CN109553665A

CN109553665A - Tobacco KC1 gene and application

Info

Publication number: CN109553665A
Application number: CN201811340505.7A
Authority: CN
Inventors: 郭玉双; 任学良; 李立芹; 鲁黎明; 王仁刚; 张洁
Original assignee: Guizhou Institute of Tobacco Science
Current assignee: Guizhou Institute of Tobacco Science
Priority date: 2018-11-12
Filing date: 2018-11-12
Publication date: 2019-04-02
Anticipated expiration: 2038-11-12
Also published as: CN109553665B

Abstract

The present invention relates to tobacco KC1 gene and applications.The sequence of the tobacco KC1 gene and its coding protein is respectively as shown in SEQ ID NO:1 and 2.The clone from tobacco obtains KC1 gene and demonstrates the biological function of the gene by yeast function complementation experiment the present invention for the first time, and tobacco KC1 gene has the function of promoting Potassium Absorption and transhipment.

Description

Tobacco KC1 gene and application

Technical field

The present invention relates to gene engineering technology fields, specifically, being related to tobacco KC1 gene and application.

Background technique

Potassium-channel is the ion channel for allowing potassium ion specifically penetrating plasma membrane, and hinders other ions penetrating, especially It is sodium ion.These channels generally consist of two parts: a part is channel region, selects and potassium ion is allowed to pass through, and is hindered Sodium ion；Another part is gate controlled switch, the switching channels according to the signal in environment.

The prior art for potassium-channel gene research in model plant arabidopsis than wide, for example, from Dreyer etc. detects the K of plant using Xenopus Oocytes⁺KC1 is had found for the first time in the bioelectrical activity of channel gene, For subsequent Geoffrey Duby etc. to finding in KC1 research, the subunit of KC1 gene expression cannot form the tetramer so as to cause KC1 The silencing of gene, but can combine and form different K by the subunit that AKT1 gene encodes⁺Channel protein, to regulate and control K⁺Suction It receives and transports, while Dietmar Geiger has also discovered KC1 in the root of arabidopsis and interacts to KAT1, to improve KAT1 is under the conditions of low potassium to K⁺Absorption.Subsequent Linda Jeanguenin etc. utilizes electrophysiological technique, successfully has found KC1 Expression regulation KAT2 gene pairs K⁺Absorption and transhipment, it was demonstrated that KC1 gene has the adjustment effect to different genes.

Tobacco is a kind of crop that consumption potassium amount is very big, and potassium content of tobacco leaf is the important indicator for measuring quality of tobacco, currently, It is less for the research of Potassium in Tobacco ion channel.

Summary of the invention

The object of the present invention is to provide tobacco KC1 gene and its protein of coding.

It is a further object of the present invention to provide the applications of tobacco KC1 gene.

In order to achieve the object of the present invention, tobacco KC1 gene provided by the invention, for the following protein (a) of coding or (b) Gene:

(a) protein that the amino acid sequence shown in SEQ ID NO:2 forms；

(b) sequence shown in SEQ ID NO:2 is substituted, lacks or adds one or several amino acid and has same function The protein as derived from (a).

The nucleotide sequence of tobacco KC1 gene of the present invention is as shown in SEQ ID NO:1, full length gene 1989bp.The present invention Clone obtains tobacco KC1 gene with the following method:

1. extracting tobacco cell total serum IgE, and invert obtained total serum IgE is extracted before carrying out the amplification of KC1 gene PCR Record is cDNA.In the present invention, the extraction of the tobacco cell total serum IgE is using extraction cell total rna commonly used in the art Trizol method specifically can be used in the embodiment of the present invention in technical solution.In the present invention, the tobacco cell is total The raw material that RNA is extracted is the fresh blade of tobacco, and the tobacco is adopted as the tobacco bred of this field routine, such as K326.

2. the tobacco cell total serum IgE reverse transcription is synthesized cDNA after extraction obtains tobacco cell total serum IgE.In this hair In bright, the synthesis of the cDNA uses the synthetic method of the cDNA of this field routine, without other particular/special requirements；Specifically The synthesis of cDNA is completed in the embodiment of the present invention using the cDNA synthetic agent box of TaKaRa company.

3. carrying out the amplification of KC1 gene PCR after obtaining cDNA, obtaining target fragment.In the present invention, the KC1 base Because the system of PCR amplification is preferably 20 μ L systems, including Premix ExTaq 10 μ L, 10 μM of forward primer 0.5 μ L, 10 μM Reverse primer 0.5 μ L, tobacco cell cDNA 1 μ L, ddH₂O 8μL.In the present invention, the PCR amplification of the KC1 gene Response procedures are preferred are as follows: 95 DEG C of initial denaturation 5min；95 DEG C of denaturation 30s；55 DEG C of annealing 30s；72 DEG C of extension 2min；35 circulations.

4. the target fragment is sequenced after KC1 gene PCR expands to obtain target fragment, KC1 gene is obtained. The present invention preferably purifies target fragment after the PCR amplification, and the present invention is to the method for the purifying without spy Different restriction is carried out using DNA purification kit well known to those skilled in the art.

5. after the completion of purifying, preferably by the target fragment after purification import in bacillus coli DH 5 alpha competent cell into Row bacterium colony PCR, is sequenced after being verified as positive colony.The present invention is after obtaining positive colony, it is preferred to use the side of bacterium colony PCR Method verifies positive colony.In the present invention, the forward primer nucleotide sequence of the bacterium colony PCR are as follows: 5'- ATGAATACAGAAACCAGTAA-3'(SEQ ID NO:3)；The nucleotides sequence of the reverse primer is classified as 5'- TTAAAAGGAAAAACAAATGG-3'(SEQ ID NO:4), the system of the bacterium colony PCR is 10 μ L, including Premix ExTaq 5 μ L, 10 μM of forward primer 0.5 μ L, 10 μM of reverse primer 0.5 μ L, ddH₂O 4μL.In the present invention, described will purifying Target fragment afterwards imports the conversion Escherichia coli that method used by bacillus coli DH 5 alpha competent cell is this field routine The method of competent cell, specific as follows: the target fragment and pMD19-T carrier are connected 10 under conditions of 16 DEG C~ 14h obtains connection product；The connection product is converted into bacillus coli DH 5 alpha competent cell, the large intestine bar after being converted Bacterium DH5 α；Bacillus coli DH 5 alpha after the conversion is inoculated on the LB plate for be coated with ampicillin and carries out screening training It supports, obtains positive colony.

6. after bacterium colony PCR verifies positive colony, it is preferred that randomly select 2~4 from the positive colony having verified that solely Vertical positive colony is sequenced, and the sequence of the tobacco KC1 gene is obtained.

The present invention also provides the biomaterial for containing the tobacco KC1 gene, the biomaterial is expression cassette, expression load Body, cloning vector, engineering bacteria or transgenic cell line.

Biomaterial the present invention also provides the tobacco KC1 gene or containing the gene is promoting plant or microorganism potassium Application in Ions Absorption and transhipment.

Plant of the present invention includes but is not limited to tobacco, arabidopsis.The microorganism includes but is not limited to yeast.

Biomaterial the present invention also provides the tobacco KC1 gene or containing the gene is in prepare transgenosis plant Using.

Application of the biomaterial in plant breeding the present invention also provides the tobacco KC1 gene or containing the gene. The purpose of the breeding is to promote plant Potassium Absorption and transhipment.

Preferably, the tobacco KC1 gene is transferred in tobacco plant, makes tobacco KC1 gene overexpression to improve The content of potassium ion in the tobacco leaf of tobacco plant.It is highly preferred that tobacco KC1 gene is transferred to tobacco using agrobacterium-mediated transformation In plant, the transgenic plant of KC1 gene overexpression is obtained.

The present invention also provides the Specific PCR primers pair for expanding tobacco KC1 gene, the nucleotides sequence of the primer pair Column are as shown in SEQ ID NO:3-4.The primer pair is with NCBI Reference Sequence:LOC104097809 for reference Sequence is obtained using the design of software primer 5.

The present invention also provides a kind of method for promoting plant Potassium Absorption and transhipment, the methods are as follows:

1) making plant includes the tobacco KC1 gene；Alternatively,

2) plant is made to be overexpressed the tobacco KC1 gene.

The method includes but be not limited to transgenosis, hybridization, backcrossing, selfing or vegetative propagation.

Clone obtains KC1 gene and demonstrates the gene by yeast function complementation experiment the present invention from tobacco for the first time Biological function, the recombination yeast after tobacco KC1 gene to be transferred to potassium native defect type yeast mutant R5421 have potassium ion It absorbs, transport function.Therefore, tobacco KC1 gene provided by the invention has the function of promoting Potassium Absorption and transhipment.

Detailed description of the invention

Fig. 1 is yeast function complementation experiment result in the embodiment of the present invention 2.Wherein, A: potassium concentration is in culture medium 20uM, B: potassium concentration is 2mM in culture medium.In figure, 1 is the recombination yeast for being transferred to tobacco KC1 gene, and 2 be negative control Group (is transferred to empty carrier), and 3 be the recombination yeast of positive controls (being transferred to arabidopsis AtAKT1 gene)；From left to right it is followed successively by bacterium Stoste, 10 times of dilutions, 100 times, growth result of 1000 times of dilutions on culture medium.

Specific embodiment

The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..Unless otherwise specified, embodiment According to conventional laboratory conditions, such as Sambrook molecular cloning experiment handbook (Sambrook J&Russell DW, Molecular Cloning:a Laboratory Manual, 2001), or according to the condition of manufacturer's specification suggestion.

The clone of 1 tobacco KC1 gene of embodiment

The fresh blade of 0.5g tobacco (tobacco bred K326) is taken, the total serum IgE of tobacco cell is extracted using Trizol method, so CDNA is synthesized using the cDNA synthetic agent box of TaKaRa company afterwards, further using Primer5.0 software design and through remarkable Work optimizes to obtain primer, and the primer includes forward primer and reverse primer, the forward primer nucleotide sequence are as follows: 5'- ATGAATACAGAAACCAGTAA-3'；The nucleotides sequence of the reverse primer is classified as 5'-TTAAAAGGAAAAACAAATGG-3', Using the cDNA of synthesis as template, PCR amplification is carried out, PCR amplification system is 20 μ L systems, including 10 μ L of Premix ExTaq, 10 μM of forward primer 0.5 μ L, 10 μM of reverse primer 0.5 μ L, tobacco cell cDNA 1 μ L, ddH₂O 8μL；The PCR The response procedures of amplification are as follows: 95 DEG C of initial denaturation 5min；95 DEG C of denaturation 30s；55 DEG C of annealing 30s；72 DEG C of extension 2min；35 are followed Ring.

After the completion of PCR amplification, the purifying of target fragment is carried out using DNA purification kit, by target fragment after purification 12h is connect with 16 DEG C of pMD19-T carrier and obtains connection product, by the obtained connection product conversion bacillus coli DH 5 alpha impression State cell converted after bacillus coli DH 5 alpha, the bacillus coli DH 5 alpha after the conversion is inoculated in and is coated with ammonia benzyl mould Screening and culturing is carried out on the LB plate of element obtains positive colony.After obtaining positive colony, verified using the method for bacterium colony PCR Positive colony, the forward primer of the bacterium colony PCR are as follows: 5'-ATGAATACAGAAACCAGTAA-3'；Reverse primer is 5'- TTAAAAGGAAAAACAAATGG-3'；The system of the bacterium colony PCR is 10 μ L, including 5 μ L of Premix ExTaq, and 10 μM just To primer 0.5 μ L, 10 μM of reverse primer 0.5 μ L, ddH₂O 4μL.Then 3 are randomly selected from the positive colony having verified that Independent positive colony is sent to biotech company and is sequenced, and obtains the sequence such as SEQ ID of tobacco KC1 gene through sequencing Shown in NO:1.

The biological function of 2 tobacco KC1 gene of embodiment is analyzed

1, experiment purpose

The biological function of tobacco KC1 gene is verified by yeast function complementation experiment.

2, experimental method

Using potassium native defect type yeast mutant R5421 as recipient bacterium.Bacterial strain R5421 can be found in Maathuis F J M and Sanders D 1996Mechanisms of potassium absorption by higher plant roots.Physiol.Plant.96,158–168.

By the T- carrier and expression vector P416 that are connected with 1 tobacco KC1 gene of embodiment, (yeast sequestered shuttling expressing is carried Body, TEF constitutive promoter, CYC1 terminator, CEN6 ARSH4 replication orgin, selection markers are URA3, large intestine bar in yeast Selection markers are Amp in bacterium.Carrier P416 can be found in Functional Expression of a ω -3Fatty Acid Desaturase Gene from Glycine max in Saccharomyces cerevisiae) double digestion is carried out respectively (restriction enzyme site is Xba I and Xho I), recycles target gene and expression vector P416, is then connected with ligase, after connection Recombinant yeast expression vector is transferred to the competent cell of bacillus coli DH 5 alpha, carries out PCR to the Escherichia coli single colonie after conversion Amplification, digestion verify whether to construct successfully.

Successful recombinant yeast expression vector will be constructed and be transferred in yeast R5421 that specific step is as follows: being taken with collarium is connect The R5421 saccharomycete of preservation lines on solid medium YPDA, 28 DEG C of culture 12h；Picking R5421 yeast single bacterium falls within Ep pipe In, add 1mL YPDA culture solution to be vortexed；Above-mentioned bacterium solution is all transferred in the triangular flask equipped with YPDA culture solution, in 30 DEG C, 250rpm shakes bacterium to OD₆₀₀=1.2,16h；It transfers, is shaken to OD by 1:10 volume ratio₆₀₀=1.0~1.2；In 28 DEG C, 1000rpm It is centrifuged 5min collection bacterium, is resuspended with the sterilizing ultrapure water of 1/2 volume；In 28 DEG C, 1000rpm is centrifuged 5min collection bacterium, blots supernatant；According to Secondary addition following ingredients (every original bacterium solution of 5mL):

Vortex 1min, mixes transformation system completely；It is placed in 30 DEG C of water-bath and incubates 30min；Place into 42 DEG C of water Thermal shock 28min in bath, cooled on ice 10min；7000rpm is centrifuged 15s, abandons supernatant；Precipitating is gently resuspended with the sterile water of 1mL； 200 μ L transformation mixtures are taken to be laid on auxotroph plate；30 DEG C are cultivated 3 days.It extracts yeast plasmid and identifies conversion results.

In the flat lining out of auxotrophy, 30 DEG C are cultivated 3 days the yeast single colonie that picking was identified；It is lacked with toothpick in nutrition Fall into plate on dip a small amount of thallus in 2mL auxotrophy liquid (Ura Minus Media 8g, glucose 20g, potassium chloride 7.5g, With NaOH tune pH value to 5.8, be settled to 1000mL) in cultivate 12h；8000rpm is centrifuged 1min and collects thallus；Supernatant is abandoned, 1mL is used Distilled water suspension thalline, 8000rpm are centrifuged 1min；Supernatant is abandoned, with 1ml distilled water resuspension, adjusts OD₆₀₀It is 0.8；It will be undiluted Bacterium solution and the bacterium solution for diluting 10 times, 100 times respectively, take 5uL on the culture medium that potassium concentration is 0,2mM respectively, 30 DEG C of trainings Result is observed after supporting 3 days.

3, experimental result

Experimental result is as shown in Figure 1, be 20uM, 2mM culture medium (AP culture medium (1L): 546 μ of phosphoric acid in potassium concentration L, L-arginine 1.742g, 1000 × vitamin solution 1mL, 1000 × trace element solution 1mL, uracil 0.77g, 100 × Ura 10mL, glucose 20g, agar powder 15g) on, negative control group saccharomycete (being transferred to P416 empty carrier) is hardly grown, and is turned The recombinant yeast of the recombination yeast and positive controls (being transferred to arabidopsis AtAKT1 gene) that enter tobacco KC1 gene can give birth to It is long.Recombinant yeast with the increase of extension rate, the recombination yeast and positive controls that are transferred to tobacco KC1 gene still may be used With growth.Result above proves that tobacco KC1 gene of the invention has potassium absorption and transport function.

Although above the present invention is described in detail with a general description of the specific embodiments, On the basis of the present invention, it can be modified or is improved, this will be apparent to those skilled in the art.Cause This, these modifications or improvements, fall within the scope of the claimed invention without departing from theon the basis of the spirit of the present invention.

Sequence table

<110>Guizhou Province Tabacco Science and Technology Institute

<120>tobacco KC1 gene and application

<130> KHP171117869.1

<160> 4

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1989

<212> DNA

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aggtgttggg agatgtttct agttgttttg gtcatatact cagcctggat ttctccattt 180

gagtttgcat tcttgtcata caaggaagat gatgctttat tcatcattga ccacattgtc 240

aactgctttt ttgctattga cattttcctt accttcttcg tcgcgtatct tcatcgagag 300

tcctatcttc ttgttgatga acctaagaaa attgcaataa ggtacttgtc aagttggttc 360

atatttgatg tatgctccac tgtaccattt caatccttga tcctcctttt cacggatcac 420

aaagaaagcg gtggagttgg cttcaagttg ctcagcatgc tcagactttg gcgtctcaga 480

cgagtcagtg ccctgtttgc aagacttgag aaggatatcc ggtttaacta cttctggaca 540

agatgcacaa aactcatatc agtaacattg tttgcagtgc actgtgctgg atgctttaac 600

tatatgattg cagatagata tcctgatcca agaaaaacat ggattggtgc tgtaaatcct 660

gatttcaaga aacaaagcgt tggggacaga tatataactt cactatattg gtctattgta 720

acgatgacaa caactggtta tggggatttg catgctgaga actcaaggga aatgttgttt 780

gacatttttt acatgttatt caacttggga ttgacttctt acatcattgg aaacatgact 840

aatcttgttg ttcattggac cagccgcacc agaaacttta gggatacagt gaaagcagcc 900

caagaatttg cgaaaaggaa tcagttgcct ccaagggtac aagatcaggt tttgtctcac 960

atatgtctca agttcagaac tgaagcattg aaacaagatg agactctcaa tggcctgccc 1020

aaagctatcc gaacaagcat tgcacatcac cttttttttc ctatagttca aaatgttcgt 1080

ttgttccaag gtgtttcacc aaacctcctt ttccaactag ttcctgaaat ggaagctgaa 1140

tacttccctc ccaagcaaga tgtgattttg cagaatgagg ctccaacaga cctgtatata 1200

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tttgctgttc gaactactga gatttctcaa attctacggc taagcaggac agcattgatg 1380

aacattcttc gcgcaaatcc agaagatgaa cgggtagtta tgaacaatct tttgctgaat 1440

ctgcagggat tcggaggctt tggttatgtg gaccaccaaa caaatggagg gccagaaatc 1500

aaaagacatc atgatacagc actgactagc atagatataa acaatttgga agctagagtt 1560

aagaagcaag agggagatga tgtacaagaa ataaacaaaa gcatgaatga tgtgtcatta 1620

aatcttgaaa ataagaagga actaaatgag cagaaagttg agctcattgg accagatgag 1680

aagggaacaa agagctgtca actgaagcct gaggtcccct gttgttctaa ctcttgtagt 1740

accatttcaa gtggttctaa agtgacaaaa tccaccaata agagagtaac cattcacatg 1800

aaaaagaacg aatcattgca aggccagttt gggaagctaa taattctacc tgattcacta 1860

gaagagctat tcagagtagc aggtcaaaga tttggaggct acaatttcaa gaaagctgta 1920

aatgcagagg atgctgaaat agatgacatt gatgtcatca gagatggcga ccatttgttt 1980

ttcctttaa 1989

<210> 2

<211> 661

<212> PRT

<213>tobacco (Nicotiana tabacum)

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Met Asn Thr Glu Thr Ser Asn Gly Phe Phe Ser Ser Asp Leu Leu Pro

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Ser Leu Gly Ala Arg Ile Asn Tyr Ala Thr Lys Leu Arg Lys Phe Ile

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Ile Ser Pro Phe Asn Pro Arg Tyr Arg Cys Trp Glu Met Phe Leu Val

35 40 45

Val Leu Val Ile Tyr Ser Ala Trp Ile Ser Pro Phe Glu Phe Ala Phe

50 55 60

Leu Ser Tyr Lys Glu Asp Asp Ala Leu Phe Ile Ile Asp His Ile Val

65 70 75 80

Asn Cys Phe Phe Ala Ile Asp Ile Phe Leu Thr Phe Phe Val Ala Tyr

85 90 95

Leu His Arg Glu Ser Tyr Leu Leu Val Asp Glu Pro Lys Lys Ile Ala

100 105 110

Ile Arg Tyr Leu Ser Ser Trp Phe Ile Phe Asp Val Cys Ser Thr Val

115 120 125

Pro Phe Gln Ser Leu Ile Leu Leu Phe Thr Asp His Lys Glu Ser Gly

130 135 140

Gly Val Gly Phe Lys Leu Leu Ser Met Leu Arg Leu Trp Arg Leu Arg

145 150 155 160

Arg Val Ser Ala Leu Phe Ala Arg Leu Glu Lys Asp Ile Arg Phe Asn

165 170 175

Tyr Phe Trp Thr Arg Cys Thr Lys Leu Ile Ser Val Thr Leu Phe Ala

180 185 190

Val His Cys Ala Gly Cys Phe Asn Tyr Met Ile Ala Asp Arg Tyr Pro

195 200 205

Asp Pro Arg Lys Thr Trp Ile Gly Ala Val Asn Pro Asp Phe Lys Lys

210 215 220

Gln Ser Val Gly Asp Arg Tyr Ile Thr Ser Leu Tyr Ser Ile Val Thr

225 230 235 240

Met Thr Thr Thr Gly Tyr Gly Asp Leu His Ala Glu Asn Ser Arg Glu

245 250 255

Met Leu Phe Asp Ile Phe Tyr Met Leu Phe Asn Leu Gly Leu Thr Ser

260 265 270

Tyr Ile Ile Gly Asn Met Thr Asn Leu Val Val His Trp Thr Ser Arg

275 280 285

Thr Arg Asn Phe Arg Asp Thr Val Lys Ala Ala Gln Glu Phe Ala Lys

290 295 300

Arg Asn Gln Leu Pro Pro Arg Val Gln Asp Gln Val Leu Ser His Ile

305 310 315 320

Cys Leu Lys Phe Arg Thr Glu Ala Leu Lys Gln Asp Glu Thr Leu Asn

325 330 335

Gly Leu Pro Lys Ala Ile Arg Thr Ser Ile Ala His His Leu Phe Phe

340 345 350

Pro Ile Val Gln Asn Val Arg Leu Phe Gln Gly Val Ser Pro Asn Leu

355 360 365

Leu Phe Gln Leu Val Pro Glu Met Glu Ala Glu Tyr Phe Pro Pro Lys

370 375 380

Gln Asp Val Ile Leu Gln Asn Glu Ala Pro Thr Asp Leu Tyr Ile Ile

385 390 395 400

Val Ser Gly Ala Val Glu Leu Ile Ala Lys Ile Glu Gly Leu Glu Gln

405 410 415

Ile Ile Gly Lys Ala Val Ala Gly Glu Leu Phe Gly Glu Ile Gly Val

420 425 430

Leu Cys Gly Arg Pro Gln Pro Phe Ala Val Arg Thr Thr Glu Ile Ser

435 440 445

Gln Ile Leu Arg Leu Ser Arg Thr Ala Leu Met Asn Ile Leu Arg Ala

450 455 460

Asn Pro Glu Asp Glu Arg Val Val Met Asn Asn Leu Leu Leu Asn Leu

465 470 475 480

Gln Gly Phe Gly Gly Phe Gly Tyr Val Asp His Gln Thr Asn Gly Gly

485 490 495

Pro Glu Ile Lys Arg His His Asp Thr Ala Leu Thr Ser Ile Asp Ile

500 505 510

Asn Asn Leu Glu Ala Arg Val Lys Lys Gln Glu Gly Asp Asp Val Gln

515 520 525

Glu Ile Asn Lys Ser Met Asn Asp Val Ser Leu Asn Leu Glu Asn Lys

530 535 540

Lys Glu Leu Asn Glu Gln Lys Val Glu Leu Ile Gly Pro Asp Glu Lys

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Gly Thr Lys Ser Cys Gln Leu Lys Pro Glu Val Pro Cys Cys Ser Asn

565 570 575

Ser Cys Ser Thr Ile Ser Ser Gly Ser Lys Val Thr Lys Ser Thr Asn

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Phe Gly Lys Leu Ile Ile Leu Pro Asp Ser Leu Glu Glu Leu Phe Arg

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Ala Glu Asp Ala Glu Ile Asp Asp Ile Asp Val Ile Arg Asp Gly Asp

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His Leu Phe Phe Leu

660

<210> 3

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atgaatacag aaaccagtaa 20

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<213>artificial sequence (Artificial Sequence)

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ttaaaaggaa aaacaaatgg 20

Claims

1. tobacco KC1 gene, which is characterized in that it is the following protein (a) of coding or gene (b):

(a) protein that the amino acid sequence shown in SEQ ID NO:2 forms；

(b) sequence shown in SEQ ID NO:2 be substituted, lack or add one or several amino acid and with same function by (a) protein derived from.

2. gene according to claim 1, which is characterized in that its nucleotide sequence is as shown in SEQ ID NO:1.

3. containing the biomaterial of gene as claimed in claim 1 or 2, the biomaterial is expression cassette, expression vector, Ke Longzai Body or engineering bacteria.

4. biomaterial described in gene as claimed in claim 1 or 2 or claim 3 is promoting plant or microorganism potassium Ions Absorption With the application in transhipment.

5. application according to claim 4, which is characterized in that the plant includes tobacco, arabidopsis, the microorganism packet Include yeast.

6. application of the biomaterial described in gene as claimed in claim 1 or 2 or claim 3 in prepare transgenosis plant.

7. application of the biomaterial described in gene as claimed in claim 1 or 2 or claim 3 in plant breeding.

8. the use as claimed in claim 7, which is characterized in that the purpose of the breeding is to promote plant Potassium Absorption and turn Fortune.

9. a kind of method for promoting plant Potassium Absorption and transhipment, which is characterized in that the method are as follows:

1) making plant includes gene as claimed in claim 1 or 2；Alternatively,

2) plant is made to be overexpressed gene as claimed in claim 1 or 2；

Wherein, the plant includes tobacco, arabidopsis.

10. according to the method described in claim 9, it is characterized in that, the method includes transgenosis, hybridization, backcrossing, selfing or Vegetative propagation.