CN1597949A - Dry resisting related gene and its coding protein and application - Google Patents
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- CN1597949A CN1597949A CN 03157198 CN03157198A CN1597949A CN 1597949 A CN1597949 A CN 1597949A CN 03157198 CN03157198 CN 03157198 CN 03157198 A CN03157198 A CN 03157198A CN 1597949 A CN1597949 A CN 1597949A
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Abstract
本发明公开了一种拟南芥抗旱相关基因及其编码蛋白与应用。本发明所提供的拟南芥抗旱相关基因ADT，是下列核苷酸序列之一：1)序列表中SEQ ID №：1的DNA序列；2)编码序列表中SEQ ID №：2蛋白质序列的多核苷酸；3)与序列表中SEQ ID№：1限定的DNA序列具有90％以上同源性，且编码相同功能蛋白质的DNA序列。本发明所提供的拟南芥抗旱相关基因ADT编码蛋白ADT，是具有序列表中SEQ ID №：2氨基酸残基序列的蛋白质，或者是将SEQ ID №：2的氨基酸残基序列经过一个或几个氨基酸残基的取代、缺失或添加且具有与SEQ ID №：2的氨基酸残基序列相同活性的由SEQ ID №：2衍生的蛋白质。本发明的基因可广泛用于培育抗旱植物品种。The invention discloses an Arabidopsis drought-resistance-related gene, its encoded protein and its application. The Arabidopsis drought-resistance-related gene ADT provided by the present invention is one of the following nucleotide sequences: 1) the DNA sequence of SEQ ID No.: 1 in the sequence listing; 2) the protein sequence of SEQ ID No.: 2 in the coding sequence listing Polynucleotide; 3) A DNA sequence that has more than 90% homology with the DNA sequence defined by SEQ ID No.: 1 in the sequence listing and encodes the same functional protein. The Arabidopsis thaliana drought-resistance-related gene ADT encoding protein ADT provided by the present invention is a protein having the amino acid residue sequence of SEQ ID No.: 2 in the sequence table, or the amino acid residue sequence of SEQ ID No.: 2 is passed through one or several A protein derived from SEQ ID No.: 2 having the same activity as the amino acid residue sequence of SEQ ID No.: 2 by substitution, deletion or addition of amino acid residues. The gene of the invention can be widely used to breed drought-resistant plant varieties.
Description
Technical field
The present invention relates to a kind of gene related to drought tolerance and proteins encoded and application in the bioengineering field, particularly a kind of gene related to drought tolerance of Arabidopis thaliana and proteins encoded and application.
Background technology
Food problem is one of several hang-ups of facing of the world today.By improve per unit area yield or enlarge cultivated area, the approach such as improve the low and medium-yield farmland of increasing investment in agriculture increases grain yield and all can run into and need overcome the adverse circumstance restriction or alleviate the problem of adverse circumstance harm.Therefore, the understanding plant is improved the resistance of plant to the reaction mechanism of adverse circumstance, has become the further important foundation research of volume increase of agricultural, and extremely countries in the world government and scientist's concern also is the focus of current life science.
Along with the mankind's Economic development and population expansion, the shortage of water resources phenomenon is on the rise, and has directly caused the expansion of arid area and increasing the weight of of climate arid degree, and climate arid trend has become the problem that the whole world is paid close attention to.In the face of serious day by day global climate arid trend, seek plant particularly farm crop drought resisting approach be very necessary.
Arabidopis thaliana is a kind of typical model plant, and its effect is suitable with experimental mouse, is widely used in plant genetics, developmental biology and molecular biological research.Most of genes of Arabidopis thaliana can both find in other plant, and any discovery of relevant Arabidopis thaliana can both be applied to other plant research, and the expert points out, will help scientist to find the method for raising crop yield to the research of Arabidopis thaliana.
Arabidopis thaliana has about 1.3 hundred million base pairs, 2.5 ten thousand genes.The function of most gene is not also known at present, utilizes mutating technology research gene function to become a kind of effective ways.For dicotyledons (mainly being Arabidopis thaliana), it is main path that T-DNA inserts sudden change, and has obtained a large amount of T-DNA insertion mutant.By research, known the function of some adverse circumstance genes, as AB13 (increase frost resistance) Steponkus to mutant, 1998), COR15a (cold induced gene) (Steponkus, 1998), CBF (resistance to cold) (Jaglo-Ottosen, 1998), DREB (increasing anti-morning, anti-cold, salt resistance) (Kasuga, 1999), Apx1 (thermotolerance) (Shi, 2001), AtNHX1 (anti-sylvite) (Yokoi, 2002) etc.
At present, also do not find effective anti-drought gene, in the face of serious day by day arid problem, seeking of anti-drought gene and gene related to drought tolerance is imperative.
The innovation and creation content
The purpose of this invention is to provide a kind of gene related to drought tolerance and proteins encoded thereof.
Gene related to drought tolerance provided by the present invention, name is called ADT (Arabidopsis Drought Tolerance), derives from the ecotypic Arabidopis thaliana of Colombia (Columbia), is one of following nucleotide sequences:
1) SEQ ID № in the sequence table: 1 dna sequence dna;
2) SEQ ID № in the code sequence tabulation: the polynucleotide of 2 protein sequences;
3) with sequence table in SEQ ID №: 1 dna sequence dna that limits has 90% above homology, and the identical function protein DNA sequence of encoding.
The dna sequence dna of sequence 1 is by 1164 based compositions in the sequence table, and the reading frame of this gene is from 5 ' end the 1st to the 1164th bit base, does not contain intron.
Arabidopis thaliana gene related to drought tolerance ADT proteins encoded ADT, be to have SEQ ID № in the sequence table: the protein of 2 amino acid residue sequences, or with SEQ ID №: 2 amino acid residue sequence is through replacement, disappearance or the interpolation of one or several amino-acid residue and have the № with SEQ ID: 2 amino acid residue sequence is identical active by SEQID №: 2 deutero-protein.
The protein that sequence 2 amino acid residue sequences are made up of 387 amino-acid residues in the sequence table.
Contain expression carrier of the present invention and clone and all belong to protection scope of the present invention.
Utilize any carrier that can guide foreign gene in plant, to express, the antisense of ADT gene provided by the present invention is imported vegetable cell, perhaps in other plant, find with ADT homologous gene and knock out then, perhaps by the method for RNAi the ADT gene is kept silent, plant just shows as drought resisting.For the ease of transgenic plant cells or plant being identified and screening, can process employed carrier, as the antibiotic marker thing that adds the alternative mark of plant or have resistance.By the plant transformed host both can be monocotyledons, also can be dicotyledons.Gene of the present invention can be widely used in the cultivation drought resistant plant variety.
The invention will be further described below in conjunction with specific embodiment.
Description of drawings
Fig. 1 is fluorescence real-time quantitative PCR (Real-Time PCR) histogram of ADT
Fig. 2 is the RT-PCR electrophoretogram of wild-type and mutant S29
Fig. 3 is the Southern analytical results electrophoretogram of wild-type and mutant S29
Fig. 4 is the Northern analytical results electrophoretogram of wild-type and mutant S29
Fig. 5 a is the excised leaf photo of wild-type and mutant S29
Fig. 5 b is wild-type and the photo of mutant S29 in the MS+2.5% glycerin medium
Fig. 5 c is the photo under wild-type and the potted plant normal condition of mutant S29
Fig. 5 d is the photo under wild-type and the potted plant drought condition of mutant S29
Embodiment
The clone of embodiment 1, ADT
Arabidopsis mutant body (what have is heterozygote, and what have is homozygote) and Colombia's wild type seeds process sterilization that T-DNA after 60 screenings inserts, 4 ℃ of vernalization are seeded in after three days in the soil (nutrition soil and sole of the foot stone are even with 1: 1 mixed).At 22 ℃ of temperature, intensity of illumination 120umol/m
2/ s (light dark period 16h/8h), relative humidity remain under the 60%-70% condition of (seedling keeps the relative humidity of 80-90% period) and cultivate, during water with the PEG+B5 nutritive medium always, phenotype is observed in the back all around.Found that mutant S29 compares with wild-type, shows as drought resisting.The corresponding gene of mutant S29 is further verified (Real-Time PCR, RT-PCR, Southern, Northern, phenotype are observed), last called after ADT.
Expression analysis one fluorescence real-time quantitative PCR (Real-Time PCR) of embodiment 2, ADT
Used Arabidopis thaliana material is at substratum MS+0 among the embodiment, MS+3.0%PEG, MS+3.5%PEG, (PEG-6000 is as the osmotic stress agent for each 100-200mg of seedling around the last growth of MS+4.0%PEG, but the people is for causing drought environment) (Plant Physiol, 1973), extract total RNA, detect the integrity of RNA through 1% agarose electrophoresis with Plant RNeasy test kit (QIAGEN).Use TaqMan Reverse TranscriptionRegents test kit (Applied Biosystems) to carry out reverse transcription, it is 1 that resultant cDNA sample dilutes respectively, 0.5, with three concentration gradients of 0.1ng/ μ l, respectively get a microlitre sample and detect, each concentration is established three repetitions in experiment.PRIMEREXPRESS software (Applied Biosystems) is used in the design of gene specific primer.SYBR Green Master mix and ABI 7900 sequence detection systems (Applied Biosystems) are used in PCR reaction and detection thereof respectively.Comparative C is adopted in the processing of data
TMethod (User Bulletin #2, ABI PRISM 7700 sequence detection systems).As internal reference data are carried out normalization method with 18S rRNA.
The result as shown in Figure 1, as can be seen from the figure, along with PEG concentration increases, the expression of ADT reduces gradually, compares with contrast and reduces by 2.52 times, 9.81 times, 22.07 times and 43.44 times respectively, illustrates that ADT is a kind of suppressor gene.
The RT-PCR of embodiment 3, ADT and mutant
1) preparation of mutant S29
According to ordinary method, behind T-DNA insertion wild-type Arabidopis thaliana ADT gene, the name of the homozygous mutation body Arabidopis thaliana that obtains is called S29.
2) RT-PCR of wild-type and mutant S29
Each 100-200mg of seedling around the growth of MS substratum is a material with wild-type Arabidopis thaliana and two kinds of mutant S29 Arabidopis thaliana, extracts total RNA of Arabidopis thaliana respectively as embodiment 1.Check the integrity of RNA through 1.0% agarose gel electrophoresis.Ss cDNA's (single strand cDNA, strand cDNA) is synthetic according to SMART
TMThe method of PCR cDNA Library Construction Kit (CLONTECH), synthesizing single-stranded (single strand) cDNA.
With 10 times of synthetic strand cDNA dilutions, as the template of following PCR reaction: 20 μ l systems include 10 * PCR damping fluid, 2 μ l, 2.5mM dNTP mix 1.6 μ l, each 1.0 μ l of the primer 1 of 5 μ M and primer 2, TAQ enzyme (15U/ μ l) 0.1 μ l.PE9600 9700 or MJ PCR instrument on increase: 94 ℃ of pre-sex change 3min, 94 ℃ of 1min, 55 ℃ of 1min, 72 ℃ of 1min amount to 34 circulations; 72 ℃ are extended 10min, with the PCR product that obtains, make 1% agarose gel electrophoresis.Primer 1:5 ' acc-cta-ttc-ttc-ata-tcg-ttc-c3 ', primer 2: 5 ' agg-gtt-aca-tat-cgc-cga-gac 3 '.The result shows that the ADT gene expresses as shown in Figure 2 in wild-type, size is 496bp, and does not express among the mutant S29, and among Fig. 2, "+" expression adds ThermoScript II, and "-" expression does not add ThermoScript II.
The Southern analytical results of embodiment 4, wild-type and mutant S29
The extraction of wild-type Arabidopis thaliana and mutant S29 arabidopsis thaliana genomic dna is undertaken by the method for Paterson et al (1993).Get 1 μ g genomic dna, use BamHI and HindIII (Takara product) enzyme to cut respectively.Enzyme is cut product through 0.7% agarose gel electrophoresis (30-50V, O/N) separate, on shaking table, handle running gel with ordinary method then: embathe 15min through 0.125N HCl, sex change liquid (1.5M NaCl, 0.5N NaOH) embathe 30min, neutralizer (1.5M NaCl, 0.5M Tris-HCl pH 7.2) embathes 30min.Use 20 * SSC that southern blotting technique is arrived HybondN at last
+On the nylon membrane.
(Jin Dongyan etc., 1996 are all carried out in prehybridization, hybridization according to a conventional method; Hybond N
+Mannual, Amersham).
Probe and mark thereof: hybridization probe is NPTII,
Primer 1:5 ' ggg-cgc-ccg-gtt-ctt-ttt-g 3 ' wherein,
Primer 2: 5 ' aca-ccc-agc-cgg-cca-cag-tcg, 3 '.Carrying out mark with the Primer-a-Gene test kit of Promega company spends the night.PCR purification kit with Qiagen company before the hybridization carries out the probe purifying.
Wash film and carry out (Hybond N with high tight method
+Mannual, Amersham): 2 * SSC, 0.1%SDS, 15min; 1 * SSC, 0.1%SDS, 65 ℃, 15min; 2 * SSC, 0.1%SDS, 65 ℃, 15min.
Preservative film parcel Hybond membrane exposes to the X-ray sheet under-70 ℃.
The result as shown in Figure 3, as can be seen from the figure, this gene is cut at the enzyme of BamHI and is hybridized to treaty 20kb band in the swimming lane; Cut at the enzyme of HindIII and to hybridize to treaty 3.3kb band in the swimming lane; Can determine that T-DNA inserts the mutant S29 that forms behind the genomic ADT gene with the form of single copy.
The Northern analytical results of embodiment 5, wild-type Arabidopis thaliana and mutant S29 Arabidopis thaliana
Used Arabidopis thaliana material is at substratum MS+0 among the embodiment, MS+3%PEG, (PEG-6000 is as the osmotic stress agent for MS+3.5%PEG, but the people is for causing drought environment) in, each 100-200mg of seedling around the growth, (QIAGEN) extracts total RNA with the PlantRNeasy test kit, check the integrity of RNA through 1.0% agarose gel electrophoresis after, combine with ultraviolet spectrophotometer and electrophoresis method and to make accurate quantification.Adjust each sample applied sample amount and be 20 μ g, carry out 1.2% denaturing formaldehyde agarose gel electrophoresis then.
The making of denaturing formaldehyde glue: 1.2%FA gel 150ml adds 15ml10 * FA gel buffer liquid configuration by the 1.5g agarose, after the fusing, cools off in 65 ℃ of water-baths in microwave oven, adds that the formaldehyde stoste of 2.7ml 37% makes again.Must be in 1 * FA damping fluid before the electrophoresis balance 30min at least.The prescription of 1 * FA damping fluid 600ml is the SDW that 50ml 10 * FA damping fluid, 10ml 37% formaldehyde stoste and 440ml DEPC handle.During last sample, per 4 parts of RNA samples add 1 part of 5 * RNA sample-loading buffer, heat 3-5min in 65 ℃ of water-baths behind the mixing, the ice bath cooling.5 * RNA sample-loading buffer 10ml includes the saturated tetrabromophenol sulfonphthalein solution of 16 μ l, 80 μ l 500mM EDTA pH8.0,720 μ l, 37% formaldehyde stoste, 2ml glycerine, 3084 μ l methane amides and 4ml10 * FA damping fluid.
After electrophoresis finishes, downcut the molecule marker swimming lane and dye with EB, the accurate indicating positions of a fluorescence chi is put on the next door.Embathe 30min on the SDW shaking table of glue with the DEPC processing of sample part, the 10 * SSC that handles with DEPC embathes 30min again.Then RNA is gone to Hybond N with 20 * SSC that DEPC handles
+On the film, film is through prehybridization, and radioactive probe hybridization after high stringency is washed film, exposes to the X-ray sheet under-70 ℃.
Probe is the RD29A gene, and this gene is a stress gene, such as produce (Plant Journal, 2003) under the coercing of environment such as arid, high salt, cold.The result as shown in Figure 4, showing does not have in the contrast that PEG coerces, RD29A genetic expression seldom in plant wild-type Arabidopis thaliana and the mutant S29 Arabidopis thaliana; After 3.0%PEG coerced, the two is all expressed but difference is not obvious; After 3.5%PEG coerced, RD29A expression of gene amount was apparently higher than wild-type in the mutant S29 Arabidopis thaliana.
The phenotype observations of embodiment 6, wild-type Arabidopis thaliana and mutant S29 Arabidopis thaliana
The observations of a, wild-type Arabidopis thaliana and mutant S29 Arabidopis thaliana excised leaf
The excised leaf percentage of water loss is relevant with drought resistance.The blade percentage of water loss can be used as that plant drought is identified fast, easily row, easy physical signs (Zhao Wei flat 1993, Liu Xue justice 1995).In the research of percentage of water loss and drought resistance, find that 6h is the material time of percentage of water loss turnover, the percentage of water loss of 6h drought resisting type in the past is lower than responsive type, the drought resisting type is higher than responsive type later on, and all meets this rule in the different growth stage measured result of different drought resisting types or same type.The result shows that the drought resistance of mutant S29 Arabidopis thaliana is higher than wild-type shown in Fig. 5 a.
B, wild-type Arabidopis thaliana and the observations of mutant S29 Arabidopis thaliana on the MS+2.5% glycerin medium
Wild-type Arabidopis thaliana and mutant S29 Arabidopis thaliana seed are transferred to (glycerine can cause drought environment) (Plant Physiol, 2003) on the MS+2.5% glycerin medium after sprouting on the MS substratum and growing up to four leaf phase seedling.The result shows that wild-type Arabidopis thaliana plant is dead shown in Fig. 5 b, mutant S29 Arabidopis thaliana plant is because drought resisting and normal growth.
Observations under c, wild-type Arabidopis thaliana and the potted plant normal condition of mutant S29 Arabidopis thaliana
Under normal operation, drought-resistant variety and not drought-resistant variety comparison, drought-resistant variety shows as late flower, and life cycle is long.The result shows that the wild-type Arabidopis thaliana plant knot of blooming already is tending towards dead shown in Fig. 5 c, mutant S29 Arabidopis thaliana plant since drought resisting and on normal growth and the substratum result coincide.
Observations under d, wild-type Arabidopis thaliana and the potted plant drought condition of mutant S29 Arabidopis thaliana
The result shows that the result of potted plant arid test conforms to fully with measurement result trend under plant excised leaf percentage of water loss, the potted plant normal condition shown in Fig. 5 d.Mutant S29 Arabidopis thaliana is low because of percentage of water loss, the strong blade of moisture holding capacity is wilted occurs lately, and it is light to be injured.
Sequence table
<160>2
<210>1
<211>1164
<212>DNA
＜213〉Arabidopis thaliana (Columbia)
<400>1
atgaaatcac?ggcgacagaa?tgtgtccgtg?gctcgacaaa?ccatccttgg?acgcgacgaa 60
aactttgaac?caatcccaat?tgatctcgtt?atcgagatat?tctcaaggtc?gcctgtgaag 120
tctatagcaa?gatgtcgttg?cgtatcaaag?ctttgggcct?ccatactccg?cctaccctat 180
ttcacggagt?tgtacttgac?caaatcttgt?gctcgcccga?ggctcttgtt?cgcctgccaa 240
aaacacagag?agttgttctt?cttctcgaca?cctcagcctc?ataatcctaa?tgagagctcg 300
tctcctttag?ctgccagttt?tcatatgaaa?attccctttg?atggtcgctt?taatattatc 360
agtcctatcg?gtggccttgt?ctttgttaga?tatgaacaga?tcttaaaggg?aaggaaaact 420
ccagaatttg?tctcggcgat?atgtaaccct?agcacgggac?aatccttaac?cttaccaaaa 480
cctaagacaa?ggaagaggat?ttggggtaca?agccattttg?ggtatgatcc?tattgagaaa 540
caattcaagg?tattgtcaat?gaatataggt?gatggggtct?ataaagagca?ttatgttctg 600
acattaggaa?ctgagaacct?ctcttggaga?aggatcgaat?gttctatacc?ccatgttcat 660
ggttctaaag?ggatatgcat?caatggtgtt?ttgtattatc?gagcaaaggc?tgacatgttt 720
tcaggtactt?taatgatagt?ttgctttgat?gttaggtttg?agaagttcag?ctatattaaa 780
atcttgaaac?ctacaacaac?tctgattagc?tacaacggta?aattggcttc?actagtgtgg 840
gaagggccta?gttatatttg?tggaaaacgt?tttgaaatgt?gggttttagg?agaccccgaa 900
aaacatgaat?ggttgaagca?tacttacgaa?ttgcgtcctc?ggtggcagaa?tgtacttgga 960
gaggacttgt?taatttttgc?tggaatgact?ggtacaaatg?aaattgtgtt?gtcgccaaag 1020
tatccatctc?accctttcta?tgttttctac?tacaatttgg?agaggaatac?tatcagaaga 1080
gttgaaatcc?aaggaatggg?agcgtttaag?gttaatgaag?attacatctt?tctagaccat 1140
gtagaggatg?tgaagcttat?ataa 1164
<210>1
<211>387
<212>PRT
＜213〉Arabidopis thaliana (Columbia)
<400>1
Met?Lys?Ser?Arg?Arg?Gln?Asn?Val?Ser?Val?Ala?Arg?Gln?Thr?Ile
1 5 10 15
Leu?Gly?Arg?Asp?Glu?Asn?Phe?Glu?Pro?Ile?Pro?Ile?Asp?Leu?Val
20 25 30
Ile?Glu?Ile?Phe?Ser?Arg?Ser?Pro?Val?Lys?Ser?Ile?Ala?Arg?Cys
35 40 45
Arg?Cys?Val?Ser?Lys?Leu?Trp?Ala?Ser?Ile?Leu?Arg?Leu?Pro?Tyr
50 55 60
Phe?Thr?Glu?Leu?Tyr?Leu?Thr?Lys?Ser?Cys?Ala?Arg?Pro?Arg?Leu
65 70 75
Leu?Phe?Ala?Cys?Gln?Lys?His?Arg?Glu?Leu?Phe?Phe?Phe?Ser?Thr
80 85 90
Pro?Gln?Pro?His?Asn?Pro?Asn?Glu?Ser?Ser?Ser?Pro?Leu?Ala?Ala
95 100 105
Ser?Phe?His?Met?Lys?Ile?Pro?Phe?Asp?Gly?Arg?Phe?Asn?Ile?Ile
110 115 120
Ser?Pro?Ile?Gly?Gly?Leu?Val?Phe?Val?Arg?Tyr?Glu?Gln?Ile?Leu
125 130 135
Lys?Gly?Arg?Lys?Thr?Pro?Glu?Phe?Val?Ser?Ala?Ile?Cys?Asn?Pro
140 145 150
Ser?Thr?Gly?Gln?Ser?Leu?Thr?Leu?Pro?Lys?Pro?Lys?Thr?Arg?Lys
155 160 165
Arg?Ile?Trp?Gly?Thr?Ser?His?Phe?Gly?Tyr?Asp?Pro?Ile?Glu?Lys
170 175 180
Gln?Phe?Lys?Val?Leu?Ser?Met?Asn?Ile?Gly?Asp?Gly?Val?Tyr?Lys
185 190 195
Glu?His?Tyr?Val?Leu?Thr?Leu?Gly?Thr?Glu?Asn?Leu?Ser?Trp?Arg
200 205 210
Arg?Ile?Glu?Cys?Ser?Ile?Pro?His?Val?His?Gly?Ser?Lys?Gly?Ile
215 220 225
Cys?Ile?Asn?Gly?Val?Leu?Tyr?Tyr?Arg?Ala?Lys?Ala?Asp?Met?Phe
230 235 240
Ser?Gly?Thr?Leu?Met?Ile?Val?Cys?Phe?Asp?Val?Arg?Phe?Glu?Lys
245 250 255
Phe?Ser?Tyr?Ile?Lys?Ile?Leu?Lys?Pro?Thr?Thr?Thr?Leu?Ile?Ser
260 265 270
Tyr?Asn?Gly?Lys?Leu?Ala?Ser?Leu?Val?Trp?Glu?Gly?Pro?Ser?Tyr
275 280 285
Ile?Cys?Gly?Lys?Arg?Phe?Glu?Met?Trp?Val?Leu?Gly?Asp?Pro?Glu
290 295 300
Lys?His?Glu?Trp?Leu?Lys?His?Thr?Tyr?Glu?Leu?Arg?Pro?Arg?Trp
305 310 315
Gln?Asn?Val?Leu?Gly?Glu?Asp?Leu?Leu?Ile?Phe?Ala?Gly?Met?Thr
320 325 330
Gly?Thr?Asn?Glu?Ile?Val?Leu?Ser?Pro?Lys?Tyr?Pro?Ser?His?Pro
335 340 345
Phe?Tyr?Val?Phe?Tyr?Tyr?Asn?Leu?Glu?Arg?Asn?Thr?Ile?Arg?Arg
350 355 360
Val?Glu?Ile?Gln?Gly?Met?Gly?Ala?Phe?Lys?Val?Asn?Glu?Asp?Tyr
365 370 375
Ile?Phe?Leu?Asp?His?Val?Glu?Asp?Val?Lys?Leu?Ile
380 385 387
Claims (7)
1, a kind of Arabidopis thaliana gene related to drought tolerance ADT is one of following nucleotide sequences:
1) SEQ ID № in the sequence table: 1 dna sequence dna;
2) SEQ ID № in the code sequence tabulation: the polynucleotide of 2 protein sequences;
3) with sequence table in SEQ ID №: 1 dna sequence dna that limits has 90% above homology, and the identical function protein DNA sequence of encoding.
2, gene according to claim 1 is characterized in that: described Arabidopis thaliana gene related to drought tolerance ADT is SEQ ID № in the sequence table: 1.
3, Arabidopis thaliana gene related to drought tolerance ADT proteins encoded ADT, be to have SEQ ID № in the sequence table: the protein of 2 amino acid residue sequences, or with SEQ ID №: 2 amino acid residue sequence is through replacement, disappearance or the interpolation of one or several amino-acid residue and have the № with SEQ ID: 2 amino acid residue sequence is identical active by SEQ ID №: 2 deutero-protein.
4, protein according to claim 3 is characterized in that: described Arabidopis thaliana gene related to drought tolerance ADT proteins encoded ADT is SEQ ID № in the sequence table: 2.
5, contain the described expression carrier of claim 1.
6, the clone that contains the described gene of claim 1.
7, the application of the described gene of claim 1 in cultivating drought resistant plant variety.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1293095C (en) * | 2005-04-25 | 2007-01-03 | 中国农业大学 | Drought resistant correlative protein and coded gene of plant and application |
| CN1324137C (en) * | 2005-07-12 | 2007-07-04 | 中国科学技术大学 | Salt resistant and drought resistant gene of Thellugielkla halophila and its coded protein and use |
| CN101525379B (en) * | 2009-04-02 | 2011-12-21 | 中国农业大学 | Plant drought-enduring associated protein, encoding gene and application thereof |
-
2003
- 2003-09-18 CN CNB031571980A patent/CN1333075C/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1293095C (en) * | 2005-04-25 | 2007-01-03 | 中国农业大学 | Drought resistant correlative protein and coded gene of plant and application |
| CN1324137C (en) * | 2005-07-12 | 2007-07-04 | 中国科学技术大学 | Salt resistant and drought resistant gene of Thellugielkla halophila and its coded protein and use |
| CN101525379B (en) * | 2009-04-02 | 2011-12-21 | 中国农业大学 | Plant drought-enduring associated protein, encoding gene and application thereof |
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|---|---|
| CN1333075C (en) | 2007-08-22 |
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