CN102532287A - Stress-resistant protein PpLEA3-17 of bryophyte as well as encoding gene and application thereof - Google Patents
Stress-resistant protein PpLEA3-17 of bryophyte as well as encoding gene and application thereof Download PDFInfo
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Abstract
The invention discloses stress-resistant protein PpLEA3-17 of bryophyte as well as an encoding gene and an application thereof. The protein related to the plant stress resistance and provided by the invention is named as PpLEA3-17, is from physcomitrella patens and is the protein in 1) or 2): (1) the protein consisting of amino acid sequences shown by the sequence 2 in a sequence table; and 2) the protein which is obtained by the amino acid residue sequence of the sequence 2 in the sequence table through substitution and/or deletion and/or addition of one or more amino acid residues, is related to the plant stress resistance and is derived by 1). The experiment for introducing the encoding gene of the PpLEA3-17 disclosed by the invention into wild type rice (Nipponbare) can prove that the survival rate (80.4%, 68.1%, 91.8% and 69.4%) and the growing state of four T2-substituted trans-PpLEA3-17-gene plant (17-1, 17-2, 17-3 and 17-4) in the drought stress experiment are obviously better than those of the control plant.
Description
Technical field
The present invention relates to the anti-contrary albumen PpLEA3-17 of bryophyte and encoding sox and application.
Background technology
The factors of coercing such as the arid in the physical environment, saline and alkaline, low temperature have material impact to growth and development of plant, can cause the extensive underproduction of farm crop when serious, and cultivating the resistance of reverse crop is one of major objective of plant husbandry.Current, the crop varieties that has resistance of reverse through the genetic engineering breeding acquisition is a kind of efficient ways.And the technical bottleneck problem of most critical is the screening and the function discovery of effective adversity gene in this method.
Bryophyte is the land pioneer plant that occur the ancient ordovician period before 500,000,000 years, and the gametophyte with " cormus " in its life history is main vegetative growth phase." cormus " blade is made up of monolayer, only locates to be made up of several confluent monolayer cells at its " middle rib ", and the weave construction of regulation and control such as no transfusion tissue and pore water metabolism is keeping the significantly characteristic of waterplant.Owing to lack water conduction and regulator control system, therefore, when original " bryophyte " dried up landing, must at first coerce greatly: water deficit, temperature shock in the face of two.Huge selective pressure forces bryophyte to be evolved out to be different from the adverse circumstance coping mechanism of " vascular plant " (fern, spermatophyte).The fact shows: be present in the more intracellular genes of bryophyte and bearing the responsibility of tackling serious adverse circumstance, they can protect cell to avoid the adverse circumstance injury.
Summary of the invention
The purpose of this invention is to provide a kind of albumen relevant and encoding sox thereof with stress resistance of plant.
The albumen relevant with stress resistance of plant provided by the present invention, name is called PpLEA3-17, derive from small liwan moss (Physcomitrella patens), is following 1) or 2) protein:
1) protein of forming by the aminoacid sequence shown in the sequence in the sequence table 2;
2) with the amino acid residue sequence of sequence in the sequence table 2 through the replacement of one or several amino-acid residue and/or disappearance and/or interpolation and relevant with stress resistance of plant by 1) deutero-protein.
In order to make 1) in PpLEA3-17 be convenient to purifying, label as shown in table 1 on proteinic N-terminal that can the aminoacid sequence shown in the sequence 2 is formed in by sequence table or C-terminal connect.
The sequence of table 1. label
| Label | Residue | Sequence |
| Poly-Arg | 5-6 (being generally 5) | RRRRR |
| Poly-His | 2-10 (being generally 6) | HHHHHH |
| FLAG | 8 | DYKDDDDK |
| Strep-tag?II | 8 | WSHPQFEK |
| c- |
10 | EQKLISEEDL |
Above-mentioned 2) but in the PpLEA3-17 synthetic, also can synthesize its encoding sox earlier, carry out biology again and express and to obtain.Above-mentioned 2) encoding sox of the PpLEA3-17 in can be through lacking sequence in the sequence table 1 codon of one or several amino-acid residue in the dna sequence dna shown in the 5 ' terminal 24-479 bit base; And/or carry out the missense mutation of one or several base pair, and/or obtain at the encoding sequence that its 5 ' end and/or 3 ' end connects the label shown in the table 1.
Above-mentioned and encoding sox plant adversity resistance related protein also belongs to protection scope of the present invention.
Specifically can be following 1 with the encoding sox of plant adversity resistance related protein)-4) in arbitrary described gene:
1) its encoding sequence is the deoxyribonucleotide from 5 ' terminal 24-479 position of sequence 1 in the sequence table;
2) its nucleotide sequence is the sequence 1 in the sequence table;
3) under stringent condition with 1) or 2) gene recombination and the gene of encoding said proteins;
4) with 1) or 2) gene have homology and the gene of encoding said proteins more than 90%.
Sequence 1 in the sequence table is by 583 based compositions, and its open reading frame (ORF) is from 5 ' terminal 24-479 bit base, and coding has the PpLEA3-17 albumen of the aminoacid sequence of sequence 2 in the sequence table.
Above-mentioned stringent condition can be that (or 0.1 * SSC), the solution of 0.1%SDS is hybridized under 65 ℃ and washed film with 0.1 * SSPE in DNA or RNA hybrid experiment.
Increase above-mentioned PpLEA3-17 full length gene or its arbitrary segmental primer to also belonging to protection scope of the present invention.
Contain above-mentioned and recombinant vectors, transgenic cell line and reorganization bacterium the plant adversity resistance related protein encoding sox and also belong to protection scope of the present invention.
Available existing plant expression vector construction contains the recombinant expression vector of PpLEA3-17 gene.Said plant expression vector comprises double base agrobacterium vector and the carrier etc. that can be used for the plant micropellet bombardment, like pCAMBIA3301, pCAMBIA1300, pBI121, pBin19, pCAMBIA2301, pCAMBIA1301-UbiN or other plant expression vector of deriving.
When using the gene constructed recombinant expression vector of PpLEA3-17; Can before its transcription initiation Nucleotide, add any enhancement type, composing type, organizing specific type or inducible promoter; Like cauliflower mosaic virus (CAMV) 35S promoter, ubiquitin (Ubiquitin) gene promoter (pUbi) etc., they can use separately or be used in combination with other plant promoter; In addition; When using gene constructed plant expression vector of the present invention; Also can use enhanser, comprise translational enhancer or transcriptional enhancer, these enhanser zones can be ATG initiator codon or neighboring region initiator codon etc.; But must be identical with the reading frame of encoding sequence, to guarantee the correct translation of whole sequence.The source of said translation wave and initiator codon is widely, can be natural, also can be synthetic.Translation initiation region can be from transcription initiation zone or structure gene.
For the ease of transgenic plant cells or plant being identified and screening; Can process used plant expression vector, as be added in the plant to express and to produce the enzyme of colour-change or the gene of luminophor (gus gene, GFP gene, luciferase genes etc.), have antibiotic marker thing (qingfengmeisu qiong affinity tag, kantlex affinity tag etc.) or the anti-chemical reagent marker gene (like anti-weedkiller gene) of resistance etc.From the security consideration of transgenic plant, can not add any selected marker, directly with adverse circumstance screening transformed plant.
Said recombinant expression vector specifically can be the plasmid pC1390 that is inserted with corn Ubiquitin promotor MCS between insert the above-mentioned recombinant expression vector that obtains with encoding sox plant adversity resistance related protein.
Another object of the present invention provides a kind of method of cultivating the transgenic plant of resistance raising.
The method of the transgenic plant that cultivation resistance provided by the present invention improves is that above-mentioned encoding sox PpLEA3-17 with plant adversity resistance related protein is imported in the purpose plant, obtains the transgenic plant that resistance improves.
Said encoding sox PpLEA3-17 with plant adversity resistance related protein imports in the plant through said recombinant expression vector.
Above-mentioned resistance specifically can be drought tolerance.
Conventional biological methods such as the plant expression vector that carries of the present invention and plant adversity resistance related protein encoding sox PpLEA3-17 can lead through Ti-plasmids, Ri plasmid, plant viral vector, directly DNA conversion, microinjection, electricity, agriculture bacillus mediated are transformed in the plant.Said plant is dicotyledons or monocotyledons, preferably paddy rice.
The present invention can obtain arid tolerance enhanced transgenic cell line and transfer-gen plant.
The present invention has found that through multiple molecular biology research technology a gene and expressing protein PpLEA3-17 thereof have significant degeneration-resistant border ability from bryophyte.PpLEA3-17 is a member in Late Embryogenesis Abundant protein (LEA) family.LEA gene after deliberation all plays important effect in different vital processes, the key protein molecule when being organism antagonism environment-stress.
The experiment that PpLEA3-17 gene of the present invention is imported wild-type paddy rice (Japan is fine) can prove: T
24 of generation are changeed PpLEA3-17 gene plant (17-1,17-2,17-3,17-4), and in the drought stress experiment, survival rate (80.4%, 68.1%, 91.8% and 69.4%) and growth conditions all will obviously be better than wild-type contrast and empty carrier adjoining tree.Explain that PpLEA3-17 gene provided by the invention and its encoded protein can significantly improve the drought tolerance of plant.To cultivating the plant with adverse resistance kind; The new variety such as crop, woods grass of particularly cultivating anti-abiotic stress (drought-enduring and/or salt tolerant) have important theory and practical significance, can be used for the cultivation and the evaluation of the required resistance of reverse plant variety of husbandry and ecological environment treatment.
Description of drawings
Fig. 1 is the proteinic two dimensional gel electrophoresis collection of illustrative plates of small liwan moss cormus, and arrow indication PpLEA3-17 albumen abundance in the plant that arid is handled significantly increases the adjoining tree that A. handles without arid; B. arid is handled 3 days plant.
Fig. 2 is the structure of plant expression vector pCU-PpLEA3-17.
Fig. 3 identifies that for the PCR of the rice plant of commentaries on classics PpLEA3-17 gene WT is the wild-type adjoining tree; 17-1,17-2,17-3,17-4 are 4 transgenic paddy rice strain systems.
Fig. 4 analyzes for RT-PCR and the Western that changes PpLEA3-17 trans-genetic hybrid rice plant, and A:RT-PCR analyzes; B:Western analyzes.
Fig. 5 is for changeing the plant height of PpLEA3-17 trans-genetic hybrid rice plant strain growth in the time of 60 days.
Fig. 6 changes the survival rate of PpLEA3-17 trans-genetic hybrid rice plant after arid is handled.
Fig. 7 changes the upgrowth situation of PpLEA3-17 gene plant in the drought tolerance experiment; A. before arid is handled, the transfer-gen plant situation; B. arid was handled after 10 days, the transfer-gen plant situation; C. after the arid rehydration, the transfer-gen plant situation.
Wherein among Fig. 3-7, WT is the wild-type adjoining tree; 17-1,17-2,17-3,17-4 are 4 transgenic paddy rice strain systems.
Embodiment
Below in conjunction with specific embodiment the present invention is described further, but the present invention is not limited to following examples.
Among the following embodiment,, be ordinary method like no specified otherwise.
The screening of embodiment 1, liver moss stress tolerance correlative protein PpLEA3-17 encoding sox PpLEA3-17 and cDNA clone thereof.
The small liwan moss arid is handled: will be grown in small liwan moss (Physcomitrella patens) (Geng Xuke etc., 2008 plant genes target practice---the application of model plant small liwan moss on the BCD substratum.The biology circular, 43 (4): 13-15; The public can obtain from Capital Normal University) cormus is transferred on the wetted filter paper, puts into the 500ml beaker, seal with thieving paper.24 ± 1 ℃ of temperature, carry out arid in the culturing room of relative humidity 30 ± 5% and handle.
After 3 days, the cormus that takes out arid is used for proteins extraction; The cormus of growing under the normal condition is done contrast.
The proteins extraction of small liwan moss cormus adopts the phenol extraction process.From under the normal condition with drought condition under protein separate with fluorescence difference gel electrophoresis (DIGE) through classical IEF/SDS-PAGE two-dimensional electrophoresis (2DE).Concrete steps are carried out according to the experimental implementation guide that Amersham Bioscience provides.
The protein pattern that obtains passes through ImageMaster 2D Platinum (Amersham Bioscience) and DeCyder 2D Software respectively, and Version 6.5 (Amersham Bioscience) analyzes.Abundance changes the separated acquisition of protein spots above 2 times under drought condition.All differential protein spots are used trypsin treatment respectively, and institute's polypeptide that obtains is identified with mass spectrograph (MALDI TOF/TOF MS) and DB (NCBI, small liwan moss genome).We find, under drought condition, have a protein on the two-dimensional electrophoresis (2DE) of classics and fluorescence difference gel electrophoresis (DIGE) gel, raised respectively 8.2 times and 12.3 times (Figure 1A, B).This protein is named as PpLEA3-17.
According to gene order design primer, to obtain the cDNA of total length.At first extract the total RNA of small liwan moss cormus after above-mentioned arid is handled, RNA is synthesized cDNA with reversed transcriptive enzyme.The CDS sequences Design primer of the gene that obtains according to above-mentioned retrieval is following: 5 '-GG
GGTACCGTAGTGTTGGAATCGGAG-3 ' and 5 '-GG
ACTAGTATGGTAGCCTCTGTGTC-3 '.The cDNA that obtains with reverse transcription is a template, carries out pcr amplification, the PCR product is carried out 0.8% agarose gel electrophoresis detect, and obtains the band that molecular weight is about 580bp, conforms to expected results.Reclaim test kit (TIANGEN) with sepharose and reclaim this fragment.Should reclaim fragment is connected with pMD19-T Simple (Takara); To connect product transformed into escherichia coli DH5 α competent cell; According to the carboxylic Bian penicillin resistance label screening positive colony on the pMD19-T Simple carrier, obtain containing the segmental recombinant plasmid of recovery.Universal primer M13 with on this recombinant plasmid vector carries out nucleotide sequencing to it; Sequencing result shows that the gene that increases (name PpLEA3-17) is made up of 583 deoxyribonucleotides; Its ORFs (ORF) for sequence 1 in the sequence table from 5 ' terminal the 24th to 479 deoxyribonucleotide, encoding amino acid sequence is the protein of sequence 2 in the sequence table.
Embodiment 2, commentaries on classics PpLEA3-17 gene are cultivated drought-enduring plant
1) structure of PpLEA3-17 expression vector pCU-PpLEA3-17
(with AAGCTTTCTAGTGCAGTGCAGCGTGAC and CTGCAGCCTCTAGTGCAGAAGTAACACCA is primer with the corn Ubiquitin promotor shown in the sequence in the sequence table 3; From corn gene group DNA, increase and obtain) (this carrier can be available from Cambia to insert plant binary expression vector pC1390; Between HindIII Queensland.Australia) and Kpn I site, constitute intermediate carrier pC1390-Ubi.The cDNA fragment of the PpLEA3-17 gene that amplification is obtained is with SpeI and Kpn I double digestion, between the Spe I and Kpn I site of forward insertion intermediate carrier pC1390-Ubi (gene of insertion just be positioned at the Ubiquitin promotor after), obtains recombinant vectors.The recombinant vectors called after pCU-PpLEA3-17 of the PpLEA3-17 gene of deoxyribonucleotide from 5 ' terminal 1-583 position that will contain sequence 1 in the ordered list.(as shown in Figure 2).This carrier has Ka Na and two selection markers of Totomycin.
2) acquisition and the evaluation of commentaries on classics PpLEA3-17 gene plant
The mode of pCU-PpLEA3-17 carrier with electric shock changed among the Agrobacterium LBA4404.28 ℃ of cultivations of picking positive colony.Treat that it is between 0.120～0.140 the time that Agrobacterium concentration reaches the OD600 value, infect paddy rice (Japan is fine) callus, infect 30min with this bacterium liquid, during shake with have gentle hands frequently.Callus is transferred on the common culture medium, and lucifuge is cultivated 3～4d altogether.Cultivate altogether finish after, callus transferred to selects to cultivate on the substratum 10～15d follow-up generation once.The new HYG resistant calli that picking comes out from former callus surface growth is transferred on the presorting substratum, presorting cultivation 7d under 28 ℃ of dark.Select creamy white, ganoid callus is transferred on the division culture medium, 28 ℃ of differentiation culture: under dark, cultivate 3d earlier, under lasting cold light shines, cultivate 15～20d then.To transfer on the root induction substratum from the seedling that resistant calli bears again, continuous light is cultivated 15d, and the seedling that upgrowth situation is good is directly transferred to field planting.Gather in the crops transgenic paddy rice seed then autumn.
After the seed results, be sowed on the MS screening culture medium that contains Totomycin (30mg/L) and screen.Treat that T1 moves on in the land for growing field crops when growing to the 4-6 leaf for plant and grow.For after the individual plant results, each single-strain seed is sowed respectively with T1, continues screening to observe the T2 separation case in generation with Totomycin, and so repeat number generation is until obtaining stable commentaries on classics PpLEA3-17 strain system.
PpLEA3-17 expresses 4 high strains systems in the commentaries on classics PpLEA3-17 strain system of selection genetic stability, carries out PCR respectively, RT-PCR and Western checking.
PCR checking: extract oryza sativa genomic dna with the CTAB method, use forward primer (5 '-GTAGTGTTGGAATCGGAG-3 ') and reverse primer (5 '-ATGGTAGCCTCTGTGTC-3 ') carry out pcr amplification.All can amplify the purpose band (Fig. 3) of 583bp in above-mentioned 4 strains system.
RT-PCR checking: extract paddy rice RNA with the Trizol method.The cDNA that obtains after the reverse transcription is used for being RT-PCR and analyzes, and primer is 5 '-GTAGTGTTGGAATCGGAG-3 ' and 5 '-ATGGTAGCCTCTGTGTC-3 ', result such as Fig. 4 A.
Western checking: use classification extraction process (Cui etc., 2005) to extract the paddy rice whole protein, the anti-Bo Ermai company that is prepared in of one in the Western trace.The result shows that the T of PpLEA3-17 gene is changeed in this 4 strain
2For plant 17-1,17-2,17-3 and 17-4, the PpLEA3-17 degree of gene expression is similar, and adjoining tree then fails to detect expression, shown in Fig. 4 B.
The drought tolerance of 3) changeing PpLEA3-17 gene strain system is identified
With in the empty carrier pC1390-Ubi importing wild-type paddy rice, prepare the empty carrier contrast according to the method that obtains to change PpLEA3-17 strain system.
This step is provided with empty carrier contrast and wild-type contrast
Under normal operation, begin to observe the phenotype of commentaries on classics PpLEA3-17 plant from the seedling phase.Change the extraordinary growth consistence that the PpLEA3-17 plant shows, but plant height is than empty carrier contrast and wild-type adjoining tree about 19.5% (Fig. 5, empty carrier contrasts consistent with wild-type adjoining tree phenotype, has omitted the empty carrier map) that descend; Strain shape, heading period and empty carrier adjoining tree and wild-type contrast be significant difference not.
With 4 T that change PpLEA3-17 gene strain system (17-1,17-2,17-3 and 17-4)
2For plant and the T that changes the empty carrier adjoining tree
2Plant in commercially available nutrition soil for plant and wild-type contrast, after the growth all around, stop to water.Rehydration is recovered experiment after 10 days.Statistics seedling survival number after 15 days.Three repetitions are established in experiment altogether, and in repeating, each strain of being tested is that plant is respectively 50 strains at every turn.The result shows: the survival rate of empty carrier adjoining tree and wild-type contrast is 0; And above-mentioned 4 survival rates of changeing the strain system of PpLEA3-17 gene are respectively 80.4%, 68.1%, 91.8% and 69.4% (Fig. 6, Fig. 7; The empty carrier contrast is consistent with wild-type adjoining tree phenotype, has omitted the empty carrier map).Explain that the drought tolerance of changeing the PpLEA3-17 gene plant obviously is superior to empty carrier contrast and wild-type adjoining tree.
Claims (10)
1. an albumen is following 1) or 2) protein:
1) protein of forming by the aminoacid sequence shown in the sequence in the sequence table 2;
2) with the amino acid residue sequence of sequence in the sequence table 2 through the replacement of one or several amino-acid residue and/or disappearance and/or interpolation and relevant with stress resistance of plant by 1) deutero-protein.
2. the said proteic encoding sox of claim 1.
3. encoding sox according to claim 2 is characterized in that: said proteic encoding sox is following 1)-4) in arbitrary described gene:
1) its encoding sequence is the deoxyribonucleotide from 5 ' terminal 24-479 position of sequence 1 in the sequence table;
2) its nucleotide sequence is the sequence 1 in the sequence table;
3) under stringent condition with 1) or 2) gene recombination and the said proteic gene of coding claim 1;
4) with 1) or 2) gene have the homology 90% or more and the said proteic gene of claim 1 of encoding.
4. the recombinant vectors, transgenic cell line or the reorganization bacterium that contain claim 2 or 3 said genes.
5. recombinant vectors according to claim 4 is characterized in that: said recombinant vectors is for inserting the recombinant expression vector that claim 2 or 3 said genes obtain between the MCS of the plasmid pC1390 that is inserted with corn Ubiquitin promotor.
6. total length or its arbitrary segmental primer of amplification claim 2 or 3 said genes are right.
7. a method of cultivating transgenic plant is that claim 2 or 3 described encoding soxs are changed in the purpose plant, obtains the resistant transgenic plant.
8. method according to claim 7 is characterized in that: claim 2 or 3 described encoding soxs are to import in the plant through claim 4 or 5 described recombinant expression vectors.
9. according to claim 7 or 8 described methods, it is characterized in that: said plant is dicotyledons or monocotyledons, preferably paddy rice.
10. according to claim 7 or 8 or 9 described methods, it is characterized in that: said resistant transgenic plant is drought-enduring transgenic plant.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102659936A (en) * | 2012-04-12 | 2012-09-12 | 中国农业科学院生物技术研究所 | Plant-stress-tolerance related protein, its encoding gene and application |
| CN102964438A (en) * | 2012-11-29 | 2013-03-13 | 中国农业科学院生物技术研究所 | Stress-resistance-related protein PpLEA3-23 of plant as well as coding gene and application of protein |
| CN103012570A (en) * | 2012-11-29 | 2013-04-03 | 中国农业科学院生物技术研究所 | Plant adverse resistance related protein PpLEA3-21 and coding gene and application thereof |
| CN104726488A (en) * | 2015-03-29 | 2015-06-24 | 河北科技大学 | Method for culturing stress-resistance herbicide-resistance transgenic aerobic rice |
| CN105801679A (en) * | 2016-04-15 | 2016-07-27 | 首都师范大学 | Application of protein PpLEA3-2 in regulating and controlling plant stress resistance |
| CN108192913A (en) * | 2018-01-26 | 2018-06-22 | 上海市农业生物基因中心 | A kind of method for improving plant abiotic stress resistance |
| CN113528535A (en) * | 2021-07-13 | 2021-10-22 | 丽水市润生苔藓科技有限公司 | Orphan gene PpDRO for improving stress resistance of plants and application thereof |
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| CN103911385B (en) * | 2014-04-24 | 2016-03-30 | 中国科学院西北高原生物研究所 | Medicago ruthenica late embryo generation Abundant protein gene and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102659936A (en) * | 2012-04-12 | 2012-09-12 | 中国农业科学院生物技术研究所 | Plant-stress-tolerance related protein, its encoding gene and application |
| CN102964438A (en) * | 2012-11-29 | 2013-03-13 | 中国农业科学院生物技术研究所 | Stress-resistance-related protein PpLEA3-23 of plant as well as coding gene and application of protein |
| CN103012570A (en) * | 2012-11-29 | 2013-04-03 | 中国农业科学院生物技术研究所 | Plant adverse resistance related protein PpLEA3-21 and coding gene and application thereof |
| CN102964438B (en) * | 2012-11-29 | 2014-05-07 | 中国农业科学院生物技术研究所 | Stress-resistance-related protein PpLEA3-23 of plant as well as coding gene and application of protein |
| CN104726488A (en) * | 2015-03-29 | 2015-06-24 | 河北科技大学 | Method for culturing stress-resistance herbicide-resistance transgenic aerobic rice |
| CN105801679A (en) * | 2016-04-15 | 2016-07-27 | 首都师范大学 | Application of protein PpLEA3-2 in regulating and controlling plant stress resistance |
| CN105801679B (en) * | 2016-04-15 | 2019-04-09 | 首都师范大学 | Application of the protein PpLEA3-2 in regulation stress resistance of plant |
| CN108192913A (en) * | 2018-01-26 | 2018-06-22 | 上海市农业生物基因中心 | A kind of method for improving plant abiotic stress resistance |
| CN108192913B (en) * | 2018-01-26 | 2021-03-12 | 上海市农业生物基因中心 | Method for improving abiotic stress resistance of plants |
| CN113528535A (en) * | 2021-07-13 | 2021-10-22 | 丽水市润生苔藓科技有限公司 | Orphan gene PpDRO for improving stress resistance of plants and application thereof |
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