CN114835787A - Quercus variabilis QsSRO1 gene and application of encoding protein thereof in plant stress resistance - Google Patents
Quercus variabilis QsSRO1 gene and application of encoding protein thereof in plant stress resistance Download PDFInfo
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Abstract
The invention belongs to the technical field of genetic engineering, and particularly relates to quercus variabilis QsSRO1 gene and application of encoding protein thereof in plant stress resistance. The nucleotide sequence of the quercus variabilis QsSRO1 gene is shown in SEQ ID No.1, and the amino acid sequence of the coded quercus variabilis QsSRO1 protein is SEQ ID No.2, and is a cell nucleus protein. Through constructing a recombinant expression vector containing quercus variabilis QsSRO1 gene, then transforming Arabidopsis thaliana by using agrobacterium, and screening and culturing the obtained transformed QsSRO1 Arabidopsis thaliana plant seeds, the performance is good in a salt resistance test. The invention proves that the over-expression of the quercus variabilis QsSRO1 gene can improve the salt resistance of plants, and lays theoretical and production practice foundation for cultivating new varieties of salt-resistant and drought-resistant transgenic crops.
Description
Technical Field
The invention belongs to the technical field of genetic engineering, and particularly relates to quercus variabilis QsSRO1 gene and application of encoding protein thereof in plant stress resistance.
Background
Quercus suber (Quercus suber) is an important broad-leaved evergreen tree species that grows slowly and has an extremely long life. Quercus suber is a major source of industrial cork, a material of very high commercial value.
Climate change is one of the major challenges and global issues facing the 21 st century. In recent decades, as global air temperatures have risen, a series of more intense, more frequent and longer lasting climatic extremes, particularly severe drought and soil salinization, have been initiated. It has been reported that various global changes such as soil salinization lead to a severe decrease in the number of quercus variabilis. Although tree species can adapt to new environmental conditions, little is known about the processes involved. Conventional crossbreeding is a method of obtaining individuals with recombined parental genes by mating between individuals of different genotypes. New genes can not be generated in the hybridization process, and the filial generation has character separation, the breeding process is slow, and the process is complex. The plant genetic engineering breeding has the characteristics of directional change of the plant, no limitation of species, short breeding period and the like. At present, modern breeding technologies such as genetic engineering and the like are urgently needed to carry out quercus variabilis so as to ensure the sustainability of economic tree species.
Disclosure of Invention
The invention aims to provide an application of quercus variabilis QsSRO1 gene and a coded protein thereof in plant stress resistance, and the stress resistance of plants is improved.
In order to achieve the above purpose, the invention provides the following scheme:
the invention provides quercus variabilis QsSRO1 protein, the amino acid sequence of which is shown as SEQ ID No.2, and the amino acid sequence (SEQ ID No.2) of quercus variabilis QsSRO1 protein is as follows:
MEANIAKASDRSKRVVLDLKRKRATQLATYLNEVRPTWDSLQNRLDKR RKLNGCQRKDMSYGPSGRSLLKCYSNYVKTGTPKRLMFYQNGEWIDFPQSV VDVVREDFQVKKSAVEVEFNGHRFMLDFLHMSRVDLKTTLQEPIAWIDEADS CFFPETFDCHQPETMEYQDPVLEEPYGPQEIKLLLEIEINGVDQSKLTECSGES NDLVKQIQINSKPASNCYAIDVENSCSRESDAKMDEDFQENKQIPANLVIAPVS ENEEFNCDSVQKLFLVGMGASGRPDILEIYRCESTSLQARFELFQKQAELTQK CRGDANVQYAWLACSKGELPTILTHGLGHCGPSTIKSMYGSGVHLAAAICSY TSANFCDVDENGVQHLVLCQVIMGKMEVVHSGSRQNLPSCKDYDSGVDDLQ NPKIYIVWTMNMNTHIYPEFVVSFKISSKTEGVTSCQVRKHQQLESSAVDLSV SQPVSDSGRSEGKAPSLGSSNTRAPKSPWMPFPMLFAAISEKVSSGVMEKINE HYELFRTKKIGRDEFIKKLRLIVGDALLRSTITNLQCQLPLRSKCEPEVLQPNLE KEKVQHSFN
in a second aspect, the invention provides a Quercus variabilis QsSRO1 gene encoding the Quercus variabilis QsSRO1 protein, which is derived from Quercus variabilis (Quercus suber). The nucleotide sequence of the quercus variabilis QsSRO1 gene is shown in SEQ ID No. 1.
The invention also provides an overexpression vector, a host cell and an engineering bacterium containing the quercus variabilis QsSRO 1.
In a third aspect, the invention provides application of the quercus variabilis QsSRO1 gene in regulation and control of plant salt resistance.
Further, the method for improving the stress resistance of the plants comprises the following steps: constructing a recombinant expression vector containing the quercus variabilis QsSRO1 gene; transforming the quercus variabilis QsSRO1 gene into a plant using agrobacterium-mediated transformation; screening and culturing positive plants.
Furthermore, the plant comprises arabidopsis thaliana, and T4 generation seeds of the transgenic arabidopsis thaliana plant have salt resistance.
Compared with the prior art, the invention has the advantages and beneficial effects that: the invention provides quercus variabilis QsSRO1 gene and a coding protein thereof, and according to the salt resistance test result of a transgenic T4 Arabidopsis plant of QsSRO1, the over-expression of the QsSRO1 gene is helpful for reducing the damage of salt to the plant when the plant is treated in a salt solution, so that the transgenic Arabidopsis of QsSRO1 has stronger salt tolerance. Therefore, the quercus variabilis QsSRO1 gene provided by the invention is suitable for improving the stress resistance of arabidopsis thaliana, especially the salt stress resistance, the functional analysis and identification of the quercus variabilis QsSRO1 gene also provide a theoretical basis for obtaining new plant varieties, the stress-resistant plant with production and application values can be cultured by utilizing the gene to carry out molecular breeding, and the theoretical and production practice basis is laid for culturing new salt-resistant and drought-resistant transgenic crop varieties.
Drawings
FIG. 1 shows the expression of the QsSRO1 gene after the quercus variabilis tissues and salt flank.
FIG. 2 shows the expression level of the QsSRO1 gene with time after quercus variabilis was stressed with salt.
FIG. 3 shows the results of molecular examination of QsSRO 1-transformed Arabidopsis thaliana.
FIG. 4 is an observation that QsSRO 1-transgenic Arabidopsis and wild-type Arabidopsis seeds germinated for 7 days in 100mM NaCl.
FIG. 5 shows the statistics of root length of seeds of QsSRO1 transgenic Arabidopsis thaliana and wild Arabidopsis thaliana germinated for 7 days in 100mM NaCl.
FIG. 6 shows subcellular localization of transiently transformed QsSRO1 protein.
Detailed Description
In order to make those skilled in the art better understand the technical solutions in the present application, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention.
Example 1: quercus variabilis QsSRO1 gene and obtaining method thereof
Method for obtaining Quercus variabilis QsSRO1
Extracting RNA from Quercus variabilis (lour.) Kuntze, and using TaKaRa PrimeScript (TM) as template RT The kit is reversely transcribed into cDNA by a reverse transcription kit method. Then, the cDNA was used as a reaction template for amplifying a target gene sequence, and PCR amplification was carried out using 5'-ATGGCGAATCTACCCCAATCGC-3' and 5'-TCAGTTTCCGGGCTCTGGTTTC-3' as primers to obtain the CDS sequence of the QsSRO1 gene.
Secondly, the tissue expression condition of the QsSRO1 gene is analyzed by fluorescent quantitative PCR
Respectively extracting the RNA of the root, stem and flower of quercus variabilis, and carrying out reverse transcription to synthesize cDNA; then, PCR amplification was carried out using cDNA of each tissue as a template and 5'-AGACGGGACGAGGGTTGT-3' and 5'-TGGATGATGTGGGCTTGG-3' as primers, and the expression of the QsSRO1 gene in different tissues of Quercus suberectus was examined. Meanwhile, qACTIN gene is used as an internal reference, and 5' primers for amplifying the qACTIN gene are as follows: 5'-GCGGATAGAATGAGCAAGGAA-3', the 3 ' primer is: 5'-GGGCCGGACTCATCATACTC-3' are provided. RT-qPCR reaction conditions: pre-denaturation at 94 ℃ for 1 min; then 30 cycles of 98 ℃ 10sec, 58 ℃ 30sec, 72 ℃ 1 min; extension was then carried out at 72 ℃ for 3 min.
The expression of quercus variabilis QsSRO1 gene in each tissue is shown in FIG. 1. The results showed that quercus variabilis QsSRO1 gene was expressed in all tissues, with the lowest expression in the stem and the highest expression in the root.
The expression of QsSRO1 gene under the salt stress condition of Quercus variabilis QsSRO1 gene is shown in FIG. 2. The result shows that the quercus variabilis QsSRO1 gene has increased expression level after the quercus variabilis is subjected to salt stress, and the expression level reaches the maximum level in 8 hours.
Example 2: application of QsSRO1 gene in improving stress resistance of plants
1. Construction of QsSRO 1-transgenic Arabidopsis thaliana
Obtaining Quercus variabilis QsSRO1 gene, preparing the following primer pairs:
primer 1: 5'-ATGGCGAATCTACCCCAATCGC-3', respectively;
primer 2: 5'-TCAGTTTCCGGGCTCTGGTTTC-3' are provided.
Taking cDNA of quercus suberectus as a template, carrying out PCR amplification by using the primer 1 and the primer 2 to obtain a PCR product with the size of 1755bp, and sequencing the PCR product. The obtained PCR product is quercus variabilis QsSRO1 gene, the sequencing result of which shows that the nucleotide sequence is shown as SEQ ID NO.1, and the PCR product can encode QsSRO1 protein with the amino acid sequence shown as SEQ ID NO. 2. Bioinformatics analysis shows that the protein encoded by the QsSRO1 gene contains 585 amino acid residues, the relative molecular mass is 66.04kDa, and the theoretical isoelectric point is 5.85. The protein QsSRO1 contains a Trp-Trp-Glu domain at the N-terminal, a polyadenylate polymerase structure in the middle of the sequence and a TATA box binding protein-associated factor 4 at the C-terminal.
The QsSRO1 gene sequence (SEQ ID NO.1) is as follows, the total length is 1755 bp:
ATGGAAGCAAATATCGCAAAGGCATCGGATAGAAGTAAGAGAGTTGTG CTCGACTTAAAAAGAAAGCGGGCAACCCAGCTTGCTACATATTTGAATGAA GTTAGGCCCACTTGGGATTCATTGCAAAATAGGCTTGACAAACGGAGGAAA TTGAATGGGTGCCAAAGGAAGGACATGAGCTATGGGCCTAGTGGGAGATC TTTGCTCAAATGTTATTCAAATTATGTGAAAACTGGGACACCAAAGCGTTTA ATGTTTTATCAGAATGGCGAATGGATTGATTTCCCCCAGAGTGTTGTTGATG TGGTTAGGGAAGATTTTCAGGTTAAGAAGTCTGCTGTAGAGGTTGAGTTCA ATGGCCATCGCTTTATGCTTGATTTTCTGCATATGTCTCGAGTGGACTTGAA AACAACCTTACAGGAACCCATTGCTTGGATTGATGAAGCAGACAGCTGCTT CTTTCCTGAAACTTTTGACTGCCATCAACCTGAAACTATGGAATATCAAGAC CCAGTGTTGGAGGAGCCTTATGGGCCTCAAGAGATCAAGCTGCTGTTGGA AATTGAAATAAATGGAGTGGATCAATCCAAGCTGACGGAATGTAGTGGGGA GTCAAATGATCTAGTTAAGCAGATACAAATCAATAGTAAACCTGCTAGCAA CTGCTATGCTATAGATGTTGAGAATAGTTGTAGTAGAGAGTCTGATGCAAAA ATGGATGAAGATTTTCAGGAAAATAAACAGATACCTGCAAATTTAGTCATAG CGCCTGTATCTGAAAATGAAGAATTTAATTGTGATTCTGTGCAGAAGTTGTT TCTTGTGGGTATGGGTGCTTCAGGCAGACCCGACATTCTTGAAATATACCGT TGCGAAAGCACTTCATTGCAAGCTCGATTTGAGCTTTTTCAGAAGCAGGCT GAACTAACCCAAAAATGTCGTGGGGATGCAAATGTTCAATATGCTTGGCTT GCTTGTTCTAAAGGGGAGCTGCCTACAATATTGACACATGGGCTTGGTCATT GTGGACCTTCCACAATTAAGTCCATGTATGGTAGTGGTGTTCATCTTGCAGC TGCTATCTGTTCTTACACCAGTGCAAATTTTTGTGATGTTGACGAAAATGGG GTACAACACTTGGTGTTGTGCCAAGTGATAATGGGAAAAATGGAGGTTGTT CATTCTGGCTCTAGACAAAACCTTCCCAGTTGCAAGGACTATGATAGTGGA GTGGATGATCTTCAAAATCCAAAGATTTATATAGTCTGGACTATGAATATGA ACACTCACATCTATCCAGAATTTGTTGTTAGTTTCAAGATCTCTTCCAAAAC TGAAGGGGTTACATCTTGTCAGGTGCGTAAGCACCAACAATTAGAGTCTTC TGCAGTTGATTTGAGCGTGAGTCAACCAGTTTCAGATTCTGGGAGATCTGA GGGGAAAGCTCCCAGTCTGGGTTCAAGCAATACAAGAGCTCCTAAATCTCC TTGGATGCCTTTTCCCATGTTGTTTGCTGCCATTTCAGAGAAAGTTTCTTCT GGGGTCATGGAGAAGATTAATGAACATTATGAGTTGTTTAGGACAAAGAAG ATAGGTCGTGATGAGTTTATTAAAAAGTTGAGACTGATAGTTGGGGATGCTT TATTGAGGTCTACAATAACAAATCTGCAATGCCAGTTACCACTGAGATCTAA GTGTGAACCGGAAGTTCTACAGCCTAACCTAGAAAAAGAAAAGGTTCAGC ACTCCTTCAAT
2. obtaining of recombinant expression vectors
And (3) connecting the PCR product obtained in the step (1) with the pCAMBIA1300 vector after restriction enzyme digestion to obtain a connecting product. The ligation products were then transformed into E.coli DH 5. alpha. competent cells and plated on LB plates containing 25. mu.g/ml ampicillin overnight. Selecting a white single colony, culturing the white single colony in an LB liquid culture medium containing 50 mu g/ml kanamycin for overnight, and carrying out colony PCR identification; meanwhile, plasmid DNA is extracted by an alkaline method for sequence determination.
The sequencing result shows that the recombinant expression vector is inserted into the QsSRO1 gene sequence shown in SEQ ID NO. 1; the recombinant expression vector was named pCAMBIA1300-QsSRO 1.
3. Acquisition and molecular detection of QsSRO 1-transformed arabidopsis thaliana
(1) Obtaining of QsSRO 1-transferred Arabidopsis thaliana
The recombinant expression vector pCAMBIA1300-QsSRO1 prepared in step 2 is used for transforming competent cells of Agrobacterium GV3101 (purchased from Biotechnology engineering (Shanghai) Co., Ltd.) to obtain recombinant engineering bacteria GV3101/pCAMBIA1300-QsSRO1 (extracted plasmid is sent for sequencing to obtain the recombinant vector pCAMBIA1300-QsSRO 1).
The GV3101/pCAMBIA1300-QsSRO1 engineering bacteria were inoculated in a single clone in YEB liquid medium containing 50mg/L chloramphenicol, and cultured with shaking at 28 ℃ for two days. The culture was centrifuged at 4000rpm/min for 5 minutes and the resulting Agrobacterium pellet was resuspended in an infection containing 5% sucrose and 0.03% Silwet L-77.
Columbia ecotype wild type Arabidopsis thaliana (Col-0) (purchased from Arabidopsis Biological Resource Center) was transformed by the floral dip method, seeds inoculated to the current generation of transgenic Arabidopsis plants (T1 generation) were harvested, and germinated seeds were screened in MS medium containing 50. mu.g/ml kanamycin (kanamycin). Seedlings from T1 generations germinated on the above medium were transferred to culture soil, seeds were harvested (T2 generations), and then homozygous transgenic QsSRO1 Arabidopsis plants (T4 generations) seeds were obtained through the same screening process.
(2) Molecular detection
The genomic DNA of wild type Arabidopsis thaliana and T4 generation plants was used as a final work, and amplification was performed using primer 1 and primer 2 in example 2. The results of PCR amplification are shown in FIG. 3(Col-0 is wild type). The results show that the transgenic T4 generation seeds can be amplified to a band of about 1800bp, but the wild Arabidopsis can not be amplified to a band of about 1800bp, so the transgenic T4 generation seeds are identified as T4 generation homozygous transgenic QsSRO1 Arabidopsis.
4. Functional study of QsSRO 1-transformed Arabidopsis thaliana
(1) Salt resistance test of seeds
Taking seeds of wild type arabidopsis (White) and a transformed QsSRO1 arabidopsis plant (T4 generation), and carrying out seed germination experiments on an MS culture medium containing 100mM NaCl under the conditions that the photoperiod is 16 hours of light and 8 hours of darkness; the illumination intensity is 300- -2 s -1 (ii) a The culture temperature under illumination condition is 21-24 deg.C, and relative humidity is 80%; the culture temperature under dark condition is 21-24 deg.C, and relative humidity is 80%.
As shown in FIGS. 4-5, Col-0 is a control group of wild type Arabidopsis, and both OE1 and OE2 are QsSRO1 Arabidopsis plants. Seeds of the QsSRO1 arabidopsis plants (T4 generation) were significantly longer in root length than wild type arabidopsis at 100mM NaCl treatment for 100mM NaCl treatment.
The results show that the QsSRO1 protein (SEQ ID NO.2) coded by the QsSRO1 gene has the function of improving the salt resistance of plant seeds.
(2) Subcellular localization of the QsSRO1 protein
10ml of enzymolysis liquid is prepared. Taking 1mth leaf age healthy growing and unopened Arabidopsis thaliana rosette leaves, cutting the leaves into about 1mm leaf strips by a blade, immersing the leaf strips in enzymolysis liquid, and carrying out enzymolysis for three hours at 60rpm under the conditions of room temperature and darkness. And (4) sieving the enzymolysis liquid with a 200-mesh sieve, and subpackaging the enzymolysis liquid with 30ml centrifuge tubes. The centrifuge was refrigerated at 4 ℃ and 120rpm, centrifuged for five minutes, the supernatant discarded, and the protoplasts were resuspended in a pre-chilled W5 solution (150mM NaCl, 125mM CaCl2, 5mM KCl, 2mM MES, pH 5.8). The cells were washed 3 times with pre-cooled W5 solution, centrifuged at 120rpm for two minutes, and placed on ice for half an hour. Resuspend at room temperature using MaMg solution. Add 10. mu.g of plasmid to 150. mu.l of protoplast, shake gently, add 120. mu.l of PEG/Ca solution, and blow gently. Shaken at 50rpm for ten minutes, 0.5ml of W5 solution was added and gently mixed. Centrifuge for five minutes at room temperature, 100 g. Add 1.5ml W5 heavy suspension protoplast, room temperature, 100rpm, centrifugal for one minute, discard the supernatant, add 200 u l W5 solution heavy suspension protoplast, room temperature incubation twelve hours. After the above steps are completed, the observation is carried out by using a fluorescence confocal microscope.
The results are shown in FIG. 6, which shows that the protein encoded by the gene QsSRO1 is present in the nucleus and belongs to a nuclear protein, on a scale of 5 μm.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions.
Sequence listing
<110> Qingdao Standd environmental technology research institute Co., Ltd
<120> quercus suber QsSRO1 gene and application of encoding protein thereof in plant stress resistance
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1755
<212> DNA
<213> Quercus suber
<400> 1
atggaagcaa atatcgcaaa ggcatcggat agaagtaaga gagttgtgct cgacttaaaa 60
agaaagcggg caacccagct tgctacatat ttgaatgaag ttaggcccac ttgggattca 120
ttgcaaaata ggcttgacaa acggaggaaa ttgaatgggt gccaaaggaa ggacatgagc 180
tatgggccta gtgggagatc tttgctcaaa tgttattcaa attatgtgaa aactgggaca 240
ccaaagcgtt taatgtttta tcagaatggc gaatggattg atttccccca gagtgttgtt 300
gatgtggtta gggaagattt tcaggttaag aagtctgctg tagaggttga gttcaatggc 360
catcgcttta tgcttgattt tctgcatatg tctcgagtgg acttgaaaac aaccttacag 420
gaacccattg cttggattga tgaagcagac agctgcttct ttcctgaaac ttttgactgc 480
catcaacctg aaactatgga atatcaagac ccagtgttgg aggagcctta tgggcctcaa 540
gagatcaagc tgctgttgga aattgaaata aatggagtgg atcaatccaa gctgacggaa 600
tgtagtgggg agtcaaatga tctagttaag cagatacaaa tcaatagtaa acctgctagc 660
aactgctatg ctatagatgt tgagaatagt tgtagtagag agtctgatgc aaaaatggat 720
gaagattttc aggaaaataa acagatacct gcaaatttag tcatagcgcc tgtatctgaa 780
aatgaagaat ttaattgtga ttctgtgcag aagttgtttc ttgtgggtat gggtgcttca 840
ggcagacccg acattcttga aatataccgt tgcgaaagca cttcattgca agctcgattt 900
gagctttttc agaagcaggc tgaactaacc caaaaatgtc gtggggatgc aaatgttcaa 960
tatgcttggc ttgcttgttc taaaggggag ctgcctacaa tattgacaca tgggcttggt 1020
cattgtggac cttccacaat taagtccatg tatggtagtg gtgttcatct tgcagctgct 1080
atctgttctt acaccagtgc aaatttttgt gatgttgacg aaaatggggt acaacacttg 1140
gtgttgtgcc aagtgataat gggaaaaatg gaggttgttc attctggctc tagacaaaac 1200
cttcccagtt gcaaggacta tgatagtgga gtggatgatc ttcaaaatcc aaagatttat 1260
atagtctgga ctatgaatat gaacactcac atctatccag aatttgttgt tagtttcaag 1320
atctcttcca aaactgaagg ggttacatct tgtcaggtgc gtaagcacca acaattagag 1380
tcttctgcag ttgatttgag cgtgagtcaa ccagtttcag attctgggag atctgagggg 1440
aaagctccca gtctgggttc aagcaataca agagctccta aatctccttg gatgcctttt 1500
cccatgttgt ttgctgccat ttcagagaaa gtttcttctg gggtcatgga gaagattaat 1560
gaacattatg agttgtttag gacaaagaag ataggtcgtg atgagtttat taaaaagttg 1620
agactgatag ttggggatgc tttattgagg tctacaataa caaatctgca atgccagtta 1680
ccactgagat ctaagtgtga accggaagtt ctacagccta acctagaaaa agaaaaggtt 1740
cagcactcct tcaat 1755
<210> 2
<211> 585
<212> PRT
<213> Quercus suber
<400> 2
Met Glu Ala Asn Ile Ala Lys Ala Ser Asp Arg Ser Lys Arg Val Val
1 5 10 15
Leu Asp Leu Lys Arg Lys Arg Ala Thr Gln Leu Ala Thr Tyr Leu Asn
20 25 30
Glu Val Arg Pro Thr Trp Asp Ser Leu Gln Asn Arg Leu Asp Lys Arg
35 40 45
Arg Lys Leu Asn Gly Cys Gln Arg Lys Asp Met Ser Tyr Gly Pro Ser
50 55 60
Gly Arg Ser Leu Leu Lys Cys Tyr Ser Asn Tyr Val Lys Thr Gly Thr
65 70 75 80
Pro Lys Arg Leu Met Phe Tyr Gln Asn Gly Glu Trp Ile Asp Phe Pro
85 90 95
Gln Ser Val Val Asp Val Val Arg Glu Asp Phe Gln Val Lys Lys Ser
100 105 110
Ala Val Glu Val Glu Phe Asn Gly His Arg Phe Met Leu Asp Phe Leu
115 120 125
His Met Ser Arg Val Asp Leu Lys Thr Thr Leu Gln Glu Pro Ile Ala
130 135 140
Trp Ile Asp Glu Ala Asp Ser Cys Phe Phe Pro Glu Thr Phe Asp Cys
145 150 155 160
His Gln Pro Glu Thr Met Glu Tyr Gln Asp Pro Val Leu Glu Glu Pro
165 170 175
Tyr Gly Pro Gln Glu Ile Lys Leu Leu Leu Glu Ile Glu Ile Asn Gly
180 185 190
Val Asp Gln Ser Lys Leu Thr Glu Cys Ser Gly Glu Ser Asn Asp Leu
195 200 205
Val Lys Gln Ile Gln Ile Asn Ser Lys Pro Ala Ser Asn Cys Tyr Ala
210 215 220
Ile Asp Val Glu Asn Ser Cys Ser Arg Glu Ser Asp Ala Lys Met Asp
225 230 235 240
Glu Asp Phe Gln Glu Asn Lys Gln Ile Pro Ala Asn Leu Val Ile Ala
245 250 255
Pro Val Ser Glu Asn Glu Glu Phe Asn Cys Asp Ser Val Gln Lys Leu
260 265 270
Phe Leu Val Gly Met Gly Ala Ser Gly Arg Pro Asp Ile Leu Glu Ile
275 280 285
Tyr Arg Cys Glu Ser Thr Ser Leu Gln Ala Arg Phe Glu Leu Phe Gln
290 295 300
Lys Gln Ala Glu Leu Thr Gln Lys Cys Arg Gly Asp Ala Asn Val Gln
305 310 315 320
Tyr Ala Trp Leu Ala Cys Ser Lys Gly Glu Leu Pro Thr Ile Leu Thr
325 330 335
His Gly Leu Gly His Cys Gly Pro Ser Thr Ile Lys Ser Met Tyr Gly
340 345 350
Ser Gly Val His Leu Ala Ala Ala Ile Cys Ser Tyr Thr Ser Ala Asn
355 360 365
Phe Cys Asp Val Asp Glu Asn Gly Val Gln His Leu Val Leu Cys Gln
370 375 380
Val Ile Met Gly Lys Met Glu Val Val His Ser Gly Ser Arg Gln Asn
385 390 395 400
Leu Pro Ser Cys Lys Asp Tyr Asp Ser Gly Val Asp Asp Leu Gln Asn
405 410 415
Pro Lys Ile Tyr Ile Val Trp Thr Met Asn Met Asn Thr His Ile Tyr
420 425 430
Pro Glu Phe Val Val Ser Phe Lys Ile Ser Ser Lys Thr Glu Gly Val
435 440 445
Thr Ser Cys Gln Val Arg Lys His Gln Gln Leu Glu Ser Ser Ala Val
450 455 460
Asp Leu Ser Val Ser Gln Pro Val Ser Asp Ser Gly Arg Ser Glu Gly
465 470 475 480
Lys Ala Pro Ser Leu Gly Ser Ser Asn Thr Arg Ala Pro Lys Ser Pro
485 490 495
Trp Met Pro Phe Pro Met Leu Phe Ala Ala Ile Ser Glu Lys Val Ser
500 505 510
Ser Gly Val Met Glu Lys Ile Asn Glu His Tyr Glu Leu Phe Arg Thr
515 520 525
Lys Lys Ile Gly Arg Asp Glu Phe Ile Lys Lys Leu Arg Leu Ile Val
530 535 540
Gly Asp Ala Leu Leu Arg Ser Thr Ile Thr Asn Leu Gln Cys Gln Leu
545 550 555 560
Pro Leu Arg Ser Lys Cys Glu Pro Glu Val Leu Gln Pro Asn Leu Glu
565 570 575
Lys Glu Lys Val Gln His Ser Phe Asn
580 585
Claims (9)
1. Quercus variabilis QsSRO1 protein is characterized in that: the amino acid sequence is shown in SEQ ID No. 2.
2. A quercus variabilis QsSRO1 gene encoding the quercus variabilis QsSRO1 protein of claim 1.
3. The quercus variabilis QsSRO1 gene of claim 2, wherein: the nucleotide sequence is shown in SEQ ID No. 1.
4. A vector containing the quercus variabilis QsSRO1 gene of claim 2 or 3.
5. A host cell comprising the vector of claim 4.
6. An engineered bacterium containing quercus variabilis QsSRO1 gene according to claim 2 or 3.
7. The use of the quercus variabilis QsSRO1 gene in the regulation of stress tolerance in plants according to claim 2 or 3, characterized in that the quercus variabilis QsSRO1 gene is overexpressed in plants.
8. Use according to claim 7, characterized in that: the method for improving the stress resistance of the plants comprises the following steps: constructing the vector of claim 4; transforming quercus variabilis QsSRO1 gene into plants using agrobacterium mediation; screening and culturing positive plants.
9. Use according to claim 7 or 8, wherein the plant comprises Arabidopsis thaliana.
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| US20070124833A1 (en) * | 2005-05-10 | 2007-05-31 | Abad Mark S | Genes and uses for plant improvement |
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