theory significance and reaches practice significance widely.
Description of drawings
Fig. 1 is a DMY-DREB plant expression vector physical map
Fig. 2 is the PCR checking photo of DMY-DREB transgene tobacco
Fig. 3 is the cold-resistant experimental result of DMY-DREB transgene tobacco
Embodiment
Method among the following embodiment is ordinary method if no special instructions.
The acquisition of embodiment 1, DMY-DREB and encoding gene thereof
With CTAB method (Steiner JJ, Poklemba CJ, Fjellstrom RG, Elliott LF., A rapidone-tube genomic DNA extraction process for PCR and RAPD analyses.NucleicAcids Res.1995 Jul 11; 23 (13): 2569-70.) extract rice grass genomic dna, concrete grammar is as follows: get rice blade of grass sheet, with liquid nitrogen grinding, add 5 milliliters in the vegetable material of per 1 gram through the CTAB of 68 ℃ of preheatings solution (with before adding the 10mM mercaptoethanol), 60 ℃ were heated 30 minutes.Add the equal-volume chloroform then: primary isoamyl alcohol, centrifugal 15 minutes of 15000g gets supernatant.The Virahol that in supernatant liquor, adds 2/3 times of volume, mixing stirs out the DNA silk with the entry needle needle point and to place new pipe, adds 75% ethanol again, and washing precipitation and centrifugal tube wall blot ethanol then, and air is dried remaining ethanol, adding TE dissolving DNA.And in-20 ℃ of preservations.With PCR (polymerase chain reaction) method amplification DMY-DREB gene DNA fragment.Homologous region design degenerate primer at the DREB proteinoid, be listed as follows: F1:5 '-ACCGC (T/C) GAGATGGC (G/A/C) GC (T/G) C-3 ' R1: 5 '-CC (A/G) AG (A/G) CGAGTC (A/C) G (G/C) GAA-3 ' 25ul PCR reaction system is: 1 μ g genomic dna, 1.5mM MgCl
2, 20mM Tris-HCl (pH8.4), 50mM KCl, 0.8mM dNTP mixture, 1 μ M primers F 1 and 1 μ M primer R1, and the Taq polysaccharase of 1 unit (Shen, Shanghai can lottery industry biotech company).In PCR-thermal cycler (Eppendorf company, Germany), carry out PCR circulation with amplification DMY-DREB gene DNA fragment according to following scheme: 94 ℃ of pre-sex change 5 minutes; Again 94 ℃ 1 minute, 60 ℃ 1 minute, 72 ℃ 1 minute, totally 30 circulations; Last 72 ℃ 10 minutes.About 90bpDNA fragment that amplification is obtained reclaims this fragment with the scheme that gel reclaims test kit (sky, Beijing latitude Time Technology company limited) and provides by test kit, with (Promega company of pGEM-Teasy vector system, the U.S.) scheme that provides by this system is cloned this fragment, order-checking (three rich polygala root biotech companies).Sequential analysis shows this sequence and DREB proteinoid gene conservative district fragment height homology.Auele Specific Primer according to this conservative section design 3 ' RACE.Get the total RNA of 1 μ g and carry out 3 ' RACE.The RACE program is carried out according to the specification sheets of 3 ' the RACE test kit that precious biotechnology (Dalian) company limited provides.5 ' the end primer of 3 ' RACE is: GSP1:5 '-CGTGCCTCAACTTCGCGGACT-3 ', 3 ' end primer is provided by test kit.The PCR condition is 94 ℃ of 5min of elder generation; 94 ℃ of 30s → 54 ℃ 30s → 72 ℃ of 30s, totally 30 circulations then; Last 72 ℃ of 10min.3 ' RACE product synthesizes 5 ' RACE primers through cloning and sequencing.Get the total RNA of 1 μ g and carry out 5 ' RACE, the RACE program is carried out according to the specification sheets of 5 ' the RACE test kit that precious biotechnology (Dalian) company limited provides.Wherein, 5 ' the phosphorylation primer of 5 ' RACE is: 5 ' CCCGACGAACCTCCC-3 ', the primer that is used for pcr amplification is respectively S1:5 '-ATCCGCTCCGCTGTGCGTCTCG-3 ', S2:5 '-CCTCGACGGCCACATTGCTC-3 ', A1:5 '-GGACTCGCCTCGGCTGCTCA-3 ', A2:5 '-CAGAGCAATGTGGCCGTCGAG-3 '.With 5 ' the phosphorylation primer carries out reverse transcription reaction, and be that primer carries out first round amplification with S1 and A1, and then make template with the first round PCR product that dilutes 100 times, be that primer carries out second and takes turns pcr amplification with S2 and A2.Wherein, first round PCR reaction conditions: 94 ℃ of 5min of elder generation; 94 ℃ of 30s → 56 ℃ 30s → 72 ℃ of 1min, totally 30 circulations then; Last 72 ℃ of 10min.Second takes turns the PCR reaction conditions: 94 ℃ of pre-sex change 5min of elder generation; 94 ℃ of 30s then, 58 ℃ of 30s, 72 ℃ of 1min, totally 30 circulations; Last 72 ℃ of 10min.
Total RNA extracts as follows: with 4 ℃ of cold pre-treatment rice blade of grass sheets of-70 ℃ of preservations of liquid nitrogen grinding, add 1ml TRIZ0L RNA extracting solution (ancient cooking vessel state biotech company) in the blade of every 0.1g, carry out the extraction of total RNA by the scheme that supplier provides.The RNA that obtains is used the DNase enzyme, and (Prmega America) digests by the scheme that supplier provides, to remove remaining DNA.
Gained 5 '-RACE and 3 '-RACE product is through cloning and sequencing, synthetic primer, is template with reverse transcription test kit (precious biotechnology (Dalian) company limited) by the cDNA that the method reverse transcription of test kit obtains with the total RNA of 1 μ g rice blade of grass sheet, the cDNA of clone DMY-DREB.The synthetic primer sequence is as follows: F2:5 '-GTCGACATGGACATCGGCGCGTTC-3 '; R2:5 '-TCAGTAGCTCCAGAGCCTGA-3 '.The 25ulPCR reaction system contains 1 μ g cDNA, 1.5mM MgCl
2, 20mM Tris-HCl (pH8.4), 50mM KCl, 0.8mM dNTP mixture, 1 μ M primers F 2 and 1 μ M primer R2, and the Pfu enzyme of 1 unit (Shen, Shanghai can lottery industry biotech company).In PCR-thermal cycler (Eppendorf company, Germany), carry out PCR circulation with the cDNA:94 ℃ of pre-sex change of amplification DMY-DREB 5 minutes according to following scheme; Again 94 ℃ 1 minute, 55 ℃ 1 minute, 72 ℃ 1 minute, totally 30 circulations; Last 72 ℃ 10 minutes.
The fragment that amplification is obtained reclaims this fragment with the scheme that gel reclaims test kit (sky, Beijing latitude Time Technology company limited) and provides by test kit, and the EcoR V site of inserting pBluescript II KS (-) (STRATAGENE company), obtain plasmid pBS::dmydreb.Order-checking (three rich polygala root biotech companies), sequencing result shows that this clip size is 669bp, has the nucleotide sequence of sequence 2 in the sequence table; Sequential analysis shows that this cDNA's has the DMY-DREB of the amino acid residue sequence of sequence 1 from the 5 ' 1st to 669 deoxynucleoside acid encodings.The the 157th to the 333rd Nucleotide of 5 ' end from sequence 2 is the encoding sequence of AP2/EREBP conserved domain, 59 amino-acid residues of encoding; DMY-DREB's is the AP2/EREBP conserved domain from the 53rd of the aminoterminal of sequence 1 to the 111st amino acids residue.
The structure of embodiment 2, DMY-DREB plant expression vector and the experiment of the resistance of reverse of transgene tobacco
To extract arabidopsis thaliana genomic dna with the CTAB method is template, with the promoter sequence of Rd29A-S and Rd29A-A PCR method amplification Arabidopis thaliana rd29A gene.Wherein, Rd29A-S:5 '-TCTAGACGCATGATTTGATGGAGGAG-3 ', Rd29A-A:5 '-TCCAAAGATTTTTTTCTTTCCAA-3 '.25ul PCR reaction system contains 100ng genomic dna, 1.5mM MgCl
2, 20mM Tris-HCl (pH8.4), 50mM KCl, 0.8 mM dNTP mixture, 1 μ M Rd29A-S and 1 μ M Rd29A-A, the Pfu enzyme of 1U (Shanghai Shen You biotech company).In PCR-thermal cycler (Eppendorf company, Germany), carry out PCR circulation according to following scheme: earlier 94 ℃ 5 minutes; Again 94 ℃ 1 minute, 54 ℃ 1 minute, 72 ℃ 1 minute, totally 30 circulations; Last 72 ℃ 10 minutes.The long dna fragmentation of the about 944bp that amplification is obtained reclaims this fragment with the scheme that gel reclaims test kit (sky, Beijing latitude Time Technology company limited) and provides by test kit, insert the EcoRV site of pBluescript IIKS (-) (STRATAGENE company), order-checking (three rich polygala root biotech companies), this fragment that shows sequencing result contains the full length sequence of the promotor of Arabidopis thaliana rd29A gene, will contain this segmental plasmid called after pBS::Prd29A.
To extract arabidopsis thaliana genomic dna with the CTAB method is template, with 3 ' end sequence of OS Trps16g2-S and OS Trps16g2-APCR method amplifying rice small subunit ribosome s16 gene.Wherein, OS Trps16g2-S:5 '-ATCGATTCGGAGCGGTGTTCTCT-3 ', OS Trps16g2-A:5 '-GATGGGCATCTGAACTGAGAAT-3 '.25ul PCR reaction system contains 100ng genomic dna, 1.5mM MgCl
2, 20mM Tris-HCl (pH8.4), 50mM KCl, 0.8mM dNTP mixture, 1 μ M OS Trps16g2-S and 1 μ M OS Trps16g2-A, the Pfu enzyme of 1U (Shanghai Shen You biotech company).In PCR-thermal cycler (Eppendorf company, Germany), carry out PCR circulation according to following scheme: earlier 94 ℃ 5 minutes; Again 94 ℃ 1 minute, 54 ℃ 1 minute, 72 ℃ 1 minute, totally 30 circulations; Last 72 ℃ 10 minutes.The long dna fragmentation of the about 764bp that amplification is obtained reclaims this fragment with the scheme that gel reclaims test kit (sky, Beijing latitude Time Technology company limited) and provides by test kit, insert the EcoR V site of pBluescript II II KS (-) (STRATAGENE company), order-checking (three rich polygala root biotech companies), this fragment that shows sequencing result contains 3 ' end sequence of paddy rice small subunit ribosome s16 gene, will contain this segmental plasmid called after pBS::Trps16g2.
Plasmid pBS::Trps16g2 with the ClaI site that the ClaI enzyme downcuts 3 ' the end sequence fragment insertion pBS::dmydreb that contains paddy rice small subunit ribosome s16 gene, is obtained plasmid pBS::dmydrebA-Trps16g2.
Plasmid pBS::dmydrebA-Trps16g2 is downcut the Xho I site that contains dmydrebA-Trps16g2 sequence fragment insertion pBS::Prd29A with Sal I enzyme, obtain plasmid pBS::Prd29A-dmydrebA-Trps16g2.
The dmydrebA expression cassette that will obtain with Xba I digested plasmid pBS::Prd29A-dmydrebA-Trps16g2, be cloned between the Xba I enzyme recognition site of pCAMBIA2300 (available from CAMBIA company), obtain the plant expression vector pC2300::Prd29A-dmydrebA of DMY-DREB, pC2300::Prd29A-dmydrebA plasmid figure as shown in Figure 1, DMY-DREB gene (dmydrebA) is subjected to the regulation and control of rd29A promotor in this carrier.This carrier is transformed agrobacterium tumefaciens lba4404, obtain containing the agrobacterium strains of pC2300::Prd29A-dmydrebA, called after TpC2300::Prd29A-dmydrebA, plasmid pCAMBIA2300 is transformed agrobacterium tumefaciens lba4404, obtain containing the bacterial strain TpCAMBIA2300 of pCAMBIA2300 empty carrier.Tobacco aseptic seedling blade is cut into the leaf dish of the about 1cm of four length of sides with sterile razor blade, be to soak 10 minutes in 1 TpC2300::Prd29A-dmydrebA and TpCAMBIA2300 (changeing the empty carrier contrast) the bacterium liquid in the O.D. value respectively, switching goes into to be total to culture medium, and (the MS minimum medium adds 6-benzyl aminopurine 1.0mg/L, indolylacetic acid 0.1mg/L, sucrose 30g/L, pH 5.2), in 25 ℃ of dark cultivations 3 days, (the MS minimum medium adds 6-benzyl aminopurine 1.0mg/L to change subculture medium again over to, indolylacetic acid 0.1mg/L, Pyocianil 250mg/L, sucrose 30g/L, pH 5.8) in illumination cultivation three days, (the MS minimum medium adds 6-benzyl aminopurine 1.0mg/L to change screening culture medium over to, indolylacetic acid 0.1mg/L, Pyocianil 250mg/L, kantlex 75mg/L, sucrose 30 g/L, pH 5.8); Continue to cultivate 7 days, change regeneration culture medium (the MS minimum medium adds 6-benzyl aminopurine 1.0mg/L, Pyocianil 250mg/L, kantlex 75mg/L, sucrose 30g/L, and pH 5.8) again over to.When treating that blade edge grows the regeneration bud of growing thickly to 2-3cm, regeneration bud is downcut, and (the MS minimum medium adds kantlex 50mg/L, sucrose 20g/L, and pH 5.8 to change root media over to scalper.) the middle cultivation 20 days, obtaining 15 transfer-gen plants and 10 strains altogether changes the empty carrier plant.
With CTAB method (Steiner JJ, Poklemba CJ, Fjellstrom RG, Elliott LF., A rapidone-tube genomic DNA extraction process for PCR and RAPD analyses.NucleicAcids Res.1995 Jul 11; 23 (13): 2569-70.) extract commentaries on classics pC2300::Prd29A-dmydrebA and pCAMBIA2300 tobacco gene group DNA, the genomic dna that extracts with 100ng is that template is carried out the PCR reaction.With primers F among the embodiment 12 and R2 is primer, and 25ul PCR reaction system contains 100ng genomic dna, 1.5mMMgCl
2, 20mM Tris-HCl (pH8.4), 50mM KCl, 0.8mM dNTP mixture, 1 μ M F2 and 1 μ MR2, the Taq polysaccharase of 1U (Shanghai Shen You biotech company).In PCR-thermal cycler (Eppendorf company, Germany), carry out PCR circulation according to following scheme: earlier 94 ℃ 5 minutes; Again 94 ℃ 1 minute, 58 ℃ 1 minute, 72 ℃ 1 minute, totally 30 circulations; Last 72 ℃ 10 minutes.Amplified production is carried out gel electrophoresis, the result as shown in Figure 2, changeing pC2300::Prd29A-dmydrebA tobacco amplified production has band (arrow shows) clearly at the 669bp place that estimates, and the no PCR product band of commentaries on classics pCAMBIA2300 tobacco (empty carrier contrast).Among Fig. 2, swimming lane 1 is for changeing the pcr amplification product of pCAMBIA2300 tobacco; Swimming lane 2 is the pcr amplification product of pC2300::Prd29A-dmydrebA; Swimming lane 3 is for changeing the pcr amplification product of pC2300::Prd29A-dmydrebA tobacco; Swimming lane M is the 1kbDNA molecular weight standard.
Will (the MS minimum medium adds kantlex 50mg/L, sucrose 20g/L, pH5.8 at root media.) in cultivate 20 days commentaries on classics pC2300::Prd29A-dmydrebA tobacco and change pCAMBIA2300 (empty carrier contrast) tobacco and change 4 ℃ of deepfreezes respectively over to after 12 hours, place-10 ℃ to place 2 hours, taking-up is after cultivating 3 days under 25 ℃ of conditions, it is good to change the pC2300::Prd29A-dmydrebA tobacco growing, and it is dead to change pCAMBIA2300 contrast tobacco, as shown in Figure 3, the left side tobacco is for changeing the pC2300::Prd29A-dmydrebA tobacco among Fig. 3, and the right side is for changeing the pCAMBIA2300 tobacco.
Sequence table
<160>2
<210>1
<211>223
<212>PRT
＜213〉rice grass (Spartina anglica)
<400>1
Met?Asp?Ile?Gly?Ala?Phe?Ser?Ser?Asp?Tyr?Ser?Ser?Gly?Thr?Pro?Ser
1 5 10 15
Pro?Val?Gly?Gly?Asp?Asp?Gly?Ser?Ser?Pro?Ser?Tyr?Met?Thr?Val?Ser
20 25 30
Ser?Ala?Pro?Pro?Lys?Arg?Arg?Ala?Gly?Arg?Thr?Lys?Phe?Lys?Glu?Thr
35 40 45
Arg?His?Pro?Val?Tyr?Lys?Gly?Val?Arg?Arg?Arg?Asn?Pro?Gly?Arg?Trp
50 55 60
Val?Cys?Glu?Val?Arg?Glu?Pro?His?Gly?Lys?Gln?Arg?Ile?Trp?Leu?Gly
65 70 75 80
Thr?Phe?Glu?Thr?Ala?Glu?Met?Ala?Ala?Arg?Ala?His?Asp?Val?Ala?Ala
85 90 95
Met?Ala?Leu?Arg?Gly?Arg?Gly?Ala?Cys?Leu?Asn?Phe?Ala?Asp?Ser?Pro
100 105 110
Arg?Leu?Leu?Arg?Val?Pro?Pro?Val?Gly?Ala?Gly?His?Asp?Glu?Ile?Arg
115 120 125
Arg?Ala?Ala?Ala?Glu?Ala?Ala?Glu?Gln?Phe?Arg?Pro?Ala?Pro?Asp?Gln
130 135 140
Ser?Asn?Val?Ala?Val?Glu?Glu?Pro?Val?Ala?Thr?Leu?Pro?Asp?Pro?Phe
145 150 155 160
Ser?Ser?Glu?Thr?His?Ser?Gly?Ala?Asp?His?Arg?Pro?Tyr?Cys?Gly?Met
165 170 175
Val?Asp?Asp?Arg?Phe?Asp?Leu?Gly?Met?Gln?Gly?Tyr?Leu?Asp?Met?Ala
180 185 190
Gln?Gly?Met?Leu?Ile?Asp?Pro?Pro?Pro?Met?Gly?Gly?Ser?Ser?Gly?Asn
195 200 205
Gly?Gly?Asp?Asp?Asp?Asp?Gly?Gly?Glu?Val?Arg?Leu?Trp?Ser?Tyr
210 215 220
<210>2
<211>669
<212>DNA
＜213〉rice grass (Spartina anglica)
<400>2
atggacatcg?gcgcgttcag?cagcgactac?tcctcgggca?cgccgtcccc?ggtcggcggc 60
gacgatggct?cctccccctc?ctacatgacg?gtgtcgtcgg?cgccgcccaa?gcggcgcgcc 120
ggccggacca?agttcaagga?gacgcggcac?ccggtgtaca?agggcgtgcg?ccggcgtaac 180
cctgggcggt?gggtctgcga?ggtgcgggag?ccgcacggca?agcagcgaat?ttggctcggg 240
acattcgaga?ccgccgagat?ggcggcgcgc?gcgcatgacg?tcgccgcgat?ggcgctccga 300
gggcgcggcg?cgtgcctcaa?cttcgcggac?tcgcctcggc?tgctcagggt?cccgcccgtg 360
ggcgcagggc?acgacgagat?tcgcagggcc?gcggccgagg?cggccgagca?gttccgcccc 420
gcgcccgatc?agagcaatgt?ggccgtcgag?gagccggtcg?cgacactacc?ggacccgttt 480
tctagcgaga?cgcacagcgg?agcggatcac?cgcccgtatt?gcggcatggt?agacgacagg 540
ttcgacttgg?ggatgcaggg?gtacctcgac?atggcgcaag?ggatgctcat?tgacccgcca 600
ccgatgggag?gttcgtcggg?gaatggtggt?gacgatgatg?acggcggtga?ggtcaggctc 660
tggagctac 669
Claims (10)
1, a kind of plant DREB transcription factor is the protein with one of following amino acid residue sequences:
1) the SEQ ID № in the sequence table: 1;
2) with SEQ ID № in the sequence table: 1 amino acid residue sequence is through replacement and/or disappearance and/or the interpolation and the amino acid residue sequence relevant with plant stress tolerance of one or several amino-acid residue.
2, plant DREB transcription factor according to claim 1 is characterized in that: described protein has the SEQ ID № in the sequence table: 1 amino acid residue sequence.
3, the encoding gene of claim 1 or 2 described plant DREB transcription factors.
4, gene according to claim 3 is characterized in that: the cDNA gene of described plant DREB transcription factor has one of following nucleotide sequence:
1) SEQ ID № in the sequence table: 2 dna sequence dna;
2) SEQ ID № in the code sequence tabulation: the polynucleotide sequence of 1 protein sequence;
3) under the rigorous condition of height can with SEQ ID № in the sequence table: the nucleotide sequence of the 2 dna sequence dnas hybridization that limit.
5, contain claim 3 or 4 described expression carrier.
6, the clone that contains claim 3 or 4 described genes.
7, the host bacterium that contains claim 3 or 4 described genes.
8, contain claim 3 or 4 described expression of gene boxes.
9, claim 1 or 2 described plant DREB transcription factors and encoding gene thereof the application in cultivating the resistance of reverse plant.
10, application according to claim 9 is characterized in that: described resistance of reverse is a winter hardiness.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005101177300A CN1313486C (en) | 2005-11-08 | 2005-11-08 | Plant DREB transcription factor and its coding gene and use |
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|---|---|---|---|
| CNB2005101177300A CN1313486C (en) | 2005-11-08 | 2005-11-08 | Plant DREB transcription factor and its coding gene and use |
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| Publication Number | Publication Date |
|---|---|
| CN1775803A CN1775803A (en) | 2006-05-24 |
| CN1313486C true CN1313486C (en) | 2007-05-02 |
Family
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| CN101684150B (en) * | 2009-07-24 | 2011-12-21 | 山东省农业科学院高新技术研究中心 | DREB transcription factor for regulating peanut stress tolerance and application thereof |
| CN103483436B (en) * | 2013-08-27 | 2015-01-07 | 西北农林科技大学 | Common buckwheat DREB (Dehydration Responsive Element Binding protein) transcription factor |
| CN106749572B (en) * | 2016-12-08 | 2019-11-12 | 中国农业科学院蔬菜花卉研究所 | Participate in the transcription factor and its application of regulation muskmelon bitter principle synthesis |
| CN112111508A (en) * | 2019-06-20 | 2020-12-22 | 新疆农业科学院核技术生物技术研究所（新疆维吾尔自治区生物技术研究中心） | Cotton stress-tolerant gene GhCBF and coding protein and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1472222A (en) * | 2002-07-29 | 2004-02-04 | 清华大学 | DREB transcription factor of corn and its encoding genes and use |
| CN1478893A (en) * | 2002-08-29 | 2004-03-03 | 中国科学院遗传与发育生物学研究所 | Paddy rice anti-reverse transcripfactor and its coding gene and application |
| CN1491960A (en) * | 2002-08-29 | 2004-04-28 | 清华大学 | Rice DREB transcription factor and its encoding gene and use |
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2005
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1472222A (en) * | 2002-07-29 | 2004-02-04 | 清华大学 | DREB transcription factor of corn and its encoding genes and use |
| CN1478893A (en) * | 2002-08-29 | 2004-03-03 | 中国科学院遗传与发育生物学研究所 | Paddy rice anti-reverse transcripfactor and its coding gene and application |
| CN1491960A (en) * | 2002-08-29 | 2004-04-28 | 清华大学 | Rice DREB transcription factor and its encoding gene and use |
Non-Patent Citations (4)
| Title |
|---|
| DREB转录因子及其在植物抗逆中的作用 王少峡，王振英，彭永康，植物生理学通迅，第40卷第1期 2004 * |
| DREB转录因子及其在植物抗逆中的作用 王少峡，王振英，彭永康，植物生理学通迅，第40卷第1期 2004;KYamaguch-Shinoxaki,KShinozaki DREB 转录因子在提高植物抗逆性中的作用 刘强，赵南明，科学通报，第45卷第1期 2000;Prd29A及DREB1A的克隆和干旱诱导型植物表达载体的构建与鉴定 押辉远 秦广雍 霍裕平，植物生理学通讯，第41卷第3期 2005 * |
| KYamaguch-Shinoxaki,KShinozaki DREB 转录因子在提高植物抗逆性中的作用 刘强，赵南明，科学通报，第45卷第1期 2000 * |
| Prd29A及DREB1A的克隆和干旱诱导型植物表达载体的构建与鉴定 押辉远 秦广雍 霍裕平，植物生理学通讯，第41卷第3期 2005 * |
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