CN103554240B - Protein GhKT2 related to potassium ion absorption capacity of plant as well as coding gene and application thereof - Google Patents

Protein GhKT2 related to potassium ion absorption capacity of plant as well as coding gene and application thereof Download PDF

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CN103554240B
CN103554240B CN201310534743.2A CN201310534743A CN103554240B CN 103554240 B CN103554240 B CN 103554240B CN 201310534743 A CN201310534743 A CN 201310534743A CN 103554240 B CN103554240 B CN 103554240B
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田晓莉
徐娟
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China Agricultural University
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Abstract

The invention discloses a protein GhKT2 related to potassium ion absorption capacity of a plant as well as a coding gene and application thereof. The protein provided by the invention is (a) or (b), wherein (a) is a protein composed of an amino acid sequence as shown in a sequence 1 in a sequence list, (b) is a protein formed by the way that substitution and/or deletion and/or addition of one or more than one amino acid residues are carried out on an amino acid sequence as shown in the sequence 1 and derivation related to low-potassium adversity stress resistance reaction of the plant, the potassium ion absorption capacity of the plant and development of the plant is carried out on the sequence 1. The invention discloses a new protein and gene, and after the gene is introduced into the plant, the low-potassium resistance of the plant can be obviously improved; the protein GhKT2 has a significant value on a new plant variety, especially a new cotton variety.

Description

The Protein G hKT2 relevant to the ability of plant absorption potassium ion and encoding gene thereof and application
Technical field
The present invention relates to a kind of Protein G hKT2 relevant to the ability of plant absorption potassium ion and encoding gene thereof and application.
Background technology
Potassium is one of required three large nutritive elements of plant-growth, all closely related with enzymic activity, protein synthesis, photosynthesis, oil synthesis, stomatal movement, ionic equilibrium and resistance.Potassium nutrition shortage causes plant strain growth slow, and plant is short and small and the bad even generation of root system development is rotten, and resistance, disease resistance reduce.
In recent years along with the popularization of China's high-yield variety, the increase of nitrogen, Phosphorus fertilizer usage and the raising to quality of agricultural product requirement, the scope of China's potassium deficient soil expands gradually.Have soil potassium deficiency or the severe potassium deficiency of 1/4 to 1/3 in Chinese arable land, the many 60kg of potash fertilizer that the potassium element that the annual crop of per hectare soil takes away comparatively is used, and this situation is also in continuous deterioration.In addition, China's Potash mineral resources is deficient, and the importation dependence of potash fertilizer is more than 70%.Soil potassium deficiency and Potassic fertilizer resources shortage have become the important limiting factor of restriction China, even world agriculture development.Therefore, improving the utilising efficiency of crop to potassium is the effective way solved the problem.
Cotton is the important cash crop of China and strategic materials, occupies an important position in national economy, and since the eighties in 20th century, China has become Cotton Production state maximum in the world, and Cotton Industry is significant in national economy.Cotton has the biological nature that growth cycle is long, biomass is large, cotton boll potassium content is high, makes it very high to the total demand of potassium.But cotton is on the low side to the receptivity of potassium element in soil, so cotton is more more responsive to the low potassium of soil than other field crops.In recent years, along with the progressively raising of output of cotton level, and the Huanghe valley, cotton region, Yangtze valley transgenic cotton against pests cultivated area continuous expansion, Cotton in China produce in potassium deficiency phenomenon more and more general, potassium deficiency degree is more and more serious, becomes one of major reason of current cotton premature senescence.
Summary of the invention
The object of this invention is to provide a kind of Protein G hKT2 relevant to the ability of plant absorption potassium ion and encoding gene thereof and application.
Protein provided by the invention, available from cotton variety " cotton No. 17 of the Liao Dynasty ", called after GhKT2 albumen is following (a) or (b) or (c) or (d): the protein that (a) is made up of the aminoacid sequence shown in sequence in sequence table 1; B the aminoacid sequence of sequence 1 is coerced the relevant protein derived by sequence 1 to plant Tolerance to low-K stress through the replacement of one or several amino-acid residue and/or disappearance and/or interpolation by (); (c) by the aminoacid sequence of sequence 1 through the replacement of one or several amino-acid residue and/or disappearance and/or interpolation and the protein that by sequence 1 derived relevant to the ability of plant absorption potassium ion; (d) by the aminoacid sequence of sequence 1 through the replacement of one or several amino-acid residue and/or disappearance and/or interpolation and the protein that by sequence 1 derived relevant to development of plants.
In order to make the protein in (a) be convenient to purifying, the N-terminal of the protein that the aminoacid sequence shown in sequence 1 forms or C-terminal label as shown in table 1 can be connected in by sequence table.
The sequence of table 1 label
Label Residue Sequence
Poly-Arg 5-6(is generally 5) RRRRR
Poly-His 2-10(is generally 6) HHHHHH
FLAG 8 DYKDDDDK
Strep-tag II 8 WSHPQFEK
c-myc 10 EQKLISEEDL
Protein in above-mentioned (b) or (c) or (d) can synthetic, also can first synthesize its encoding gene, then carries out biological expression and obtain.The encoding gene of the protein in above-mentioned (b) or (c) or (d) is by the codon by lacking one or several amino-acid residue in the DNA sequence dna shown in sequence in sequence table 2, and/or carry out the missense mutation of one or several base pair, and/or the encoding sequence connecting the label shown in table 1 is held to obtain at its 5 ' end and/or 3 '.
The gene (GhKT2 gene) of code for said proteins also belongs to protection scope of the present invention.
Described gene specifically can be the DNA molecular of (1) or (2) or (3) or (4) or (5) or (6) or (7) as follows: the DNA molecular shown in sequence 2 in (1) sequence table; (2) DNA sequence dna limited with (1) is under strict conditions hybridized and the DNA molecular of coded plant Tolerance to low-K stress Stress Related Protein; (3) DNA sequence dna limited with (1) has more than 90% homology and the DNA molecular of coded plant Tolerance to low-K stress Stress Related Protein; (4) to hybridize to the DNA sequence dna that (1) limits under strict conditions and the DNA molecular of the albumen relevant with the ability of plant absorption potassium ion of encoding; (5) to (1) or the DNA sequence dna that limits, there is more than 90% homology and the DNA molecular of the albumen relevant with the ability of plant absorption potassium ion of encoding; (6) DNA sequence dna limited with (1) is under strict conditions hybridized and the DNA molecular of coded plant development associated protein; (7) DNA sequence dna limited with (1) has more than 90% homology and the DNA molecular of coded plant development associated protein.Above-mentioned stringent condition can be in the solution of 6 × SSC, 0.5%SDS, hybridizes under 65oC, then uses 2 × SSC, 0.1%SDS and 1 × SSC, 0.1%SDS respectively to wash film once.
Expression cassette containing described GhKT2 gene, recombinant vectors, transgenic cell line or recombinant bacterium all belong to protection scope of the present invention.
Available existing plant expression vector construction contains the recombinant vectors of described gene.Described plant expression vector comprises double base agrobacterium vector and can be used for the carrier etc. of plant micropellet bombardment.Described plant expression vector also can comprise 3 ' end untranslated region of foreign gene, namely comprises the DNA fragmentation of polyadenylation signals and any other participation mRNA processing or genetic expression.The bootable polyadenylic acid of described polyadenylation signals joins 3 ' end of mRNA precursor.When using described gene constructed recombinant vectors, can add any one enhancement type promotor or constitutive promoter before its transcription initiation Nucleotide, they can be used alone or are combined with other plant promoter; In addition, when using gene constructed recombinant vectors of the present invention, also can use enhanser, comprise translational enhancer or transcriptional enhancer, but must be identical with the reading frame of encoding sequence, to ensure the correct translation of whole sequence.The source of described translation control signal and initiator codon is widely, can be natural, also can be synthesis.Translation initiation region can from transcription initiation region or structure gene.For the ease of identifying transgenic plant cells or plant and screening, can process plant expression vector used, the coding can expressed in plant as added can produce enzyme or the gene of luminophor, the antibiotic marker thing with resistance or the chemical resistance reagent marker gene etc. of colour-change.From the security consideration of transgenic plant, any selected marker can not be added, directly with adverse circumstance screening transformed plant.Described recombinant vectors specifically can be the recombinant plasmid multiple clone site that described GhKT2 gene inserts pBI121 carrier obtained.
The present invention also protects a kind of method of cultivating transgenic plant, is by described GhKT2 channel genes object plant, obtains Tolerance to low-K stress and coerce the transgenic plant of ability higher than described object plant.It is less than 100 μMs that described low potassium specifically can be potassium concentration.
The present invention also protects a kind of method of cultivating transgenic plant, is by described GhKT2 channel genes object plant, and the ability of the potassium ion that is absorbed is higher than the transgenic plant of described object plant.
The present invention also protects a kind of method of cultivating transgenic plant, is by described GhKT2 channel genes object plant, obtains the transgenic plant of potassium accumulation volume higher than described object plant.
The present invention also protects a kind of method of cultivating transgenic plant, is by described GhKT2 channel genes object plant, obtains the transgenic plant of biomass higher than described object plant.
Above in arbitrary described method, the plant tissue of conversion by using Ti-plasmids, Ri plasmid, plant viral vector, directly delivered DNA, microinjection, conductance, conventional biology methods transformed plant cells or the tissue such as agriculture bacillus mediated, and is cultivated into plant by the expression vector carrying described GhKT2 gene.Described GhKT2 gene specifically imports in described object plant by described recombinant vectors.Described object plant is monocotyledons or dicotyledons.Described dicotyledons is specially Arabidopis thaliana, as Columbia ecotype Arabidopis thaliana.
By the further research to vegetable cell potassium-channel and potassium transporter function and Regulation Mechanism, progressively will illustrate the molecular regulation mechanism that plant potassium nutrition efficient absorption utilizes, the potassium nutrition character for Crop Improvement is also provided important theoretical foundation by this.
The invention discloses new albumen and gene, and prove after described gene transfered plant, plant Ability of bearing low potassium can be significantly improved.The present invention is for new variety of plant, and particularly new cotton variety has substantial worth.
Accompanying drawing explanation
Fig. 1 is the Multiple Sequence Alignment result in embodiment 1.
Fig. 2 is the relative expression quantity result of GhKT2 gene in embodiment 1.
Fig. 3 is the Subcellular Localization result of GhKT2 albumen in embodiment 1.
Fig. 4 is the structural representation of recombinant plasmid pBI121-GhKT2.
Fig. 5 is the relative expression quantity result of GhKT2 gene in embodiment 2.
Fig. 6 is the phenotype photo (being inverted cultivation after 10 days) in the step one of embodiment 2.
Fig. 7 is the phenotype photo (being inverted cultivation after 26 days) in the step one of embodiment 3.
Fig. 8 is in the step 2 of embodiment 3, when adopting MS substratum, and the dry matter result of Columbia ecotype Arabidopis thaliana, K27 strain, K6 strain, K14 strain.
Fig. 9 is in the step 2 of embodiment 3, when adopting low potassium substratum, and the dry matter result of Columbia ecotype Arabidopis thaliana, K27 strain, K6 strain, K14 strain.
Figure 10 is in the step 2 of embodiment 3, when adopting MS substratum, and the potassium content result of Columbia ecotype Arabidopis thaliana, K27 strain, K6 strain, K14 strain every gram of dry weight.
Figure 11 is in the step 2 of embodiment 3, when adopting low potassium substratum, and the potassium content result of Columbia ecotype Arabidopis thaliana, K27 strain, K6 strain, K14 strain every gram of dry weight.
Figure 12 is in the step 2 of embodiment 3, the potassium accumulation volume result of Columbia ecotype Arabidopis thaliana, K27 strain, K6 strain, K14 strain.
Figure 13 is in the step 2 of embodiment 3, when adopting low potassium substratum, and the potassium accumulation volume result of the plant of Columbia ecotype Arabidopis thaliana, K27 strain, K6 strain, K14 strain.
Embodiment
Following embodiment is convenient to understand the present invention better, but does not limit the present invention.Experimental technique in following embodiment, if no special instructions, is ordinary method.Test materials used in following embodiment, if no special instructions, is and purchases available from routine biochemistry reagent shop.Quantitative test in following examples, all arranges and repeats experiment for three times, results averaged.
Columbia ecotype Arabidopis thaliana (representing with " WT "): Salk institute genome analysis laboratory (Salk Institute Genomic Analysis Laboratory).Plant binary transformation vector pBI121(is also known as pBI121 carrier): Chinese plasmid vector strain cell pnca gene preservation center.Agrobacterium strains EHA105: sky, Beijing bounties Gene Tech. Company Limited.Cotton variety " cotton No. 17 of the Liao Dynasty ": reference: Hu Yushu, to the reviews and prospects of our province short season breeding for disease resistance, " science and wealth " magazine 12 phases in 2010.
The discovery of embodiment 1, GhKT2 albumen and encoding gene thereof, expression analysis and Subcellular Localization
One, the discovery of GhKT2 albumen and encoding gene thereof
Utilize database retrieval and compare the related gene sequence of other species, splice and verify the est sequence of cotton, carry out 5 ' RACE and 3 ' RACE, a new albumen is obtained from cotton variety " cotton No. 17 of the Liao Dynasty ", by its called after GhKT2 albumen, as shown in the sequence 1 of sequence table (being made up of 792 amino-acid residues).Be GhKT2 gene by the unnamed gene of coding GhKT2 albumen, the open reading frame of its cDNA is as shown in the sequence 2 of sequence table (being made up of 2379 Nucleotide).
In order to analyze the difference of the Homologous gene sequences of GhKT2 gene and other species, first utilizing clustalXversion1.83 to build the Multiple Sequence Alignment structure of EBP1 sequence, using the default parameters of software.Use MEGA4 to carry out evolutionary tree verification according to the Multiple Sequence Alignment result of albumen and generate EBP1 unrooted evolutionary tree, evolutionary tree generates and adopts adjacent method (neighbor joining).The similarity that Multiple Sequence Alignment the results are shown in Figure the VvKUP2 albumen of 1, GhKT2 protein sequence and grape is 87.8%, is 83.4% with the similarity of Arabidopis thaliana AtKT2 albumen, is 80.5% with the similarity of ice plant McHAK2 albumen.
Two, the expression analysis of GhKT2 albumen and encoding gene thereof
By fluorescence real-time quantitative PCR, the expression pattern of GhKT2 gene in cotton variety " the Liao Dynasty cotton No. 17 " is analyzed: fluorescence real-time quantitative PCR instrument used is ABI 7500 Fast(Applied Biosystem), primer pair used is " 5 '-TCTTGGAGGACCTCACTGCT-3 ' " and " 5 '-CATCTGCTCGAAGAACCACA-3 ' ", be the normal cDNA obtained for the total serum IgE reverse transcription at each position of potassium (2.5mM) level cotton in lower tri-leaf period for detecting the sample of tissue specific expression, be the cDNA that the RNA reverse transcription of cotton root obtains after 0.03mM low potassium process different time for detecting the sample of low potassium inducing properties, PCR program: 94 DEG C of sex change 30s, 94 DEG C of sex change 5s, 60 DEG C of annealing 35s, 40 circulations, relative expression quantity adopts 2 -Δ Δ Ctmethod calculates, with cotton UBQ7 gene in contrast (primer pair for the identification of cotton UBQ7 gene is: 5 '-AAGAAGAAGACCTACACCAAGCC-3 ' and 5 '-GCCCACACTTACCGCAATA-3 ').The relative expression quantity of normal potassium supply level cotton in lower tri-leaf period each position GhKT2 gene is shown in Fig. 2 A, and GhKT2 gene all has certain expression amount in cotton different tissues, and expression amount in leaf is relatively high.After 0.03mM low potassium process different time, the relative expression quantity of cotton root GhKT2 gene is shown in Fig. 2 B, and after low potassium process 24h, GhKT2 gene expression amount reaches maximum, illustrates that GhKT2 gene pairs low potassium stress has response.
Three, the Subcellular Localization of GhKT2 albumen
Adopt protein trans-membrane region analysis tool TMpred to predict GhKT2 protein structure, find that its optimum topological framework contains 14 cross-film districts.The transgenic arabidopsis root that this step utilizes genetic stability to transform is to study the Subcellular Localization of GhKT2 albumen.Connect the recombinant vectors of GFP gene according to plant binary transformation vector pCAMBIA3300(, this carrier by China Agricultural University's agronomy and Biotechnology Institute grandson pass Puritanism award be so kind as to give) multiple clone site and the coding region sequence of GhKT2 gene design forward and the reverse primer of the whole coding region of GhKT2 gene of increasing.Obtain pCAMBIA3300-GhKT2-GFP recombinant vectors, and proceeded to Agrobacterium EHA105.
Forward primer: 5 '-C gAGCTCaTGGATCTTGAGTTTGGGAAGT-3 ';
Reverse primer: 5 '-GC tCTAGAcACCACATAAACCATGCCA-3 '.
With OlympusFV1000 type microscopic examination reporter gene.The Arabidopis thaliana that control vector transforms, its fluorescence is mainly positioned at the positions (Fig. 3 A) such as the nucleus of cell and cytolemma.The Arabidopis thaliana root cell fluorescence localization that recombinant vectors transforms is on cytolemma or on cell walls (Fig. 3 B).In order to get rid of the possibility that KT2-GFP expresses on cell walls, the tip of a root is utilized plasmolysis after 500mM treatment with mannitol 10min, as shown in Figure 3 C, in tip of a root plasmolysis cell, fluorescent signal is extremely faint on cell walls.As can be seen here, KT2-GFP fusion rotein is positioned on cytolemma, instead of on cell walls.
The acquisition of embodiment 2, transgenic plant
One, the structure of recombinant plasmid
1, the total serum IgE of cotton variety " the Liao Dynasty cotton No. 17 " blade is extracted and reverse transcription is cDNA.
2, the cDNA obtained with step 1, for template, carries out pcr amplification with the primer pair of F1 and R1 composition, obtains pcr amplification product.
F1：5’-GC TCTAGAATGGATCTTGAGTTTGGGAAGT-3’
R1：5’-C GAGCTCTTACACCACATAAACCATGCCA-3’
3, with the pcr amplification product that restriction enzyme XbaI and SacI double digestion step 2 obtain, digestion products is reclaimed.
4, with restriction enzyme XbaI and SacI double digestion pBI121 carrier, the carrier framework of about 12000bp is reclaimed.
5, the digestion products of step 3 is connected with the carrier framework of step 4, obtains recombinant plasmid pBI121-GhKT2.The structural representation of recombinant plasmid pBI121-GhKT2 is shown in Fig. 4.According to sequencing result, structrual description carries out to recombinant plasmid pBI121-GhKT2 as follows: the double chain DNA molecule shown in sequence 2 inserting sequence table between XbaI and the SacI restriction enzyme site of carrier pBI121.
Two, the acquisition of transgenic plant
1, recombinant plasmid pBI121-GhKT2 is imported agrobacterium strains EHA105, obtain recombinational agrobacterium.
2, treat Columbia ecotype Arabidopsis plant Post flowering, cut off major branch top, promote side shoot development; In 6 days after beta pruning, the bacteria suspension of recombinational agrobacterium step 1 obtained dips on the inflorescence that wet Arabidopis thaliana do not show money or valuables one carries unintentionally, then the Arabidopis thaliana black plastic bag being full of gas is encased, keep flat, plastics bag is removed after light culture 24h, recover illumination, cultivate plant to solid according to a conventional method, results mature T 1for seed.
3, the MS culture medium culturing T containing 50mg/L kantlex is adopted 1positive plant (positive plant shows as: true leaf health is in deep green, and root is stretched in substratum) is also therefrom selected for seed.
4, positive plant selfing step 3 obtained obtains T 2for seed.
5, the MS culture medium culturing T containing 50mg/L kantlex is adopted 2also positive plant (screening criteria is the same) is therefrom selected for seed.
For a certain T 1for plant, if its T 2positive plant is, this T for plant 1be a transgenic line isozygotied for plant and self progeny thereof.
5, T is cultivated 2for plant and selfing obtain T 3for seed.
6, T is got 3for the blade of plant, extract total serum IgE reverse transcription is cDNA, with the expression amount of the primer pair qualification GhKT2 gene of F2 and R2 composition, with the expression amount of primer pair qualification AtActin gene (reference gene) of F3 and R3 composition.
F2：5’-TCTTGGAGGACCTCACTGCT-3’；
R2：5’-CATCTGCTCGAAGAACCACA-3’。
F3：5’-GGCAAGTCATCACGATTGG-3’；
R3：5’-CAGCTTCCATTCCCACAAAC-3’。
By the transgenic line isozygotied higher for the relative expression quantity of three GhKT2 genes called after K27 strain, K6 strain, K14 strain respectively.
The T of Columbia ecotype Arabidopis thaliana, K27 strain 3for the T of plant, K6 strain 3for the T of plant, K14 strain 3for in plant, the relative expression quantity of GhKT2 gene is shown in Fig. 5.
Three, the acquisition of empty carrier plant is turned
Replace recombinant plasmid pBI121-GhKT2 to carry out step 2 with pBI121 carrier, obtain turning empty carrier plant.
The qualification of embodiment 3, transgenic plant
MS culture medium prescription:
Macroelement: 1.65g NH 4nO 3, 1.9g KNO 3, 0.17g KH 2pO 4, 0.37 gm gsO 47H 2o, 0.44 gcaCl 22H 2o;
Trace element: 22.3mg MnSO 44H 2o, 8.6mg ZnSO 47H 2o, 0.025mg CoCl 26H 2o, 0.025mgCuSO 45H 2o, 0.025mg Na 2moO 42H 2o, 0.83mg KI, 6.2mg H 3bO 3;
Molysite: 27.8mg FeSO 47H 2o, 37.3mg Na 2eDTA;
By water-soluble to macroelement, trace element, molysite and 9g agar and be settled to 1L with water, obtain MS substratum (potassium concentration is about 19.9mM).
Low potassium culture medium prescription:
Macroelement: 2.3g NH 4nO 3, 0.37g MgSO 47H 2o, 0.144g NH 4h 2pO 4, 0.44g CaCl 22H 2o;
Trace element: with the trace element in MS substratum.
Molysite: with the molysite in MS substratum.
By water-soluble to macroelement, trace element, molysite and 9g agar and be settled to 1L with water, then add KCl, obtain K +concentration is the low potassium substratum of 100 μMs.
One, phenotype analytical
By Columbia ecotype Arabidopis thaliana seed, the T turning empty carrier plant 3for the T of seed, K27 strain 3for the T of seed, K6 strain 3for the T of seed, K14 strain 3be determined as follows respectively for seed: seed is laid in MS substratum, 4 DEG C of vernalization 3 days, then cultivate 4 days in illumination box, then individual plant is divided into two groups, moves into MS substratum respectively and be inverted cultivation 10 days and low potassium substratum inversion cultivation 26 days (in often kind of substratum, each strain arranges 4 strain individual plants).Individual plant moves into the photo of MS substratum inversion cultivation after 10 days and sees Fig. 6.Individual plant moves into the photo of low potassium substratum inversion cultivation after 26 days and sees Fig. 7.No matter be adopt normal MS culture medium culturing 10 days or adopt low potassium culture medium culturing 26 days, root and the hat of K27 strain, K6 strain, K14 strain are all greater than Columbia ecotype Arabidopis thaliana, turn empty carrier plant consistent with Columbia ecotype Arabidopis thaliana phenotype.Result shows, imports the growth potential that GhKT2 gene can improve plant.
Two, the biomass of transgenic arabidopsis, potassium content and potassium accumulation quantitative analysis
By the T of Columbia ecotype Arabidopis thaliana seed, K27 strain 3for the T of seed, K6 strain 3for the T of seed, K14 strain 3for seed and the T turning empty carrier plant 3be determined as follows respectively for seed (each strain 80 seedling): seed is laid in MS substratum, 4 DEG C of vernalization 3 days, then cultivate 4 days in illumination box; Then individual plant is divided into two groups, move into MS substratum and low potassium substratum respectively, be positioned in illumination box and cultivate 7 days, from the junction of hypocotyl and radicle, plant is divided into two portions (root and bizet), be placed in baking oven respectively to dry to constant weight in 80 DEG C, weigh (obtaining dry matter); After weighing, root and bizet are placed on respectively in crucible in retort furnace, process (first 300 DEG C of charing 1h, then 575 DEG C of ashing 5h), then be dissolved in the 0.1mol/L HCl aqueous solution, measure potassium content (potassium content in every gram of dry weight) with Z-2000 atomic absorption spectrophotometer (high and new technology company of Hitachi).
When adopting MS substratum, the dry matter of Columbia ecotype Arabidopis thaliana, K27 strain, K6 strain, K14 strain the results are shown in Figure 8.When adopting low potassium substratum, the dry matter of Columbia ecotype Arabidopis thaliana, K27 strain, K6 strain, K14 strain the results are shown in Figure 9.No matter adopt MS culture medium culturing or adopt low potassium culture medium culturing, the root of three transgenic lines and the biomass of bizet are all higher than wild-type.When adopting MS substratum, compared with Columbia ecotype Arabidopis thaliana, the biomass of the root of three transgenic lines adds 207.4%, 102.1% and 70.2% respectively, and the biomass of bizet adds 137.2%, 94.5% and 45.6% respectively.When adopting low potassium substratum, compared with Columbia ecotype Arabidopis thaliana, the biomass of the root of three transgenic lines adds 30.2%, 13.2% and 17.5% respectively, and the biomass of bizet adds 10.1%, 5.2% and 4.1% respectively.No matter be adopt MS substratum or adopt low potassium substratum, the dry matter result turning empty carrier plant does not all have significant difference with Columbia ecotype Arabidopis thaliana.
When adopting MS substratum, the potassium content of Columbia ecotype Arabidopis thaliana, K27 strain, K6 strain, K14 strain every gram of dry weight the results are shown in Figure 10.When adopting low potassium substratum, the potassium content of Columbia ecotype Arabidopis thaliana, K27 strain, K6 strain, K14 strain every gram of dry weight the results are shown in Figure 11.When adopting MS substratum, compared with Columbia ecotype Arabidopis thaliana, the potassium content of K27 strain, K6 strain, K14 strain root adds 4.6%, 12.8% and 8.9% respectively, and the potassium content of bizet adds 21.0%, 18.3% and 8.7% respectively.When adopting low potassium substratum, compared with Columbia ecotype Arabidopis thaliana, the potassium content of K27 strain, K6 strain, K14 strain root adds 10.5%, 5.3% and 12.6% respectively, and the potassium content of bizet adds 4.7%, 22.9% and 10.9% respectively.No matter be adopt MS substratum or adopt low potassium substratum, the potassium content turning empty carrier plant every gram of dry weight does not all have significant difference with Columbia ecotype Arabidopis thaliana.
Potassium accumulation volume (potassium content in each plant) reacts the ability that plant absorbs potassium.When adopting MS substratum, the potassium accumulation volume of Columbia ecotype Arabidopis thaliana, K27 strain, K6 strain, K14 strain is shown in Figure 12.When adopting low potassium substratum, the potassium accumulation volume of Columbia ecotype Arabidopis thaliana, K27 strain, K6 strain, K14 strain is shown in Figure 13.No matter adopt MS substratum or low potassium substratum, in three transgenic lines, the potassium accumulation volume of root and bizet is all significantly higher than Columbia ecotype Arabidopis thaliana.When adopting low potassium substratum, compared with Columbia ecotype Arabidopis thaliana, the potassium content of K27 strain, K6 strain, K14 strain root adds 180%, 94.8% and 63.8% respectively, and the potassium content of bizet adds 187.3%, 126.0% and 58.7% respectively.No matter be adopt MS substratum or adopt low potassium substratum, the potassium accumulation volume turning empty carrier plant does not all have significant difference with Columbia ecotype Arabidopis thaliana.

Claims (12)

1. a protein, the protein be made up of the aminoacid sequence shown in sequence in sequence table 1.
2. the gene of protein described in coding claim 1.
3. gene as claimed in claim 2, is characterized in that: described gene is the DNA molecular shown in sequence in sequence table 2.
4. the expression cassette containing gene described in Claims 2 or 3.
5. the recombinant vectors containing gene described in Claims 2 or 3.
6. the transgenic cell line containing gene described in Claims 2 or 3.
7. the recombinant bacterium containing gene described in Claims 2 or 3.
8. cultivating a method for transgenic plant, is by channel genes object plant described in Claims 2 or 3, obtains Tolerance to low-K stress and coerces the transgenic plant of ability higher than described object plant;
Described object plant is Arabidopis thaliana.
9. cultivating a method for transgenic plant, is by channel genes object plant described in Claims 2 or 3, and the ability of the potassium ion that is absorbed is higher than the transgenic plant of described object plant;
Described object plant is Arabidopis thaliana.
10. cultivate a method for transgenic plant, be by channel genes object plant described in Claims 2 or 3, obtain the transgenic plant of potassium accumulation volume higher than described object plant;
Described object plant is Arabidopis thaliana.
11. 1 kinds of methods of cultivating transgenic plant, are by channel genes object plant described in Claims 2 or 3, obtain the transgenic plant of biomass higher than described object plant;
Described object plant is Arabidopis thaliana.
12., as the method as described in arbitrary in claim 8 to 11, is characterized in that: gene described in Claims 2 or 3 imports described object plant by recombinant vectors described in claim 5;
Described object plant is Arabidopis thaliana.
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