CN102559673A - Promoter specifically induced and expressed under condition of lack of phosphorus in cabbage type rape - Google Patents
Promoter specifically induced and expressed under condition of lack of phosphorus in cabbage type rape Download PDFInfo
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Abstract
The invention belongs to the technical field of plant gene engineering and is characterized in that: a promoter pBnPR1 which is not induced under the condition of lack of elements such as nitrogen, potassium, sulfur, iron and the like and is only induced and expressed under the condition of lack of phosphorus is cloned from the cabbage type rape, and the nucleotide sequence of the promoter is shown in 1-1874 basic groups in a sequence table SEQ ID NO.1. The promoter and a marker gene GUS are fused to convert arabidopsis, and the results show that the GUS gene is only expressed under the condition of lack of phosphorus and is not expressed under normal conditions and the condition of lack of nitrogen, potassium, sulfur and iron.
Description
Technical field
The invention belongs to the plant gene engineering technology field, be specifically related to the clone and the application of a phosphorus starvation induced specific expression promoter of swede type rape.
Background technology
Phosphorus is the essential nutritive element of growth and development of plants.Plant is mainly through the form Absorption of Phosphorus of root system with phosphate radical.Because phosphate radical is easy to fixed and the conversion of soil microorganisms by soil mineral, adsorption of metal ions, so the biological effectiveness of phosphorus is very low in the soil.Scarce phosphorus becomes the principal element of restriction crop yield and quality raising.People often keep crop yield through a large amount of application of P fertilizer, and this has not only increased agriculture production cost, have also aggravated the consumption of this Nonrenewable resources of phosphorus ore, and have caused environmental problems such as body eutrophication.This has just promoted world wide implants phosphorus nutrition and has studied efficiently.Scientist's expectation is cultivated the efficient or anti-low-phosphorous crop of phosphorus nutrition through molecular biology method by genetically engineered.Experiment shows that this is an economical and effective and eco-friendly measure.L ó pez-Bucio etc. are through the transgene tobacco of the excessive generation Hydrocerol A of genetically engineered cultivation, and under low-phosphorous condition, this more blade of can growing that grows tobacco accumulates more biological yield (L ó pez-Bucio etc., 2000).Wang etc. will change in the soybean from the purple acid phosphatase gene of Arabidopis thaliana, on acid soil the every strain of transfer-gen plant bear pods the number and the seed number average significantly improve (Wang etc., 2009).Nilsson etc. are with myb transcription factor PHR1 overexpression in Arabidopis thaliana, and the result shows that transfer-gen plant has improved the absorption of phosphorus (Nilsson etc., 2007).
In the plant phosphorus nutrition genes involved of having reported transforms, generally be the 35S promoter of set of applications moulding, strongly expressed.Because constitutive promoter can both drive the expression of downstream gene under different tissues and varying environment condition, the result causes under the normal growth condition, and degeneration-resistant albumen also produces in a large number.This not only wastes energy, and influences the normal physiological metabolism of plant, makes plant-growth unusual.Therefore find the promotor of a specifically expressing under scarce phosphorus condition to have vital role to plant phosphorus nutrition genetically engineered.The research that is used to come from swede type rape and is cultivated phosphorus efficiency or anti-low-phosphorous rape by phosphor deficiency speciality induction expression promoter construction of expression vector does not also appear in the newspapers.The promotor of inducible expression is only just expressed when plant is coerced by phosphorus, can not cause degeneration-resistant proteic excess accumulation.Take all factors into consideration these factors, the present invention attempts to find one to be coerced the promotor of special abduction delivering by phosphorus, for the phosphorus nutrition genetically engineered of swede type rape and other crop provides new instrument.
Live in for a long time and lack in the phosphorus environment, plant evolution goes out a series of adaptation mechanisms to be dealt with and lacks phosphorus and coerce, and comprises changing the root configuration, increase the root gross density, inducing high affine phosphorus transporter, secretion organic acid and Phosphoric acid esterase etc.The inevitable differential expression by number of genes of the variation of physiology and phenotype mediates.Misson etc. are by biochip technology research Arabidopis thaliana short-term, mid-term and the long-term phosphorus side of body _ changes in gene expression situation under compeling, and the result shows has 612 genes to be induced respectively, and 254 genes are suppressed (Misson etc., 2005).This just provides possibility for we clone phosphorus stress-inducing expression promotor.At present, about the report of the clone of phosphorus starvation induced specific expression promoter and application seldom.Therefore, will not only help to study phosphorus stress response expression of gene regulatory mechanism to the research and the technological invention of phosphorus starvation induced expression promotor, and in the cultivation of phosphorus efficiency or anti-low-phosphorous crop varieties, have using value.The present invention utilizes biotechnology exactly, from the swede type rape genome, is cloned into the promotor that a phosphorus is coerced special abduction delivering, and this promotor and marker gene GUS are merged, and changes in the Arabidopis thaliana, has verified the specificity of this promotor phosphorus abduction delivering.
Summary of the invention
The objective of the invention is to overcome the defective of prior art, provide a kind of and express promotor from the swede type rape phosphor deficiency speciality induction.Another object of the present invention provides the application method of above-mentioned promotor.
The present invention realizes through following technical scheme:
The applicant clones not lacked by nitrogen, potassium, sulphur and ferro element of obtaining and induces from the swede type rape genome, only by the pBnPR1 promoter sequence of phosphorus starvation induced expression, its nucleotide sequence is shown in sequence table SEQ ID NO.1.Described pBnPR1 promoter sequence, it be among the sequence table SEQ ID NO.1 sequence shown in the 1-1874 bit base or with sequence homology shown in the sequence table SEQ IDNO.1 at 90% above nucleotide sequence, and can be by the homologous sequence of phosphorus starvation induced expression.This promotor under normal and nitrogen stress, potassium deficiency, a lack of sulfur, iron deficiency condition, do not express or expression amount extremely a little less than.After coerced by scarce phosphorus, this promotor can be by remarkable abduction delivering.
Being named as the BnPR1 gene is that the applicant clones a gene that expressed by phosphor deficiency speciality induction that obtains from swede type rape; Its partial nucleotide sequence is shown in 1875-2014 base among the sequence table SEQ ID NO:1; In ncbi database, retrieve sequence through bioinformatic analysis with this gene height homologous BAC, according to this BAC sequences Design primer: PG2F (5 '
AAGCTTAA GAAGTCCGTGAAGGGGTC 3 ') and P56oR (5 '
GGATCCCCACTCCGGCAA CGACTC 3 '); Simultaneously two ends are added restriction enzyme site at this primer; The promoter sequence of amplification gene BnPR1 in the swede type rape genome with the above-mentioned single band order-checking that amplifies, obtains the nucleotide sequence shown in sequence table SEQ ID NO:1.The result show this sequence can with gene BnPR1 splicing, and be positioned at the upper reaches of gene BnPR1, confirm that this gene BnPR1 is the candidate segment of promotor, the applicant is with its called after pBnPR1.The pBnPR1 fragment is replaced the 35S promoter among the plant expression vector pBI121, construct new plant expression vector with reporter gene GUS.The colored method (floral dip) of dipping in through agriculture bacillus mediated changes pBnPR1 fragment and reporter gene GUS in the Arabidopis thaliana over to.Through antibiotic-screening, PCR evaluation, many, finally obtain to isozygoty transformed plant for isozygotying.To contain the pBnPR1 fragment and reporter gene GUS transformed plant is cultivated under different condition, identify the expression characterization of this promotor candidate segment through the GUS histochemical stain.Coloration result shows: normally, under nitrogen stress, potassium deficiency, a lack of sulfur and the iron deficiency condition, this transformed plant is not colored, and explains that promotor pBnPR1 is not induced by these conditions.Under scarce phosphorus condition, the root of transformed plant and part vein are mazarine, explain that promotor pBnPR1 is efficiently expressed by phosphorus starvation induced, and mainly in root system, express (as shown in Figure 3).
More detailed technical scheme sees that " embodiment " is said.
Description of drawings
Sequence table SEQ ID NO:1 is the nucleotide sequence that phosphor deficiency speciality induction is expressed promotor in the present invention's swede type rape of cloning, and wherein 1-1874bp is a promoter sequence of the present invention, and 1875-2010 is the partial nucleotide sequence of gene BnPR1.
Fig. 1: demonstration be the electrophoretogram of amplification BnPR1 promotor from the swede type rape genomic dna, the figure right side is DL2000marker.
Fig. 2: demonstration be the pBnPR1-pBI expression vector that makes up.
Fig. 3: demonstration be the GUS histochemical stain of pBnPR1-pBI arabidopsis thaliana transformation under the different treatment condition.
Embodiment
The clone of the phosphorus starvation induced expression promotor of embodiment 1 rape pBnPR1
(1) gene BnPR1 is that the applicant clones a gene that expressed by phosphor deficiency speciality induction that obtains from swede type rape; Its partial nucleotide sequence detects less than this expression of gene in the swede type rape of cultivating under normal operation shown in 1875-2014 base among the sequence table SEQ ID NO:1.In order to identify the expression characterization of this gene promoter,, in ncbi database, retrieve AC189399 with gene BnPR1 sequence height homologous BAC clone at first through bioinformatic analysis.Sequence with AC189399 is reference, design primer PG2F (5 '
AAGCTTAA GAAGTCCGTGAAGGGGTC3 ') and P56oR (5 '
GGATCCCCACTCCGGCAA CGACTC 3 '), wherein with the restriction endonuclease sites of base for adding of underscore, primer PG2F has added a Hind III enzyme and has cut other site, and primer P56oR has added a BamH I restriction enzyme site.The genomic dna magnificent two No. 4 with swede type rape is template, utilizes Japan to spin the promotor candidate segment of the high-fidelity DNA polymerase KOD plus amplification gene BnPR1 of (Shanghai) bio tech ltd.The PCR reaction system is: DNA (50ng/ μ l) 2.0 μ l, ddH
2O 11.0 μ l, 10X PCR damping fluid 2.0 μ l, MgSO
4(25mM) 1.0 μ l, dNTP (2.0mM) 2.0 μ l, primer PG2F (10 μ M) 0.8 μ l, primer P56oR (10 μ M) 0.8 μ l, KOD-plus (1U/ μ l) 0.4 μ l.Wherein, PCR damping fluid, MgSO
4, dNTP, KOD-plus archaeal dna polymerase all spin (Shanghai) bio tech ltd from Japan, and primer is synthetic by Beijing AudioCodes biotechnology Ltd.The condition of pcr amplification is: 94 ℃ of preparatory sex change 3 minutes, and 94 ℃ of sex change 30 seconds, 57 ℃ of annealing 30 seconds, 68 ℃ were extended 2 minutes 30 seconds, 35 circulations, last 68 ℃ of insulations 5 minutes.The PCR product is electrophoresis in 0.8% sepharose, and the result is single band (see figure 1).
(2) order-checking and sequential analysis PCR product are through purifying; End adds " dA ", is connected with
-T Easy carrier (containing bio tech ltd available from clean ocean, Wuhan).Transformed into escherichia coli DH5 α then; The warp evaluation contains promotor pBnPR1 fragment and the correct positive colony of direction of insertion send the order-checking of Wuhan order-checking portion of the big Gene science limited-liability company of China; Its sequence is shown in sequence table SEQ ID NO:1, and sequence length is 2014bp.The sequence of this sequence and gene BnPR1 has 140bp overlapping, and is positioned at the upper reaches of this BnPR1 gene, thinks candidate's promotor of gene BnPR1, and the applicant is with its called after pBnPR1.The sequence of this promotor is shown in 1-1874bp among the SEQ ID NO:1; Sequence length is 1874 bases; In the downstream of this promotor be gene BnPR1 partial nucleotide sequence (promptly; 140 bases of 5 ' end), be transcription initiation site at the 1875th bit base " A " of SEQ ID NO:1, the 1843rd bit base is the TATA frame.
The structure of embodiment 2 plant expression vectors
For confirming the expression characterization of promotor pBnPR1, make up the plant expression vector that contains promotor pBnPR1 and marker gene GUS.
(1) selects the right-on clone of promoter sequence among the embodiment 1, in the LB substratum that contains 50 μ g/ml ammonia benzyls, shake bacterium in a large number, utilize a small amount of plasmid extraction kit of Dao Pu company to extract plasmid (method is referring to Dao Pu company specification sheets).Plasmid and the pB1121 carrier (crop genetic improvement National Key Laboratory of Hua Zhong Agriculture University write legal document for others peach be so kind as to give) that contain promotor pBnPR1 with restriction enzyme Hind III and BamH I double digestion.It is following that enzyme is cut system: plasmid or pBI121 carrier 33.0 μ l, 10X BamH I damping fluid 4.0 μ l, restriction endonuclease BamH I 1.0 μ l, restriction endonuclease Hind III2.0 μ l (restriction enzyme and damping fluid are from MBI company).37 ℃ of enzymes were cut 10 hours.Enzyme is cut product electrophoresis in 0.8% sepharose, downcuts target stripe with knife blade, with the gel recovery test kit recovery (working method is referring to the test kit specification sheets of Dao Pu company) of Dao Pu company.
(2) connect the recovery product, linked system is following: pBnPR1 promoter fragment 8.0 μ l, pBI121 fragment 8.0 μ l, 10X T4 damping fluid 2.0 μ l, T4 ligase enzyme 2.0 μ l (ligase enzyme and damping fluid are from MBI company).The mixing linked system, 4 ℃ connect 16 hours.
(3) will connect product is transformed in the bacillus coli DH 5 alpha.Converted product is coated on the LB solid medium that contains 50 μ g/ml kantlex, cultivated 16 hours for 37 ℃.Choose mono-clonal in the LB liquid nutrient medium that contains 50 μ g/ml kantlex 37 ℃ cultivated 10 hours.With primer PG2F and P56oR are detected bacterium liquid.The PCR reaction system is: bacterium liquid 2.0 μ l, ddH
2O13.9 μ l; 10X PCR damping fluid 2.0 μ l, dNTP (10mM) 0.4 μ l, primer PG2F (10 μ M) 0.8 μ l; Primer P56oR (10 μ M) 0.8 μ l; RTaq (5U/ μ l) 0.1 μ l (rTaq polysaccharase wherein, the PCR damping fluid is available from precious biotechnology Dalian ltd, dNTP is from Roche company).The condition of pcr amplification is: 94 ℃ of preparatory sex change 3 minutes, and 94 ℃ of sex change 30 seconds, 60 ℃ of annealing 30 seconds, 72 ℃ were extended 2 minutes 30 seconds, 35 circulations, last 72 ℃ of insulations 5 minutes.The PCR product is electrophoresis in 0.8% sepharose, and clip size is identical with the pBnPR1 promotor, and the clone who is single band thinks positive colony.Select this positive colony, shake bacterium, the upgrading grain further identifies with HindIII and BamH I double digestion whether insert clip size meets.Enzyme is cut system and method with (1) step among the embodiment 2.With the plant expression vector called after pBnPR1-pBI that builds.
Embodiment 3 usefulness expression vector pBnPR1-pBI arabidopsis thaliana transformations
The double base plant expression vector pBnPR1-pBI that embodiment 2 is made up transforms Agrobacterium LBA4404 through freeze-thaw method.
(1) the single bacterium colony of the competent preparation of Agrobacterium inoculation Agrobacterium (yeast extract 1.0g/L, sucrose 5.0g/L, sal epsom 0.5g/L transfers pH to 7.0 with NaOH for beef extract 5.0g/L, Tryptones 5.0g/L) in 50ml YEB nutrient solution.28 ℃, 200 rev/mins of shaking culture are to OD
260Be about 0.5, ice bath 30 minutes.5000 rev/mins of centrifugal 5 minutes collection thalline are outwelled supernatant, use 10ml 0.5mol/l NaCl suspension thalline then.Recentrifuge, thalline are suspended in 1ml 20mmol/l CaCl
2In the solution.Get 100 μ l bacterium liquid branches and install in the freezing 1.5ml centrifuge tube, liquid nitrogen flash freezer, and be stored in-80 ℃ of refrigerators.
(2) freeze-thaw method transforms Agrobacterium LBA4404 and gets 1.0 μ g plant expression vector pBnPR1-pBI, adds in the competence Agrobacterium that 100 μ l thaw on ice, mixes ice bath 30 minutes gently.In liquid nitrogen, froze rapidly 1 minute then, 37 ℃ of water-baths were melted 5 minutes, and ice bath 2 minutes adds the 1mlYEB nutrient solution.28 ℃, 100 rev/mins of wave and culture 3 hours.5000 rev/mins centrifugal 3 minutes, remove most of supernatant, the resuspended thalline in the remaining 100 μ l left and right sides.Be coated on the YEB solid medium that contains 50 μ g/ml Rifampins and 50 μ g/ml kantlex, cultivated 2 days for 28 ℃.Picking list bacterium colony is in the YEB nutrient solution that contains 50 μ g/ml Rifampins and 50 μ g/ml kantlex, and 28 ℃, 200 rev/mins are shaken bacterium.PG2F and P56oR are confirmed the existence of promotor pBnPR1 with the special primer among the embodiment 2.The PCR reaction conditions is referring to embodiment 2.
(3) arabidopsis thaliana transformation.Inflorescence soaking method (floral dip) is adopted in the conversion of Arabidopis thaliana, carries out with reference to (1998) reported method such as clough and somewhat modified.The single bacterium colony of Agrobacterium that picking contains the pBnPR1-pBI plasmid inserts in the 3ml YEB liquid nutrient medium (50 μ g/ml Rifampins and 50 μ g/ml kantlex).28 ℃, 200 rpms were shaken bacterium 1 day, the bacterium liquid that obtains were inserted in the 150ml YEB liquid nutrient medium (50 μ g/ml Rifampins and 50 μ g/ml kantlex) by 2% (volume ratio) again, and 28 ℃, 200 rpms of joltings make the concentration of Agrobacterium reach OD
600Be about 1.8.Centrifugal 15 minutes of 4 ℃ of following 5000g, outwell supernatant, thalline is resuspended in 5% sucrose solution that contains 0.02%Silwet-77 (available from LEHEL SEEDS company), the OD that solution is final
600Be about 0.8.Well-grown Arabidopis thaliana inflorescence is immersed in 5-10 second in the bacterium liquid, upright also with black plastic bag parcel, preserved moisture 24 hours.Remove plastics bag, recover the normal cultured plant, gather in the crops seed after about one month.
(4) screening of positive seedling with the T0 of results for the Arabidopis thaliana seed with 70% alcohol disinfecting 1 minute; 84 thimerosals of 50% concentration (commercially available thimerosal product commonly used; Beijing wash the precious sterilized articles therefrom http://www.beijingxidebao.com.cn/ of ltd) sterilization 5 minutes; Use sterilization washed with de-ionized water 5 times again, the 1/2MS that evenly is layered on 0.6% agar cultivates the enterprising row filter of base (additional 50mg/L kantlex).Can distinguish in about 10 days and transform seedling and wild-type seedling (being non-conversion seedling).It is thus clear that transform the shoot root well-grown, it is green that blade is; And the wild-type shoot root almost can not be grown, and blade is yellow-white.The conversion transplantation of seedlings that filters out is grown in nutrition soil (volume ratio vermiculite and peat mix at 1: 1).(5) PCR that transforms seedling identifies that seedling length to be transformed arrives a certain size; Get its blade, (get in the fresh and tender blade and extract genomic dna, concrete grammar is with reference to Li Jia etc. to extract genomic dna; The method of the total DNA of a kind of effective extraction rape leaf; Hua Zhong Agriculture University's journal, 1994,13 (5): the 521-523 reported method).PG2F and P56oR are confirmed exist (the PCR reaction conditions is referring to embodiment 2) of promotor pBnPR1 with the special primer among the embodiment 2.Whether utilize primer that GU1F (5 ' GCGTTTCGATGCGGTCAC 3 ') and GU1R (5 ' GCGAGGTACGGTAGGAGTTGG 3 ') are detected GUS exists.The PCR system is: transform DNA (50ng/ μ l) the 2.0 μ l of seedling, ddH
2O 13.9 μ l, 10X PCR damping fluid 2.0 μ l, dNTP (10mM) 0.4 μ l; Primer GU1F (10 μ M) 0.8 μ l; Primer GU1R (10 μ M) 0.8 μ l, and rTaq (5U/ μ l) 0.1 μ l (rTaq polysaccharase wherein, the PCR damping fluid is available from precious biotechnology Dalian ltd; DNTP is available from Roche company, and primer is synthetic by Beijing AudioCodes biotechnology Ltd).The condition of pcr amplification is: 94 ℃ of preparatory sex change 3 minutes, and 94 ℃ of sex change 30 seconds, 60 ℃ of annealing 30 seconds, 72 ℃ were extended 1 minute, 35 circulations, last 72 ℃ of insulations 5 minutes.The plant that has only promotor pBnPR1 and GUS all to exist is just thought real conversion seedling, normal cultured, results seed.Warp too much is commissioned to train to support with antibiotic-screening and is obtained to isozygoty the conversion seedling.
The GUS tissue chemical analysis of embodiment 4 arabidopsis thaliana transformations
With the T3 that isozygotys that obtains for arabidopsis thaliana transformation cultivation in 1/2 intensity Hoagland and Arnon nutritive medium (Li Hesheng etc., 2002), wait to grow 6 lotus throne leaves after; A part of plant is lacked phosphorus, nitrogen stress, potassium deficiency; A lack of sulfur and iron deficiency are handled, and another part material normal cultured is as contrast.Handle and carry out GUS dyeing after 4 days.Method is at first plant to be put into the centrifuge tube that 5ml contains staining fluid, in the whole immersion of the assurance tissue solution.The composition and the concentration of staining fluid are following: the X-Gluc of 1mg/ml, the phosphate buffered saline buffer of 100mmol/l, the Na of 10mmol/l
2EDTA, the K of 0.5mmol/l
3[Fe (CN)
6] and K
4[Fe (CN)
6], 20% methyl alcohol, 0.1% Triton X-100.37 ℃ of lucifuges dyeed 12 hours then.With the ethanol decolorization of 70% concentration 3 times, the ethanol decolorization of using 100% concentration again is white in color to the blade of contrast for several times.In time take a picture, preserve experimental result.The result shows that lacking phosphorus handles whole of Arabidopis thaliana and part vein and be blue, show lack phosphorus after promotor pBnPR1 drive the GUS great expression; And each histoorgan of the Arabidopis thaliana of growing under nitrogen stress, potassium deficiency, a lack of sulfur, iron deficiency and the normal condition is white in color, and shows that GUS is not by the abduction delivering (see figure 3) under these conditions.
The main reference document
1, Li Hesheng etc. modern plants physiology. Beijing: Higher Education Publishing House, 2002;
2、Clough?S?J，Bent?A?F(1998)Floral?dip：a?simplified?method?for?Agrobacterium-mediated?transformation?ofArabidopsis?thaliana.Plant?J，16：735～743；
3、L6pez-Bucio?J，de?Ia?Vega?OM，Guevara-García?A，Herrera-Estrella?L(2000)Enhanced?phosphorus?uptake?intransgenic?tobacco?plants?that?overproduce?citrate.Nat?Biotech?18：450-453；
4、Misson，J.，et?al.(2005)A?genome-wide?transcriptional?analysis?using?Arabidopsis?thaliana?Affymetrix?genechips?determined?plant?responses?to?phosphate?deprivation.Proc.Natl.Acad.Sci.USA?102：11934-11939；
5、Nilsson?L，Muller?R，Nielsen?T?H(2007)Increased?expression?of?the?MYB-related?transcription?factor，PHR1，leads?to?enhanced?phosphate?uptake?inArabidopsis?thaliana.Plant?Cell?Environ?30：1499-1512；
6、Wang?X，Wang?Y，Tian?J，Lim?BL，Yan?X，Liao?H(2009)Overexpressing?AtPAP?15enhances?phosphorusefficiency?in?soybean.Plant?Physiol?151：233-240；
7, Li Jia etc., the method for the total DNA of a kind of effective extraction rape leaf, Hua Zhong Agriculture University's journal, 1994,13 (5): 521-523.
Claims (3)
1. one kind by nitrogen stress, potassium deficiency, a lack of sulfur and iron deficiency stress-inducing, and only by the pBnPR1 promoter sequence of phosphorus starvation induced expression, its nucleotide sequence is shown in 1-1874bp among the sequence table SEQ ID NO:1.
2. the plant expression vector that contains the described promotor nucleotide sequence of claim 1.
3. the application of the described promotor of claim 1 in cultivating phosphorus efficiency or anti-low-phosphorous crop.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103194447A (en) * | 2012-01-09 | 2013-07-10 | 华中农业大学 | Brassica napus phosphorus deficiency inducible expression promoter |
| CN104774850A (en) * | 2015-04-03 | 2015-07-15 | 华中农业大学 | BnNRT1-3 gene overexpression for improving rape nitrogen utilization rate |
| CN106834345A (en) * | 2016-12-27 | 2017-06-13 | 河南大学 | A kind of method that polygenes superposition cotransformation improves rape synthesis resistance |
| CN108424912A (en) * | 2018-02-01 | 2018-08-21 | 山西省农业科学院作物科学研究所 | The promoter of corn low-phosphorus stress induced expression and its application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101050462A (en) * | 2007-04-02 | 2007-10-10 | 中国科学院遗传与发育生物学研究所 | Induction gene lack of phosphor from Arabidopsis thaliana, coded protein, and application |
| CN101298616A (en) * | 2008-06-20 | 2008-11-05 | 中国科学院遗传与发育生物学研究所 | Promoter for expressing phosphor deficiency speciality induction in root and use thereof |
| CN101365794A (en) * | 2005-08-12 | 2009-02-11 | 巴斯福植物科学有限公司 | Nucleic acid sequences encoding proteins associated with abiotic stress response and plant cells and plants with increased tolerance to environmental stress |
-
2010
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101365794A (en) * | 2005-08-12 | 2009-02-11 | 巴斯福植物科学有限公司 | Nucleic acid sequences encoding proteins associated with abiotic stress response and plant cells and plants with increased tolerance to environmental stress |
| CN101050462A (en) * | 2007-04-02 | 2007-10-10 | 中国科学院遗传与发育生物学研究所 | Induction gene lack of phosphor from Arabidopsis thaliana, coded protein, and application |
| CN101298616A (en) * | 2008-06-20 | 2008-11-05 | 中国科学院遗传与发育生物学研究所 | Promoter for expressing phosphor deficiency speciality induction in root and use thereof |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103194447A (en) * | 2012-01-09 | 2013-07-10 | 华中农业大学 | Brassica napus phosphorus deficiency inducible expression promoter |
| CN103194447B (en) * | 2012-01-09 | 2014-04-16 | 华中农业大学 | Brassica napus phosphorus deficiency inducible expression promoter |
| CN104774850A (en) * | 2015-04-03 | 2015-07-15 | 华中农业大学 | BnNRT1-3 gene overexpression for improving rape nitrogen utilization rate |
| CN106834345A (en) * | 2016-12-27 | 2017-06-13 | 河南大学 | A kind of method that polygenes superposition cotransformation improves rape synthesis resistance |
| CN108424912A (en) * | 2018-02-01 | 2018-08-21 | 山西省农业科学院作物科学研究所 | The promoter of corn low-phosphorus stress induced expression and its application |
| CN108424912B (en) * | 2018-02-01 | 2021-07-13 | 山西省农业科学院作物科学研究所 | Promoter for low-phosphorus stress induced expression of corn and application thereof |
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