CN114835787B - Application of Quercus suberectus QsSRO1 gene and encoding protein thereof in plant stress resistance - Google Patents
Application of Quercus suberectus QsSRO1 gene and encoding protein thereof in plant stress resistance Download PDFInfo
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Abstract
The application belongs to the technical field of genetic engineering, and particularly relates to application of a cork oak QsSRO1 gene and a coding protein thereof in plant stress resistance. The nucleotide sequence of the cork oak QsSRO1 gene is shown as SEQ ID No.1, and the coded cork oak QsSRO1 protein has the amino acid sequence of SEQ ID No.2, and is a cell nuclear protein. The recombinant expression vector containing the Quercus suberectus QsSRO1 gene is constructed, the agrobacterium is utilized to transform the arabidopsis, and the QsSRO 1-transformed arabidopsis plant seeds obtained after screening and cultivation perform well in a salt resistance test. The application proves that the over-expression of the QsSRO1 gene of the cork oak can improve the salt resistance of plants, and lays a theoretical and production practice foundation for cultivating new varieties of transgenic crops with salt resistance and drought resistance.
Description
Technical Field
The application belongs to the technical field of genetic engineering, and particularly relates to an application of a Quercus suberectus QsSRO1 gene and a coding protein thereof in plant stress resistance.
Background
Quercus suber is an important broad-leaved evergreen tree species that grows slowly and has an extremely long life. Cork oak is a major source of industrial cork, a material of great commercial value.
Climate change is one of the major challenges and global problems faced in the 21 st century. In recent decades, as the global air temperature rises, a series of more intense, frequent and long lasting climatic extreme events, in particular severe drought and soil salinization, are caused. Various global changes such as soil salinization have been reported to result in a serious decrease in the number of cork oaks. Although tree species can adapt to new environmental conditions, little is known about the processes involved. Conventional cross breeding is a method of obtaining individuals with a recombination of parental genes by mating between individuals of different genotypes. The new gene is not generated in the hybridization process, the character separation of the filial generation occurs, the breeding process is slow, and the process is complex. The plant genetic engineering breeding has the characteristics of directionally changing plants, being not limited by 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 cork oak so as to ensure the sustainability of economic tree species.
Disclosure of Invention
The application aims to provide an application of a Quercus suberectus 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 object, the present application provides the following solutions:
the first aspect of the application provides a Quercus suberectus 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 the Quercus suberectus QsSRO1 protein is as follows:
MEANIAKASDRSKRVVLDLKRKRATQLATYLNEVRPTWDSLQNRLDKR RKLNGCQRKDMSYGPSGRSLLKCYSNYVKTGTPKRLMFYQNGEWIDFPQSV VDVVREDFQVKKSAVEVEFNGHRFMLDFLHMSRVDLKTTLQEPIAWIDEADS CFFPETFDCHQPETMEYQDPVLEEPYGPQEIKLLLEIEINGVDQSKLTECSGES NDLVKQIQINSKPASNCYAIDVENSCSRESDAKMDEDFQENKQIPANLVIAPVS ENEEFNCDSVQKLFLVGMGASGRPDILEIYRCESTSLQARFELFQKQAELTQK CRGDANVQYAWLACSKGELPTILTHGLGHCGPSTIKSMYGSGVHLAAAICSY TSANFCDVDENGVQHLVLCQVIMGKMEVVHSGSRQNLPSCKDYDSGVDDLQ NPKIYIVWTMNMNTHIYPEFVVSFKISSKTEGVTSCQVRKHQQLESSAVDLSV SQPVSDSGRSEGKAPSLGSSNTRAPKSPWMPFPMLFAAISEKVSSGVMEKINE HYELFRTKKIGRDEFIKKLRLIVGDALLRSTITNLQCQLPLRSKCEPEVLQPNLE KEKVQHSFN
in a second aspect, the application provides a cork oak QsSRO1 gene encoding said cork oak QsSRO1 protein, derived from Quercus suber. The nucleotide sequence of the Quercus suberectus QsSRO1 gene is shown as SEQ ID No. 1.
The application also provides an over-expression vector, a host cell and engineering bacteria containing the quercus variabilis QsSRO1.
The third aspect of the application provides application of the Quercus suberectus QsSRO1 gene in regulation and control of plant salt resistance.
Further, the method for improving stress resistance of the plant comprises the following steps: constructing a recombinant expression vector containing the Quercus suberectus QsSRO1 gene; transforming the Quercus suberect QsSRO1 gene into a plant using Agrobacterium mediation; and (5) screening and culturing positive plants.
Further, the plants include arabidopsis thaliana, and T4 generation seeds of transgenic arabidopsis thaliana plants are salt resistant.
Compared with the prior art, the application has the advantages that: the application provides a QsSRO1 gene of cork oak and a coded protein thereof, and the salt resistance test result of transgenic T4 arabidopsis plants of QsSRO1 shows that when the cork oak is treated in a salt solution, the excessive expression of the QsSRO1 gene is helpful for reducing the damage of salt to plants, so that the transgenic arabidopsis of QsSRO1 has stronger tolerance to salt. Therefore, the cork oak QsSRO1 gene provided by the application is suitable for improving the stress resistance of arabidopsis thaliana, especially the salt stress resistance, and functional analysis and identification of the cork oak QsSRO1 gene also provide a theoretical basis for obtaining new plant varieties, and the use of the gene for molecular breeding can be used for culturing stress-resistant plants with production application value, so that theoretical and production practice bases are laid for culturing new transgenic crop varieties with salt resistance and drought resistance.
Drawings
FIG. 1 shows the expression of the QsSRO1 gene after the treatment of various tissues and saline hypochondriacs of Quercus suberectus.
FIG. 2 shows the change of the expression level of QsSRO1 gene with time after salt stress of Quercus suberectus.
FIG. 3 shows the results of molecular detection of Arabidopsis thaliana transformed with QsSRO1.
FIG. 4 is the observation that Arabidopsis thaliana transformed with QsSRO1 gene and wild type Arabidopsis thaliana seeds germinate for 7 days under 100mM NaCl treatment.
FIG. 5 is a root length statistic of 7 days of germination of Arabidopsis thaliana and wild type Arabidopsis thaliana seeds transformed with QsSRO1 under 100mM NaCl treatment.
FIG. 6 shows subcellular localization of transient transformed QsSRO1 proteins.
Detailed Description
In order to enable those skilled in the art to better understand the technical solution of the present application, the technical solution of the present application will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present application. Unless otherwise defined, 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 application belongs. The test materials used in the examples described below, unless otherwise specified, were purchased from conventional biochemical reagent stores. 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 application.
Example 1: quercus suberectus QsSRO1 gene and obtaining method thereof
1. Method for obtaining QsSRO1 of Quercus suberectus
Extracting RNA of Quercus suberectus, and using TaKaRa PrimeScriptTM with RNA as template RT The kit is reversely transcribed into cDNA by a reverse transcription kit method. And then, taking cDNA as a reaction template for amplifying a target gene sequence, and carrying out PCR (polymerase chain reaction) amplification by taking 5'-ATGGCGAATCTACCCCAATCGC-3' and 5'-TCAGTTTCCGGGCTCTGGTTTC-3' as primers to obtain a CDS sequence of the QsSRO1 gene.
2. Analysis of tissue expression of QsSRO1 Gene by fluorescent quantitative PCR
Respectively extracting RNA of root, stem and flower of Quercus suberectus, and performing reverse transcription to synthesize cDNA; and then respectively carrying out PCR amplification by taking cDNA of each tissue as a template and 5'-AGACGGGACGAGGGTTGT-3' and 5'-TGGATGATGTGGGCTTGG-3' as primers, and detecting the expression condition of the QsSRO1 gene in different tissues of the cork oak. Meanwhile, the qACTIN gene is taken as an internal reference, and the 5' primer for amplifying the qACTIN gene is as follows: 5'-GCGGATAGAATGAGCAAGGAA-3',3' primers are: 5'-GGGCCGGACTCATCATACTC-3'. RT-qPCR reaction conditions: firstly, pre-denaturing at 94 ℃ for 1min; then 98℃for 10sec,58℃for 30sec,72℃for 1min, 30 cycles total; and then extending at 72 ℃ for 3min.
The expression of the Quercus suberectus QsSRO1 gene in each tissue is shown in FIG. 1. The results show that the Quercus suberectus QsSRO1 gene is expressed in all tissues, wherein the expression quantity in stems is the lowest, and the expression quantity in roots is the highest.
The QsSRO1 gene expression of the QsSRO1 gene under the condition of salt stress is shown in figure 2. The results show that the expression level of the QsSRO1 gene of the cork oak is increased after the cork oak is subjected to salt stress, and the expression level is highest in 8 hours.
Example 2: application of QsSRO1 gene in improving stress resistance of plants
1. Construction of QsSRO 1-transformed Arabidopsis thaliana
Obtaining the QsSRO1 gene of Quercus suberectus, and preparing the following primer pairs:
primer 1:5'-ATGGCGAATCTACCCCAATCGC-3';
primer 2:5'-TCAGTTTCCGGGCTCTGGTTTC-3'.
The cDNA of the Quercus suberectus is used as a template, the primer 1 and the primer 2 are adopted for PCR amplification, a PCR product with the size of 1755bp is obtained, and the sequencing is carried out. The obtained PCR product is a QsSRO1 gene of Quercus suberectus, the nucleotide sequence shown by the sequencing result is shown as SEQ ID NO.1, and the PCR product can code 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 QsSRO1 protein contains a Trp-Trp-Glu domain at the N end, a polyadenylation polymerase structure is arranged in the middle of the sequence, and a TATA box binding protein related factor 4 is arranged at the C end.
The QsSRO1 gene sequence (SEQ ID NO. 1) is as follows, with a full length of 1755bp:
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 recombinant expression vectors
And (3) connecting the PCR product obtained in the step (1) and the pCAMBIA1300 vector after restriction enzyme digestion to obtain a connecting product. The ligation product was then transformed into E.coli DH 5. Alpha. Competent cells and plated on LB plates containing 25. Mu.g/ml ampicillin and cultured overnight. White single colonies were picked up and cultured overnight in LB liquid medium containing 50. Mu.g/ml kanamycin and colony PCR identified; and extracting plasmid DNA by an alkaline method to perform sequence determination.
Sequencing results show 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-QsSRO1.
3. Acquisition and molecular detection of QsSRO 1-transformed Arabidopsis thaliana
(1) Acquisition of transgenic QsSRO1 Arabidopsis thaliana
The recombinant expression vector pCAMBIA1300-QsSRO1 prepared in the step 2 is transformed into competent cells of agrobacterium GV3101 (purchased from biological engineering (Shanghai) Co., ltd.) to obtain recombinant engineering bacteria GV3101/pCAMBIA1300-QsSRO1 (the extracted plasmid is sent to be sequenced and is the recombinant vector pCAMBIA1300-QsSRO 1).
GV3101/pCAMBIA1300-QsSRO1 engineering bacteria are inoculated in YEB liquid culture medium containing 50mg/L chloramphenicol in a single clone, and are cultured for two days at 28 ℃ in a shaking way. The culture broth was centrifuged at 4000rpm/min for 5 minutes and the resulting agrobacterium pellet was resuspended in bacterial cells using an infestation solution containing 5% sucrose and 0.03% silwet l-77.
Columbia ecological wild type Arabidopsis thaliana (Col-0) (purchased from Arabidopsis Biological Resource Center) was transformed by the cotton seed dip method, seeds (T1 generation) from current generation transgenic Arabidopsis plants were harvested, and germinated seeds were selected in MS medium containing 50. Mu.g/ml kanamycin (kanamycin). And transferring the T1 generation seedlings germinated on the culture medium into culture soil, harvesting seeds (T2 generation), and obtaining homozygous QsSRO1 arabidopsis plant (T4 generation) seeds through the same screening process.
(2) Molecular detection
The genomic DNA of the wild type Arabidopsis and T4 generation plants was used as the final strand, and the primers 1 and 2 in example 2 were used for amplification. The experimental results of PCR amplification are shown in FIG. 3 (Col-0 is wild type). The result shows that the transgenic T4 generation seeds can be amplified to about 1800bp bands, but the wild arabidopsis thaliana cannot be amplified to about 1800bp bands, so that the transgenic T4 generation seeds are identified as T4 generation homozygous transgenic QsSRO1 arabidopsis thaliana.
4. Functional study of Arabidopsis thaliana transformed to QsSRO1
(1) Salt resistance test of seeds
Taking seeds of wild arabidopsis thaliana (White) and transgenic arabidopsis thaliana plants (generation T4) of QsSRO1, and performing a seed germination experiment on an MS medium containing 100mM NaCl, wherein the experimental condition is that the photoperiod is 16 hours of illumination and 8 hours of darkness; the illumination intensity is 300-400 mu mol m -2 s -1 The method comprises the steps of carrying out a first treatment on the surface of the The culture temperature under the illumination condition is 21-24 ℃, and the relative humidity is 80%; the culturing temperature under dark condition is 21-24deg.C, and the relative humidity is 80%.
As shown in FIGS. 4 to 5, col-0 is a control group of wild type Arabidopsis thaliana, and OE1 and OE2 are both QsSRO 1-transformed Arabidopsis thaliana plants. Seeds of QsSRO 1-transformed Arabidopsis plants (T4 generation) were significantly longer in root length than wild-type Arabidopsis at 100mM NaCl concentration.
The result shows that the QsSRO1 protein (SEQ ID NO. 2) encoded by the QsSRO1 gene has the function of improving the salt resistance of plant seeds.
(2) Subcellular localization of QsSRO1 proteins
Preparing 10ml of enzymolysis liquid. Taking 1mth healthy and not yet flowering arabidopsis rosette leaves, cutting the leaves into about 1mm strips, immersing the strips in an enzymolysis solution, and carrying out enzymolysis for three hours at 60rpm under the conditions of room temperature and darkness. And (5) screening the enzymolysis liquid by a 200-mesh sieve, and sub-packaging the enzymolysis liquid into a 30ml centrifuge tube. The cryocentrifuge was centrifuged at 4 ℃,120rpm for five minutes, the supernatant was discarded, and the protoplasts were resuspended with a pre-chilled W5 solution (150mM NaCl,125mM CaCl2, 5mM KCl,2mM MES,pH =5.8). The solution was rinsed 3 times with pre-chilled W5 solution, centrifuged at 120rpm for two minutes and placed on ice for half an hour. At normal temperature, maMg solution was used for resuspension. Mu.g of plasmid was taken and added to 150. Mu.l of protoplasts, mixed gently and shaken well, 120. Mu.l of PEG/Ca solution was added and gently blown. Shaking at 50rpm for ten minutes, adding 0.5ml of W5 solution, and gently mixing. At room temperature, 100g, and centrifuged for five minutes. The protoplasts were resuspended by adding 1.5ml of W5, at room temperature, 100rpm, centrifuging for one minute, discarding the supernatant, and re-suspending the protoplasts by adding 200. Mu. l W5 solution, incubating for twelve hours at room temperature. After the above steps are completed, observation is performed by using a fluorescence confocal microscope.
The results are shown in FIG. 6, which shows the scale bars of 5 μm, and the results indicate that the encoded protein of the QsSRO1 gene exists in the nucleus and belongs to a nuclear protein.
The above embodiments are only for illustrating the technical solution of the present application, and are not limiting; although the application has been described in detail with reference to the foregoing embodiments, it will be apparent to one skilled in the art that modifications may be made to the technical solutions described in the foregoing embodiments, or equivalents may be substituted for some of the technical features thereof; such modifications and substitutions do not depart from the spirit and scope of the corresponding technical solutions.
Sequence listing
<110> Qingdao Stanny Heterol environmental technology institute Co., ltd
<120> application of Quercus suberectus QsSRO1 gene and encoding protein thereof in plant stress resistance
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1755
<212> DNA
<213> Quercus suber (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 (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 suberect QsSRO1 protein, its characterized in that: the amino acid sequence is shown as SEQ ID No. 2.
2. A cork oak qsro 1 gene encoding a cork oak qsro 1 protein of claim 1.
3. The quercus variabilis QsSRO1 gene according to claim 2, characterized in that: the nucleotide sequence is shown as SEQ ID No. 1.
4. A vector comprising the quercus variabilis QsSRO1 gene of claim 2 or 3.
5. A host cell comprising the vector of claim 4.
6. An engineered bacterium comprising the QsSRO1 gene of Quercus suberecta according to claim 2 or 3.
7. Use of the cork oak QsSRO1 gene according to claim 2 or 3 for regulating plant stress resistance, wherein the cork oak QsSRO1 gene is overexpressed in plants, the stress resistance being resistance to salt stress.
8. The 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 the Quercus suberectus QsSRO1 gene into plants by using agrobacterium mediation; and (5) screening and culturing positive plants.
9. The use according to claim 7 or 8, wherein the plant comprises arabidopsis thaliana.
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