CN110357956A - A kind of preparation method and applications of rice auxin output albumen OsPIN1b antibody - Google Patents
A kind of preparation method and applications of rice auxin output albumen OsPIN1b antibody Download PDFInfo
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- CN110357956A CN110357956A CN201910652459.2A CN201910652459A CN110357956A CN 110357956 A CN110357956 A CN 110357956A CN 201910652459 A CN201910652459 A CN 201910652459A CN 110357956 A CN110357956 A CN 110357956A
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- ospin1b
- auxin
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- 101000691855 Oryza sativa subsp. japonica Probable auxin efflux carrier component 1b Proteins 0.000 title claims abstract description 70
- 235000007164 Oryza sativa Nutrition 0.000 title claims abstract description 67
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/06—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies from serum
- C07K16/065—Purification, fragmentation
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/16—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from plants
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/415—Assays involving biological materials from specific organisms or of a specific nature from plants
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- Engineering & Computer Science (AREA)
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Abstract
A kind of preparation method of rice auxin output albumen OsPIN1b antibody, include the following steps: to export albumen OsPIN1b by bioinformatics method analyzing rice auxin, the polypeptide sequence that one section of sequence is QSSRNPTPRGSSFNC is chosen from OsPIN1b amino acid sequence, according to polypeptide sequence synthesis polypeptide, again polypeptide antigen will be obtained after polypeptide and carrier conjugation, polypeptide antigen is injected into New Zealand White Rabbit, after initial immunity, 3 booster immunizations are carried out, menses, which isolate and purify clearly, obtains rice auxin output albumen OsPIN1b polyclonal antibody.The polyclonal antibody of this method preparation can export albumen OsPIN1b by auxin in immunoblot assay specific recognition rice, the polar translocation that carrier protein PIN family regulation auxin is exported to disclose auximone is provided fundamental basis with development of plants mechanism, has extensive use in the detection and Function Identification of auxin output albumen PIN1.
Description
Technical field
The present invention relates to field of biotechnology, and in particular to a kind of system of rice auxin output albumen OsPIN1b antibody
Preparation Method and its application.
Background technique
Auxin is the Plant Hormone being found earliest, and the characteristic of polar translocation is that it is different from various plants hormone
Exclusive feature.Currently, it has been recognized that uneven distribution of the auxin input and output carrier on cytoplasma membrane realizes
The polar translocation of auxin, and auxin is established in the intracorporal gradient distribution of plant.The polar translocation of auxin takes part in plant
The many important growth and development processes of object, including the branch of plant, tropism growth and growth and development of root system etc..Auxin
And its polar translocation not only influences the morphological feature of plant, but also between absorption, transport and the distribution of mineral element there is
Close connection.
PIN protein family is to study more auxin output carrier at present, and PIN albumen is a kind of transmembrane protein, it
Polarity orientation on cell membrane determines the spatial distribution of auxin.In arabidopsis, it is cloned into from PIN gene family altogether
8 genes；The homologous gene of 12 PIN is predicted in rice.OsPIN1 is expressed in root restriction and vascular tissue, silencing
In the transgenic plant of OsPIN1, the generation and development of adventitious root are suppressed, and illustrate that OsPIN1 is relying on the indefinite of auxin
It plays an important role in root generating process.The present invention is that detection rice auxin output albumen OsPIN1b determines in cell membrane
Position, distribution and cell endocytosis, research are prepared for the polyclonal antibody of rice auxin output albumen OsPIN1b a kind of, are into one
Step is carried out and analyzing rice auxin output albumen OsPIN1b function lays the foundation.
The a variety of rice varieties of rice auxin output vector gene OsPIN1b and successful conversion are obtained currently, having cloned,
Its high expression in paddy rice root tip；By adjusting paddy rice root tip tissue activity, the transport of auxin and the elongation of root restriction are influenced.
There is research to speculate that OsPIN1b gene may participate in the geotropism of rice root.
Rice auxin output carrier OsPIN1b antibody is prepared, is conducive to further through immunohistochemistry, immunoblotting etc.
Method observes the cell membrane localization of auxin output albumen OsPIN1b in rice and other plants, discloses Polar Transport of Auxin
The mechanism of action of regulating growth of plants furthers elucidate the inner link of plant hormone interaction and development of plants, from
And experiment basis is established using auxin regulating growth of plants.
Present invention firstly discloses the preparation methods and the antibody of the antibody of rice auxin output albumen OsPIN1b
Immunohistochemistry application.
Summary of the invention
It is an object of the invention to fill up the blank of rice auxin output albumen OsPIN1b detection field, one is provided
Preparation method of the kind for the polyclonal antibody of rice auxin output albumen OsPIN1b.
The problem to be solved by the invention is to provide the polyclonal antibodies of rice auxin output albumen OsPIN1b a kind of
Preparation method, the invention adopts the following technical scheme:
A kind of preparation method of the polyclonal antibody of rice auxin output albumen OsPIN1b, includes the following steps:
1) pass through bioinformatic analysis rice Oryza sativa subsp.japonica (Nipponbare) gene
The albumen OsPIN1b of OsPIN1b (LOC_Os02g50960 or Os02g0743400) expression.
2) a segment polypeptide sequence QSSRNPTPRGSSFNC is chosen from antibody coding albumen OsPIN1b, method particularly includes:
According to peptide chain structure complexity, oxidizable degree, synthesis difficulty, amino acid classification and distribution etc. finally determine antibody coding
Amino acid sequence of the position 15 the albumen 244-258 amino acid as synthesis polypeptide, wherein the 258th amino acids are by H → C, sequence
It is classified as QSSRNPTPRGSSFNC.
3) according to the polypeptide sequence synthesis polypeptide in step 2；
4) polypeptide antigen will be obtained after polypeptide and carrier conjugation；
5) polypeptide antigen is injected into New Zealand White Rabbit, after initial immunity 4 weeks, every 1 booster immunization of progress in 2 weeks, altogether
Three times, menses, which isolate and purify clearly, obtains rice auxin output albumen OsPIN1b polyclonal antibody, and the IgG antibody is dense for processing
Degree is 0.51mg/mL；
6) indirect elisa method evaluates the potency of polyclonal antibody.
After tested, when antibody is diluted with 1:20000 times, detected value 0.718, for qualified rabbit polyclonal antibody.
Above-mentioned rice auxin output albumen OsPIN1b antibody is also applied to western blot test by the present invention.
The present invention also by above-mentioned rice auxin output albumen OsPIN1b antibody be applied to detection rice, corn, rape,
The cellular localization and the regularity of distribution of auxin output albumen OsPIN1b in potato, wheat and soybean tip of a root epidermal cell.
Above-mentioned rice auxin output albumen OsPIN1b antibody is also applied to detection rice auxin and exports egg by the present invention
The cell endocytosis of white OsPIN1b.
The present invention carries out analysis comparison by the antibody coding albumen OsPIN1b to rice and chooses specific polypeptide sequence
QSSRNPTPRGSSFNC is synthesized to obtain polypeptide according to this segment polypeptide sequence, and the polypeptide and BSA are coupled to obtain polypeptide antigen,
New Zealand White Rabbit is repeatedly immunized with the polypeptide antigen, obtains the polyclonal of the auxin output albumen OsPIN1b of specificity for the first time
Antibody.Using method provided by the present invention prepare OsPIN1b polyclonal antibody, can specific recognition rice auxin it is defeated
Albumen OsPIN1b out, the cell membrane that can be realized auxin output albumen OsPIN1b in paddy rice root tip epidermal cell and embryo are fixed
Position, and the distribution and cell endocytosis situation of identification rice auxin output albumen OsPIN1b, export albumen PIN1 in auxin
Detection and Function Identification, research in have extensive use.In addition, this antibody can also to rape, corn, potato, wheat and
Auxin output albumen PIN1 carries out cell membrane localization in the root and embryo of soybean.
Detailed description of the invention
Fig. 1 is immunoblotting analysis figure (Western blot)；
Fig. 2 is immunohistochemistry figure: big with the antibody A nti-rabbit OsPIN1b antibody detection six of preparation
The cell membrane localization of PIN1 albumen in cereal crops rice, corn, rape, potato, wheat and the soybean tip of a root, Bar=10 μm；
Fig. 3 is immunohistochemistry figure: detecting grain with the antibody A nti-rabbit OsPIN1b antibody of preparation
The cell membrane localization of PIN1 albumen in crop rice, corn, wheat and soybean blastocyte, Bar=10 μm；
Fig. 4 is immunohistochemistry figure: utilizing the antibody A nti-rabbit OsPIN1b antibody of preparation, detection two
The cell endocytosis of OsPIN1b albumen in kind rice varieties LTH and Nipponbare tip of a root epidermal cell, Bar=10 μm.
Specific embodiment
Embodiment 1:
A kind of preparation method of the polyclonal antibody of rice auxin output albumen OsPIN1b provided in this embodiment, packet
Include following steps:
1) pass through bioinformatic analysis rice Oryza sativa subsp.japonica (Nipponbare) gene
OsPIN1b (LOC_Os02g50960 or Os02g0743400) expresses albumen OsPIN1b.
2) a segment polypeptide sequence QSSRNPTPRGSSFNC is chosen from antibody coding albumen OsPIN1b, method particularly includes:
According to polypeptide structure complexity, oxidizable degree, synthesis difficulty, amino acid classification and distribution etc. finally determine antibody coding
Amino acid sequence of the position 15 the albumen 244-258 amino acid as synthesis polypeptide, wherein the 258th amino acids are by H → C, sequence
It is classified as QSSRNPTPRGSSFNC.
3) according to the polypeptide sequence in step 2, Peptide synthesizer synthesis polypeptide, specific procedure are utilized are as follows: 1. de- Fmoc are protected
Resin is protected, it is made to expose amino.Deprotecting regent is the 20%PIP prepared with DMF；2. amino acid is dissolved with DMF；3. journey
Sequence successively independently extracts each 4mL of amino acid solution；4. the amino acid solution of each loading 4mL, program extraction carboxyl activator
0.4M HBTU (is prepared) with DMF, amino carboxyl condensing agent 0.8M DIEA (being prepared with DMF), so that the activated exposure of the latter
The amino acid of carboxyl and the previous amino acid for exposing amino by deprotection carry out condensation out, and result in formation of two
Peptide.It recycles by this method, up to the last one amino acid condensation to the end of peptide chain.Each step in synthesis process requires to use
DMF, DCM elution blocking group simultaneously elute the amino acid for having neither part nor lot in reaction residual from reaction system.
4) be coupled the polypeptide of synthesis with BSA with SPDP connection method: 4.6mg SPDP is dissolved in 740uL DMSO, eventually
Concentration is 20mM.0.1008g BSA is dissolved in 2mL PBS-EDTA solution, is stored at room temperature 1h.It is extra to be eluted using desalting column
SPDP.Ambient temperature overnight in the BSA-SPDP system being coupled is added in 4mg polypeptide.By carboxyl or amino-terminal residue by polypeptide and
BSA polypeptide coupling carrier couples, and obtains polypeptide antigen；
5) polypeptide antigen is injected into healthy new zealand white rabbit, after initial immunity 4 weeks, exempted from every 1 reinforcement of progress in 2 weeks
Epidemic disease, three times, injecting pathway is identical as initial immunity for coprocessing, ear source venous blood collection, separation serum, and Protein G post separation is pure
Change IgG antibody: PBS (pH7.4) balances Protein G affinity column, with 0.1M Gly-HCl (pH3.0) elution, PBS (pH7.4)
Dialysis.It obtains rice auxin and exports albumen OsPIN1b polyclonal antibody, the IgG antibody concentration is 0.51mg/mL；
6) indirect elisa method evaluates the potency of polyclonal antibody, the specific steps are as follows:
A. it is coated with: by antigen OsPIN1b with the concentration of 1 μ g/mL, 200 μ L is added with the every hole of liquid-transfering gun on ELISA Plate,
4 DEG C overnight；
B. it closes: 1% 200 μ L of BSA solution is added, is incubated 1 hour under the conditions of 37 DEG C；
C. it dilutes: by rabbit polyclonal antibody respectively with 1:200,1:1000,1:5000,1:10000,1:20000,1:
60000,1:240000 dilutions；
D. it is loaded: after 50 hole μ L/ of rabbit polyclonal antibody diluted is added, being placed in insulating box and incubated under the conditions of 37 DEG C
1 hour；
E. it washs: with detergent TBST with 200 hole μ L/ washing hole 2 times；
Plus ELIAS secondary antibody f.: being added in each hole is made using the diluted goat-anti rabbit HRP liquid of 1:5000 as ELIAS secondary antibody
With, and washed 3 times with TBST with 200 holes μ L/ after incubating 45min under the conditions of 37 DEG C；
G. it develops the color: 100 μ L TMB developing solutions being added in each hole, 37 DEG C are developed the color 10 minutes；
I. it terminates: the 50 microlitres/hole of sulfuric acid of 2 mol/Ls is added, microplate reader reads each hole light absorption value.
Testing result: when antibody is diluted with 1:20000 times, detected value 0.718, for qualified rabbit polyclonal antibody.
Embodiment 2:
Application of the OsPIN1b antibody in immunoblotting:
Testing result: as shown in Figure 1, Western blot detects expression of the OsPIN1b albumen in rice root tissue, water
1:500 times of sample of rice root tissue dilutes, and albumen shows target stripe at 65 KD.
Embodiment 3:
Positioning of the OsPIN1b antibody in six kinds of different cereal crops tip of a root epidermal cells.
1. trial crops: rice, corn, rape, potato, wheat and soybean；
2. experimental material: Microcloth, 4% paraformaldehyde, PBS buffer solution (pH7.4), Triton X-100, collapse
Enzyme Driselase, DMSO, Nonidet-P40, BSA, sterile water, mountant.Primary antibody: Anti-OsPIN1b antibody；
Secondary antibody: Donkey Anti-rabbit IgG (H+L)-Alexafluor 488coupled.
3. experimental procedure
A. the acquisition of material: six generalized grain crop of water planting 7 days after crop sends out roots, takes tip of a root 1cm or so, use is sterile
Water cleans up.It is placed in the inside to be surrounded by 24 orifice plates of Microcloth, then three times with PBS purging.
B. to addition 4% paraformaldehyde fixer of 1mL in organized hole is filled, room temperature fixes 1h under vacuum state；With
1mL PBS+0.5%Triton X-100 is purged 5 times, then is purged 2 times with distilled water, and purging is 10min every time；Finally by group
It knits and is soaked in distilled water, 37 DEG C of placement 12min.
C. with 2% driselase of 1mL (Driselase) in 37 DEG C of processing 1h, with the PBS+0.5%Triton X-100 of 1mL
Purging 5 times, each 12min；1mL penetrating fluid (containing 10%DMSO and 2%Nonidet-P40) is added, room temperature handles 1.5h, PBS
Purging 6 times, each 10min.
D. plus 1mL 2%BSA solution, in 37 DEG C of closing 1h；It is added 1mL primary antibody (1:200 times dilutes), was placed in 4 DEG C
Night；It is purged 5 times with 1mL PBS+0.5%Triton X-100, each 12min.
E. 1mL secondary antibody (1:500 times dilutes) is added, in 37 DEG C of placement 4h；It is blown with 1mL PBS+0.5%Triton X-100
It washes 5 times, each 12min.
F. mountant is dripped on glass slide, and the material of processing is put into wherein, it is micro- with laser co-focusing after tabletting
Sem observation.
4. experimental result: OsPIN1b antibody can be well with rice Os PIN1b protein binding as can be seen from Figure 2, this is anti-
The binding ability of body and PIN1 albumen in six generalized grain crops is different, and corn, the binding ability of potato are stronger, wheat and big
Beans take second place, and can hardly generate immune response with rape, it was demonstrated that the antibody is simultaneously to rice, corn, potato, wheat, soybean
Deng applicable.Furthermore it is possible to which exporting albumen to PIN1 by the binding site of OsPIN1b antibody and PIN1 albumen carries out crop root
In cell membrane localization.
Embodiment 4
Experimental material is derived from four kinds of rice, corn, wheat and soybean cereal crops embryos: by the seed water logging of four kinds of crops
Bubble overnight, removes kind of a shell with scalpel in second day, takes out the embryo in seed, other experimental procedures are same as Example 3.
Experimental result: from figure 3, it can be seen that OsPIN1b antibody also can be with the PIN1 albumen on rice, wheat cell membrane
In conjunction with, and it is preferable with the PIN1 protein binding capacity on wheat cell membrane, green fluorescence can be clearly seen that on cell membrane.
Illustrate that the OsPIN1b antibody of preparation is applicable not only to detection root PIN1 albumen positioning, it may also be used for detection PIN1 albumen is not
With the cellular localization in plant embryos, this is the polarity fortune for disclosing auximone output carrier protein PIN family regulation auxin
Defeated and development of plants mechanism based theoretical.
Embodiment 5:
Utilize the cell of OsPIN1b albumen in OsPIN1b antibody test difference rice varieties LTH and the Nipponbare tip of a root
Endocytosis: before immunohistochemical experiment, first being handled paddy rice root tip with Protein transport inhibitor brefeldin a (BFA),
OsPIN1b albumen in different paddy rice root tip epidermal cells is observed after the completion assembles the BFA corpusculum to be formed.
1. material and reagent:
Oryza sativa l. TH, rice Nipponbare, 0.25 μM of BFA solution, Microcloth, 4% paraformaldehyde, PBS buffering
Liquid (pH7.4), Triton X-100, driselase Driselase, DMSO, Nonidet-P40, BSA, sterile water, mountant；
Primary antibody: Anti-rabbit OsPIN1b antibody；
Secondary antibody: Donkey Anti-rabbit IgG (H+L)-Alexafluor 488coupled.
2. experimental method:
A. before immunohistochemical experiment, first paddy rice root tip is carried out with Protein transport inhibitor brefeldin a (BFA)
Processing, observes OsPIN1b albumen in different rice epidermal cells after the completion and assembles the BFA corpusculum to be formed.
The tip of a root is put into containing 1mL by the radicle tip of a root 1cm that rice seedlings are b. cut with shaver blade with tweezers are careful
ddH2It in the hole of 24 orifice plates of O, is carefully purged with 1mL liquid-transfering gun, is careful not to hurt root；Sucking liquid adds
1mL ddH2The tip of a root is impregnated 15min or so by O, it is ensured that the culture medium on tip of a root surface is cleaned, and is not influenced medicament and is entered root
Tip Cells；
C. sucking liquid adds 0.25 μM of 1mL of BFA solution, at dark [being wrapped 24 orifice plates with tinfoil]
Manage 90min；After the completion of processing, paddy rice root tip is taken out, the inside is placed in and is surrounded by 24 orifice plates of Microcloth, purged with PBS
Three times.
D. 4% paraformaldehyde fixer of 1mL is added, room temperature fixes 1h under vacuum state；With the PBS+0.5% of 1mL
Triton X-100 solution purges 5 times, then is purged 2 times with distilled water, and purging is 10min every time；Finally tissue is soaked in
Distilled water, 37 DEG C of placement 12min.
E. with driselase (Driselase) solution of 1mL 2% in 37 DEG C of processing 1h, with the PBS+0.5%Triton of 1mL
X-100 solution purges 5 times, each 12min；1mL penetrating fluid (containing 10%DMSO and 2%Nonidet-P40), room temperature processing is added
1.5h, PBS are purged 6 times, each 10min.
F. 1mL 2%BSA is added, in 37 DEG C of closing 1h；It is added 1mL primary antibody (1:200 times dilutes), is stood overnight in 4 DEG C；
It is purged 5 times with the PBS+0.5%Triton X-100 solution of 1mL, each 12min.
G. 1mL secondary antibody (1:500 times dilutes) is added, in 37 DEG C of placement 4h；With the PBS+0.5%Triton X-100 of 1mL
Solution purges 5 times, each 12min.
H. mountant is dripped on glass slide, and the paddy rice root tip tissue of processing is put into wherein, it is total with laser after tabletting
Focusing microscope observation.
3. experimental result:
Fig. 4 can be seen that Lijiang xintuanheigu LTH, two kinds of rice roots of OryzasativaLcv.Nipponbare Nipponbare inhibit in Protein transport
Under agent BFA effect, the OsPIN1 protein binding in the intracellular OsPIN1b antibody of rice root and rice forms BFA corpusculum.Experiment
As a result being successfully authenticated OsPIN1b antibody can be used to detect well the thin of the output of auxin in different rice albumen OsPIN1b
The experiment of born of the same parents' endocytosis exports carrier and plant growth and development to facilitate the polar translocation mechanism of research auxin for auxin
Relationship is provided fundamental basis.
Claims (5)
1. a kind of preparation method of rice auxin output albumen OsPIN1b antibody, which comprises the steps of:
1) pass through bioinformatic analysis rice Oryza sativa subsp.japonica (Nipponbare) gene
The albumen OsPIN1b of OsPIN1b (LOC_Os02g50960 or Os02g0743400) expression；
2) a segment polypeptide sequence QSSRNPTPRGSSFNC is chosen from coding albumen OsPIN1b be used to prepare antibody；
3) according to the polypeptide sequence synthesis polypeptide in step 2；
4) polypeptide antigen will be obtained after polypeptide and carrier conjugation；
5) polypeptide antigen is injected into New Zealand White Rabbit, after initial immunity, carries out 3 booster immunizations, menses isolate and purify obtain clearly
It obtains rice auxin and exports albumen OsPIN1b polyclonal antibody, the IgG antibody concentration is 0.51mg/mL；
6) indirect elisa method evaluates the potency of polyclonal antibody.
2. a kind of preparation method of rice auxin output albumen OsPIN1b antibody according to claim 1, feature exist
In all or part of amino acid sequence of the polypeptide sequence QSSRNPTPRGSSFNC is used for the preparation of OsPIN1b antibody.
3. a kind of rice auxin output albumen OsPIN1b antibody according to claim 1 is applied to immunoblotting.
4. a kind of rice auxin according to claim 1 exports albumen OsPIN1b antibody, answered by ImmunohistochemistryMethods Methods
For detect arabidopsis, rice, corn, rape, potato, in wheat and soybean auxin output albumen PIN1 in the tip of a root, embryo
And cellular localization and the regularity of distribution in epidermal leaf cells and cell membrane.
5. a kind of rice auxin according to claim 1 exports albumen OsPIN1b antibody, answered by ImmunohistochemistryMethods Methods
For detect arabidopsis, rice, corn, rape, potato, in wheat and soybean auxin output albumen PIN1 cell born of the same parents
It gulps down.
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Citations (3)
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|---|---|---|---|---|
| US20020170088A1 (en) * | 2000-11-03 | 2002-11-14 | The Regents Of The University Of California | Novel auxin binding proteins and uses thereof |
| CN101228279A (en) * | 2005-07-19 | 2008-07-23 | 巴斯福植物科学有限公司 | Yield increase in plants overexpressing the MTP genes |
| CN105219783A (en) * | 2015-10-20 | 2016-01-06 | 南京林业大学 | A kind of hybridized Chinese tuliptree PIN1 gene and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20020170088A1 (en) * | 2000-11-03 | 2002-11-14 | The Regents Of The University Of California | Novel auxin binding proteins and uses thereof |
| CN101228279A (en) * | 2005-07-19 | 2008-07-23 | 巴斯福植物科学有限公司 | Yield increase in plants overexpressing the MTP genes |
| CN105219783A (en) * | 2015-10-20 | 2016-01-06 | 南京林业大学 | A kind of hybridized Chinese tuliptree PIN1 gene and application thereof |
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| TARAS PASTERNAK等: "Protocol: an improved and universal procedure for whole-mount immunolocalization in plants", 《PLANT METHODS》 * |
| 邹纯雪等: "生长素的外输载体PIN蛋白家族研究进展", 《中国细胞生物学学报》 * |
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