CN106676128A - Paddy rice OsWOX11 protein and application of paddy rice OsWOX11 protein for coding gene - Google Patents
Paddy rice OsWOX11 protein and application of paddy rice OsWOX11 protein for coding gene Download PDFInfo
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Abstract
The invention relates to paddy rice OsWOX11 protein and an application of the paddy rice OsWOX11 protein for coding gene. The invention concretely relates to a recombinant expression vector, the recombinant expression vector can over-express WOX protein in plant cells, preferably the paddy rice WOX11 protein; the recombinant expression vector contains transgene cell line or engineering bacteria of recombinant expression vector, and the invention also provides the application of the recombinant expression vector or engineering bacteria. The overexpression of the WOX gene can influence the callus generation capability of an explant in a plant tissue culture. The protein provides a novel direction for plant tissue culture and variety breeding, has high practical application value, and has wide application prospect.
Description
Technical field
The invention belongs to technical field of biological genetic engineering, and in particular to rice Os WOX11 albumen and its coding
The application of gene.
Background technology
Plant Tissue Breeding is referred under aseptic and manually operated environmental condition, using appropriate culture medium pair
In vitro plant organ histocyte and protoplast is cultivated so as to regenerative cell or whole plant
Technology.The type of its research mainly includes the training of tissue cultures, organ culture, cell culture and protoplast
Support.Tissue cultures have been widely used in crop breeding, the production of secondary metabolites, the quick breeding of plant
Deng field.
Callus is depended on using technologies such as the genetic transformation based on tissue cultures, breed of variety and quick breedings
The formation of tissue.Traditionally, it is believed that callus is a dedifferentiation, the cell mass of Disordered Splitting.With
Research deepen continuously, it has been found that callus similar to an orderly root restriction, its formed more like
Be a transdifferentiation process (Sugimoto, K., Jiao, Y. and Meyerowitz, E.M., 2010,
Arabidopsis regeneration from multiple tissues occurs via a root development
Pathway, Dev Cell 18,463-471；Liu,J.、Sheng,L.、Xu,Y.、Li,J.、Yang,Z.、
Huang, H. and Xu, L., 2014, WOX11and 12are involved in the first-step cell fate
Transition during de novo root organogenesis in Arabidopsis, Plant Cell 26,
1081-1093〕.Plant is considered to have very strong power of regeneration, and its adult stem cell is distributed in each tissue
In, and the cell derived of callus be exactly these potential adult stem cells (Sugimoto, K. etc., ibid；
Liu, J. etc., ibid).Plant tissue training is improved by using the regulatory mechanism of adult stem cell in plant body
Callus generating ability is strengthening the new method of Callus formation ability, more from source during supporting
Plus the meaning with universality.
WOX (WUSCHEL related homeobox) is the distinctive transcription factor of a class plant, is contained
Homeodomain, play a very important role in plant growth and development process (van der Graaff,
E., Laux, T. and Rensing, S.A., 2009, The WUS homeobox-containing (WOX)
Protein family, Genome Biology 10,248).Current research shows the usual table of WOX genes
Up in stem cell niche, and with maintaining the function of cell dryness, such as WUS, WOX5 and WOX4
Respectively in the OC (organize center) of shoot apical meristem, the merismatic QC of roots and tops
Express at (quiescent center) and forming layer, the function with the attribute for maintaining its peripheral stem.
In sum, callus originates from the stem cell in plant body, and WOX genes generally all have maintenance
The function of cell dryness, and the function all very guards in each species, and this is established for our research
Theoretical foundation.
The content of the invention
It is an object of the invention to provide the training method of new plant callus and WOX albumen and its volume
Code gene regulates and controls the application in Callus formation in tissue cultures.
More specifically, it is an object of the invention to provide the training method of new Rice Callus and paddy rice
WOX albumen or its coded sequence regulate and control the application in Callus formation in tissue cultures.
More specifically, as one embodiment, the WOX of homeodomain provided by the present invention
Albumen, entitled OsWOX11, from paddy rice (Oryza sativa ssp.japonica), amino acid sequence
Row such as SEQ ID NO：Shown in 1.
Therefore, first aspect present invention provides a kind of recombinant expression carrier, and the recombinant expression carrier can plant
Overexpression WOX albumen in thing cell.
In a specific embodiment, the recombinant expression carrier can in plant cell overexpression paddy rice
WOX albumen.
In a specific embodiment, the recombinant expression carrier can in plant cell overexpression paddy rice, jade
The WOX11 albumen of rice, willow, tomato or arabidopsis.
In a specific embodiment, the amino acid of WOX11 (i.e. OsWOX11) albumen of the paddy rice
Sequence such as SEQ ID NO：Shown in 1.
In a specific embodiment, the recombinant expression carrier contains SEQ ID NO:Code sequence shown in 4
Row.
Second aspect present invention provides a kind of Agrobacterium, and the Agrobacterium has proceeded to the recombinant expressed load of the present invention
Body.
In a specific embodiment, the Agrobacterium is Agrobacterium tumefaciems.
Third aspect present invention provides plant WOX albumen or its coded sequence, energy overexpression plant WOX
The recombinant expression carrier of albumen and the Agrobacterium containing the recombinant expression carrier regulate and control (outstanding in tissue cultures
It is to improve or strengthen) application in Callus formation.
Fourth aspect present invention provides plant WOX albumen or its coded sequence, energy overexpression plant WOX
The recombinant expression carrier of albumen and the Agrobacterium containing the recombinant expression carrier are in callus is formed
Using.
Fifth aspect present invention provides a kind of method for forming callus, and methods described includes：
(1) recombinant expression carrier of overexpression plant WOX albumen is proceeded in plant；
(2) tissue or organ of the plant of the acquisition overexpression WOX albumen；With
(3) tissue or organ are cultivated under conditions of callus is suitably formed；
So as to form callus.
Sixth aspect present invention provides a kind of method for producing the enhanced plant of callus ability, methods described
Including：
(1) with the recombinant expression carrier transformed plant of overexpression plant WOX；
(2) propagating materials of the plant is harvested；With
(3) the propagating materials regeneration plant is made；
So as to produce the enhanced plant of callus ability.
Seventh aspect present invention provides a kind of method for improving vegetable regeneration capacity, and methods described includes used table
Up to plant WOX recombinant expression carrier transformed plant the step of.
It is in tissue cultures, the over-express vector conversion of above-mentioned rice Os WOX11 gene is angiospermous
Pattern species arabidopsis (Arabidopsis thaliana Col-0), can obtain producing the increasing of callus ability
Strong plant, improves the power of regeneration of plant.Therefore a Fineness gene is provided for plant genetics and breeding,
Simultaneously also to the deep molecule mechanism for understanding Callus formation process, further with understand plant regeneration
Process has very important significance.The callus induction mode of the present invention, albumen and its encoding gene
With higher actual application value, have a extensive future.
Description of the drawings
Fig. 1 shows the result of the overexpression amount that OsWOX11 genes are detected in transgenic arabidopsis.OX1,
OX2 and OX3 are respectively three independent transformation strains.WT is non-transgenic wild type control.
Fig. 2 shows the phenotype of OsWOX11 overexpression Arabidopsis leaf explant bulk-growth callus.Material
Material is incubated at callus inducing medium upper 8 day.A, non-transgenic wildtype Arabidopsis thaliana.B, transgenosis
Overexpression OsWOX11 (pCAMBIA1300-35S:OsWOX11) arabidopsis.
Fig. 3 shows alignment's figure of the WOX11 family protein homeodomains of different plant species.
Specific embodiment
The present invention provides the training method and WOX albumen and its encoding gene of new plant callus and exists
Regulate and control the application in Callus formation in tissue cultures.
WOX (WUSCHEL related homeobox) is the distinctive transcription factor of a class plant, is contained
Homeodomain, plays a very important role in plant growth and development process.WOX includes
WUS, WOX5, WOX4, WOX11 etc..
The invention particularly relates to from the WOX11 of paddy rice (Oryza sativa ssp.japonica), i.e.,
OsWOX11.Specifically, the amino acid sequence of OsWOX11 of the present invention such as amino acid sequence such as SEQ ID
NO：Shown in 1.
The present invention also includes SEQ ID NO:The mutant of amino acid sequence shown in 1.These mutant include：
With SEQ ID NO:1 has at least 80%, preferably at least 85%, preferably at least 90%, preferably at least 95
%, preferably at least 97% sequence thereto simultaneously retains SEQ ID NO:The amino acid sequence of 1 BA
Row.The sequence thereto that the BLASTp of such as NCBI can be adopted to calculate between the sequence of two comparisons.This
Class mutant includes the WOX11 albumen from different plant species, such as from corn, willow, tomato, plan
The WOX11 albumen of southern mustard etc..Preferably, from the WOX11 family proteins and OsWOX11 of other plants
(SEQ ID NO:1) homeodomain shown in 18-85 positions has more than 80%, more than 85%, 90%
More than, more than 95%, even more than 98% sequence thereto.It is further preferred that the present invention includes and SEQ
ID NO:Amino acid sequence shown in 1 has at least 90%, preferably at least 95%, more preferably at least 97%
More than sequence thereto, (the i.e. 18-85 amino acids sequences of WOX11 albumen while homeodomain
Row) between sequence thereto more than 90%, preferably more than 95%, more preferably more than 97% sequence
The mutant of homogeny.
Mutant also includes：In SEQ ID NO:Have in sequence shown in 1 one or several mutation (insertion,
Disappearance or replace), while still retaining SEQ ID NO:The amino acid sequence of 1 BA.The number
Individual mutation is often referred within 1-10, such as 1-8,1-5 or 1-3.Replace and preferably protect
Keeping property replaces.For example, in the art, when carrying out conservative replaces with similar nature or similar amino acid,
The function of protein or polypeptide will not generally be changed." similar nature or similar amino acid " is included for example,
The family of the amino acid residue with similar side chain, these families include the amino acid (example with basic side chain
Such as lysine, arginine, histidine), amino acid (such as aspartic acid, paddy ammonia with acid side-chain
Acid), amino acid with uncharged polar side chain (for example glycine, asparagine, glutamine,
Serine, threonine, tyrosine, cysteine), amino acid (such as the third ammonia with non-polar sidechain
Acid, valine, leucine, isoleucine proline, phenylalanine, methionine, tryptophan), tool
There is the amino acid (such as threonine, valine, isoleucine) of β-branched building block and with aromatic side chain
Amino acid (such as tyrosine, phenylalanine, tryptophan, histidine).Therefore, in polypeptide of the present invention
One or several sites are replaced with another amino acid residue from the same side chain class, substantial shadow is will not be in
Ring its activity.
Additionally, it is as well known to those skilled in the art, in gene cloning operation, it is often necessary to design suitable enzyme
Enzyme site, this certainly will introduce one or more incoherent residues in expressed albumen end, and this is not
Affect the activity of destination protein.And for example in order to construction of fusion protein, promote recombinant protein expression, be derived from
The dynamic recombinant protein or the purifying beneficial to recombinant protein being secreted into outside host cell, it is often necessary to by some ammonia
Base acid is added in other appropriate areas to the N- ends of recombinant protein, C- ends or the albumen, for example,
Including but not limited to, suitable joint peptide, signal peptide, leader peptide, end extension, glutathione S-transfer
Enzyme (GST), maltose E binding protein, the label of albumin A, such as 6His or Flag, or Xa factor
Or the proteolytic enzyme site of fibrin ferment or enterokinase.It should be understood that the presence of these amino acid sequences will not shadow
Ring the activity to gained polypeptide.Therefore, the C-terminal and/or N for being also included within polypeptide of the present invention of the invention is last
End addition one or several amino acid (such as aforementioned joint peptide, signal peptide, leader peptide, end extension, GST,
The label of maltose E binding protein, such as albumin A, 6His or Flag, or Xa factor or fibrin ferment or intestines
Proteolytic enzyme site of kinases etc.) obtained by polypeptide, these polypeptides still with biology as herein described live
Property.
The present invention includes the polynucleotides sequence of coding WOX albumen, especially OsWOX11 albumen and its mutant
Row.The coded sequence example of OsWOX11 just like SEQ ID NO:Shown in 4.The polynucleotides of the present invention can
Being DNA form or rna form.DNA form includes cDNA, genomic DNA or artificial synthesized
DNA.DNA can be single-stranded or double-strand.DNA can be coding strand or noncoding strand.Compile
The coding region sequence of code mature polypeptide can be with SEQ ID NO:Coded sequence shown in 4 is identical or letter
And variant." variant of degeneracy " refers to that in the present invention coding has SEQ ID NO:1 amino
Acid sequence, but with SEQ ID NO:The differentiated nucleotide sequence of coded sequence shown in 4.
Coding SEQ ID NO:The polynucleotides of 1 mature polypeptide include：The coding of encoding mature polypeptide
Sequence；The coded sequence of mature polypeptide and various additional coding sequences；Mature polypeptide coded sequence (and appoint
The additional coding sequence of choosing) and non-coding sequence.
Herein, the polynucleotides of coded polypeptide can be the polynucleotides for including coding said polypeptide, also may be used
Being the polynucleotides for also including additional code and/or non-coding sequence.
The invention further relates to have at least 50% with above-mentioned polynucleotide sequence hybridization and between two sequences,
Preferably at least 70%, more preferably at least 80%, more preferably at least 90%, most preferably at least 95% homogeny
Polynucleotides.The present invention be more particularly directed to interfertile with polynucleotides of the present invention many under strict conditions
Nucleotides.In the present invention, " stringent condition " is referred to：(1) compared with LIS and higher temperature
Under hybridization and wash-out, such as 0.2 × SSC, 0.1%SDS, 60 DEG C；Or added with denaturant during (2) hybridization,
Such as 50% (v/v) formamide, 0.1% calf serum/0.1%Ficoll, 42 DEG C etc.；Or (3) are only at two
Homogeny between sequence just hybridizes at least more than 90% when more preferably more than 95%.Also, can
The polypeptide of the polynucleotide encoding of hybridization and SEQ ID NO:Polypeptide shown in 1 has identical biological function
And activity.
Although it should be understood that the present invention is by taking OsWOX11 as an example, from the base with OsWOX11 of other plants
Because very high homology (as having more than 50%, such as more than 60%, more than 70%, more than 80%, more than 85%,
More than 90%, more than 95%, even more than 98% sequence thereto) other genes also the present invention consideration
Within the scope of.The Method and kit for of aligned sequences homogeny is also well known in the art, such as BLAST.
As an example, shown other plants include corn, willow, tomato and arabidopsis.Wherein, the WOX11 of corn
(ZmWOX11) the number of logging in is EU954172；The WOX11 of willow includes PtrWOX11/12a, gene
Number：Potri.013G066900；And PtrWOX11/12b, gene number：Potri.019G040800；Tomato
(SlWOX11) the number of logging in is XP_004242129.1；The gene number of arabidopsis (AtWOX11)：
AT3G03660；And the gene number of paddy rice (OsWOX11)：Os07g48560.
The nucleotides full length sequence of the WOX albumen of the present invention can generally use PCR TRAPs, recombination method or people
The method of work synthesis is obtained.For PCR TRAPs, can according to relevant nucleotide sequence disclosed in this invention,
Especially open reading frame sequence to be designing primer, and with commercially available cDNA storehouses or by those skilled in the art
CDNA storehouses prepared by known conventional method expand and obtain relevant sequence as template.When sequence it is longer
When, it is often necessary to carry out twice or multiple PCR is expanded, the fragment for then again amplifying each time is by correct time
Sequence is stitched together.
At present, it is already possible to obtain encoding completely by chemical synthesis albumen of the present invention (or its fragment, or
Its derivative) DNA sequence dna.Then the DNA sequence dna can be introduced as known in the art various existing
In DNA molecular (or such as carrier) and cell.Additionally, be able to also will be mutated by chemical synthesis introducing the present invention
In protein sequence.
The present invention also relates to include the carrier of the polynucleotides of the present invention, and with carrier Jing genes of the invention
The host cell that engineering is produced.The carrier of the present invention is preferably over-express vector.
In the present invention, the polynucleotide sequence of coding WOX albumen can be inserted into recombinant expression carrier.Art
Language " recombinant expression carrier " refers to bacterial plasmid well known in the art, bacteriophage, yeast plasmid, plant cell
Virus, mammalian cell virus or other carriers.In a word, as long as can replicate in host's body and stable,
Any plasmid and carrier can be used.One key character of expression vector is to usually contain replication orgin, open
Mover, marker gene and translation control element.
Method well-known to those having ordinary skill in the art can be used to build the DNA sequence dna containing coding WOX albumen and conjunction
The expression vector of suitable transcription/translation control signal.These methods include that recombinant DNA technology in vi, DNA are closed
Into technology, In vivo recombination technology etc..Described DNA sequence dna can be effectively connected to suitably opening in expression vector
On mover, to instruct mRNA to synthesize.Expression vector also including translation initiation ribosome bind site and
Transcription terminator.
Additionally, expression vector preferably includes one or more selected markers, to provide for selecting
The dihyrofolate reductase of the phenotypic character of the host cell of conversion, such as eukaryotic culture, neomycin resist
Property and green fluorescent protein (GFP), or for colibacillary kanamycins or amicillin resistance.
When the polynucleotides of the present invention are expressed in higher eucaryotic cells, if inserting enhancer sequence in the carrier
When row transcription will be strengthened.Enhancer is the cis-acting factors of DNA, generally about has 10 to arrive
300 base-pairs, act on promoter to strengthen the transcription of gene.
Persons skilled in the art are aware that how to select appropriate carrier, promoter, enhancer and host
Cell.The method for building recombinant expression carrier is also well known in the art.
The present invention provides a kind of Agrobacterium, and the Agrobacterium has proceeded to the recombinant expression carrier of the present invention.Generally,
Agrobacterium is Agrobacterium tumefaciems.The method for proceeding to is also known, for example, can adopt electric shocking method.
The present invention provides plant WOX albumen or its coded sequence, the weight of energy overexpression plant WOX albumen
Group expression vector and the Agrobacterium containing the recombinant expression carrier regulate and control (especially to improve in tissue cultures
Or strengthen) application in Callus formation, and plant WOX albumen or its coded sequence, can mistake table
Up to the recombinant expression carrier of plant WOX albumen and the Agrobacterium containing the recombinant expression carrier is healed in formation
Application in injured tissue.
The present invention provides a kind of method for forming callus, and methods described includes：
(1) by the recombinant expression carrier of overexpression plant WOX albumen or agriculture bar containing the recombinant expression carrier
Bacterium is proceeded in plant；
(2) tissue or organ of the plant of the acquisition overexpression WOX albumen；With
(3) tissue or organ are cultivated under conditions of callus is suitably formed；
So as to form callus.
There is also provided a kind of method for producing the enhanced plant of callus ability, methods described includes：
(1) Agrobacterium with the recombinant expression carrier of overexpression plant WOX or containing the recombinant expression carrier turns
Change plant；
(2) propagating materials of the plant is harvested；With
(3) the propagating materials regeneration plant is made；
So as to produce the enhanced plant of callus ability.
There is also provided a kind of method for improving vegetable regeneration capacity, methods described includes using overexpression plant
The step of recombinant expression carrier of WOX or the Agrobacterium-mediated Transformation plant containing the recombinant expression carrier.
As used herein, described " plant " includes crops and gardening plant, including but not limited to：
Cruciferae, grass family, the rose family.Such as, described " plant " includes but is not limited to：Cruciferae
The arabidopsis (Arabidopsis thaliana) of Brassica genus, paddy rice gramineous and corn, rosaceous cherry
Peach, additionally including tobacco, melon fruits and vegetables, rape etc..
With reference to specific embodiment, the present invention is expanded on further.It should be understood that these embodiments are only used for
The bright present invention rather than restriction the scope of the present invention.The experiment side of unreceipted actual conditions in the following example
Method, generally according to normal condition such as Sambrook et al., molecular cloning：Lab guide (New York：
Cold Spring Harbor Laboratory Press, 1989) described in condition, or according to manufacture
Condition proposed by manufacturer.Unless otherwise indicated, otherwise percentage and number are calculated by weight.
Unless otherwise defined, all specialties used in text are ripe with one skilled in the art institute with scientific words
The meaning known is identical.Additionally, any similar to described content or impartial method and material all can be applicable to
In the present invention.Preferable implementation described in text only presents a demonstration with material and is used.The primer is prompt by the English Weihe River
Base (Shanghai) trade Co., Ltd provides, and examining order is completed by Shanghai Bo Shang bio tech ltd.
Embodiment 1, the transgenic Arabidopsis plants for obtaining overexpression OsWOX11
First, the structure of OsWOX11 gene overexpressions carrier
From rice genome database (http://rice.plantbiology.msu.edu) obtain OsWOX11
(Os07g48560) CDS sequences (the SEQ ID NO of gene:4), set according to 5 ' and 3 ' end sequences
Meter pair of primers, primer sequence is respectively F ends：5’-
cgcggatccATGGACGGCGGCCACAGCCCGGAC-3’(SEQ ID NO:, and R ends 5)
5’-acgcgtcgacTCGCTCAACTCGATCAAGACG-3’(SEQ ID NO:6)。
Extract the total serum IgE (using TRIzol, Invitrogen) that paddy rice spire is cultivated two days on inducing culture
Extracting), the total serum IgE with paddy rice as template, with RevertAid Reverse reverse transcriptase (Thermo
Scientific cDNA) is synthesized (according to User Guide:RevertAid Reverse Transcriptase, Thermo
The user's manual of Scientific is carried out).With cDNA as template, the OsWOX11 bases of PCR amplifying rices
The total length CDS sequence of cause.PCR reaction systems include (50 μ l)：The μ l of template 0.5, high-fidelity enzyme KOD
The μ l of plus (TOYOBO) 1, the μ l of 10 × buffer solution 5,2.5 μM of dNTP 5 μ l, 25mM MgSO45 μ l,
The each 2.5 μ l of 5 ' and 3 ' primers of 10 μ l, the μ l of water 28.5.Reaction condition is：94 DEG C of denaturations 5 minutes；
94 DEG C of denaturation 30 seconds, 55 DEG C are annealed 30 seconds, and 68 DEG C extend 1 minute, totally 35 circulations.Reaction terminates
Afterwards, PCR primer is carried out into 0.2% agarose gel electrophoresis detection, reclaims and purify the amplification of about 800bp
Fragment, is connected to carrier pCAMBIA1300-35S and is carried after being reclaimed using BamHI enzymes and SalI digestions
(transformed by pCAMBIA1300) on body, form OsWOX11 gene overexpression carriers
pCAMBIA1300-35S:OsWOX11。
2nd, detection of expression of the overexpression OsWOX11 in arabidopsis
By the OsWOX11 gene overexpressions carrier for building (i.e.
pCAMBIA1300-35S:OsWOX11), Agrobacterium GV3101 is converted using electric shocking method, is reused often
The flower-dipping method arabidopsis thaliana transformation of rule.After arabidopsis seed maturity, seed is collected, be dried, using 75%
Ethanol sterilize 20 minutes, uniformly sow seed in containing hygromycin (25mg/L) 1/2MS culture mediums on,
Wherein 1/2MS culture mediums compound method is：MS (being purchased from phytoTechology Laboratories) 2.2g/L；
Sucrose 10g/l；MES (being purchased from the two-way western Bath company in Shanghai) 0.5g/L；Agar 1% (pH 5.8).
Transformation seedlings are obtained after about 3 weeks, by conversion transplantation of seedlings into soil, and clip blade, using preceding method
Extract RNA and carry out reverse transcription, by masterplate of resulting cDNA real-time fluorescence quantitative PCR inspection is carried out
Survey, PCR system (20 μ l) is：The μ l of template 0.5, the μ l of sterilized water 7.9, the F ends of OsWOX11 are quantitative
Primer：5’-ATCTCCTCCGACTGCTTCGTC-3’(SEQ ID NO:7) determine at 0.8 μ l, and R ends
Amount-the CTCGAGATGGCGAACAGGTC-3 ' of primer 5 ' (SEQ ID NO:8) 0.8 μ l, Sybrgreen
PCR Mix 10μl.Reaction condition is：95 DEG C of denaturations 3 minutes；95 DEG C of denaturation 10 seconds, 60 DEG C of annealing
30 seconds, 72 DEG C extended 30 seconds, totally 40 circulations.Real time fluorescent quantitative result is as shown in figure 1, knot
Fruit is displayed in Arabidopsis leaf the high expression for occurring in that OsWOX11.
The phenotypic evaluation of embodiment 2, overexpression OsWOX11 in arabidopsis
After transgenic arabidopsis knot, seed is collected, be dried, cultivated 12 days on 1/2MS culture mediums
Afterwards, it is in callus inducing medium and CIM culture medium compound methods by explant of blade：B5 (purchases
From phytoTechology Laboratories) 3.21g/L；MES 0.5g/L；Sucrose 20-40g/L；2,4-D
2.2 μM of (being purchased from SIGMA)；0.2 μM of Kinetin (being purchased from SIGMA)；Agar 0.8% (pH 5.8)
Upper induction 7 days, as a result as shown in Figure 2.Compared with wild type, transgenic arabidopsis form callus
Ability is remarkably reinforced.
Claims (10)
1. a kind of recombinant expression carrier, the recombinant expression carrier can in plant cell overexpression WOX eggs
In vain, the WOX11 albumen of preferred paddy rice, corn, willow, tomato and arabidopsis.
2. recombinant expression carrier as claimed in claim 1, it is characterised in that
The amino acid sequence of the paddy rice WOX11 albumen such as SEQ ID NO：Shown in 1；Or
The recombinant expression carrier contains SEQ ID NO:Nucleotide sequence shown in 4.
3. a kind of transgenic cell line or engineering bacteria, the transgenic cell line or Agrobacterium proceed to right will
Seek the recombinant expression carrier any one of 1-2.
4. transgenic cell line as claimed in claim 3 or engineering bacteria, it is characterised in that described to turn base
Because clone is plant cell or zooblast, the engineering bacteria is Agrobacterium, preferably Agrobacterium tumefaciems.
5. plant WOX albumen or its coded sequence, can overexpression plant WOX albumen recombinant expressed load
Body and the Agrobacterium containing the recombinant expression carrier regulate and control answering in Callus formation in tissue cultures
With, or the application in callus is formed.
6. application as claimed in claim 5, it is characterised in that the recombinant expression carrier such as right will
Ask any one of 1-2.
7. it is a kind of formed callus method, it is characterised in that methods described includes：
(1) by the recombinant expression carrier of overexpression plant WOX albumen or the recombinant expression carrier has been proceeded to
Agrobacterium is proceeded in plant；
(2) tissue or organ of the plant of the acquisition overexpression WOX albumen；With
(3) tissue or organ are cultivated under conditions of callus is suitably formed；
So as to form callus.
8. a kind of method for producing the enhanced plant of callus ability, methods described includes：
(1) with the recombinant expression carrier of overexpression plant WOX or the agriculture bar of the recombinant expression carrier has been proceeded to
Bacterium transformed plant；
(2) propagating materials of the plant is harvested；With
(3) the propagating materials regeneration plant is made；
So as to produce the enhanced plant of callus ability.
9. a kind of method for improving vegetable regeneration capacity, methods described includes the weight with overexpression plant WOX
The step of group expression vector transformed plant.
10. method as claimed in any one of claims 7-9, it is characterised in that the plant is selected from：
Cruciferae, grass family and rosaceous plant, including arabidopsis, paddy rice, corn, cherry, tobacco, melon
Fruits and vegetables and rape.
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| WO2018224001A1 (en) * | 2017-06-07 | 2018-12-13 | Institute Of Crop Science, Chinese Academy Of Agricultural Sciences | Method for improving transformation efficiency of plant and method for transforming plant |
| CN109355297A (en) * | 2018-11-19 | 2019-02-19 | 浙江农林大学 | Dendrobium candidum DcWOX4 gene and its application in raising axis tiller |
| CN109608527A (en) * | 2018-11-09 | 2019-04-12 | 中国中医科学院中药研究所 | For detecting specific gene PgWOX11 and its detection method and the application of ginseng adventitious root stem cell |
| CN113388619A (en) * | 2021-07-01 | 2021-09-14 | 中国农业科学院蔬菜花卉研究所 | Cloning method of lily bulbil formation regulation gene LlWOX11 and application thereof |
| CN115851755A (en) * | 2022-07-11 | 2023-03-28 | 内蒙古农业大学 | Meng nong hybrid wheatgrass tillering angle regulation factor and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104450777A (en) * | 2014-10-13 | 2015-03-25 | 南京农业大学 | Method for improving potassium absorption efficiency of plant and resisting against potassium deficiency stress and recombinant expression vector used therein |
-
2015
- 2015-11-09 CN CN201510756266.3A patent/CN106676128A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104450777A (en) * | 2014-10-13 | 2015-03-25 | 南京农业大学 | Method for improving potassium absorption efficiency of plant and resisting against potassium deficiency stress and recombinant expression vector used therein |
Non-Patent Citations (2)
| Title |
|---|
| JINGCHUN LIU等: ""WOX11 and 12 Are Involved in the First-Step Cell Fate Transition during de Novo Root Organogenesis in Arabidopsis"", 《THE PLANT CELL》 * |
| YU ZHAO等: "The WUSCHEL-Related Homeobox Gene WOX11 Is Required to Activate Shoot-Borne Crown Root Development in Rice", 《THE PLANT CELL》 * |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| EP3619225A4 (en) * | 2017-06-07 | 2020-12-30 | Institute of Crop Science, Chinese Academy of Agricultural Sciences | Method for improving transformation efficiency of plant and method for transforming plant |
| WO2018224001A1 (en) * | 2017-06-07 | 2018-12-13 | Institute Of Crop Science, Chinese Academy Of Agricultural Sciences | Method for improving transformation efficiency of plant and method for transforming plant |
| US11447784B2 (en) | 2017-06-07 | 2022-09-20 | Institute Of Crop Science, Chinese Academy Of Agricultural Sciences | Method for improving transformation efficiency of plant and method for transforming plant |
| CN111278849A (en) * | 2017-06-07 | 2020-06-12 | 中国农业科学院作物科学研究所 | Method for improving transformation efficiency of plant and method for transforming plant |
| CN108085319A (en) * | 2017-12-25 | 2018-05-29 | 中国科学院遗传与发育生物学研究所 | Plant tillering angle GAP-associated protein GAP and its encoding gene and application |
| CN109608527A (en) * | 2018-11-09 | 2019-04-12 | 中国中医科学院中药研究所 | For detecting specific gene PgWOX11 and its detection method and the application of ginseng adventitious root stem cell |
| CN109608527B (en) * | 2018-11-09 | 2024-01-05 | 中国中医科学院中药研究所 | Specific gene PgWOX11 for detecting ginseng adventitious root stem cells, and detection method and application thereof |
| CN109355297B (en) * | 2018-11-19 | 2021-04-13 | 浙江农林大学 | Dendrobium officinale DcWOX4 gene and application thereof in improving plant stem tillering |
| CN109355297A (en) * | 2018-11-19 | 2019-02-19 | 浙江农林大学 | Dendrobium candidum DcWOX4 gene and its application in raising axis tiller |
| CN113388619A (en) * | 2021-07-01 | 2021-09-14 | 中国农业科学院蔬菜花卉研究所 | Cloning method of lily bulbil formation regulation gene LlWOX11 and application thereof |
| CN113388619B (en) * | 2021-07-01 | 2022-08-02 | 中国农业科学院蔬菜花卉研究所 | Cloning method of lily bulbil formation regulation gene LlWOX11 and application thereof |
| CN115851755A (en) * | 2022-07-11 | 2023-03-28 | 内蒙古农业大学 | Meng nong hybrid wheatgrass tillering angle regulation factor and application thereof |
| CN115851755B (en) * | 2022-07-11 | 2024-05-24 | 内蒙古农业大学 | Mongolian hybrid agro-grass tillering angle regulating factor and application thereof |
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