CN1583789A - Soybean transcripting factor, its coding gene and use thereof - Google Patents
Soybean transcripting factor, its coding gene and use thereof Download PDFInfo
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- CN1583789A CN1583789A CN 03153943 CN03153943A CN1583789A CN 1583789 A CN1583789 A CN 1583789A CN 03153943 CN03153943 CN 03153943 CN 03153943 A CN03153943 A CN 03153943A CN 1583789 A CN1583789 A CN 1583789A
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- 125000000539 amino acid group Chemical group 0.000 claims abstract description 13
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- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 13
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- 230000002103 transcriptional effect Effects 0.000 claims description 5
- GOJUJUVQIVIZAV-UHFFFAOYSA-N 2-amino-4,6-dichloropyrimidine-5-carbaldehyde Chemical group NC1=NC(Cl)=C(C=O)C(Cl)=N1 GOJUJUVQIVIZAV-UHFFFAOYSA-N 0.000 claims description 4
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Abstract
The invention relates to a soybean transcriptional control factor and coded protein and application. The soybean transcriptional control factor GmDREBa has the protein of amino acid residue sequence in sequence map's sequence 2, or is making sequence 2's amino acid residue sequence through one or several amino acid residue sequence substituted or lacking or adding, and has the same activity with sequence 2's amino acid residue sequence and it's derived from sequence 2. This invention also provide soybean transcriptional control factor GmDREBa's coded gene. The soybean transcriptional control of factorGmDREBa's coded gene is important for cultivating adverse tolerance plant breed variety especially for adverse tolerance soybean breed variety, and for increasing output of crop especially soybean.
Description
Technical field
The present invention relates to the EREBP/AP2 class transcription factor relevant and encoding gene and application in the plant genetic engineering field, particularly derive from the EREBP/AP2 class transcription factor relevant and encoding gene and the application of soybean with resistance of reverse with resistance of reverse.
Background technology
The variation of physical chemical factor in the environment, the factors of coercing such as for example arid, saline and alkaline, low temperature have material impact to growth and development of plant, can cause the extensive underproduction of farm crop when serious, and cultivating the resistance of reverse crop is one of major objective of plant husbandry.At present, the using gene engineering breeding has become one of important method that strengthens the crop resistance of reverse.Higher plant cell has number of ways to reply various environment stresses in the environment.Wherein transcription factor plays a part the expression of the regulation and control effector of anti-the retrocorrelation.
Soybean is important oil crops, is the main source of plant protein, understands fully its anti-contrary mechanism, and then improves its resistance of reverse, has important theory and realistic meaning.
The innovation and creation content
The purpose of this invention is to provide a kind of soybean transcriptional regulator and the encoding gene thereof relevant with resistance of reverse.
Soybean transcriptional regulator name provided by the present invention is called GmDREBa, be protein, or the amino acid residue sequence of sequence 2 is passed through replacement, disappearance or the interpolation of one or several amino-acid residue and has identical active by sequence 2 deutero-protein with the amino acid residue sequence of sequence 2 with sequence 2 amino acid residue sequences in the sequence table.
The protein that sequence 2 amino acid residue sequences are made up of 158 amino-acid residues in the sequence table is the EREBP/AP2 class transcription factor in the soybean.Containing conservative AP2 sequence from the nitrogen end to carbon teminal 13-76 amino acids residue in the sequence 2, is the conservative territory of EREBP/AP2 function of GmDREBa encoding gene.
The encoding gene of GmDREBa is one of following nucleotide sequences:
1) dna sequence dna of sequence 1 in the sequence table;
2) dna sequence dna of sequence 2 protein sequences in the code sequence tabulation;
3) with sequence table in the dna sequence dna that limits of sequence 1 have 90% above homology, and the identical function protein DNA sequence of encoding.
The dna sequence dna of sequence 1 is by 846 based compositions in the sequence table, the reading frame of this gene is to hold the 140th to the 616th bit base from 5 ', comprise 477 base pairs, have 230 base pair non-translational regions, its expression to be subjected to inducing of ABA, low temperature, high salt and arid at 3 ' end.
Utilize any carrier that can guide foreign gene to express in plant, the encoding gene importing vegetable cell with GmDREBa provided by the present invention can obtain transgenic cell line and transfer-gen plant that the environment stress tolerance is enhanced.When using gene constructed plant expression vector of the present invention, before its transcription initiation Nucleotide, can add any enhancing promotor or inducible promoter.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.From the security consideration of transgenic plant, can not add any selected marker, directly with adverse circumstance screening transformed plant.Carry GmDREBa of the present invention expression vector 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 the plant transformed tissue cultivating is become plant.By the plant transformed host both can be monocotyledons, also can be dicotyledons, as: paddy rice, wheat, corn, cucumber, tomato, willow, turfgrass, lucerne place etc.Gene pairs of the present invention is cultivated the particularly anti-contrary soybean varieties of plant with adverse resistance kind, and particularly soybean yields is significant to improve farm crop.
The present invention will be further described below in conjunction with drawings and Examples.
Description of drawings
Fig. 1 is that the similar proteic similarity with other plant of GmDREBa albumen compares
Fig. 2 is the Northern results of hybridization
Embodiment
The screening of embodiment 1, soybean GmDREBa encoding gene and the clone of cDNA thereof
Carry out the BLAST retrieval in the soybean est database, screen 3 DREB genoids, these genes all do not contain intron.From big pulse family rich 1, extract total DNA with ordinary method,, obtain 3 dna fragmentations through the PCR method amplification respectively, show the DREB proteinoid that they are all encoded and contain the EREBP/AP2 conserved structure through sequential analysis according to 3 pairs of primers of sequences Design of 3 dreb genes.GmDREBa is that primer amplification obtains with 5 '-CCACAATAAGGGGGATGGAT and 5 '-CGGAACGTACTATCTACTGA.GmDREBa is made up of 846 base pairs, reading frame is 5 ' end the 140th to 616 bit base, the protein of being made up of 158 amino-acid residues of encoding is wherein from the conservative territory of EREBP/AP2 function that 13-76 amino-acid residue conserved domain of nitrogen end is this gene.
Similar proteic comparison in embodiment 2, GmDREBa and the plant
The similarity of table 1 GmDREBa and isoformgene and diversity percentage ratio
| ??GmDREBa | ???AtDREB2A | ??AtDREB2B | ??GmDREB1 | ???OsDREB2 | |
| ?GmDREBa | ??*** | ???45.3 | ??39.6 | ??28.3 | ??42.1 |
| ?AtDREB2A | ??76.5 | ???*** | ??45.2 | ??30.5 | ??33.9 |
| ?AtDREB2B | ??83.5 | ???51.7 | ??*** | ??26.4 | ??31 |
| ?GmDREB1 | ??83.1 | ???124 | ??0 | ??*** | ??29.3 |
| ?OsDREB2 | ??77.9 | ???106.2 | ??114.7 | ??112.6 | ??*** |
Same proteinoid OsDREB2, AtDREB2A, AtDREB2B and the GmDREB1 of GmDREBa aminoacid sequence and other plant such as paddy rice, Arabidopis thaliana and soybean are compared, the result shows that its similarity is respectively 42.1,45.3,39.6 and 28.3% shown in Fig. 1 and table 1.
Embodiment 3, the GmDREBa expression characteristic under environment stress
Beans 23 seed kinds in soybean Shanxi in basin, are got seedling after 2 week of growth and are carried out the following various processing of coercing respectively:
Salt is handled: soybean seedling is moved in the 250mM NaCl solution.
ABA handles: soybean seedling is moved in the 100uM ABA solution.
Osmotic stress is handled: soybean seedling is moved in the 20%PEG solution.
Subzero treatment: soybean seedling is moved in 4 ℃ of aqueous solution.
Illumination cultivation, and respectively in the sampling in 0,0.5,1,3,6,12 and 24 hour of various processing back.Collect fresh blade 1g and grind in liquid nitrogen, be suspended from the 4mol/L sulphur hydracid guanidine, mixture adds dehydrated alcohol and precipitates total RNA with acid phenol, chloroform extracting in the supernatant, water-soluble afterwards, obtains total RNA.Be that probe carries out the Northern analysis according to a conventional method with GmDREBa DNA respectively.The result shows that GmDREB1 is induced by 100uM ABA, 250mM NaCl, low temperature (4 ℃) and osmotic stress obviously as shown in Figure 2.A handles for 250mM NaCl among Fig. 2; B is that osmotic stress is handled; C is a subzero treatment; D is that 100uMABA handles.
Sequence table
<160>2
<210>1
<211>846
<212>DNA
＜213〉Glycine soybean (Glycine max (L.) Merrill)
<400>1
ccacaataag?ggggatggat?ctaagtccct?ggccgataca?ctggcgaaat?ggaaagaata?????60
taatgcctgg?ctggagtcta?acaatgaagc?tgagaagccg?gttaggaagg?tccctgccaa????120
gggatcaaag?aagggatgta?tgaaaggcaa?aggaggacct?gagaacttgc?gctgtaatta????180
cagaggagtt?aggcaaagga?catggggaaa?atgggttgct?gaaatccgag?agccaaacag????240
aggaagtagg?ctctggttgg?gtacttttcc?tactgccatt?agcgctgctc?ttgcttatga????300
tgaagcagcg?atggcaatgt?atggtttctg?tgcacgcctc?aactttccca?atgttcaagt????360
ttcaactttt?tccgaggaac?cgtctagaaa?ttctccagct?gctgcttacc?agtcaagaaa????420
ttctccatct?gctaaagaat?ccggttctgc?gttggtgata?ttagagaggt?ctgagtgcat????480
gatgttgtgg?aacaattctg?gtggagatgc?agcagaggat?gatggcatgg?aagacctttc????540
cttatcctta?agtgtgaaac?atgaggaagg?ggaggatgaa?tcagggacca?gttcttccta????600
tctttcattg?tcttgatgta?tggtttgcat?actctgattt?ggccgtggct?ggaaatcata????660
gccttcatag?aggtggatga?ttagcttagg?atgaacgaat?cttgatatta?gtacctggag????720
attagctgtt?gtaaaattga?cttggttgag?aagtgttcca?ttcttcagga?attgacctaa????780
tgcaatctgg?atatccagtc?aagttgtaag?atgtgaaatg?tattttgcct?tgcatgatag????840
atgact???????????????????????????????????????????????????????????????846
<210>2
<211>158
<212>PRT
＜213〉Glycine soybean (Glycine max (L.) Merrill)
<400>2
Met?Lys?Gly?Lys?Gly?Gly?Pro?Glu?Asn?Leu?Arg?Cys?Asn?Tyr?Arg?Gly
1???????????????5???????????????????10??????????????????15
Val?Arg?Gln?Arg?Thr?Trp?Gly?Lys?Trp?Val?Ala?Glu?Ile?Arg?Glu?Pro
20??????????????????25??????????????????30
Asn?Arg?Gly?Ser?Arg?Leu?Trp?Leu?Gly?Thr?Phe?Pro?Thr?Ala?Ile?Ser
35??????????????????40??????????????????45
Ala?Ala?Leu?Ala?Tyr?Asp?Glu?Ala?Ala?Met?Ala?Met?Tyr?Gly?Phe?Cys
50??????????????????55??????????????????60
Ala?Arg?Leu?Asn?Phe?Pro?Asn?Val?Gln?Val?Ser?Thr?Phe?Ser?Glu?Glu
65??????????????????70??????????????????75??????????????????80
Pro?Ser?Arg?Asn?Ser?Pro?Ala?Ala?Ala?Tyr?Gln?Ser?Arg?Asn?Ser?Pro
85??????????????????90??????????????????95
Ser?Ala?Lys?Glu?Ser?Gly?Ser?Ala?Leu?Val?Ile?Leu?Glu?Arg?Ser?Glu
100?????????????????105?????????????????110
Cys?Met?Met?Leu?Trp?Asn?Asn?Ser?Gly?Gly?Asp?Ala?Ala?Glu?Asp?Asp
115?????????????????120?????????????????125
Gly?Met?Glu?Asp?Leu?Ser?Leu?Ser?Leu?Ser?Val?Lys?His?Glu?Glu?Gly
130?????????????????135?????????????????140
Glu?Asp?Glu?Ser?Gly?Thr?Ser?Ser?Ser?Tyr?Leu?Ser?Leu?Ser
145?????????????????150?????????????????155
Claims (10)
1, soybean transcriptional regulator GmDREBa, be protein, or the amino acid residue sequence of sequence 2 is passed through replacement, disappearance or the interpolation of one or several amino-acid residue and has identical active by sequence 2 deutero-protein with the amino acid residue sequence of sequence 2 with sequence 2 amino acid residue sequences in the sequence table.
2, soybean transcriptional regulator GmDREBa according to claim 1 is characterized in that: it is the protein with sequence 2 amino acid residue sequences in the sequence table.
3, soybean transcriptional regulator GmDREBa according to claim 2 is characterized in that: in the described sequence 2, be the conservative territory of EREBP/AP2 function of described GmDREBa encoding gene from the 13rd at nitrogen end to the 76th amino acids residue conserved domain.
4, the encoding gene of GmDREBa is one of following nucleotide sequences:
1) dna sequence dna of sequence 1 in the sequence table;
2) dna sequence dna of sequence 2 protein sequences in the code sequence tabulation;
3) with sequence table in the dna sequence dna that limits of sequence 1 have 90% above homology, and the identical function protein DNA sequence of encoding.
5, gene according to claim 4 is characterized in that: the encoding gene of described GmDREBa is the dna sequence dna of sequence 1 in the sequence table.
6, gene according to claim 5 is characterized in that: the reading frame of described dna sequence dna is for holding the 140th to the 616th bit base from 5 '.
7, contain the described expression carrier of claim 4.
8, the clone that contains the described gene of claim 4.
9, the application of the described gene of claim 4 in cultivating the plant with adverse resistance kind.
10, application according to claim 9 is characterized in that: the application of described gene in cultivating anti-contrary soybean varieties.
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| CN 03153943 CN1286849C (en) | 2003-08-21 | 2003-08-21 | Soybean transcripting factor, its coding gene and use thereof |
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| Application Number | Priority Date | Filing Date | Title |
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| CN 03153943 CN1286849C (en) | 2003-08-21 | 2003-08-21 | Soybean transcripting factor, its coding gene and use thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100349916C (en) * | 2005-11-22 | 2007-11-21 | 中国科学院遗传与发育生物学研究所 | PHD transcription factor of soybean and its coding gene and usage |
| CN100419080C (en) * | 2005-02-25 | 2008-09-17 | 中国科学院遗传与发育生物学研究所 | Soyabean thermal excited transcryption factor, its coding gene and application |
| CN1919867B (en) * | 2006-08-09 | 2010-06-09 | 中国科学院遗传与发育生物学研究所 | Soybean Trihelix transcription factor, encode gene and application thereof |
| CN1919866B (en) * | 2006-08-09 | 2010-06-09 | 中国科学院遗传与发育生物学研究所 | Soybean Trihelix transcription factor, encode gene and application thereof |
| CN102234321A (en) * | 2010-04-27 | 2011-11-09 | 中国农业科学院作物科学研究所 | Plant stress-tolerant associated protein GmNF-YB1 and encoding gene and application thereof |
| CN102234319A (en) * | 2010-04-27 | 2011-11-09 | 中国农业科学院作物科学研究所 | Plant stress tolerance related protein GmTPRPK1, encoding gene thereof, and application thereof |
| CN103588867A (en) * | 2012-08-16 | 2014-02-19 | 中国科学院遗传与发育生物学研究所 | Soybean transcription factor GmMYB174a, and coding gene and applications thereof |
-
2003
- 2003-08-21 CN CN 03153943 patent/CN1286849C/en not_active Expired - Fee Related
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100419080C (en) * | 2005-02-25 | 2008-09-17 | 中国科学院遗传与发育生物学研究所 | Soyabean thermal excited transcryption factor, its coding gene and application |
| CN100349916C (en) * | 2005-11-22 | 2007-11-21 | 中国科学院遗传与发育生物学研究所 | PHD transcription factor of soybean and its coding gene and usage |
| CN1919867B (en) * | 2006-08-09 | 2010-06-09 | 中国科学院遗传与发育生物学研究所 | Soybean Trihelix transcription factor, encode gene and application thereof |
| CN1919866B (en) * | 2006-08-09 | 2010-06-09 | 中国科学院遗传与发育生物学研究所 | Soybean Trihelix transcription factor, encode gene and application thereof |
| CN102234321A (en) * | 2010-04-27 | 2011-11-09 | 中国农业科学院作物科学研究所 | Plant stress-tolerant associated protein GmNF-YB1 and encoding gene and application thereof |
| CN102234319A (en) * | 2010-04-27 | 2011-11-09 | 中国农业科学院作物科学研究所 | Plant stress tolerance related protein GmTPRPK1, encoding gene thereof, and application thereof |
| CN102234321B (en) * | 2010-04-27 | 2013-05-29 | 中国农业科学院作物科学研究所 | Plant stress-tolerant associated protein GmNF-YB1 and encoding gene and application thereof |
| CN102234319B (en) * | 2010-04-27 | 2013-09-18 | 中国农业科学院作物科学研究所 | Plant stress tolerance related protein GmTPRPK1, encoding gene thereof, and application thereof |
| CN103588867A (en) * | 2012-08-16 | 2014-02-19 | 中国科学院遗传与发育生物学研究所 | Soybean transcription factor GmMYB174a, and coding gene and applications thereof |
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| Publication number | Publication date |
|---|---|
| CN1286849C (en) | 2006-11-29 |
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