CN103160516B - Rape stress resistance gene and application - Google Patents
Rape stress resistance gene and application Download PDFInfo
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- CN103160516B CN103160516B CN201310074873.2A CN201310074873A CN103160516B CN 103160516 B CN103160516 B CN 103160516B CN 201310074873 A CN201310074873 A CN 201310074873A CN 103160516 B CN103160516 B CN 103160516B
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Abstract
The invention discloses a rape stress resistance gene and an application. The Brassica napus stress resistance gene DUF1645, a nucleotide sequence of the gene is shown as SEQ ID NO:1. Through over expression of Brassica napus stress resistance gene DUF1645 in arabidopsis thaliana, the invention found that the gene can greatly enhance the drought resistance and waterlogging tolerance of the plants, the plants enable normal growth in 15 days of drought stress or 10 days of waterlogging stress, and the yield and the quality are not influenced, the gene provides guarantee of yield enhancement of crops under drought and waterlogging adverse situation, so that the gene provided by the invention has good application prospect in breeding for stress tolerance of crops.
Description
Technical field
The invention belongs to biology field.Specifically, more specifically relate to a kind of rape adversity gene DUF1645, also relate to the application of this gene in crop degeneration-resistant (comprising drought resisting and resistance to stain) breeding simultaneously, the overexpression of rape adversity gene DUF1645 in Arabidopis thaliana shows, this gene can significantly strengthen drought resistance and the waterlogging tolerance of plant, for improving the output of crop under arid and water stain adverse circumstance, gives security.
Background technology
Under field conditions (factors), plant often can be subjected to many abiotic stresses as the injury of arid, water stain, salt marsh, low temperature etc. in growth and development process, and output of the growing of plant, crop etc. is exerted an influence.Wherein arid and water stain be the adverse circumstance factor the most general in various environment-stress.Arid and the water stain threat to agriculture production is a global problem, is subject to the impact of Global climate change, and the frequency and the intensity of arid, precipitation and continuing precipitation event significantly increase, and the agricultural arid causing thus and accumulated water harm are on the rise.According to statistics, in the total losses causing in all kinds of natural disasteies, the loss that meteorologicdisasters causes accounts for 85%, and the disaster that wherein arid factor causes has accounted for again the more than 50% of Loss Caused by Meteorological Disasters.According to incompletely statistics, over nearly more than 50 years, China every year on average arid area suffered from drought reaches 2,217 ten thousand hectares.Enter the nineties in 20th century, average annual area suffered from drought rises to 2,711 ten thousand hectares, 2000 of severe drought and calendar year 2001 are respectively up to 4054 and 3,847 ten thousand hectares, national area suffered from drought in 2000 for since the establishment of the nation, wherein have no harvest 8,000,000 hectares, because of nearly 60,000,000,000 kilograms of drought loss grain, 51,000,000,000 yuan of cash crop losses, its impact has surpassed 3 years natural disasteies of 1959-1961.And the agricultural losses that accumulated water harm causes are only second to arid, according to Food and Argriculture OrganizationFAO's statistics, show, only developing country has at least 1,200 ten thousand hectares of high-yielding grain fields to reduce its throughput because of accumulated water problem in recent years, the crop production reduction causing because of accumulated water harm is every year up to 15%-80%, as corn 25%-30%, soybean 50%-60%, wheat 39%-44%.
In long-term evolutionary process, higher plant develops the system that has produced a set of impression and conduction moisture stress signal gradually, and the mechanism that forms a series of physiology or growth carrys out this coercing in response environment, alleviate to greatest extent arid or the water stain injury causing.This biological adaptation phenomenon of plant is very complicated, in form, relates to many different cellular fories and structural character; On physiology, relate to many important biochemical metabolism approach; On molecular biology, relate to and comprise the serial genes expression that is transmitted to genetic transcription control from signal.Along with the development of modern molecular biology and biotechnology, how plant experiences adverse circumstance signal by cell, and conduction adverse circumstance stimulates, activate a series of molecular pathways and regulate and control related gene expression and physiological response to adapt to adverse circumstance, become the focus of scientist's research.In plant, the gene of water stress induction is divided into two classes: a class is the protein gene working in the resistance of plant, to maintain the various Physiological and Biochemical Metabolism activities of cell, normally carries out.The synthase gene that comprises osmotic factor (as proline(Pro), trimethyl-glycine etc.); directly Cell protection is avoided functional protein (as Lea albumen, ooze and adjust albumen, aquaporin, the ionophorous protein etc.) gene of water stress injury, and toxicity degrading enzyme (as glutathione s-transferase, soluble epoxide hydrolase, superoxidase, catalase and ascorbate peroxidase enzyme etc.) gene.Another kind of be the protein gene playing regulatory role in signal conduction and genetic expression process, the transcription factor gene that mainly comprises transmission of signal and regulate gene expression, the protein kinase gene of induction and transduction stress signal, and the proteinase gene playing an important role in signal transduction.Closely during the last ten years, the research of plant responding water stress is obtaining many new discoveries and breakthrough aspect gene expression regulation, protein modified and interaction, signal delivery network.Especially several large class transcription factors such as DREB1/CBF, DREB2, AREB/ABF, NAC, ZFHD have played vital role in the gene expression regulation of response Water Stress.
Plant anti-adversity associated protein DUF1645 and the encoding gene thereof in the present invention, mentioned, so far, be included in the report that does not all also have any correlation function in model plant Arabidopis thaliana, is albumen and the gene of a unknown function.
Summary of the invention
The object of the invention is to be to provide a kind of swede type rape (Brassica napus) adversity gene DUF1645, the nucleotides sequence of this gene is classified as shown in SEQ ID NO:1, comprises the gene order that has 70% homology with the DNA sequence dna shown in SEQ ID NO:1 at least; Be also included within the mutant allele or the derivative that add, replace, insert or delete one or more Nucleotide and produce.By the overexpression in Arabidopis thaliana by rape adversity gene DUF1645, find that this gene can significantly strengthen drought resistance and the waterlogging tolerance of plant, drought stress 15 days with interior or water stain coerce 10 days normal with interior plant strain growth, and output, quality are substantially unaffected, for improving the output of crop under arid and water stain adverse circumstance, give security.
A further object of the invention is to be to provide the application of the degeneration-resistant gene DUF1645 of a kind of swede type rape in plant stress-resistance (drought resisting and resistance to stain) breeding, improved the tolerance of plant to adverse circumstance (arid and water stain), the disadvantageous effect of environment stress is avoided or be subject to less to the yield and quality that has guaranteed crop.
In order to realize foregoing invention object, the present invention adopts following technical measures:
The degeneration-resistant gene DUF1645 of swede type rape, its preparation process is:
A kind of rape adversity gene DUF1645 and with the acquisition of the Arabidopis thaliana DUF1645 of rape 70% homology:
1) the rape strain zy036 and the 51070(Hu Z Y that in a pair of resistance (being mainly drought resisting and waterlogging tolerance), there are differences, Wang XF, Zhan GM, Liu GH, Hua W, Wang HZ.2009.Unusually large oilbodies are highly correlated with lower oil content in Brassica napus.Plant Cell Reports, 28:541-549.) in poor subtracting obtain this gene est sequence, the Arabidopis thaliana DUF1645 gene order (At5G14730) that retrieval has been delivered in ncbi database, and with this coding region sequence BLAST NCBI est database, find the est sequence of the rape DUF1645 gene of homology with it, and design the both sides primer (forward primer: 5 '-ATGCAAGATTTTTCATATTCTACGG-3 ' of gene coded sequence, reverse primer: 5 '-TTATCGACTGAGTCTACTCATTC-3 ').Arabidopis thaliana DUF1645 gene is according to the primers in database: (forward primer: 5 '-ATGAGTACAATGCAAGATTTATC-3 '; Reverse primer: 5 '-TTAATTACGGAATCTACCCATCC-3 ').
2) take rape, Arabidopis thaliana cDNA the first chain carries out RT-PCR amplification as template, the fragment that amplification is obtained checks order, obtain rape and Arabidopis thaliana DUF1645 gene order, the degeneration-resistant gene DUF1645 of a kind of swede type rape, its sequence is shown in SEQ ID NO:1, arabidopsis gene sequence with in database, delivered identical.
By aforesaid method, swede type rape and Arabidopis thaliana adversity gene DUF1645 have been obtained, adopt a kind of transgenic arabidopsis that improves DUF1645 expression activity, it is characterized in that transgenic plant show as DUF1645 content and increase, than the resistance of acceptor contrast (non-transgenic plant), strengthen to some extent.
The application of the degeneration-resistant gene DUF1645 of swede type rape in Arabidopis thaliana degeneration-resistant (drought resisting and resistance to stain) breeding, its application process is:
1) rape clone being obtained, Arabidopis thaliana adversity gene DUF1645 respectively with PCR8/GW/TOPO(invitrogen company) plasmid is connected, called after topo-BnDUF1645 and topo-AhDUF1645, utilize PCR8/GW/TOPO plasmid and plasmid expression vector Pearleygate100(invitrogen company) characteristic that can vitro recombination is transferred to expression vector by degeneration-resistant protein gene DUF1645, called after Pearleygate100-BnDUF1645 and Pearleygate100-AhDUF1645;
2) by step 1) in carrier Pearleygate100-BnDUF1645 and the Pearleygate100-AhDUF1645 of preparation proceed to agrobacterium tumefaciens GV3101(invitrogen company), then import in Arabidopis thaliana plant;
3) screening positive plant.
The seed that plantation Arabidopis thaliana T0 generation gathers in the crops, treat 10 days left and right spraying herbicide (Liberty, Invitrogen company) for screening positive plant, leaf DNA is extracted laggard performing PCR and is identified, finally confirms the positive plant that gene has entered.
In the present invention, term " transgenic plant " used refers to the gene that contains importing the genetic expression that stably enhancer or inhibitor imports and produces the plant with specific biological character.
The plant of mentioning in the present invention comprises that paddy rice, wheat, corn etc. are subject to arid and water stain adverse circumstance to affect larger food crop, also comprises the cash crop such as rape, soybean, cotton, also comprises the vegetable crops such as cucumber, tomato of summer growth.
The method of degeneration-resistant protein gene DUF1645 described in clone the present invention is the method often adopting in this area.Extracting plant leaf DNA is conventional Protocols in Molecular Biology, the method of extracting mRNA also has multiple proven technique, test kit (TRIzol Reagent) can obtain (Invitrogen company) from commercial channels, and construction cDNA library is also conventional Protocols in Molecular Biology.Build the vector construction described in the present invention and by carrier be transfected into that plant enzyme used is cut, connection, inflorescence are infected etc., and method is also common technology in this area.Related plasmid (entry vector PCR8/GW/TOPO, plasmid expression vector Pearleygate100) wherein, media for transfection (if agrobacterium tumefaciens GV3101 and agents useful for same composition are as sucrose, 6-BA etc.) can obtain from commercial channels.
Compared with prior art, the present invention has the following advantages:
The present invention is rape adversity gene DUF1645 sequence openly first both at home and abroad, and it belongs to the degeneration-resistant albumen of tenuigenin I micromolecular, and the function that its overexpression can improve stress resistance of plant does not at present have relevant report.Experimental result of the present invention shows that the content of transgenic arabidopsis DUF1645 contrasts (non-transgenic plant) and compares all and have significant improvement with acceptor, and therefore its resistance be also enhanced.Therefore the present invention proposes to utilize the overexpression of DUF1645 to strengthen stress resistance of plant for use in the anti-adversity seed selection of farm crop and vegetables, slows down thus the injury that arid and water stain adverse circumstance cause yield of commercial crops and quality.
Accompanying drawing explanation
Fig. 1 is a kind of plant expression vector schematic diagram.
By DUF1645 coding sequence and PCR8/GW/TOPO plasmid screening after connection forward carrier, obtain PCR8/GW/TOPO-DUF1645, then PCR8/GW/TOPO-DUF1645 plasmid and expression vector Pearleygate100 recombinate, and screening positive clone obtains Pearleygate100-DUF1645.
Fig. 2 is a kind of herbicide screening transfer-gen plant schematic diagram later.
In plant results seed T0 generation after Arabidopis thaliana inflorescence infects, sow 2 weeks rear spraying herbicide screening positive plants.
Fig. 3 is a kind of DUF1645 gene Arabidopis thaliana T1 PCR qualification result schematic diagram in generation that turns.
123 for turning rape adversity gene, and ABCD is for turning Arabidopis thaliana adversity gene, and WT is contrast Wild plant.
Comparison schematic diagram after water stain, the arid processing of Fig. 4 transfer-gen plant and adjoining tree.
A in Fig. 4: accumulated water is coerced contrast, left side wild-type, the right turns DUF1645 plant; B in Fig. 4: accumulated water is coerced 10 days, left side wild-type blade flavescence, the right turns DUF1645 plant leaf without considerable change; C in Fig. 4: drought stress contrast, above wild-type, turn DUF1645 plant below; D in Fig. 4: the drought stress 15 days of not watering, above wild-type blade purpling wilt, turn DUF1645 plant leaf below and change not obvious; E in Fig. 4: spray 6%PEG drought stress 15 days, above wild-type blade purpling, turn DUF1645 plant leaf below and change not obvious.
Embodiment
Conventionally according to normal condition as people such as Sambrook, molecular cloning: laboratory manual (New York:Cold Sprin g Harbor Laboratory Press, 1989), or people (the Blackwell Science Press such as Draper, 1988) described condition, or the condition of advising according to agents useful for same manufacturer.
Embodiment 1: the acquisition of rape and Arabidopis thaliana adversity gene DUF1645:
The Arabidopis thaliana DUF1645(At5G14730 that retrieval has been delivered in ncbi database) gene order, BLAST is to rape est sequence and splice rape DUF1645 coding region total length.Both sides primer (the forward primer: 5 '-ATGCAAGATTTTTCATATTCTACGG-3 ' of design gene coded sequence; Reverse primer: 5 '-TTATCGACTGAGTCTACTCATTC-3 ').Arabidopis thaliana DUF1645 gene is according to the primers in database: (forward primer: 5 '-ATGAGTACAATGCAAGATTTATC-3 '; Reverse primer: 5 '-TTAATTACGGAATCTACCCATCC-3 ')
1, extract rape, Arabidopis thaliana mRNA.
The extraction of RNA (TRIZOL TM Kit extracts RNA)
Liquid nitrogen grinding 100mg material
A. add 1ml TRIZOL, room temperature (20-25 ℃, lower same) is placed 5min.
B. add 200ul chloroform, thermal agitation 30s, room temperature is placed 2min.
C.12000g, 15min, 4 ℃, get supernatant to new pipe, add 500ul Virahol, mix rear room temperature and place 15min.
D.12000g, 15min, 4 ℃, remove supernatant, add the volume ratio of 1ml70%(raw spirit and H2O) ethanol.
E.7500g, 7min,, removes supernatant, dry air by 4 ℃.
F.DEPC-H
2o dissolves.
2, the reverse transcription of cDNA the first chain adopts RevertAid H Minus First Strand cDNA Synthesis Kit (Ferme ntas), and operation is carried out with reference to used kit explanation.
3, take cDNA carries out pcr amplification as template, has obtained a kind of rape adversity gene DUF1645, and its nucleotides sequence is classified as shown in SEQ ID NO:1.A protein for rape adversity gene DUF1645 coding, its sequence is the aminoacid sequence shown in SEQ ID NO:2.
This gene overexpression can significantly strengthen drought resistance and the waterlogging tolerance of plant, drought stress 15 days with interior or water stain coerce 10 days normal with interior assurance plant strain growth, and output, quality are substantially unaffected, for improving the output of crop under arid and water stain adverse circumstance, give security.
The 20 μ l reaction systems of rape and Arabidopis thaliana PCR are as follows:
Time and the temperature of reaction are as follows:
94℃3min
94℃ 45s，
59℃ 45s
72℃ 2min 30s，30cycles
72℃ 5min
Embodiment 2:
The application of the degeneration-resistant gene DUF1645 of swede type rape in plant stress-resistance (drought resisting and resistance to stain) breeding, its application process is:
2.1 structures of DUF1645 expression vector and the conversion of Arabidopis thaliana:
By the rape adversity gene DUF1645 obtaining by pcr amplification in embodiment 1 and TOPO entry vector PCR8/GW/TOPO(invitrogen company) be connected (linked system: PCR product 4 μ l, TOPO carrier 1 μ l, Buffter1 μ l, room temperature connects 5 minutes) after, be transformed in competent cell DH5 α (invitrogen company), the screening of grand enzyme element, carrier primer (T7 primer) identifies that with rape DUF1645 gene amplification forward inserts clone (pcr amplification condition is with embodiment 1), plasmid is through preparing in a small amount Hou Yu Pearleygate100(invitrogen company) (the restructuring system: recombinase 0.5 μ l of recombinating, TOPO carrier 1 μ l, Pearleygate100 carrier 1 μ l, 25 ℃ connect 1h), and be transformed into competent cell DH5 α (with reference to people such as Sambrook, molecular cloning: laboratory manual.New York:Cold Spring Harbor Laboratory Press, 1989) in, kantlex screening, its Insert Fragment is identified (pcr amplification condition is with embodiment 1) through carrier primer (35S promoter aligning primer) with rape DUF1645 gene downstream primer PCR, Fig. 1 is shown in by schematic diagram.PCR extracts plasmid after identifying, transform Agrobacterium.
The conversion process of Arabidopis thaliana:
Reagent preparation
Infiltration substratum (1L): 1/2xMurashige-Skoog; 5%(mass ratio) sucrose; 0.5 gram of MES; With KOH, be adjusted to pH5.7; Add again: the 6-BA mother liquor of 10 microlitre 1mg/ml; 200 microlitre Silwet L-77
Step of converting
(1) prepare Agrobacterium (agrobacterium tumefaciens GV3101) the bacterium liquid 10ml that has transformed corresponding plasmid, transforming evening before that day, proceed to large bottle overnight incubation, second day takes out agrobacterium liquid O.D600 while using and works as between 1.2 to 1.6.
(2) room temperature 5000rpm is centrifugal 15 minutes.
(3) abandon supernatant, Agrobacterium precipitation is suspended in the infiltration substratum of respective volume, make O.D600 in 0.8 left and right.
(4) whole plant is directly dipped to agrobacterium suspension 30s.
(5) lucifuge overnight incubation, is then normally cultured to knot.
The screening of 2.2 transgenic arabidopsis and checking
The screening of transformant
By the Arabidopis thaliana seed of vernalization in watering the Artificial Soil of supersaturation PNS nutritive medium, and with on preservative film cover.Manual simulation's natural condition illumination two days later (illumination 16 hours, dark 8 hours), took off film after three days.
Artificial culture chamber condition: relative humidity 80%, constant temperature 20-240C, intensity of illumination 80-200umol/M2/S, periodicity of illumination is 8h Hei An ﹑ 16h illumination cultivation.One week left and right, spray herbicide screening positive plant.
PCR identifies
(1) for the extraction of the total DNA of transformed plant of PCR
A.70%(volume ratio) ethanol is cleaned blade, takes about 100mg
B. add 600ul extraction buffer (0.2MTris-Cl, 0.25 NaCl, 25mM EDTA, 0.5% SDS, pH7.5), room temperature is ground fast.
C.1.5ml Ependorff pipe mesoscale eddies mixes 5-10s.
D.12000rpm, 25min, room temperature.Get supernatant, add equal-volume Virahol ,-20 degrees Celsius of precipitations are spent the night.
E.12000rpm, 15min, room temperature.Add 70% ethanol 200ul foam washing DNA precipitation.
F.12000rpm, 15min, room temperature.Remove ethanol.Be inverted on paper handkerchief, treat that ethanol volatilization is clean.
G. add sterilized water 100ul dissolving and slightly put forward DNA precipitation.With spectrophotometric determination or electrophoresis, estimate its concentration.
H. take total DNA as template, carry out PCR.
(2) PCR program
The proportioning of PCR reaction mixture is identified with plasmid PCR, according to goal gene in plant expression vector and upstream 35S promoter sequence and rape DUF1645 gene downstream primer 5 '-TTATCGACTGAGTCTACTCATTC-3 ', the time of reaction and temperature are done as follows:
94℃3min
94℃ 45s，
59℃ 45s
72℃ 2min 30s，30cycles
72℃ 5min
Detected result shows, most of transformed plants all can amplify the big or small electrophoretic band of expection, and negative control does not have, and shows to have contained in transgenic arabidopsis genome foreign gene DNA fragmentation, and result is shown as Fig. 3.
2.3 Arabidopis thaliana resistance experiments
To turning DUF1645 plant and wild-type material, carry out drought stress and accumulated water Stress treatment respectively, as seen from Figure 4, accumulated water after 10 days wild-type material blade mostly start flavescence, and converting material has no significant effect.Drought stress has been done two processing, and the 6%PEG that do not water and spray for 15 days processes, and process the most purpling look of wild-type material blade wilting after 15 days, and the variation of converting material blade is not obvious.Therefore illustrate that transfer-gen plant drought resistance and waterlogging tolerance all obviously strengthen.
SEQUENCE LISTING
<110> Inst. of Oil Crops, Chinese Academy of Agriculture
<120> rape adversity gene and application thereof
<130> rape adversity gene and application thereof
<160> 2
<170> PatentIn version 3.1
<210> 1
<211> 750
<212> DNA
<213> rape
<400> 1
atgcaagatt tttcatattc tacggagtcc tttagcccta gcttcagttt ttatgtctcc 60
ggtgacggtg agatcgtcga agccgccgtt agagttgtcc gtgagtctta cggcgacgac 120
accgctgagt tcgagttcga aacattaccg ttacacgaag agaacttctt ccacttccca 180
acgacgacgg agacgaaatc tttcgatagc tcacacaagg aggaagaaga cgatgacgtg 240
gcagcggatg gagtcacctc gaaggatctt ttctacggtg gtggatggat gcttgatcag 300
tctatatcct cgccgtctca gtcgccttca tcttccgagt cttcagactc ggaggatctt 360
tccccgagag gatacaactg tttctggagc ccgacccgat ctccggcgag agttgatagc 420
tccaagagta aaacttcgag cgcaccgaaa cggtgccgta ttaaggatct tttaaggaga 480
agccacagcg acggagctgt ttcgaccacg agcgagccta aacgatgtcg tttcaaggat 540
cttctgagga gaagtcacag cgacggcaga ggaagtgggt ctttagtttc gccgagcggg 600
aacagctcac cagcagtaag agggagaaac aaaacgacgt cgaataagta tactaatact 660
ggggaagtga atatgagaag aaagacttat ttaccttaca ggcaggattt gatcggcgtt 720
ttcgccggaa tgagtagact cagtcgataa 750
<210> 2
<211> 249
<212> PRT
<213> rape
<400> 2
Met Gln Asp Phe Ser Tyr Ser Thr Glu Ser Phe Ser Pro Ser Phe Ser
1 5 10 15
Phe Tyr Val Ser Gly Asp Gly Glu Ile Val Glu Ala Ala Val Arg Val
20 25 30
Val Arg Glu Ser Tyr Gly Asp Asp Thr Ala Glu Phe Glu Phe Glu Thr
35 40 45
Leu Pro Leu His Glu Glu Asn Phe Phe His Phe Pro Thr Thr Thr Glu
50 55 60
Thr Lys Ser Phe Asp Ser Ser His Lys Glu Glu Glu Asp Asp Asp Val
65 70 75 80
Ala Ala Asp Gly Val Thr Ser Lys Asp Leu Phe Tyr Gly Gly Gly Trp
85 90 95
Met Leu Asp Gln Ser Ile Ser Ser Pro Ser Gln Ser Pro Ser Ser Ser
100 105 110
Glu Ser Ser Asp Ser Glu Asp Leu Ser Pro Arg Gly Tyr Asn Cys Phe
115 120 125
Trp Ser Pro Thr Arg Ser Pro Ala Arg Val Asp Ser Ser Lys Ser Lys
130 135 140
Thr Ser Ser Ala Pro Lys Arg Cys Arg Ile Lys Asp Leu Leu Arg Arg
145 150 155 160
Ser His Ser Asp Gly Ala Val Ser Thr Thr Ser Glu Pro Lys Arg Cys
165 170 175
Arg Phe Lys Asp Leu Leu Arg Arg Ser His Ser Asp Gly Arg Gly Ser
180 185 190
Gly Ser Leu Val Ser Pro Ser Gly Asn Ser Ser Pro Ala Val Arg Gly
195 200 205
Arg Asn Lys Thr Thr Ser Asn Lys Tyr Thr Asn Thr Gly Glu Val Asn
210 215 220
Met Arg Arg Lys Thr Tyr Leu Pro Tyr Arg Gln Asp Leu Ile Gly Val
225 230 235 240
Phe Ala Gly Met Ser Arg Leu Ser Arg
245
Claims (4)
1. the degeneration-resistant gene of swede type rape, its sequence is shown in SEQ ID NO:1.
2. a protein for the degeneration-resistant gene translation of swede type rape claimed in claim 1, its sequence is shown in SEQ ID NO:2.
3. the application of the degeneration-resistant gene of a kind of swede type rape claimed in claim 1 in Arabidopis thaliana Drought-resistant Breeding.
4. the application of the degeneration-resistant gene of a kind of swede type rape claimed in claim 1 in the anti-accumulated water breeding of Arabidopis thaliana.
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| CN107889642A (en) * | 2017-12-09 | 2018-04-10 | 湖南科技大学 | The method that rape drought-resistance is improved using plant extracts |
| CN109207487B (en) * | 2018-11-22 | 2021-12-03 | 中国农业科学院油料作物研究所 | Rape stain-resistant gene BnalPP1, and preparation method and application thereof |
| CN110804614B (en) * | 2019-10-08 | 2021-11-09 | 湖南农业大学 | Cabbage type rape drought-resistant gene BnatZF1A, primer, expression vector and application thereof, and method for improving drought resistance |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101736012A (en) * | 2008-11-27 | 2010-06-16 | 上海市农业科学院 | Stress resistance ERF transcription factor gene derived from Brassica napus |
| CN102094025A (en) * | 2009-12-11 | 2011-06-15 | 上海市农业科学院 | Brassica napus AP2/ERF family transcription factor gene and preparation method thereof and application thereof |
-
2013
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101736012A (en) * | 2008-11-27 | 2010-06-16 | 上海市农业科学院 | Stress resistance ERF transcription factor gene derived from Brassica napus |
| CN102094025A (en) * | 2009-12-11 | 2011-06-15 | 上海市农业科学院 | Brassica napus AP2/ERF family transcription factor gene and preparation method thereof and application thereof |
Non-Patent Citations (10)
| Title |
|---|
| Bateman et al.DUFs: families in search of function.《Acta Crystallographica Section F》.2010,第66卷全文. |
| DUFs: families in search of function;Bateman et al;《Acta Crystallographica Section F》;20100305;第66卷;全文 * |
| Hansen et al.Plant glycosyltransferases beyond CAZy:aperspective on DUF families.《Frontiers in Plant Science》.2012,第3卷全文. |
| HU et al.Unusually large oilbodies are highly correlated with lower oil content in Brassica napus.《Plant Cell Rep》.2009,第28卷(第4期),全文. |
| Lee et al.Transcriptome Analysis in Brassica rapa under the Abiotic Stresses Using Brassica 24K Oligo Microarray.《Molecules and Cells》.2008,第26卷(第6期),全文. |
| Plant glycosyltransferases beyond CAZy:aperspective on DUF families;Hansen et al;《Frontiers in Plant Science》;20120328;第3卷;全文 * |
| Transcriptome Analysis in Brassica rapa under the Abiotic Stresses Using Brassica 24K Oligo Microarray;Lee et al;《Molecules and Cells》;20081231;第26卷(第6期);全文 * |
| Unusually large oilbodies are highly correlated with lower oil content in Brassica napus;HU et al;《Plant Cell Rep》;20090430;第28卷(第4期);全文 * |
| 丛野等.甘蓝型油菜发芽种子耐湿性的主基因+多基因遗传分析.《作物学报》.2009,第35卷(第8期),全文. |
| 甘蓝型油菜发芽种子耐湿性的主基因+多基因遗传分析;丛野等;《作物学报》;20091231;第35卷(第8期);全文 * |
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