Nothing Special   »   [go: up one dir, main page]

CN105017393B - With the relevant protein B hDNAJC2 of stress resistance of plant and its encoding gene and application - Google Patents

With the relevant protein B hDNAJC2 of stress resistance of plant and its encoding gene and application Download PDF

Info

Publication number
CN105017393B
CN105017393B CN201410176146.1A CN201410176146A CN105017393B CN 105017393 B CN105017393 B CN 105017393B CN 201410176146 A CN201410176146 A CN 201410176146A CN 105017393 B CN105017393 B CN 105017393B
Authority
CN
China
Prior art keywords
plant
resistance
sequence
bhdnajc2
protein
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201410176146.1A
Other languages
Chinese (zh)
Other versions
CN105017393A (en
Inventor
邓馨
陈世璇
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Institute of Botany of CAS
Original Assignee
Institute of Botany of CAS
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Institute of Botany of CAS filed Critical Institute of Botany of CAS
Priority to CN201410176146.1A priority Critical patent/CN105017393B/en
Priority to PCT/CN2015/079960 priority patent/WO2015165425A1/en
Publication of CN105017393A publication Critical patent/CN105017393A/en
Application granted granted Critical
Publication of CN105017393B publication Critical patent/CN105017393B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C12N15/8273—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for drought, cold, salt resistance

Landscapes

  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Cell Biology (AREA)
  • Botany (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Peptides Or Proteins (AREA)

Abstract

The invention discloses a kind of and relevant protein B hDNAJC2 of stress resistance of plant and its encoding gene and applications.Albumen provided by the present invention be it is following a) or b)：A) protein that amino acid sequence forms shown in sequence in sequence table 1；B) amino acid sequence of protein defined by a) by the substitution of one or several amino acid residues and/or is lacked and ored add, and with the relevant protein of stress resistance of plant.It is demonstrated experimentally that the encoding gene of albumen is imported wildtype Arabidopsis thaliana, transgenic arabidopsis is obtained, compared with wildtype Arabidopsis thaliana, drought resisting, salt resistance and alkali resistance significantly improve.The new varieties such as crop, the woods grass that the present invention improves cultivation drought resisting, salt resistance and alkali resistance have important theory and practical significance, the cultivation and identification of the resistance plant kind that can be used for needed for farming and animal husbandry and ecological environment treatment.

Description

With the relevant protein B hDNAJC2 of stress resistance of plant and its encoding gene and application
Technical field
The invention belongs to genetic engineering fields, are related to a kind of and the relevant protein B hDNAJC2 of stress resistance of plant and its coding Gene and application.
Background technology
With the shortage of global water resources and the growth of extreme weather events, influence of the adverse circumstance to the yield and quality of crop Increasingly significant.Therefore, resistance day show important by molecule manipulation technology raising crop.
After organism is by drought stress or other poor environment factors, a series of stress reaction is will produce, wherein hot Shock protein (heat shock protein, HSP) on the one hand can promote nascent protein fold and the albumen of false folding again just It really folds, denaturation collectin on the other hand can also be made to degrade, while maintaining a series of stability of enzymes and cell membrane.Further The study found that the content of HSPs is proportionate with the resistance to coercive of organism, and the stress ability of organism can be improved and inverse Survival rate in border.
Resurrection plant can endure the condition of Extreme drought, can restore normal activities quickly again when moisture content abundance. Resurrection plant in known angiosperm is seldom, and is mainly distributed on South Africa, South America and Australia.Revolve capsule lettuce tongue (Boea Hygrometrica it is) a kind of Gesneriaceae resurrection plant being distributed in China, the plant is in the karst based on limestone Heavy alkali area growth with high salt is vigorous, and blade has very strong drought-enduring recovery ability, in room temperature, the item that relative air humidity is 0 After being grown 72 hours under part, leaf r elative water content is reduced to 3% or so, blade area shrinkage to former blade area 1/3 hereinafter, Photosynthesis stops substantially.As long as water supply again, blade can absorb water stretching, extension, and revert to it is untreated before the apparent shape of blade State and physiological status (including photosynthetic recovery).
Invention content
The object of the present invention is to provide a kind of with the relevant protein B hDNAJC2 of stress resistance of plant and its encoding gene with answer With.
Protein provided by the present invention, entitled BhDNAJC2 derive from the rotation capsule lettuce tongue (Boea of Gesneriaceae Hygrometrica), it is following (a) or (b)：
(a) protein that amino acid sequence forms shown in sequence in sequence table 1；
(b) by the amino acid sequence of protein defined by (a) by one or several amino acid residues substitution and/or Lack and or add, and with the relevant protein of stress resistance of plant.
For the ease of the purifying of BhDNAJC2 albumen, the amino acid residue sequence of sequence 1 can be formed in by sequence table The upper label as shown in the table of amino terminal or carboxyl terminal connection of protein.
Table：The sequence of label
Tag residues sequence
Poly-Arg 5-6 (being usually 5) RRRRR
Poly-His 2-10 (being usually 6) HHHHHH
FLAG 8 DYKDDDDK
Strep-tag II 8 WSHPQFEK
c-myc 10 EQKLISEEDL
Protein in above-mentioned (b) can be artificial synthesized, also can first synthesize its encoding gene, then carries out biological expression and obtain. The encoding gene of protein in above-mentioned (b) can be one or several by will be lacked in DNA sequence dna shown in sequence in sequence table 2 The codon of amino acid residue, and/or carry out the missense mutation of one or several base-pairs.
The nucleic acid molecules for encoding the BhDNAJC2 albumen also belong to protection scope of the present invention.
The nucleic acid molecules can be DNA, such as cDNA, genomic DNA or recombinant DNA；The nucleic acid molecules can also be RNA, such as mRNA, hnRNA or tRNA.
In one embodiment of the invention, the nucleic acid molecules are specially the gene for encoding the BhDNAJC2 albumen (being named as BhDNAJC2)；The BhDNAJC2 genes can be following 1) to any DNA molecular in 5)：
1) coded sequence is DNA molecular shown in 193-696 of sequence 2 in sequence table；
2) DNA molecular shown in 181-721 of sequence 2 in sequence table；
3) DNA molecular shown in sequence 2 in sequence table；
4) under strict conditions with 1) -3) in the DNA molecular of any restriction hybridize and the DNA of code for said proteins points Son；
5) with 1) -4) in the DNA molecular of any restriction there are the DNA of 90% or more homology and code for said proteins points Son.
Above-mentioned stringent condition can be with 6 × SSC, and the solution of 0.5%SDS hybridizes at 65 DEG C, then with 2 × SSC, It is primary that 0.1%SDS and 1 × SSC, 0.1%SDS respectively wash film.
Wherein, sequence 2 is made of 913 nucleotide, and 193-696 are ORF, in polynucleotide shown in sequence 1 BhDNAJC2 albumen.
Recombinant vector, expression cassette, transgenic cell line or recombinant bacterium containing above-mentioned nucleic acid molecules also belong to the present invention's Protection domain.
The recombinant vector can be recombinant expression carrier or recombinant cloning vector.
The recombinant expression carrier can use existing plant expression vector construction.The plant expression vector includes double base agriculture Bacillus carrier and the carrier etc. that can be used for plant micropellet bombardment, as pGreen0029, pCAMBIA3301, pCAMBIA1300, The derivative plant expression vector of pBI121, pBin19, pCAMBIA2301, pCAMBIA1301-UbiN or other.The plant expression Carrier also may include 3 ' end untranslated regions of foreign gene, that is, include that polyadenylation signals and any other participation mRNA are processed Or the DNA fragmentation of gene expression.The bootable polyadenylic acid of polyadenylation signals is added to 3 ' ends of mRNA precursor.Using institute When stating gene constructed recombinant expression carrier, enhanced any type, composing type, tissue can be added before its transcription initiation nucleotide Idiotype or inducible promoter, such as cauliflower mosaic virus (CAMV) 35S promoter, ubiquitin gene Ubiquitin start Sub (pUbi), stress induced promoter rd29A etc., they can be used alone or are used in combination with other plant promoters； In addition, when using gene constructed recombinant expression carrier of the invention, enhancer, including translational enhancer or transcription also can be used to increase Hadron, these enhancer regions can be ATG initiation codon or neighboring region initiation codon etc., but required and coded sequence Reading frame it is identical, to ensure the correct translation of entire sequence.The source of the translation control signal and initiation codon is wide General, it can be natural, can also be synthesis.Translation initiation region can come from transcription initiation region or structural gene. For the ease of transgenic plant cells or plant are identified and screened, recombinant expression carrier used can be processed, such as The coding that being added can express in plant can generate the enzyme of color change or the gene of luminophor, resistant antibiotic Marker or anti-chemical reagent marker gene etc..Also it can be not added with any selected marker, directly screened and converted with adverse circumstance Plant.
In an embodiment of the present invention, start the promoter of the BhDNAJC2 genetic transcriptions in the recombinant expression carrier Specially pA35S promoters；The terminator for terminating the BhDNAJC2 genetic transcriptions is specially pA35S terminators.
More specifically, the recombinant expression carrier is interleaving in the recombination site attR1 and attR2 of pLeela carriers Enter the recombinant plasmid that the BhDNAJC2 genes obtain.
The expression cassette is by that can start the promoters of the BhDNAJC2 gene expressions, the BhDNAJC2 genes, with And transcription terminator composition.
The BhDNAJC2 albumen or the nucleic acid molecules or recombinant expression carrier, expression cassette or recombinant bacterium are such as Under it is any in application also belong to protection scope of the present invention：
(a1) regulate and control stress resistance of plant；
(a2) plant variety that selection and breeding resistance improves.
In the present invention, the regulation and control stress resistance of plant is embodied in：In the plant, if the BhDNAJC2 genes Expression quantity it is higher, then the resistance of the plant is stronger.
In the present invention, the method for the plant variety that the selection and breeding resistance improves, specifically may include will be described The step of higher plant of BhDNAJC2 gene expression amounts hybridizes as parent.
It is a further object to provide a kind of methods for the genetically modified plants that cultivation resistance improves.
The method provided by the present invention for cultivating the genetically modified plants that resistance improves, specifically may include following steps：
A) encoding gene that the BhDNAJC2 albumen is imported into purpose plant obtains expressing turning for the encoding gene Gene plant；
B) it is obtained compared with the purpose plant from genetically modified plants obtained by step a), the transgenosis that resistance improves is planted Object.
Expression quantity of the BhDNAJC2 albumen in the genetically modified plants is higher than the purpose plant.Described in coding The gene (the i.e. described BhDNAJC2 genes) of BhDNAJC2 albumen concretely following 1) to 5) in any DNA molecular：
1) coded sequence is DNA molecular shown in 193-696 of sequence 2 in sequence table；
2) DNA molecular shown in 181-721 of sequence 2 in sequence table；
3) DNA molecular shown in sequence 2 in sequence table；
4) under strict conditions with 1) -3) in the DNA molecular of any restriction hybridize and the DNA of code for said proteins points Son；
5) with 1) -4) in the DNA molecular of any restriction there are the DNA of 90% or more homology and code for said proteins points Son.
Above-mentioned stringent condition can be with 6 × SSC, and the solution of 0.5%SDS hybridizes at 65 DEG C, then with 2 × SSC, It is primary that 0.1%SDS and 1 × SSC, 0.1%SDS respectively wash film.
The BhDNAJC2 genes can specifically be imported by any of the above-described recombinant expression carrier in the purpose plant, Obtain the genetically modified plants.It specifically can be by using Ti-plasmids, Ri plasmids, plant viral vector, directly delivered DNA, micro- The recombinant expression carrier is converted plant cell or group by the conventional biology methods such as injection, conductance, agriculture bacillus mediated, particle gun It knits, and the plant tissue of conversion is cultivated into plant.Agriculture bacillus mediated equal biological methods are transformed into plant cell or tissue.
In above application or method, the resistance is concretely at least one of following：Drought resistance, salt resistance Property, alkali resistance.
In above application or method, the plant both can be dicotyledon or monocotyledon.
In one embodiment of the invention, the plant is dicotyledon, specially arabidopsis, such as the quasi- south of wild type Mustard Col-0.
The primer pair for expanding the BhDNAJC2 full length genes or its any segment also belongs to protection scope of the present invention.It is real Verify bright, the present invention screens one by drought-induced expression from resurrection plant rotation capsule lettuce tongue (Boea hygrometrica) BhDNAJC2 genes the channel genes wildtype Arabidopsis thaliana is obtained into BhDNAJC2 transgenic arabidopsis, with the quasi- south of wild type Mustard is compared, and drought resisting, salt resistance and alkali resistance significantly improve, and illustrates that BhDNAJC2 is and the relevant egg of plant drought, salt resistance and alkali resistant In vain.The new products such as crop, the woods grass that therefore BhDNAJC2 albumen and its encoding gene improve cultivation drought resisting, salt resistance and alkali resistance Kind has important theory and practical significance, the cultivation for the resistance plant kind that can be used for needed for farming and animal husbandry and ecological environment treatment With identification.
Description of the drawings
Fig. 1 is expression feelings of the BhDNAJC2 genes in the arid resuscitation process of rotation capsule lettuce tongue (Boea hygrometrica) Condition.In figure, significant difference (p is indicated between different lowercases<0.05).
Fig. 2 is T3 for BhDNAJC2 genes in BhDNAJC2 transgenic arabidopsis homozygous lines OE4-5, OE7-7, OE8-7 The RT-PCR testing results of expression.
Fig. 3 is that T3 is identified for the drought-resistant ability of BhDNAJC2 transgenic arabidopsis homozygous lines OE4-5, OE7-7, OE8-7. Wherein, A is each plant phenotype in 1/2MS (pH5.6) control medium；B is each plant phenotype on 1/2MS+20%PEG culture mediums C is root long statistical result；D is Ion leakage result；E is Fv/Fm results.In figure, difference is indicated between different lowercases Significantly (p<0.05).
Fig. 4 is that T3 is identified for the saline-alkaline tolerance of BhDNAJC2 transgenic arabidopsis homozygous lines OE4-5, OE7-7, OE8-7. Wherein, A is each plant phenotype in 1/2MS (pH5.6) control medium；B is 1/2MS+150mmolL-1It is each in NaCl culture medium Plant phenotype；C is root long statistical result；D is Ion leakage result；E is Fv/Fm results.In figure, between different lowercases Indicate significant difference (p<0.05).
Fig. 5 is that T3 is identified for the alkali resistant ability of BhDNAJC2 transgenic arabidopsis homozygous lines OE4-5, OE7-7, OE8-7. Wherein, A is each plant phenotype in 1/2MS (pH5.6) control medium；B is respectively to be planted on 1/2MS culture mediums (pH9.0) culture medium Strain phenotype；C is root long statistical result；D is Ion leakage result；E is Fv/Fm results.In figure, table between different lowercases Show significant difference (p<0.05).
Specific implementation mode
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
RNase inhibitor (is purchased from Takara companies).Reverse transcriptase SuperScript TM III (are purchased from Invitrogen Company).PENTR/D-TOPO carriers (are purchased from Invitrogen companies).Phusion High-Fidelity DNA Polymerase (is purchased from NEB companies).
Plant expression vector pLeela：It is recorded in " A novel role for histone methyltransferase KYP/SUVH4in the control of Arabidopsis primary seed dormancy, New Phytologist (2012)193:A 605-616 " texts, the public can obtain from Institute of Botany, Chinese Academy of Sciences.
Agrobacterium Gv3101 (Agrobacterium tumefaciens strain GV3101)：It is recorded in " Binary Agrobacterium vectors for plant transformation,M Bevanin,Nucleic Acids Research(1984)12(22):A 8711-8721 " texts, the public can obtain from Institute of Botany, Chinese Academy of Sciences.
Wildtype Arabidopsis thaliana Col-0 (ecotype Columbia-0, Arabidopsis thaliana)：It is recorded in “Arabidopsis,a useful weed.Meyerowitz EM,Cell(1989)56:The text of 263-270. " one, the public can be from Institute of Botany, Chinese Academy of Sciences obtains.
Embodiment 1, the clone of BhDNAJC2 genes and expression
Extraction handles 8 hours rotation capsule lettuce tongue (Boea hygrometrica) blade total serum IgE through drought stress, utilizesThe construction cDNA library library construction Kit (Stratagene, La Jolla, CA).Therefrom select at random 4800 genes are sequenced, and find wherein 1 coding DNA J type heat shock protein.Them are analyzed to lure in normal condition and arid Lead the dynamic change of rear gene expression.As a result, it has been found that the gene by drought-induced up-regulation, obtains the method for the gene specifically such as Under：
The total serum IgE of rotation capsule lettuce tongue (Boea hygrometrica) blade handled through drought stress is extracted, and using it as mould Plate carries out reverse transcription.Refer to Zhennan Zhang, Bo Wang, Dongmei Sun*＆Xin Deng*.2013, Molecular cloning and differential expression of sHSP gene family members from the resurrection plant Boea hygrometrica in response to abiotic stresses.Biologia68(4):Mono- texts of 651-661.
(1) DNA digests (10 μ l)：2 μ g of total serum IgE；10×DNase buffer1μl；DNaseI(50U/μl)0.4μl； 0.1 μ l of RNase inhibitor；DEPC-H2O complements to 10 μ l.37℃30min.
(2) reverse transcription (25 μ l)：10 μ l of total serum IgE by DnaseI digestion process；OligodT1μl；DEPC-H2O2.5μ l；70 DEG C of incubation 5min, place 5min on ice；Then 5 × buffer5 μ l are added；2mM dNTP5μl；0.5 μ of RNase inhibitor l；Reverse transcriptase SuperScript TM III1 μ l.42 DEG C of incubation 1h.
The cDNA that reverse transcription obtains dilutes 10 times and is used as template, using BhDNAJC2-F and BhDNAJC2-R as primer, uses Phusion High-Fidelity DNA Polymerase carry out the amplification of full length gene.Reaction condition is：First 98 DEG C of pre- changes Property 45sec；Then 98 DEG C denaturation 10sec, 55 DEG C annealing 20sec, then 72 DEG C extension 1.5min, totally 40 cycle；Last 72 DEG C Extend 10min.
BhDNAJC2-F：5’-CACCTTTGATTTCTGAATGGAGTTC-3 ' (is to be carried with pENTR/D-TOPO at underscore The matched sequence of body indentation, there, sequence thereafter are 181-201 of sequence 2)；
BhDNAJC2-R：(697-721 s' of sequence 2 is reversed by 5 '-TTGTGAA GCTGGTACACTC GAATTT-3 ' Complementary series).
After reaction, PCR product is detected into row agarose gel electrophoresis, it is 541bp's or so as a result to obtain size PCR product recycles the PCR product band.
The PCR product is connect with pENTR/D-TOPO carriers, correct plasmid will be sequenced and be denoted as intermediate carrier BhDNAJC2-pENTR.The structure of intermediate carrier BhDNAJC2-pENTR is described as：By the 181-721 of sequence in sequence table 2 The recombinant plasmid that DNA sequence dna shown in position obtains after being connect with pENTR/D-TOPO carriers.
It is BhDNAJC2 genes by unnamed gene shown in sequence 2, the code area of the gene is in sequence 2 from 5 ' ends the The albumen of the gene code is named as BhDNAJC2 albumen by 193-696 nucleotide, and the amino acid sequence of the albumen is sequence Sequence 1 in table.
The expression study of embodiment 2, BhDNAJC2 genes in revolving capsule lettuce dried lactuca drought resuscitation process
2 kinds of different degrees of Osmotic treatments are carried out to rotation capsule lettuce tongue (Boea hygrometrica), respectively：
Fresh Plants (F)：Normal watering daily；
In Fresh Plants soil at a slow speed arid 5 days (S5D)：Continuous 5 days without watering；
In Fresh Plants soil at a slow speed arid 14 days (S14D)：Continuous 14 days without watering.
In Fresh Plants soil at a slow speed arid 14 days after rehydration 3 days (A)：Continuous 14 days without carrying out 3 days just after watering Often watering recovery.
The plant number each handled is at least 5 plants.
The total serum IgE of 3 kinds of processing rotation capsule lettuce tongue blades is extracted respectively, reverse transcription obtains cDNA.Use gene-specific primer BhDNAJC2-f and BhDNAJC2-r carries out Q-PCR amplifications.And using 18S rRNA genes as internal reference, amplimer be 18S-f and 18S-r。
BhDNAJC2-f：5 '-GTTCTTGGTGTTCGCTCGTAC-3 ' (229-249 of sequence 2)；
BhDNAJC2-r：5 '-AGCAGAGCCGTACAATCCAG-3 ' (419-438 reverse complemental sequences of sequence 2 Row).
18S-f：5’-CTTAGTTGGTGGAGCGATTTG-3’；
18S-r：5’-CCTGTTATTGCCTCAAACTTCC-3’.
Reaction system is：CDNA1 μ l, 10 μ l2 × SYBR Green Master Mix, each 0.5 μ l of upstream and downstream primer, ddH2O8.5μl.Response procedures are：95 DEG C of pre-degenerations 30 seconds；95 DEG C are denaturalized 5 seconds, and 55 DEG C are annealed 30 seconds, and 72 DEG C extend 30 minutes, Totally 40 cycles.
The results are shown in Figure 1, it can be seen that the expression quantity of BhDNAJC2 genes is significantly higher than F groups in S14D groups.As it can be seen that BhDNAJC2 genes are obviously by drought-induced.
Embodiment 3, the acquisition of BhDNAJC2 transfer-gen plants and resistance identification
One, the acquisition of BhDNAJC2 transgenic arabidopsis
1, the acquisition of recombinant expression carrier pLeela-BhDNAJC2
By the intermediate carrier BhDNAJC2-pENTR built in embodiment 1, by LR reactions, (LR clone enzymes, are purchased from Invitrogen companies) by the PCR product importing plant expression vector pLeela of embodiment 1, homologous recombination obtains recombinant vector.
It will show to insert embodiment 1 between attR1 the and attR2 recombination sites of plant expression vector pLeela through sequencing The recombinant vector of PCR product (i.e. DNA fragmentation shown in 181-721 of sequence 2) be named as pLeela-BhDNAJC2.
In recombinant expression carrier pLeela-BhDNAJC2, the promoter for starting the BhDNAJC2 genetic transcriptions is PA35S promoters, the terminator for terminating the BhDNAJC2 genetic transcriptions are pA35S terminators.
2, the acquisition of recombinational agrobacterium
Above-mentioned recombinant expression carrier pLeela-BhDNAJC2 is transformed into Agrobacterium Gv3101, with containing 50 μ gmL-1 Kanamycin sulfate, 50 μ gmL-1Gentamicin and 50 μ gmL-1The YEB culture mediums of rifampin are screened, and are recombinated Agrobacterium.
The plasmid of extraction recombinant bacterium sends to sequencing, and as a result the plasmid is that (sequencing shows in pLeela pLeela-BhDNAJC2 The recombinant plasmid of DNA fragmentation shown in 181-721 of sequence 2 is inserted between attR1 the and attR2 recombination sites of carrier), it says Bright is positive restructuring bacterium, and by recombinant bacterium, it is named as Gv3101/pLeela-BhDNAJC2.
It tests while the control for being transferred to pLeela empty carriers into Agrobacterium Gv3101 is set, gained Agrobacterium is named as Gv3101/pLeela。
3, the acquisition and identification of BhDNAJC2 transgenic arabidopsis
(1) acquisition of BhDNAJC2 transgenic arabidopsis
It is wild that the conversion of inflorescence infusion method is respectively adopted in recombinant bacterium Gv3101/pLeela-BhDNAJC2 and Gv3101/pLeela Raw type arabidopsis Col-0 (ecotype Columbia-0, Arabidopsis thaliana) obtains T0 and turns for BhDNAJC2 is turned In gene arabidopsis and T0 generations, are transferred to the arabidopsis of pLeela empty carriers.
It takes T0 for BhDNAJC2 transgenic arabidopsis seeds, is all equably seeded in containing 10mgL-1The 1/ of phosphine oxamate On 2MS culture mediums, resistant surviving seedling (T1 generations) is moved to hot-house culture, and (22 DEG C of cultivation temperature, the photoperiod 16/8 is small When), collect seeds of the T1 for BhDNAJC2 transgenic arabidopsis.
100 T1 are taken to be seeded in containing 10mgL for BhDNAJC2 transgenic arabidopsis seeds respectively-1The 1/2MS of phosphine oxamate It is 3 by segregation ratio in culture medium：1 T2 moves to greenhouse training for the surviving seedling (5-10) of BhDNAJC2 transgenic arabidopsis It supports, collects seeds of the T2 for BhDNAJC2 transgenic arabidopsis.
100 T2 are taken to be seeded in containing 10mgL for BhDNAJC2 transgenic arabidopsis seeds respectively-1The 1/2MS of phosphine oxamate In culture medium, the survival seedling (10 or so) that there will be no separation T3 for BhDNAJC2 transgenic arabidopsis moves to hot-house culture, Collect seeds (homozygote) of the T3 for BhDNAJC2 transgenic arabidopsis.
From the T3 obtained for randomly selecting 3 plants in BhDNAJC2 transgenic arabidopsis homozygous lines, it is denoted as respectively：OE4-5、 OE7-7、OE8-7。
(2) identification of BhDNAJC2 transgenic arabidopsis
Respectively extract 10 days seedling ages above-mentioned T3 for BhDNAJC2 transgenic arabidopsis homozygous lines OE4-5, OE7-7, The total serum IgE of OE8-7 and wildtype Arabidopsis thaliana Col-0, and using it as template, with BhDNAJC2-F and BhDNAJC2-R, (sequence is shown in Embodiment 1) it is that primer carries out RT-PCR.PCR amplification program is：95 DEG C of pre-degeneration 4min；94 DEG C are denaturalized 30 seconds, 55 DEG C of annealing 30 Second, 72 DEG C extend 2 minutes, and totally 30 recycle；Finally again 72 DEG C extend 10 minutes.
With 18S rRNA genes as internal reference, amplimer is 18S-f and 18S-r for experiment (sequence is shown in embodiment 2).PCR Amplification program is：95 DEG C of pre-degeneration 4min；94 DEG C are denaturalized 30 seconds, and 55 DEG C are annealed 30 seconds, and 72 DEG C extend 30 seconds, and totally 30 recycle； Finally again 72 DEG C extend 10 minutes.
Experimental setup using the wildtype Arabidopsis thaliana Col-0 of non-transgenosis and be transferred to the arabidopsis of pLeela empty carriers as pair According to.
After reaction, PCR product is detected into row agarose gel electrophoresis, every group of sample carries out 2 repetitions.
The results are shown in Figure 2, it can be seen that does not have BhDNAJC2 in the wildtype Arabidopsis thaliana Col-0 (WT) of non-transgenosis Gene amplifies band, and T3 all has size in BhDNAJC2 transgenic arabidopsis homozygous lines OE4-5, OE7-7, OE8-7 The about target fragment of 541bp, it is seen that T3 is sun for BhDNAJC2 transgenic arabidopsis homozygous lines OE4-5, OE7-7, OE8-7 Property.And the wildtype Arabidopsis thaliana Col-0 (WT) for being transferred to the Arabidopsis plants of pLeela empty carriers and non-transgenosis is unanimously, does not expand Go out the target fragment that size is about 541bp.
Two, the resistance identification of BhDNAJC2 transgenic arabidopsis
1, experimental method
By the positive T3 of step 1 identification for BhDNAJC2 transgenic arabidopsis homozygous lines OE4-5, OE7-7, OE8-7, It is transferred to the Arabidopsis plant of pLeela empty carriers, and the wildtype Arabidopsis thaliana Col-0 of non-transgenosis, is sowed at 1/2MS culture mediums (pH5.6) it on, after 4 DEG C of vernalization 3 days, cultivates under conditions of 22 DEG C, 50% humidity, illumination 16h and dark 8h, it will be small after 4 days Seedling is transferred to the following three kinds culture mediums for Analysis of Resistance respectively, is taken a picture after 10 days, and statistics root long, measurement ion ooze (relative conductivity) and PSII Efficiency of primary conversion of light energy (Fv/Fm) are leaked, to analyze the drought resisting of each strain, salt resistance and alkali resistant Ability.Each 6-8 plants of strain, experiment is set altogether to be repeated three times, and results are averaged.
Culture medium for analyzing drought resistance：1/2MS+20%PEG (mass percentage) culture medium, pH5.6；
Culture medium for analyzing salt-resistance：1/2MS+150mmol·L-1NaCl culture medium, pH5.6；
For analyzing alkali-resisting culture medium：1/2MS culture mediums (pH9.0).
In experiment, when each Analysis of Resistance, is respectively provided with 1/2MS (pH5.6) as a contrast.
Cell membrane has selection permeability and remains stable, but when plant is by environment-stress, cell membrane system first by It destroys, cell membrane a large amount of cavities occurs and even ruptures, and cell Dissolve things inside is largely spilt into extracellularly, leads to cell leaching liquor conductance Rate increased dramatically, index (Wang YC, the Qu GZ, Li whether relative conductivity has come to harm as measure of cell plasma membrane HY,Wu YJ,Wang C,Liu GF,Yang CP(2010).Enhanced salt tolerance of transgenic poplar plants expressing a manganese superoxide dismutase from Tamarix androssowiI.Molecular Biology Reports37,1119-1124.).Fv/Fm reflects II reaction center luminous energy of PS Transformation efficiency, the size and variation characteristic of parameter value are often used for judging and speculating that plant to the resistance of environmental factor, also has People be used as Heat Resistance of Plant, low temperature resistant, salt tolerant, resistance to strong light, drought-resistant, antipollution etc. important indicator (Yang CW, Peng CL,Duan J,Lin GZ,Chen YZ(2002).Responses of chlorophyll fluorescence and carotenoids biosynthesis to high light stress in rice seedling leaves at different leaf position.Acta Botanica Sinica44,1303-1308.)。
Wherein, relative conductivity assay method is as follows：It is measured and is planted using conductivity meter (EC215, Italian HANNA companies) The relative conductivity of object takes the seedling that complete each strain is cultivated under different disposal, with distilled water flushing 3 times, is blotted with filter paper Surface moisture respectively takes 0.1g (fresh weight), is respectively placed in the scale test tube of 6ml deionized waters, screws a lid on to be placed in and soak at room temperature Bubble processing 4h.Leaching liquor conductance A is measured with conductivity gauge, then boiling water bath heats 30min, shakes up after being cooled to room temperature, surveys again Determine leaching liquor conductance B.Relative conductivity=A/B × 100%.
Fv/Fm assay methods are as follows：It is imaged system (MAXI-Imaging-Pam, German Walz using modulated chlorophyll fluorescence Company) PSII Efficiency of primary conversion of light energy (Fv/Fm) is measured, the seedling of complete each strain is cultivated under different disposal under dark After handling 20min, chlorophyll fluorescence Fv/Fm is measured.Data are read with ImagingWin v2.32, are imported in excel and are divided Analysis.
2, experimental result
(1) Drought Resistance Analysis
The results are shown in Figure 3.
A. root long result
As shown in A-C in Fig. 3.Wherein, A is each plant phenotype observation in 1/2MS (pH5.6) control medium in Fig. 3, can To find out, the almost the same (p of root long of each plant>0.05).B is 1/2MS+20%PEG (mass percentage) culture medium in Fig. 3 Upper each plant phenotype observation, it can be seen that compared with wildtype Arabidopsis thaliana Col-0 (WT), T3 generations turn BhDNAJC2 arabidopsis strains It is the longer (p of root long of OE4-5, OE7-7, OE8-7<0.05).B is counted in root long result such as Fig. 3 shown in C, in 1/2MS+ in Fig. 3 On 20%PEG (mass percentage) culture medium, T3 for BhDNAJC2 transgenic arabidopsis homozygous lines OE4-5, OE7-7, The root long of OE8-7 is respectively 5.00cm, 5.00cm, 5.17cm, is longer than the root long 4.28cm of wildtype Arabidopsis thaliana Col-0 (WT), And with the root long of each plant in 1/2MS (pH5.6) control medium without Osmotic treatment without significant difference (p>0.05).
B. Ion leakage result
As shown in D in Fig. 3, it can be seen that on 1/2MS+20%PEG (mass percentage) culture medium, T3 generations The relative conductivity of BhDNAJC2 transgenic arabidopsis homozygous lines OE4-5, OE7-7, OE8-7 are respectively 38%, 31%, 28%, substantially less than (p<0.05) relative conductivity 46% of wildtype Arabidopsis thaliana Col-0 (WT).
C.Fv/Fm results
As shown in E in Fig. 3, it can be seen that on 1/2MS+20%PEG (mass percentage) culture medium, T3 generations The Fv/Fm of BhDNAJC2 transgenic arabidopsis homozygous lines OE4-5, OE7-7, OE8-7 are respectively 0.62,0.64,0.59, significantly Higher than (P<0.05) the Fv/Fm values 0.5 of wildtype Arabidopsis thaliana Col-0 (WT).
(2) salt-resistance is analyzed
The results are shown in Figure 4.
A. root long result
As shown in A-C in Fig. 4.Wherein, A is each plant phenotype observation in 1/2MS (pH5.6) control medium in Fig. 4, can To find out, the almost the same (p of root long of each plant>0.05).B is 1/2MS+150mmolL in Fig. 4-1It is respectively planted in NaCl culture medium Strain Phenotypic Observation, it can be seen that compared with wildtype Arabidopsis thaliana Col-0 (WT), T3 generations turn BhDNAJC2 arabidopsis strains OE4- 5, the longer (p of the root long of OE7-7, OE8-7<0.05).B is counted in root long result such as Fig. 4 shown in C, in 1/2MS+ in Fig. 4 On 150mmolL-1NaCl culture mediums, T3 is for BhDNAJC2 transgenic arabidopsis homozygous lines OE4-5, OE7-7, OE8-7 Root long is respectively 1.91cm, 1.87cm, 2.69cm, hence it is evident that is longer than (p<0.05) root long of wildtype Arabidopsis thaliana Col-0 (WT) 1.38cm。
B. Ion leakage result
As shown in D in Fig. 4, it can be seen that on 1/2MS+150mmolL-1NaCl culture mediums, T3 turns for BhDNAJC2 The Ion leakage value of gene arabidopsis homozygous lines OE4-5, OE7-7, OE8-7 are respectively 65%, 61%, 62%, substantially less than (p<0.05) the Ion leakage value 77% of wildtype Arabidopsis thaliana Col-0 (WT).
C.Fv/Fm results
As shown in E in Fig. 4, it can be seen that in 1/2MS+150mmolL-1In NaCl culture medium, T3 turns for BhDNAJC2 The Fv/Fm of gene arabidopsis homozygous lines OE4-5, OE7-7, OE8-7 are respectively 0.80,0.81,0.78, are significantly higher than (p< 0.05) the Fv/Fm values 0.23 of wildtype Arabidopsis thaliana Col-0 (WT).
(3) alkali resistance is analyzed
The results are shown in Figure 5.
A. root long result
As shown in A-C in Fig. 5.Wherein, A is each plant phenotype observation in 1/2MS (pH5.6) control medium in Fig. 5, can To find out, the almost the same (p of root long of each plant>0.05).B is that each plant phenotype is seen on 1/2MS culture mediums (pH9.0) in Fig. 5 Examine, it can be seen that compared with wildtype Arabidopsis thaliana Col-0 (WT), T3 generation turn BhDNAJC2 arabidopsis strains OE4-5, OE7-7, Longer (the p of root long of OE8-7<0.05).B is counted in root long result such as Fig. 5 shown in C, at 1/2MS culture mediums (pH9.0) in Fig. 5 On, T3 is respectively 7.32cm for the root long of BhDNAJC2 transgenic arabidopsis homozygous lines OE4-5, OE7-7, OE8-7, 6.04cm, 8.40cm, hence it is evident that be longer than (p<0.05) the root long 0.90cm of wildtype Arabidopsis thaliana Col-0 (WT).
B. Ion leakage result
As shown in D in Fig. 5, it can be seen that on 1/2MS culture mediums (pH9.0), T3 is pure for BhDNAJC2 transgenic arabidopsis The Ion leakage value for closing strain OE4-5, OE7-7, OE8-7 is respectively 26%, 25%, 20%, substantially less than (p<0.05) wild The Ion leakage value 59% of type arabidopsis Col-0 (WT).
C.Fv/Fm results
As shown in E in Fig. 5, it can be seen that on 1/2MS culture mediums (pH9.0), T3 is for BhDNAJC2 transgenic arabidopsis The Fv/Fm of homozygous lines OE4-5, OE7-7, OE8-7 are respectively 082,0.79,0.83, are significantly higher than (p<0.05) wild type is quasi- The Fv/Fm values 0.17 of southern mustard Col-0 (WT).
In addition, for above each as a result, turning empty carrier arabidopsis and non-transgenosis wildtype Arabidopsis thaliana Col-0 (WT) result Without significant difference.
In conclusion the drought resisting of BhDNAJC2 transfer-gen plants, salt resistance and alkali resistance are substantially better than the plant of non-transgenosis, Illustrating BhDNAJC2 is and the relevant albumen of plant drought, salt resistance and alkali resistant.

Claims (12)

1. protein, the protein that amino acid sequence forms shown in sequence in sequence table 1.
2. encoding the nucleic acid molecules of protein described in claim 1.
3. nucleic acid molecules according to claim 2, it is characterised in that：The nucleic acid molecules are described in coding claim 1 The gene of protein；The gene is following 1) to any DNA molecular in 3)：
1) coded sequence is DNA molecular shown in 193-696 of sequence 2 in sequence table；
2) DNA molecular shown in 181-721 of sequence 2 in sequence table；
3) DNA molecular shown in sequence 2 in sequence table.
4. the recombinant vector, expression cassette containing nucleic acid molecules described in Claims 2 or 3 or recombinant bacterium.
5. recombinant vector according to claim 4, it is characterised in that：The recombinant vector is recombinant expression carrier or recombination Cloning vector.
6. recombinant vector according to claim 5, it is characterised in that：In the recombinant expression carrier, start the gene Transcription promoter be pA35S promoters.
7. the recombination described in protein described in claim 1 or nucleic acid molecules according to claim 2 or 3 or claim 4 Carrier, expression cassette or recombinant bacterium it is following it is any in application：
(a1) regulate and control stress resistance of plant；
(a2) plant variety that selection and breeding resistance improves.
8. application according to claim 7, it is characterised in that：The resistance is at least one of following：Drought resistance, Salt-resistance, alkali resistance.
9. application according to claim 7 or 8, it is characterised in that：The plant is dicotyledon or monocotyledon.
10. the method for cultivating the genetically modified plants that resistance improves, includes the following steps：
A) encoding gene that protein described in claim 1 is imported into purpose plant obtains expressing turning for the encoding gene Gene plant；
B) it is obtained compared with the purpose plant from genetically modified plants obtained by step a), the genetically modified plants that resistance improves.
11. according to the method described in claim 10, it is characterized in that：The resistance is at least one of following：Drought resisting Property, salt-resistance, alkali resistance.
12. the method according to claim 10 or 11, it is characterised in that：The plant is that dicotyledon or unifacial leaf are planted Object.
CN201410176146.1A 2014-04-29 2014-04-29 With the relevant protein B hDNAJC2 of stress resistance of plant and its encoding gene and application Active CN105017393B (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
CN201410176146.1A CN105017393B (en) 2014-04-29 2014-04-29 With the relevant protein B hDNAJC2 of stress resistance of plant and its encoding gene and application
PCT/CN2015/079960 WO2015165425A1 (en) 2014-04-29 2015-05-27 Methods and materials for improving plant stress resistance

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201410176146.1A CN105017393B (en) 2014-04-29 2014-04-29 With the relevant protein B hDNAJC2 of stress resistance of plant and its encoding gene and application

Publications (2)

Publication Number Publication Date
CN105017393A CN105017393A (en) 2015-11-04
CN105017393B true CN105017393B (en) 2018-10-02

Family

ID=54358196

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201410176146.1A Active CN105017393B (en) 2014-04-29 2014-04-29 With the relevant protein B hDNAJC2 of stress resistance of plant and its encoding gene and application

Country Status (2)

Country Link
CN (1) CN105017393B (en)
WO (1) WO2015165425A1 (en)

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105524925B (en) * 2016-01-11 2019-03-08 北京市农林科学院 Plant inducible promoter and its application
WO2018234191A1 (en) * 2017-06-19 2018-12-27 University Of Copenhagen Increased drought resistance in plants
CN109678940B (en) * 2017-10-18 2022-05-10 中国科学院植物研究所 Protein BhDnaJ6, and coding gene and application thereof
CN110669121B (en) * 2019-11-14 2022-05-24 九圣禾种业股份有限公司 Method for improving salt tolerance of cotton by using LeDNAJ gene and application
CN112772416B (en) * 2021-01-28 2022-08-12 苏州金铁红林业科技有限公司 Tissue culture seedling raising method for phyllostachys microphylla
CN113881685B (en) * 2021-10-09 2024-03-29 江苏省农业科学院 Gene PpHSP20-like1 for promoting plant organ to produce red color and application thereof
CN116514941B (en) * 2023-06-09 2024-05-03 青岛农业大学 MsRGP protein, coding gene thereof and application of MsRGP protein in improving drought resistance and salt tolerance of plants

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008043245A1 (en) * 2006-09-11 2008-04-17 The Chinese University Of Hong Kong Abiotic stress tolerance conferred by j-domain-containing proteins
CN101921774A (en) * 2010-05-24 2010-12-22 南京大学 Application of Dnaj-like protein and encoded gene thereof
CN103361370A (en) * 2012-03-27 2013-10-23 中国科学技术大学 Desiccation tolerance and resurrection related gene BdSDMT from resurrection plant Boea densihispidula and application thereof
CN103361361A (en) * 2012-03-27 2013-10-23 中国科学技术大学 Desiccation tolerance and resurrection related gene BdBCP1 from resurrection plant Boea densihispidula and application thereof

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20110039908A (en) * 2009-10-12 2011-04-20 고려대학교 산학협력단 A method for promoting resistance against biological or abiological environmental stress in plants
CN103451193B (en) * 2013-09-25 2015-01-21 大连民族学院 Populus deltoidesx populus nigra PdHSP70 gene and application thereof

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008043245A1 (en) * 2006-09-11 2008-04-17 The Chinese University Of Hong Kong Abiotic stress tolerance conferred by j-domain-containing proteins
CN101921774A (en) * 2010-05-24 2010-12-22 南京大学 Application of Dnaj-like protein and encoded gene thereof
CN103361370A (en) * 2012-03-27 2013-10-23 中国科学技术大学 Desiccation tolerance and resurrection related gene BdSDMT from resurrection plant Boea densihispidula and application thereof
CN103361361A (en) * 2012-03-27 2013-10-23 中国科学技术大学 Desiccation tolerance and resurrection related gene BdBCP1 from resurrection plant Boea densihispidula and application thereof

Also Published As

Publication number Publication date
CN105017393A (en) 2015-11-04
WO2015165425A1 (en) 2015-11-05

Similar Documents

Publication Publication Date Title
CN105017393B (en) With the relevant protein B hDNAJC2 of stress resistance of plant and its encoding gene and application
CN104292317B (en) Drought resistant correlative protein of plant and coding gene and application thereof
US20190085355A1 (en) Drought tolerant maize
BR112019019977A2 (en) methods to increase grain yield
CN111032684A (en) Method for increasing grain productivity
CN104558128B (en) The albumen related to anti-Fusarium graminearum stem rot and its encoding gene and application
CN105566463B (en) One kind albumen relevant to Chlorophyll synthesis and its encoding gene and application
CN108642065A (en) A kind of paddy endosperm silty related gene OsSecY2 and its coding protein and application
CN110713994B (en) Plant stress tolerance associated protein TaMAPK3, and coding gene and application thereof
CN108276481B (en) Upland cotton GhLEA3 gene and application thereof in low-temperature stress resistance
CN105777882B (en) One plant resistance related protein TaWRKY35 and its encoding gene and application
CN103897047B (en) Protein B hHSP70-1 and encoding gene thereof and application
CN103570813B (en) With plant adversity resistance related protein Gh01399 and encoding gene and application
CN104945492B (en) Plant stress tolerance correlative protein TaAREB3 and its encoding gene and application
CN107325161B (en) Protein related to low-nitrogen stress and high-salt stress resistance as well as encoding gene and application thereof
CN105647940A (en) Method for improving rice yield through OsGRF6 gene, and applications thereof
CN106337041B (en) Plant stress tolerance correlative protein and its encoding gene and application
CN104892740B (en) Plant stress tolerance correlative protein GmEF13 and its encoding gene and application
CN102911262B (en) Protein related with plant tolerance and coding gene and applications thereof
CN103172714A (en) Rice leaf rolling-associated protein OsMYB103L as well as encoding gene and application thereof
CN103819544B (en) Drought tolerance in plants associated protein AtPR1 and encoding gene thereof and application
CN105950598B (en) Rice dormancy-breaking related protein and coding gene and application thereof
CN104140461B (en) The LTP albumen relevant to plant cold tolerance and encoding gene thereof and application
CN107176983B (en) Application of protein PpLEA3-3 in regulation and control of plant stress resistance
Zheng et al. Genome-wide analysis of TCP gene family in Osmanthus fragrans reveals a class I gene OfTCP13 modulate leaf morphology

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant