CN106929522B - Amino acid transport gene OsAAP1 promotes the application of paddy growth under low nitrogen - Google Patents
Amino acid transport gene OsAAP1 promotes the application of paddy growth under low nitrogen Download PDFInfo
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8201—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation
- C12N15/8202—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation by biological means, e.g. cell mediated or natural vector
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- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
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- C12Y306/00—Hydrolases acting on acid anhydrides (3.6)
- C12Y306/03—Hydrolases acting on acid anhydrides (3.6) acting on acid anhydrides; catalysing transmembrane movement of substances (3.6.3)
Abstract
The invention discloses amino acid transport genesOsAAP1The application for promoting paddy growth under low nitrogen, belongs to plant genetic engineering field.OsAAP1The amino acid and its cDNA sequence of DNA encoding the protein are respectively as shown in SEQ ID NO.1,2.The present invention passes through building riceOsAAP1The overexpression plant of gene, discovery improve under low nitrogenOsAAP1The expression of gene can be such that rice root length, radical, plant height, tiller number increases, biomass, output increased.Plant is interfered by building, discovery passes through reduction under low nitrogenOsAAP1Gene expression can be such that rice root length, radical, plant height, tiller number reduces.ThereforeOsAAP1Gene can be used for rice and promote paddy growth, Lai Tigao rice biological amount, yield under low nitrogen.OsAAP1Gene has important application in terms of the low fertilizer of rice tolerance or lean soil adaptability, in reducing farmland fertilizer usage amount and rice varieties improvement.
Description
Technical field
The invention belongs to plant genetic engineering fields, and in particular to amino acid transport geneOsAAP1Promote water under low nitrogen
The application of rice growth.
Background technique
Plant obtains nitrogen by absorbing ammonia, nitrate anion, amino acid, soluble peptide in soil etc.；The absorption of nitrogen and
Transhipment relies primarily on ammonium root transport protein (AMT), nitrate anion transport protein (NRT), amino acid transport proteins (AAT), peptide transport
The transport proteins such as albumen (PTR) complete (Williams L, Miller A. Transporters responsible for
the uptake and partitioning of nitrogenous solutes. Annu Rev Plant Biol and
Plant Mol Biol, 2001,52:659-688.).Ammonium passes through glutamine synthelase after absorbing by plant AMT again
(GS) and glutamate synthase (GOGAT) synthesizes glutamine and glutamic acid, the latter further form other amino acid (Sonoda
Y, Ikeda A, Saiki S, et al. Feedback regulation of the ammonium transporter
gene family AMT1 by glutamine in rice. Plant Cell Physiol, 2003, 44: 1396-
1402.).Plant can absorb ring by the NRT2 of high affine movement system (HATS) and the NRT1 of low affine movement system (LATS)
Nitrate in border forms ammonium by nitrate reductase (NR) and nitrite reductase (NiR) reduction, further forms amino acid
(Paungfoo-Lonhienne C, Lonhienne T G, Rentsch D, et al. Plants can use
protein as a nitrogen source without assistance from other organisms. PNAS,
2008,105:4524-4529.).
In higher plant, AAT is a kind of transmembrane protein, by amino acid from it is extracellular be transported to it is intracellular, while also in amino acid
Long distance transportation, cause of disease reaction and abiotic stress etc. play an important role (Tegeder M. Transporters
for amino acids in plant cells: some functions and many unknowns. Curr Opin
Plant Biol, 2012,15:1-7.).AAT gene is divided into two superfamilies: APC(amino acid, polyamines and choline turn
Fortune) superfamily and AAAP(amino acid/auxin permease) superfamily.APC superfamily is divided into three subfamilies: CATs(sun from
Sub- amino acid transporter) family, ACTs(amino acid/choline transport albumen) family and PHSs(polyamines, H+ cotransport egg
It is white) family.AAAP superfamily is divided into six subfamilies: AAPs(amino acid permease) family, LHTs(lysine and histidine turn
Transport albumen) family, ProTs(proline transport protein) family, GATs(gamma-amino acid butyric acid, GABA) family, AUXs(growth
Plain transport protein) family and ANTs(aromatic series and neutral amino acid transporter) family (Fischer WN, Andre B,
Rentsch D, et al. Amino acid transport in plants. Trends Plant Sci, 1998, 3:
188-195.).
85 AAT family members (Zhao H, Ma H, Yu L, et al. Genome- is found in rice genome altogether
wide survey and expression analysis of amino acid transporter gene family in
Rice (Oryza sativa L.) PLoS ONE, 2012,7:e49210.).Research is foundOsAAT5、OsAAT7、OsAAT24、OsAAT49WithOsAAT60T-DNA insertion mutation body yield of brown rice and plant dry weight decline, it was demonstrated that AAT
Nitrogen accumulation and Carbon and nitrogen allocation important role (Lu Y, Song Z, Lu K, et al. Molecular to rice
characterization, expression and functional analysis of the amino acid
transporter gene family (OsAATs) in rice. Acta physiol Plant, 2012, 34: 1943-
1962.).Research finds overexpressionOsAAP6It will increase shelf stability albumen and amino acid content in rice grain, so as to improve
Rice nutrition and flavor (Peng B, Kong H, Li Y, et al. OsAAP6 functions as an important
regulator of grain protein content and nutritional quality in rice. Nat
Commun, 2014,5:doi:10.1038.).Amino acid transporter to the Amino Acid Absorptions of the various plants such as rice,
Transhipment and storage play an important role.Report about rice AAT family member research is seldom, at present rice amino acid transport
FamilyOsAAP1The growth and development of gene pairs rice does not have any research at present.Present invention discover thatOsAAP1Gene is right under low nitrogen
The influence of rice biological amount has more important role, can be applied to the raising of plant nitrogen use efficiency, reduces fertilizer application and rice
Plant type improvement.
Summary of the invention
It is an object of the invention to solve problems of the prior art, amino acid transport gene is providedOsAAP1Low
Promote the application of paddy growth under nitrogen.
The purpose of the invention is achieved by the following technical solution:
The present invention is with the amino acid transport gene of riceOsAAP1For object, cloned from being spent in rice in 11OsAAP1's
CDNA sequence.Pass through buildingOsAAP1The overexpression vector of gene, the genetic transforming method mediated using Agrobacterium EHA105 will
Overexpression vector is imported in normal japonica rice variety and is spent in 11, is obtainedOsAAP1The overexpression plant of gene, in low nitrogen (nitrogen concentration
1mM or less) culture under, overexpress root long, radical, plant height, tiller number of plant etc. with control wild type in spend 11 compared to significantly
It improves.It constructs simultaneouslyOsAAP1The interference carrier of gene, by interference carrier import in spend in 11, obtainOsAAP1Gene is done
Plant is disturbed, under low nitrogen culture, root long, radical, plant height, tiller number etc. significantly reduce compared with spending 11 in.These result tables
It is bright, pass through raisingOsAAP1Gene expression can promote paddy growth, increase Biomass and yield；OsAAP1Gene is in rice
There is application value in terms of being resistant to low fertilizer or lean soil adaptability, it is in addition negative in reduction farmland fertilizer usage amount, mitigation environment
Also there is important application in influence；It can also be applied in rice varieties improvement by molecular breeding.
Based on present invention discover thatOsAAP1The function of gene can be used under the conditions of low nitrogen promoting paddy growth, improve
Rice biological amount, yield.It can specifically be realized by genetic engineering, i.e., overexpression improvesOsAAP1The expression of gene, makes rice root
Length, radical, plant height, tiller number etc. increase, and achieve the purpose that improve rice biological amount, yield.DescribedOsAAP1Gene coding
OsAAP1 albumen amino acid sequence as shown in SEQ ID NO.1；DescribedOsAAP1The cDNA sequence of gene is preferably such as SEQ
Shown in ID NO.2.
It is construed as, (i.e. not in the activated centre of albumen) under the premise of not influencing OsAAP1 protein active, ability
It is one or several that field technique personnel can carry out various substitutions, additions and/or deletions to amino acid sequence shown in SEQ ID NO.1
Amino acid obtains the amino acid sequence with same function.Therefore, OsAAP1 albumen further includes amino acid shown in SEQ ID NO.1
Sequence is substituted, replaces and/or increases that one or several amino acid obtain has same active protein.In addition, Ying Li
Solution, it is contemplated that the degeneracy of codon and the preferences of different plant species codon, those skilled in the art can according to need
The codon expressed using suitable particular species.
Advantages of the present invention and effect:
(1) present invention clonesOsAAP1Gene improve expression after can make under low nitrogen rice root long, radical, plant height,
Tiller number increases, explanationOsAAP1Gene pairs improves rice biological amount, yield has more apparent effect, therefore, pass through genetic engineering
Technology improvesOsAAP1The expression of gene can be improved phytomass, yield.It is not only does this facilitate and is used by reducing nitrogenous fertilizer
High-yield rice is cultivated, the breed improvement of plant can also be carried out by molecular breeding.
(2)OsAAP1The successful clone of gene further demonstrates weight of the amino acid transport family in nitrogen absorption process
It acts on, there is important meaning to the biological function for illustrating amino acid transport family, in addition to further appreciating that plant nitrogen generation
Thank to approach, improving nitrogen absorption efficiency has great impetus.
(3) although being cloned into some nitrogen nutrition approach at present influences the gene of plant growth, to plant growth
The molecular mechanism of development is still unclear.And what the present invention clonedOsAAP1Gene can be improved the biomass of rice, plant to determining
The key factor of object volume increase has great impetus.
Detailed description of the invention
Fig. 1 be spend 11 in the lower control of low nitrogen (0.5mM ammonium nitrate) culture,OsAAP1Gene overexpress 3 strains of plant andOsAAP1The whole strain phenotypic map of gene interference 3 strains of plant.
Fig. 2 is flower 11(WT in control),OsAAP1Gene overexpress 3 strains (OE1-3) of plant andOsAAP1Gene is dry
Disturb the statistics histogram of 3 strain (Ri1-3) root longs under Different nitrogen levels condition of culture of plant, data using SPSS software into
Row variable analysis (ANOVA) uses Duncan ' s to carry out significance difference analysis in 0.05 level, same concentration culture
At this concentration compared with the control, the table with significant difference is * to rice index.Low nitrogen concentration is ammonium nitrate 0.5mM, middle nitrogen
Concentration is ammonium nitrate 2mM, and high nitrogen concentration is ammonium nitrate 8mM.
Fig. 3 is flower 11(WT in control),OsAAP1Gene overexpress 3 strains (OE1-3) of plant andOsAAP1Gene is dry
Disturb the statistics histogram of 3 strain (Ri1-3) radicals under Different nitrogen levels condition of culture of plant, data using SPSS software into
Row variable analysis (ANOVA) uses Duncan ' s to carry out significance difference analysis in 0.05 level, same concentration culture
At this concentration compared with the control, the table with significant difference is * to rice index.Low nitrogen concentration is ammonium nitrate 0.5mM, middle nitrogen
Concentration is ammonium nitrate 2mM, and high nitrogen concentration is ammonium nitrate 8mM.
Fig. 4 is flower 11(WT in control),OsAAP1Gene overexpress 3 strains (OE1-3) of plant andOsAAP1Gene is dry
Disturb the statistics histogram of 3 strain (Ri1-3) plant heights under Different nitrogen levels condition of culture of plant, data using SPSS software into
Row variable analysis (ANOVA) uses Duncan ' s to carry out significance difference analysis in 0.05 level, same concentration culture
At this concentration compared with the control, the table with significant difference is * to rice index.Low nitrogen concentration is ammonium nitrate 0.5mM, middle nitrogen
Concentration is ammonium nitrate 2mM, and high nitrogen concentration is ammonium nitrate 8mM.
Specific embodiment
Below with reference to embodiment, the present invention will be further described in detail, and embodiments of the present invention are not limited thereto.
Unless otherwise specified, the conventional means that technological means used in following embodiments is well known to those skilled in the art；Used
Experimental method is conventional method, and can according to described recombinant technique (referring to molecular cloning, laboratory manual, second edition,
CSH Press, Cold SpringHarbor, New York) it completes；Material, reagent used etc., are commercially available.
Embodiment 1OsAAP1The building of gene overexpression plant
It extracts in rice and spends 11 RNA, and its reverse transcription is utilized into primer pair at cDNA:
F1:5'-AGATCTATGGGGATGGAGAGGCCGCAAGAG-3'(BglII),
R1:5'-CTTAAGTCATGAGGAGACGCTGAATGG-3'(AflII)；
Pass through PCR amplificationOsAAP1After the cDNA of gene, pass throughBglII andAflPCAMBIA-1301 is connected into after II digestion
Carrier (pCAMBIA-1301 carrier is purchased from Cambia company), is constructedOsAAP1The overexpression vector of geneOsAAP1-
p1301.The genetic transforming method mediated using Agrobacterium EHA105, overexpression vector is imported in normal rice varieties and spends 11
In.
It by the transplanting of obtained all transgenic plants in the basket with soil, periodically waters, fertilising is grown tall about to seedling
When 10cm, the hygromycin solution 48 for being 50mg/L by the concentration that 50 plants of Transgenic Rice seedlings are soaked in the preparation of 500mL distilled water
Hour, leaf is green later and diastole, growth conditions good stand are positive transgenic plant；And leaf is withered and yellow and crimps
Plant be negative plant, it is dead immediately.Positive plant single-strain planting and sowing, until T2 is molten in above-mentioned hygromycin for identifying
It is withered and yellow without any leaf and to crimp be homozygous transgenic plant to get arriving in liquidOsAAP1Gene overexpresses plant.It will surpass
Expression plant, the middle seed for spending 11 are soaked seed 3 days with distilled water on culture dish and after culture 7 days, are transferred to rice nutrition liquid culture,
Nutrient solution prescription is formulated with reference to International Rice, but ammonium nitrate is tuned into the low nitrogen of 0.5mM(), nitrogen in 2mM() and 8mM(high nitrogen),
It cultivates 40 days respectively, observes phenotype, statistics root long, radical and plant height.Plant phenotype, root long, the statistical result of radical and plant height
See Fig. 1-4, it is seen that under low nitrogen culture,OsAAP1Gene overexpress plant compared with spending 11 plant in control root long, radical and
Plant height increases, and reaches significant difference.And in middle nitrogen, there is no more significant than compareing for overexpression plant root long, radical and plant height
Increase.In high nitrogen, overexpression plant only has radical to increase up to significant difference than control, and root long and plant height be not than control
It dramatically increases.Later period,OsAAP1Tiller number, amino acid content and the last yield of gene overexpression plant are also all remarkably higher than
11 are spent in control.
Embodiment 2OsAAP1The building of gene interference plant
It extracts in rice and spends 11 RNA, and its reverse transcription is utilized into primer pair at cDNA:
F2:5'-GGTACCTGGGGATGGAGAGGCCGCAA-3'(KpnI),
R2:5'-GGATCCTTGCGCTTGCCATGGACGGGGT-3'(BamH I)；
F3:5'-ACTAGTTGGGGATGGAGAGGCCGCAA-3'(SpeI),
R3:5'-GAGCTCTTGCGCTTGCCATGGACGGGGT-3'(Sac I)；
Respective PCR amplification goes outOsAAP1The cDNA segment of gene, by connecting after above-mentioned corresponding digestion with restriction enzyme
Enter pTCK303 carrier, constructsOsAAP1The interference expression vector of geneOsAAP1-pTCK303.It is situated between using Agrobacterium EHA105
Interference expression vector is imported in normal japonica rice variety and is spent in 11 by the genetic transforming method led.
It by the transplanting of obtained all transgenic plants in the basket with soil, periodically waters, fertilising is grown tall about to seedling
When 10cm, the hygromycin solution 48 for being 50mg/L by the concentration that 50 plants of Transgenic Rice seedlings are soaked in the preparation of 500mL distilled water
Hour, leaf is green later and diastole, growth conditions good stand are positive transgenic plant；And leaf is withered and yellow and crimps
Plant be negative plant, it is dead immediately.Positive plant single-strain planting and sowing, until T2 is molten in above-mentioned hygromycin for identifying
It is withered and yellow without any leaf and to crimp be homozygous transgenic plant to get arriving in liquidOsAAP1Gene interferes plant.It will interference
Plant, the middle seed for spending 11 are soaked seed 3 days with distilled water on culture dish and after culture 7 days, are transferred to rice nutrition liquid culture, nutrition
Formula of liquid is formulated with reference to International Rice, but ammonium nitrate is tuned into the low nitrogen of 0.5mM(), nitrogen in 2mM() and 8mM(high nitrogen), distinguish
Phenotype, statistics root long, radical and plant height are observed in culture 40 days.Plant phenotype, root long, the statistical result of radical and plant height are shown in figure
1-4, it is seen that under low nitrogen, middle nitrogen and high nitrogen culture,OsAAP1Gene interferes plant root long, root compared with spending 11 plant in control
Several and plant height is reduced, and reaches significant difference.Later period,OsAAP1Gene interferes the tiller number of plant, amino acid content and most
Yield afterwards is also substantially less than in control and spends 11.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present invention,
It should be equivalent substitute mode, be included within the scope of the present invention.
SEQUENCE LISTING
<110>Wuhan Bioengineering Institute
<120>amino acid transport gene OsAAP1 promotes the application of paddy growth under low nitrogen
<130> 1
<160> 8
<170> PatentIn version 3.3
<210> 1
<211> 487
<212> PRT
<213> Oryza sativa
<400> 1
Met Gly Met Glu Arg Pro Gln Glu Lys Val Ala Thr Thr Thr Thr Ala
1 5 10 15
Ala Phe Asn Leu Ala Glu Ser Gly Tyr Ala Asp Arg Pro Asp Leu Asp
20 25 30
Asp Asp Gly Arg Glu Lys Arg Thr Gly Thr Leu Val Thr Ala Ser Ala
35 40 45
His Ile Ile Thr Ala Val Ile Gly Ser Gly Val Leu Ser Leu Ala Trp
50 55 60
Ala Ile Ala Gln Leu Gly Trp Val Ile Gly Pro Ala Val Leu Val Ala
65 70 75 80
Phe Ser Val Ile Thr Trp Phe Cys Ser Ser Leu Leu Ala Asp Cys Tyr
85 90 95
Arg Ser Pro Asp Pro Val His Gly Lys Arg Asn Tyr Thr Tyr Gly Gln
100 105 110
Ala Val Arg Ala Asn Leu Gly Val Ala Lys Tyr Arg Leu Cys Ser Val
115 120 125
Ala Gln Tyr Val Asn Leu Val Gly Val Thr Ile Gly Tyr Thr Ile Thr
130 135 140
Thr Ala Ile Ser Met Gly Ala Ile Lys Arg Ser Asn Trp Phe His Arg
145 150 155 160
Asn Gly His Asp Ala Ala Cys Leu Ala Ser Asp Thr Thr Asn Met Ile
165 170 175
Ile Phe Ala Gly Ile Gln Ile Leu Leu Ser Gln Leu Pro Asn Phe His
180 185 190
Lys Ile Trp Trp Leu Ser Ile Val Ala Ala Val Met Ser Leu Ala Tyr
195 200 205
Ser Thr Ile Gly Leu Gly Leu Ser Ile Ala Lys Ile Ala Gly Gly Ala
210 215 220
His Pro Glu Ala Thr Leu Thr Gly Val Thr Val Gly Val Asp Val Ser
225 230 235 240
Ala Ser Glu Lys Ile Trp Arg Thr Phe Gln Ser Leu Gly Asp Ile Ala
245 250 255
Phe Ala Tyr Ser Tyr Ser Asn Val Leu Ile Glu Ile Gln Asp Thr Leu
260 265 270
Arg Ser Ser Pro Ala Glu Asn Glu Val Met Lys Lys Ala Ser Phe Ile
275 280 285
Gly Val Ser Thr Thr Thr Thr Phe Tyr Met Leu Cys Gly Val Leu Gly
290 295 300
Tyr Ala Ala Phe Gly Asn Arg Ala Pro Gly Asn Phe Leu Thr Gly Phe
305 310 315 320
Gly Phe Tyr Glu Pro Phe Trp Leu Val Asp Val Gly Asn Val Cys Ile
325 330 335
Val Val His Leu Val Gly Ala Tyr Gln Val Phe Cys Gln Pro Ile Tyr
340 345 350
Gln Phe Ala Glu Ala Trp Ala Arg Ser Arg Trp Pro Asp Ser Ala Phe
355 360 365
Val Asn Gly Glu Arg Val Leu Arg Leu Pro Leu Gly Ala Gly Asp Phe
370 375 380
Pro Val Ser Ala Leu Arg Leu Val Trp Arg Thr Ala Tyr Val Val Leu
385 390 395 400
Thr Ala Val Ala Ala Met Ala Phe Pro Phe Phe Asn Asp Phe Leu Gly
405 410 415
Leu Ile Gly Ala Val Ser Phe Trp Pro Leu Thr Val Tyr Phe Pro Val
420 425 430
Gln Met Tyr Met Ser Gln Ala Lys Val Arg Arg Phe Ser Pro Thr Trp
435 440 445
Thr Trp Met Asn Val Leu Ser Leu Ala Cys Leu Val Val Ser Leu Leu
450 455 460
Ala Ala Ala Gly Ser Ile Gln Gly Leu Ile Lys Ser Val Ala His Tyr
465 470 475 480
Lys Pro Phe Ser Val Ser Ser
485
<210> 2
<211> 1464
<212> DNA
<213> Oryza sativa
<400> 2
atggggatgg agaggccgca agagaaggtg gccaccacca ccaccgccgc cttcaacctc 60
gccgagtccg gctacgccga ccgccccgac ctcgacgacg acggccgcga gaagcgcaca 120
gggacgctgg tgacggcgag cgcgcacata ataacggcgg tgatcggctc cggcgtgctg 180
tcgctggcgt gggcgatagc gcagctgggg tgggtgatcg ggccggccgt gctggtggcg 240
ttctcggtca taacctggtt ctgctccagc ctcctcgccg actgctaccg atctcccgac 300
cccgtccatg gcaagcgcaa ctacacctac ggccaagccg tcagggccaa cctaggtgtg 360
gccaagtaca ggctctgctc ggtggcacag tacgtcaatc tcgtcggcgt caccattggc 420
tacaccatca ctacggccat cagcatgggt gcgatcaaac ggtccaactg gttccatcgc 480
aacggccacg acgcagcctg cttggcatct gacacgacca acatgatcat atttgctggc 540
atccaaatcc tcctctcgca gctgccgaat tttcacaaaa tttggtggct ctccattgtc 600
gctgctgtca tgtcactggc ctactcaacc attggccttg gcctctccat tgcaaaaatt 660
gcaggtgggg cccaccccga ggcaaccctc acaggggtga ctgttggagt ggatgtgtct 720
gcaagtgaga aaatctggag aacttttcag tcacttggtg acattgcctt tgcatactcc 780
tactccaatg tcctcataga aattcaggac acgctgcggt cgagcccggc ggagaacgag 840
gtgatgaaga aggcgtcgtt catcggagtc tcgacgacga cgacgttcta catgctgtgc 900
ggcgtgctcg gctacgcggc gttcggcaac cgcgcgccgg ggaacttcct caccggcttc 960
ggcttctacg agcccttctg gctcgtcgac gtcggcaacg tctgcatcgt cgtccacctc 1020
gtcggcgcct accaggtctt ctgccagccc atctaccagt tcgccgaggc ctgggcgcgc 1080
tcgcggtggc cggacagcgc cttcgtcaac ggcgagcgcg tgctccggct gccgctcggc 1140
gccggcgact tccccgtcag cgcgctccgc ctcgtctggc gcacggccta cgtcgtgctc 1200
accgccgtcg ccgccatggc gttccccttc ttcaacgact tcctcggcct catcggcgcc 1260
gtctccttct ggccgctcac cgtctacttc cccgtccaga tgtacatgtc tcaggccaag 1320
gtccggcgat tctcgccgac gtggacgtgg atgaacgtgc tcagcctcgc ctgcctcgtc 1380
gtctccctcc tcgccgccgc cggctccatc cagggcctca tcaaatccgt cgcacattac 1440
aagccattca gcgtctcctc atga 1464
<210> 3
<211> 30
<212> DNA
<213> Artificial
<220>
<223>primers F 1
<400> 3
agatctatgg ggatggagag gccgcaagag 30
<210> 4
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<212> DNA
<213> Artificial
<220>
<223>primer R1
<400> 4
cttaagtcat gaggagacgc tgaatgg 27
<210> 5
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<212> DNA
<213> Artificial
<220>
<223>primers F 2
<400> 5
ggtacctggg gatggagagg ccgcaa 26
<210> 6
<211> 28
<212> DNA
<213> Artificial
<220>
<223>primer R2
<400> 6
ggatccttgc gcttgccatg gacggggt 28
<210> 7
<211> 26
<212> DNA
<213> Artificial
<220>
<223>primers F 3
<400> 7
actagttggg gatggagagg ccgcaa 26
<210> 8
<211> 28
<212> DNA
<213> Artificial
<220>
<223>primer R3
<400> 8
gagctcttgc gcttgccatg gacggggt 28
Claims (4)
1.OsAAP1Gene promotes the application in paddy growth under low nitrogen；It is characterized by: describedOsAAP1Gene coding
OsAAP1 albumen amino acid sequence as shown in SEQ ID NO.1.
2.OsAAP1Gene improves the application in rice biological amount under low nitrogen, it is characterised in that: the rice biological amount packet
Include rice root long, radical, plant height；DescribedOsAAP1The amino acid sequence such as SEQ ID of the OsAAP1 albumen of gene coding
Shown in NO.1.
3.OsAAP1Gene improves the application in rice yield under low nitrogen, it is characterised in that: the raising rice yield packet
It includes and achievees the purpose that improve rice yield by improving rice tillering number；DescribedOsAAP1The OsAAP1 albumen of gene coding
Amino acid sequence is as shown in SEQ ID NO.1.
4. application according to claim 1-3, it is characterised in that: describedOsAAP1The cDNA sequence of gene is such as
Shown in SEQ ID NO.2.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN109280668B (en) * | 2018-10-15 | 2021-11-23 | 贵州省烟草科学研究院 | Tobacco amino acid transporter gene NtTAT and application thereof |
| CN109486823B (en) * | 2018-12-28 | 2021-10-08 | 武汉生物工程学院 | Application of high-expression indica rice type promoter in japonica rice |
| CN110819640B (en) * | 2019-12-19 | 2022-06-21 | 中国烟草总公司郑州烟草研究院 | Tobacco NtNRP1 gene and application thereof |
| CN112029796A (en) * | 2020-09-15 | 2020-12-04 | 贵州大学 | Application of amino acid transport gene |
| CN112410309B (en) * | 2020-11-27 | 2022-02-08 | 河南农业大学 | Application of GmAAP protein and GmAAP gene in soybean breeding |
| CN114532338B (en) * | 2022-02-16 | 2023-08-22 | 贵州大学 | Rice culture solution containing gamma-aminobutyric acid, application of rice culture solution in nitrogen stress resistance of rice and rice culture method |
| CN114990151B (en) * | 2022-04-18 | 2023-04-14 | 河北省农林科学院粮油作物研究所 | Crop nitrogen utilization efficiency and grain yield collaborative improvement method based on gene editing technology |
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