CN102040656A - Rice dehydrin protein with bacteriostatic activity and application thereof - Google Patents
Rice dehydrin protein with bacteriostatic activity and application thereof Download PDFInfo
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Abstract
The invention discloses a rice dehydrin protein with bacteriostatic activity, which is a protein with one of the following amino acid residue sequences: 1) SEQ ID NO: 1; 2) and (3) mixing the amino acid sequence shown in SEQ ID NO: 1, the protein has bacteriostatic activity and is obtained by substituting and/or deleting and/or adding one or more amino acid residues; 3) SEQ ID NO: 1, or a fragment of more than 15 amino acids; the rice dehydrin protein expressed in vitro, the protein fragment and the fragment synthesized in vitro all have the bacteriostatic activity on specific gram-positive bacteria, the dehydrin protein extracted from the plant disclosed by the invention can be used for inhibiting the gram-positive bacteria, and the resistance to the biological stress can be improved by transferring the rice dehydrin protein coding gene into the plant.
Description
Technical field
The present invention relates to the genetically engineered field, be specifically related to a kind of paddy rice dehydration fibroin matter and application with bacteriostatic activity.
Background technology
Microbiotic can suppress or kill and wound pathogenic micro-organism, and it is used the guarantee human health has been brought into play important effect.Yet,, cause antibiotic curative effect to descend because antibiotic excessive use has increased drug-resistance of bacteria.Antibacterial peptide is the micromolecule polypeptide with anti-microbial activity that exists in the biology, and they are important defence lines together that biology is resisted the pathogenic micro-organism invasion.Antibacterial peptide can act on the cytolemma of pathogenic bacteria, makes film depolarize or formation " hole ", causes cell content outflow or peptide molecule to enter cell interior and target effect, and then killing bacteria.Because antibacterial peptide can act on a plurality of sites of bacterium, thus be difficult for causing the resistance of bacterium, and antibiotic spectrum width.Therefore, antibacterial peptide might become new antibacterials.In addition, except that clinical application, the Application Areas of antibacterial peptide also comprises: be used for food preservatives, fodder additives, cosmetic field and improve disease resistance of plant etc.
The antibacterial peptide of first report is cecropin (cecropins) (Boman et al, 1974), and subsequently, Magainin (Magainins), melittin (Melitins), alexin priorities such as (Defensins) are found (Shi Qiang etc., 2000).Three during the last ten years, and people differentiate and obtained thousands of antibacterial peptides that the main screening strategy that is adopted is broadly divided into following several: one is based on the screening strategy of isolation technique; The 2nd, utilization variance technique of display screening antibacterial peptide encoding gene; Three are based on homologous sequence screening antibacterial peptide encoding gene; The 4th, differentiate antibacterial peptide with bioinformatics method; The 5th, high flux screening is differentiated the strategy (Wang Haijiao etc., 2007) of antibacterial peptide.2006, one-tenth hawk etc. has been developed a kind of easy high throughput method, this method is based on the oligo expression library of various cDNA expression libraries or synthetic, upholder (as millipore filtration) is placed on the solid medium that contains suitable resistance, expression library is coated on the upholder by certain density, treat that bacterium colony is long after a certain size, upholder is forwarded to continue to cultivate dyeing then on the substratum that contains inductor; Differentiate clone or cell activity.If the abduction delivering product is harmful to the host, make the active reduction of host or dead, then dead volume is by the dyeing after stain.
Dehydration plain (Dehydrin) finds that by people such as Close this is that a class can be by Exogenous ABA, arid and salinity inductive protein the earliest, and these protein have very strong wetting ability (Close et al, 1989).After this, people have reported the dehydration quality white matter in the different plants successively.Press the classification of Dure to contrary shock protein matter, dehydration fibroin matter belongs to LEA D-II family, and molecular weight does not wait from 9kD to 200kD, generally contains following three conservative fragments: Y, K and S fragment (Dure et al.1993) in its structure.Wherein, the K fragment is rich in Methionin, and common for all Dehydrin family proteins, its conserved sequence is EKKGIMDKIKEKLPG (Close et al.1997).But also the do not dewater report of the enough bacteria growing inhibitings of fibroin mass-energy up to the present.
Utilize the high flux screening system, the inventor differentiates that from expression libraries such as paddy rice, frog having obtained tens has inhibiting clone to intestinal bacteria growths, with the growth phase situation of cell in the inducing culture not relatively, the cell extent of growth obviously descends.In order further to verify the bacteria resistance function of paddy rice dehydration fibroin matter, the present invention utilizes Escherichia coli system to express dehydration fibroin matter and fragment thereof, external fragment of having synthesized dehydration fibroin matter, and carried out the bacteriostatic activity experiment of paddy rice dehydration fibroin matter.
Summary of the invention
The purpose of this invention is to provide a kind of paddy rice dehydration fibroin matter that bacteriostatic activity arranged and be applied to antibacterial practice.
First purpose of the present invention provides a kind of paddy rice dehydration fibroin matter (English name is Dehydrin) with bacteriostatic activity, and deriving from paddy rice (Oryzae Sativa L.) is the protein with one of following amino acid residue sequences:
1) the SEQ ID NO:1 in the sequence table;
2) with the amino-acid residue of SEQ ID NO:1 in the sequence table through the replacement of one or several amino-acid residue and/or disappearance and/or interpolation and have the protein of bacteriostatic activity;
3) 15 the above fragments of amino acid of the SEQ ID NO:1 in the sequence table;
Above-mentioned encoding gene with paddy rice dehydration fibroin matter (Dehydrin) of bacteriostatic activity also belongs to protection scope of the present invention, and its nucleotide sequence meets one of following characteristics:
1) nucleotide sequence of SEQ ID No:2 in the sequence table;
2) with sequence table in the dna sequence dna of SEQ ID No:2 85% above homology is arranged, and the identical function protein DNA sequence of encoding;
3) fragment of being longer than 45bp in the dna sequence dna of SEQ ID No:2 in the sequence table, and coding identical function protein DNA sequence.
Another object of the present invention is the application of paddy rice dehydration fibroin matter in antibacterial.
Experimental results show that, after described paddy rice dehydration fibroin matter or its fragment are expressed in bacterium, its bacterial lysate or protein purification can suppress the growth of gram positive bacteriums such as bacillus pumilus, subtilis, yellow sarcina and streptococcus aureus, simultaneously, the described protein fragments of external synthetic can suppress the growth of gram positive bacteriums such as bacillus pumilus, subtilis, yellow sarcina and streptococcus aureus.
So this protein is expressed in bacterium, fungi or plant or the form of purifying and the plasmid, bacterial strain and the transgenic plant that contain the said gene sequence also can be applicable to inhibition to bacterial growth.
Beneficial effect of the present invention:
With the method for Screening of Rice cDNA expression library, obtained to have dehydration element (Dehydrin) clone of bacteriostatic activity, in bacterium, express Dehydrin protein, its lysate has bacteriostatic activity to gram-positive microorganism; With Dehydrin protein partitioned representation, show that the De-K3 fragment has the obvious suppression effect to bacillus pumilus, subtilis, Sarcina lutea, streptococcus aureus etc.; Further the external Dehydrin fragment of having synthesized also proves to have bacteriostatic activity.Dehydrin protein or the fragment expressed in bacterium, fungi or the plant are mixed in feed, the foods and cosmetics, can be used for inhibition bacterium; In addition, change Dehydrin gene or fragment over to plant, its coded protein is induced down in the particular organisms adverse circumstance express, can improve the resistivity of plant adverse circumstance;
Description of drawings
Fig. 1 is the RR46 that screening obtains from a rice cDNA library clone's of the present invention liquid culture growth curve.
Fig. 2 is the external bacteriostatic experiment result of the bacterial lysate of paddy rice dehydration fibroin matter of the present invention.1 lysate for the expression of the Dehydrin after inducing bacterium; 2 lysates for the expression of the pET28a empty carrier after inducing bacterium; 3 for not inducing the bacterial lysate of Dehydrin expression; 4 is the bacterial lysate of inductive pET28a empty carrier not.
Fig. 3 is the external bacteriostatic experiment result of De-K3 fragment lysate of the present invention.Wherein 1 is the expression of the De-K3 after inducing bacterial lysate; 2 bacterial lysates for the pET28a empty carrier after inducing; 3 for not inducing the bacterial lysate of De-K3 expression; 4 for not inducing the bacterial lysate of pET28a empty carrier;
Fig. 4 is the segmental external bacteriostatic experiment result of external synthetic Dehydrin of the present invention.Wherein the concentration of 1,2,3 used synthetic polypeptide is respectively 1mM, 0.5mM and 0.25mM.
Embodiment
Method therefor is the used ordinary method of Molecular Biology Lab among the following embodiment.
Biomaterial, main agents and instrument
Paddy rice expression cDNA library (carrier: pBluescript, recipient bacterium: SOLR), coli strain BL21-DE3-pLysS; Expression vector pET28a, indicator strain streptococcus aureus (Staphylococcus aureus), rice leaf spot bacteria (Xanthomonas oryzae pv.oryzae) are this laboratory and preserve.Bacterium indicator bacillus pumilus (Bacillus pumilus, CVCC709), subtilis (Bacillus subtilis, CVCC717), intestinal bacteria (Escherichia coli, CVCC1570), Sarcina lutea (Sarcina lutea CVCC1600) all buys from national veterinary microorganism culture presevation administrative center.Dna sequencing and Oligo are synthetic to be finished by the Huada Gene Research Center, Beijing, and polypeptide is synthetic to be finished by Shanghai Huada Tianyuan Biotechnology Co.ltd.IPTG is available from match Parkson, Beijing Bioisystech Co., Ltd; Penbritin, kantlex are given birth to the worker available from Shanghai; Millipore filtration is available from Beijing Sheng He membrane-biotechnology company; Trypan blue (Typan Blue) and tetrabromophenol sulfonphthalein are available from Sigma company; Restriction enzyme is available from TaKaRa company; Ultrasonic Cell Disruptor is available from NingBo XinZhi Biology Science Co., Ltd.
Embodiment 1, the plain clone's of dehydration acquisition and the growth curve of bacterium
Press (Cheng et al such as Cheng, 2009) screening method, use expression library to screen, after IPTG induces, select trypan blue dyeing and become blue bacterium colony from rice root, through 2 take turns abduction delivering checking after, choose 35 clones and carried out end sequencing, sequencing result carries out BLASTX to be analyzed, and finds to have 7 clones coding Dehydrin protein, wherein 4 clones are the proteinic gene of coding SK3-Dehydrin, and their numbering is respectively RR46, RR51, RR142 and RR306.
Get RR46 clone's the bacterium that spends the night, 100mL contains penbritin 100 μ g/mL LB liquid nutrient mediums in the inoculation of 1% ratio, 37 ℃ of shaking culture are during to OD600 value 0.1 left and right sides, taking out 50mL adding IPTG (final concentration 1mmol/L) induces, not inductive contrast of remaining conduct, OD600 is once surveyed in every 1h sampling, draw the growth curve of RR46 clone bacterium, the SOLR bacterium of band pBluescript empty carrier is negative contrast, from growth curve as seen, the growth of the positive colony after inducing is considerably slower than not inductive bacterial strain and control strain (Fig. 1).
The clone of embodiment 2, dehydrin gene and protein expression
With the RR46 plasmid DNA is template, amplification has obtained the dehydrin gene, be cloned into (Zhai is from Jie etc. 2009) in the pET28a expression vector, clone's pET28a-Dehydrin plasmid is transformed in e. coli bl21-DE3-pLysS bacterial strain, line picking list colony inoculation 3mL contains in the kantlex 50 μ g/mL LB liquid nutrient mediums, and 37 ℃ of shaking culture are spent the night.100mL contains kantlex 50 μ g/mL LB liquid nutrient mediums by the switching of 1% inoculum size, and 37 ℃ are cultured to OD600=0.6～0.8, takes out 50mL and adds IPTG to final concentration 1mmol/L, induces 3h, the identical time of all the other microbial culture.Centrifugal, abandon supernatant, clean thalline 3 times, each 15mL sterilized water.Use 5mL PBS damping fluid resuspended again, (4 ℃ of centrifugal 10min, supernatant are Dehydrin protein bacterial lysate for the 10min of ON/OFF=5S/5S), 12000r/min in ultrasonication on ice.Collect 250mL Dehydrin abduction delivering bacterium, the centrifugal supernatant that goes cleans 3 times, each 15mL sterilized water, and 12000r/min after the ultrasonication, 4 ℃ of centrifugal 10min, supernatant Ni column purification is collected the higher component of protein content.
The inhibition of embodiment 3, dehydration fibroin confrontation bacterial growth
Beat the about 0.7cm circular filter paper of diameter sheet with the file puncher sanction, the oven dry of sterilization back, be taped against the media surface that remain to be tried bacterium containing of prepared fresh, the liquid protein that 20 μ L are to be measured is added on the filter paper, 37 ℃ of overnight incubation, second day observation inhibition zone also taken pictures, the result shows, bacterial lysate is to bacillus pumilus, subtilis, Sarcina lutea, streptococcus aureus has tangible fungistatic effect, but to intestinal bacteria, rice leaf spot bacteria does not have visible fungistatic effect (Fig. 2), and the Dehydrin protein of purifying does not have bacteriostatic activity (data are not attached).
Embodiment 4, the segmental expression of dehydration fibroin matter
Dehydrin protein can be divided into two parts, in order to determine to be responsible for the fragment of bacteriostatic activity, has carried out the subclone experiment of Dehydrin gene.Amplification De-S fragment the primer is, 5 '-GGAATTC
CATATGGAGGATGAGAGGAACACGG-3 ' and 5 '-CG
GGATCCTTAGCCGTTGTCATCGATCACC-3 '., amplification De-K3 fragment the primer is 5 '-GGAATTC
CATATGGAGGTGATCGATGACAAC-3 ' and 5 '-CG
GGATCCTTAAGCGCTGCTCTTGTGC-3 ', wherein underscore is the recognition site of restriction enzyme Nde I and BamH I.Behind the pcr amplification with product cloning in the pET28a carrier, two fragment protein of abduction delivering after the sequence verification.With their bacterial lysate and the protein of purifying above-mentioned indicator is carried out bacteriostatic experiment respectively then, the result shows, De-K3 fragment lysate has tangible fungistatic effect (Fig. 3) to bacillus pumilus, subtilis, Sarcina lutea, streptococcus aureus, and the fragment protein of De-S fragment lysate, purifying does not have visible fungistatic effect (data are not attached).
The bacteriostatic activity of embodiment 5, external synthetic dehydration fibroin matter
In order further to verify the fungistatic effect of De-K3, external fragment of synthesizing De-K3, its sequence is KKKKGLKEKIKEKLPGHK, and carried out the bacteriostatic experiment of synthetic polypeptide, the result shows, the polypeptide solution of three kinds of concentration (1mM, 0.5mM and 0.25mM) all has fungistatic effect to bacillus pumilus, subtilis, streptococcus aureus, and Sarcina lutea is not had visible fungistatic effect (Fig. 4).
Minimum inhibition concentration of embodiment 6, polypeptide (MIC) and minimum are killed concentration (MBC) and are detected
Adopt media dilution method to detect (Woong et al, 2006), in 96 holes, add the external synthetic polypeptide of different concns, add 10 then
5The indicator of CFU/mL, making polypeptide and bacterium cumulative volume is 200 μ L.Behind 37 ℃ of cultivation 12h, observe the turbidity in 96 each hole of orifice plate, minimum peptide concentration with the invisible microorganism growth of naked eyes is minimum inhibition concentration MIC, the sample of the above-mentioned bacterial growth that is invisible to the naked eye is taken out, be coated onto LB and cultivate 12h for dull and stereotyped last 37 ℃, do not have the minimum peptide concentration of bacterial growth to be minimum and kill concentration MBC, above-mentioned test is triplicate at least.After testing, external synthetic polypeptide is 130 μ g/mL to the minimum inhibition concentration of bacillus pumilus, and minimum is killed concentration and is＞400 μ g/mL.
Reference:
●Boman?HG，Nordstrom?K，Normark?S.Characteristics?of?an?inducible?cell-free?antibacterial?reaction?in?hemolymph?of?Samia?cynthia?pupae.Infect?Immun，1974，10(1)：136-145.
● the strong Liu Fei roc of stone. the expression of antibacterial peptide clone gene and transgenic research present situation. biotechnology progress, 2000,20 (1): 37-40.
●Dure?L.Response?of?plants?to?cellular?dehydration?during?environmental?stress.Rockville：American?Society?Physiologists，1993.91-103.
●Cheng?XY，Liu?GZ，Ye?GY，Wang?HJ，Shen?XJ，Wu?KC，Xie?JH，Altosaar?I.Screening?and?cloning?of?antimicrobial?DNA?sequences?using?a?vital?staining?method.Gene，2009，430：132-139.
●Woong?SJ，Hong?KK，Ki?YL，Sun?AK，Yeon?SH，In?HL，Antifungal?activity?of?synthetic?peptide?derived?from?halocidin，antimicrobial?peptide?from?the?tunicate，Halocynthia?aurantium.Federation?of?European?Biochemical?Societies，2006，580：1490-1496.
● the tender Li Li cloud Liu state of Wang Hai shakes. the screening strategy of antibacterial peptide and applied research progress thereof. and biotechnology circular 2007,5:29-33.
●Close?TJ，Kortt?AA，Chandler?PM，A?cDNA-based?comparison?of?dehydration-induced?proteins(dehydrins)in?barley?and?corn.Plant?Molecular?Biology，1989，13：95-108.
●Close?TJ.Dehydrins：a?commonalty?in?the?response?of?plants?to?dehydration?and?low?temperature.Physiol?Plant，1997，100：291-296.
●Close?TJ.Dehydrins：emergence?of?a?biochemical?role?of?a?family?of?plant?dehydration?proteins.Physiol?Plant，1996，97：795-803.
●Campos?F，Zamudio?F，Covarrubias?AA.Two?different?late?embryogenesis?abundant?proteins?from?Arabidopsis?thaliana?contain?specific?domains?that?inhibit?Escherichia?coli?growth.Biochemical?and?Biophysical?Research?Communications，2006，342：406-413.
● Zhai shakes from the tender Li Li cloud Liu state of Jie Wang Hai.The vivoexpression and the purifying research of paddy rice dehydration fibroin matter, plant physiology and molecular biology research, 2009,206-209.
Sequence table
＜110〉Agricultural University Of Hebei
＜120〉a kind of paddy rice dehydration fibroin matter and application with bacteriostatic activity
<130>
<160>2
<170>PatentIn?version?3.5
<210>1
<211>196
<212>PRT
<213>Oryza?sativa
<400>1
Glu?Val?Ile?Asp?Asp?Asn?Gly?Glu?Val?Val?Lys?Arg?Lys?Lys?Lys?Lys
1 5 10 15
Gly?Leu?Lys?Glu?Lys?Ile?Lys?Glu?Lys?Leu?Pro?Gly?His?Lys?Asp?His
20 25 30
Ala?Gly?Glu?His?Ala?Pro?Pro?Pro?Ala?Ala?Thr?Gly?Phe?Pro?Ala?Pro
35 40 45
Ala?Pro?Pro?Ala?Ser?Val?Val?Thr?Ala?Ala?Pro?Thr?Pro?Ala?Pro?Ala
50 55 60
Pro?Val?Val?Thr?His?Gly?Asp?His?His?His?Asp?Thr?Ala?Val?Pro?Val
65 70 75 80
Glu?Lys?Ile?Glu?Gly?Asp?His?Ala?Lys?Thr?Glu?Ala?Thr?Leu?Pro?Arg
85 90 95
Ala?Pro?Glu?Glu?Glu?Lys?Lys?Gly?Phe?Leu?Asp?Lys?Ile?Lys?Glu?Lys
100 105 110
Leu?Pro?Gly?Gly?His?Lys?Lys?Pro?Glu?Asp?Ala?Thr?Ala?Val?Pro?Pro
115 120 125
Pro?Ala?Ala?Ser?Pro?Ala?Ala?Pro?Ala?Thr?Thr?Pro?Ala?Pro?Ala?His
130 135 140
Pro?Pro?Pro?Ala?Thr?Glu?Glu?Val?Ser?Ser?Pro?Asp?Gly?Lys?Glu?Lys
145 150 155 160
Lys?Gly?Ile?Leu?Gly?Lys?Ile?Met?Glu?Lys?Leu?Pro?Gly?Tyr?His?Lys
165 170 175
Gly?Ser?Gly?Glu?Glu?Asp?Lys?Thr?Ala?Ala?Ala?Ala?Thr?Gly?Glu?His
180 185 190
Lys?Ser?Ser?Ala
195
<210>2
<211>591
<212>DNA
<213>Oryza?sativa
<400>2
gaggtgatcg?atgacaacgg?cgaggtggtc?aagaggaaga?agaagaaggg?gctcaaggag 60
aagatcaagg?agaagctgcc?cggccacaag?gaccatgccg?gtgagcatgc?tcctccgccc 120
gcggcgacgg?gcttccccgc?gccggctccg?cctgcatccg?tggtgacggc?cgcgcccacg 180
ccagctcctg?ctcccgtggt?gactcacggc?gatcaccacc?acgacaccgc?cgtccccgtg 240
gagaagatcg?agggtgatca?cgccaagacg?gaggcgaccc?tgccacgtgc?acccgaggag 300
gagaagaagg?gcttcctcga?caagatcaag?gagaagctgc?ccggcggcca?caagaagccg 360
gaagacgcaa?ctgctgtgcc?gccgccggcc?gcctcaccgg?ctgctcctgc?cactactccg 420
gcgccagcac?acccaccgcc?ggctacagag?gaagtgagca?gcccggatgg?gaaggagaag 480
aagggtatac?tgggcaagat?catggagaaa?ctgcccggtt?accacaaggg?ctccggcgag 540
gaagacaaga?ccgccgccgc?cgccaccggc?gagcacaaga?gcagcgctta?a 591
Claims (6)
1. paddy rice dehydration fibroin matter with bacteriostatic activity is characterized in that described protein has one of following amino acid residue sequences:
1) the SEQ ID NO:1 in the sequence table;
2) with the amino-acid residue of SEQ ID NO:1 in the sequence table through the replacement of one or several amino-acid residue and/or disappearance and/or interpolation and have the protein of bacteriostatic activity;
3) 15 the above fragments of amino acid of the SEQ ID NO:1 in the sequence table;
2. the encoding gene with paddy rice dehydration fibroin matter of bacteriostatic activity as claimed in claim 1 is characterized in that it has one of following nucleotide sequence:
1) nucleotide sequence of SEQ ID No:2 in the sequence table;
2) with sequence table in the dna sequence dna of SEQ ID No:2 85% above homology is arranged, and the identical function protein DNA sequence of encoding;
3) fragment of being longer than 45bp in the dna sequence dna of SEQ ID No:2 in the sequence table, and coding identical function protein DNA sequence.
3. express the proteinic bacterium in the claim 1, its lysate is in the purposes that suppresses bacillus pumilus, subtilis, yellow sarcina and streptococcus aureus.
4. express the protein in the claim 1 or the bacterium of polypeptide, the purposes of its lysate in suppressing the gram positive bacterium growth.
5. the protein of external synthetic claim 1 or polypeptide fragment, its purposes in suppressing the gram positive bacterium growth.
6. the transgenic cell line that contains the described gene of claim 2 is characterized in that it is applied to the opposing of plant to biological adverse circumstance.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102492030A (en) * | 2011-12-06 | 2012-06-13 | 吉林大学 | Gene comprising dehydrin functional domain and application thereof in anti-drought alkali-resistant gene engineering |
| CN106967724A (en) * | 2017-03-24 | 2017-07-21 | 上海交通大学 | Afriocan agapanthus SK3Type dehydrin protein and its encoding gene and probe |
| CN111116722A (en) * | 2020-01-20 | 2020-05-08 | 华中农业大学 | Wild rice antibacterial peptide OrR935 and application thereof |
-
2009
- 2009-10-12 CN CN2009100756484A patent/CN102040656A/en active Pending
Non-Patent Citations (3)
| Title |
|---|
| M. M. MUTHALIF ET AL.: "Identification of Dehydrin-Like Proteins Responsive to Chilling in Floral Buds of Blueberry (Vaccinium, section Cyanococcus)", 《PLANT PHYSIOLOGY》 * |
| SANG-CHOON LEE ET AL.: "Characterization of an Abiotic Stress-inducible Dehydrin Gene,", 《MOL. CELLS》 * |
| TIMOTHY J. CLOSE ET AL.: "Dehydrins: A commonalty in the response of plants to dehydration and low temperature", 《PHYSIOLOGIA PLANTARUM》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102492030A (en) * | 2011-12-06 | 2012-06-13 | 吉林大学 | Gene comprising dehydrin functional domain and application thereof in anti-drought alkali-resistant gene engineering |
| CN106967724A (en) * | 2017-03-24 | 2017-07-21 | 上海交通大学 | Afriocan agapanthus SK3Type dehydrin protein and its encoding gene and probe |
| CN111116722A (en) * | 2020-01-20 | 2020-05-08 | 华中农业大学 | Wild rice antibacterial peptide OrR935 and application thereof |
| CN111116722B (en) * | 2020-01-20 | 2022-03-08 | 华中农业大学 | Wild rice antibacterial peptide OrR935 and application thereof |
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