CN106868017A - A kind of adjusting and controlling rice seedling stage heat resistance gene TBZ1 and application - Google Patents
A kind of adjusting and controlling rice seedling stage heat resistance gene TBZ1 and application Download PDFInfo
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Abstract
The present invention relates to a kind of adjusting and controlling rice seedling stage heat resistance gene TBZ1 and application.Gene TBZ1 open reading frame sequences are：SEQ ID NO.1, the sequence of its code area is：SEQ ID NO.2.Carrier containing described gene is plant expression vector, is suitable to be expressed in paddy rice.Convert the cell or tissue of plant with described carrier, and the plant cell that will convert or tissue cultivating are into plant.The paddy rice regulation and control seedling stage heat resistance gene TBZ1 that the present invention is provided is a new Rice Panicle response environment temperature gene, and the gene can be with adjusting and controlling rice seedling stage temperature capacity.Gene pairs of the invention cultivates heat-resisting plant new varieties, particularly cultivates heat-resisting new rice variety significant.It is that rice breeding and the research of Rice Panicle developmental mechanism provide new genetic resources, can be applied at adjusting and controlling rice seedling stage heat-resisting aspect.
Description
Technical field
The invention belongs to biology field, it is related to a kind of adjusting and controlling rice seedling stage heat resistance gene TBZ1 with application.
Background technology
With global warming, Climatic regionalization has turned into one of key constraints of plant growth.In global climate
Persistently warm, under the background that Short-Term High Temperature takes place frequently, carry out the Research on Mining of heat-resisting related gene to promoting paddy rice to continue, safety it is raw
Produce significant.
The thermal shock transcription factor and thermal shock albumen of paddy rice response high temperature signal have and compare in-depth study, but to regulating and controlling these
It is little that the upstream element of gene is also knows about, and High Temperature Stress response and such as optical signal presence of other envirment factors of plant are contacted,
By multiple regulation and control, the HTT1 genes in the African wild rice of current research discovery are used as the heat-resisting of proteasome gene energy adjusting and controlling rice
Property.Zinc finger protein is the transcription factor that a class has finger structure domain, in gene expression, cell differentiation, embryonic development, enhancing
The aspects such as resistance have important regulating and controlling effect, and the response of biological development and stress to plant is most important, is adjusted in light
It is also irreplaceable in section growth and development of plants.B-box type zinc finger protein genes belong in the protein factor of coding all containing 1 or
2 zinc finger proteins of the highly conserved B-box domains being made up of 40-47 amino acid residue.
Paddy rice heat resistance is a quantitative character, there is multiple regulatory sites, but currently known adjusting and controlling rice heat resistance
Gene seldom, the present invention probe into influence paddy rice regulation and control seedling stage heat resistance gene and its application method, be the further right mankind
Rice Panicle regulation and control seedling stage heat resistance mechanism is recognized completely, and the crucial base of heat resistance is controlled with molecular breeding technology directional transformation
Thus improve rice seedling heat resistance.
The content of the invention
It is an object of the invention to provide a kind of adjusting and controlling rice seedling stage heat resistance gene TBZ1 and application.
The technical scheme is that：A kind of adjusting and controlling rice seedling stage heat resistance gene TBZ1, open reading frame sequence is：
SEQ ID NO.1, the sequence of its code area is：SEQ ID NO.2.
A kind of B-box zinc finger proteins of adjusting and controlling rice seedling stage Holstein Cattle, with SEQ ID No：Amino acid sequence shown in 3
Row.
A kind of carrier containing described gene.It is plant expression vector, is suitable to be expressed in paddy rice.For example, having
SEQ ID No：The expression vector of the pUbi-OsTBZ1-RNAi shown in 4.
A kind of host cell, the cell is containing described gene, or contains described carrier.The cell is selected from large intestine
Bacilli-cell, agrobatcerium cell or plant cell.
It is a kind of influence plant seedling stage heat resistance method, methods described include use described in carrier convert plant cell or
Tissue, and the plant cell that will convert or tissue cultivating are into plant.The conversion passes through agrobacterium-mediated transformation or particle bombardment
Carry out.The plant is grass.
The heat-resisting factor TBZ1 of paddy rice that the present invention is identified be B-box zinc finger proteins in BBX22 genes, the gene it is normal
Expression ensure that paddy rice to a certain degree of tolerance of high temperature, and improving its expression quantity by genetic engineering transformation appropriateness can enter one
Step improves the heat resistance of paddy rice, so as to reach the purpose of volume increase.
When gene of the invention be used to change paddy rice Holstein Cattle, following methods can be used：(1) by water of the invention
The heat-resisting gene TBZ1 of rice is cloned into plant conversion carrier；(2) constructed plant conversion carrier is converted into renewable paddy rice group
Knit (or organ) and the heat-resisting gene of paddy rice of the invention is expressed in the tissue of conversion；(3) tissue (or the device that will be converted
Official) it is trained plant.
The pUbi-OsTBZ1-RNAi expression vectors that the present invention builds, the expression of suppression OsTBZ1 genes that can be special, from
And reduce the seedling heat resistance of paddy rice.
The paddy rice regulation and control seedling stage heat resistance gene TBZ1 that the present invention is provided is a new Rice Panicle response environment temperature base
Cause, the gene can be with adjusting and controlling rice seedling stage temperature capacity.Gene pairs of the invention cultivates heat-resisting plant new varieties, particularly cultivates
Heat-resisting new rice variety is significant.It is that rice breeding and the research of Rice Panicle developmental mechanism provide new genetic resources,
Can be applied at adjusting and controlling rice seedling stage heat-resisting aspect.
Brief description of the drawings
Fig. 1 pCAMBIA1390 Vector maps；
Fig. 2 pUbi-OSTBZ1-RNAi Vector maps；
Fig. 3 semiquantitive PCRs identify expressions of the T1 for OsTBZ1 in transfer-gen plant；
Fig. 4 quantitative PCRs identify expressions of the T1 for OsBBX22 in transfer-gen plant；
The relative expression quantity of Fig. 5 difference heat stress times OsTBZ1；
Wild type, the heat-resisting phenotype of transgenic line OsTBZ1-RNAi-2, Os TBZ1-RNAi-4 under Fig. 6 heat stresses；
A1, A2, A3 represent wild type, transgenic line OsTBZ1-RNAi-2, OsTBZ1-RNAi-4 under normal condition respectively；B1、
B2, B3 represent the wild type of heat stress, transgenic line OsTBZ1-RNAi-2, OsTBZ1-RNAi-4 respectively.
Specific embodiment
Below by way of specific embodiment, the present invention will be described in detail.Experimental technique in following embodiments, such as without
Specified otherwise, is conventional method.Medicinal raw material used, reagent material etc. in following embodiments, unless otherwise specified, are
Commercially available purchase product.
The OsTBZ1-RNAi vector constructions of embodiment 1
The present invention passes through NCBI (http://www.ncbi.nlm.nih.gov/) the BLAST instruments in website carry out
The homology analysis of OsTBZ1 nucleotide sequences and amino acid sequence, using Rice-map (http://www.ricemap.org/
Index.jsp) the architectural feature of web analytics OsTBZ1 genes, by with rice varieties Nipponbare (Oryz asativa L
Ssp.) genome sequence is compared, and specific regions in selection Os TBZ1 gene mRNAs, artificial synthesized OsTBZ1-RNAi passes through
Two restriction enzyme sites of speI and pstI are connected into Jin Sirui standard vectors pUC7, and what picking had synthesized contains OsTBZ1-RNAi's
The pUC57 mycelia of purpose fragment, in LB fluid nutrient mediums of the 5mL containing Amp antibiotic, overnight incubation in 37 DEG C of shaking tables.Carry again
Take pUC57 plasmids.The plasmid that will be extracted, with Spe I, I two kinds of restriction endonucleases of Pst carry out double digestion reaction.Double digestion system is shown in Table 1.
The double digestion reaction system of table 1
Mentioned reagent is sufficiently mixed, is placed in 37 DEG C in PCR instrument, reaction 2h 30min.After the completion of double digestion, by double digestion
Product carries out gel electrophoresis, the small purpose fragment of gel extraction, then carries out DNA product purifying.
[specially：The mRNAt distinguished sequence reverse complementary sequences of design and OsTBZ1, by the DNA pieces of Jin Sirui companies
The artificial synthesized technology of section obtains the special palindromic sequences of OsTBZ1 of complementary forward direction sequence+intervening sequence+complementary reverse sequence, leads to
Cross two restriction enzyme sites of speI and pstI to be connected into Jin Sirui standard vectors pUC7, by special primer (actagtgaga
gaga；Atactcctactacatgtac) PCR checkings and the sequence verification selection correct positive colony of sequencing are interfered for RNAi
(RNA interference, RNAi) vector construction.]
Purpose fragment is connected into expression vector：
Purpose fragment OsTBZ1-RNAi obtained in the previous step is connected into pCUbi1390 expression vectors as shown in Figure 1.Again
Plasmid PCR detection is carried out, reaction system is shown in Table 2.
The plasmid PCR reaction system of table 2
If it is correct to obtain purpose fragment, genes of interest OsTBZ1-RNAi is connected in pCUbi1390 expression vectors, is carried
Body is built into pUbi-OsTBZ1-RNAi expression vectors, as shown in Figure 2.
The genetic transformation of the OsTBZ1-RNAi expression vectors of embodiment 2
(1) preparation of Agrobacterium competent cell
Go bail for the Agrobacterium tumefaciems EHA105 for being stored in -80 DEG C, by it on the YEB flat boards containing rifampin (50ug/mL)
Line, is placed in 28 DEG C and cultivates 2 days.From the single bacterium colony of picking Agrobacterium on YEB flat boards, the YEB Liquid Cultures of 5mL are inoculated in
In base, shaking table 200rpm is placed in, 28 DEG C of shaken cultivations are overnight.The bacterium solution 2mL of incubated overnight is taken, the YEB liquid of 50ml is inoculated in
In culture medium, 200rpm, 28 DEG C of shaken cultivations, until OD are similarly in600Reach 0.5.During bacterium solution moved into sterile centrifugation tube,
Ice bath 30min, then 5000rpm, are centrifuged 10min, abandon supernatant, collects thalline.The thalline of collection is suspended in 10mL again,
In the NaCl of 0.15mol/L precoolings, 5000rpm is centrifuged 10min, supernatant discarded.The thalline of collection is resuspended to 1mL,
The CaCl of 20mmol/L precoolings2In solution.Agrobacterium suspension is sub-packed in 1.5mL sterile centrifugation tubes, often pipe 200uL is placed in
80 DEG C of Refrigerator stores of quick-frozen 1min in liquid nitrogen, ﹣ are with standby.
(2) expression vector is transferred to Agrobacterium competent cell
Agrobacterium EHA105 competent cells are placed on ice, the recombinant plasmid vector addition that 1uL contains genes of interest is taken
To wherein, after fully mixing, 30min is placed on ice.Snap frozen 1min in liquid nitrogen is placed on again, and water is then gone to rapidly
In bath, 37 DEG C of water-bath 3min treat that it melts.1mLYEB fluid nutrient mediums are added thereto to, after fully mixing, shaking table is put into
In, 220rpm, 28 DEG C of renewal cultivation 3h.By the bacterium solution after renewal cultivation in 5000rpm, 3min is centrifuged, collects thalline retains
200uL bacterium solutions.Bacterium solution is spread evenly across containing kanamycins (50ug/mL), the YEB solid mediums of rifampin (50ug/mL)
On, sealed after drying.Flat board is placed in incubator, is cultivated 2 days under the conditions of 28 DEG C.After Agrobacterium single bacterium colony is grown, picking
Carry out bacterium colony PCR detections.
(3) Agrobacterium bacterium colony PCR identifications
Picking Agrobacterium single bacterium colony, is detected whether correctly to be transferred in Agrobacterium tumefaciems EHA105 with colony polymerase chain reaction (PCR) method.
The agriculture bacillus mediated rice conversion of embodiment 3
(1) Fiber differentiation of Rice Callus
Choose full, without the mature seed for going mouldy, the glume of seed is removed using brown rice machine, note keeping the complete of mature embryo
Whole property.First with 70% ethanol soaking disinfection 2min, then it is soaked in 30% liquor natrii hypochloritis containing 0.1%Tween20,
It is placed in shaking table, 100rpm, vibration sterilization 30min.In superclean bench, using the seed 5 times after sterilized water cleaning and disinfecting.
Transfer them to and be covered with the culture dish of aseptic filter paper, after suck dry moisture, then mature embryo callus is inoculated in inducing culture,
In continuous light (100 μm of olm-2·s-1), cultivated under the conditions of 32 DEG C.
(2) culture of Agrobacterium
The Agrobacterium containing destination gene expression carrier of the positive will be accredited as, in the YEB culture mediums containing kana and Rif
Upper line, 28 DEG C are cultivated 2 days.The thalline that will be collected into, is washed with the appropriate AAM culture mediums for being added with AS (100 μm of ol/L).Wash
After the completion of washing, it is suspended in and is added with the AAM culture mediums of AS (100 μm of ol/L), is aggressively shaken, and adjusts bacterial concentration and be
OD600Value is between 0.3-0.5.
(3) co-cultivation of Rice Callus and Agrobacterium
Picking is induced 7 days, and the good callus of growth conditions, is put into the centrifuge tube of 50mL sterilizings.By 30ml agricultures
Bacillus soak is added thereto, and soaks 2min, and period will constantly rock centrifuge tube, bacterium solution is fully contacted with callus.Again
Callus after the completion of dip-dye is placed on aseptic filter paper, bacterium solution is blotted.Meanwhile, spread 1 on base flat board is co-cultured in advance
Aseptic filter paper is opened, and is soaked with the AAM culture mediums of 1mL, remove bubble above.The callus blotted after bacterium solution is placed in paving
Have on the co-cultivation base of filter paper, be placed under 25 DEG C of dark surrounds, cultivate 3 days.A usual expression vector needs 100 or so
Callus.Callus after co-culturing 3 days is put into the sterile centrifugation tube of 50mL.With aseptic water washing 4-5 times, until washing
Untill liquid becomes as clear as crystal.The carbenicillin solution (500mg/L) of 30mL sterilizings is added, is placed on shaking table, 100rpm shakes
Swing washing 15min.After the completion of washing, cleaning solution is outwelled, callus is placed on aseptic filter paper, blot cleaning solution.
(4) screening of resistant calli
After co-culturing, the callus of Agrobacterium wash-out is placed in the screening and culturing medium containing carbenicillin and hygromycin
On, in 32 DEG C, cultivated under continuous light, screen 14 days, every 7 days subcultures are once.2-3 circulation of screening, callus is general
In 10 days or so brownings, regrowed out resistant calli in the callus of browning.
(5) differentiation of resistant calli
The callus of the color foresythia containing resistance is transferred on the differential medium without selective agent, 28 are placed in
DEG C, cultivated under continuous light environment, break up 14 days, every 7 days subcultures are once.Typically by 15-25 days or so, kanamycin-resistant callus tissue occurs
Green bud, and then put out new shoots.Until continuing to differentiate seedling after 30-40 days.
(6) transfer-gen plant is taken root and is transplanted
By transgenic regenerated plant seedling replanting in the root media containing Basta (50mg/L), 32 DEG C are placed in, continued
Under illumination condition, cultivate 7 days.After seedling takes root and grows up to, the seedling of well developed root system is chosen, with the culture medium on warm water cleaning root
Afterwards, planted in moving to soil in greenhouse.
(7) identification of transfer-gen plant
In the extraction PCR positives T3 generations that will be obtained, turn OsTBZ1-RNAi paddy rice rice strain Ri-1, Ri-2, Ri-3 and Ri-4
Total serum IgE, reverse transcription obtains cDNA as template, and turning the primer in OsTBZ1-RNA rice strains with above T0 generations carries out sxemiquantitative
And real-time quantitative fluorescence PCR, detect expression quantity of the OsTBZ1 genes on transcriptional level in each material.Experiment is repeated 3 times, as a result
Average.It is control with wild rice (WT).Result is as shown in Figure 6, it can be seen that compared to the wild type water of non-transgenosis
In rice Nipponbare (WT), PCR positives T3 generations, turn OsTBZ1 genes in OsTBZ1-RNAi rice strains Ri-1, Ri-2, Ri-3 and Ri-4
Expression quantity significantly reduced on transcriptional level.Empty carrier paddy rice is turned using same method detection T3 generations, as a result with wild type
Paddy rice is without significant difference.
The expression of the OsTBZ1-RNAi transgenic paddy rices of embodiment 4 response heat stress and phenotypic analysis
Choose wild type, T2 ripe for homozygous transgenic mutant strain (OsTBZ1-RNAi-2, OsTBZ1-RNAi-4)
Seed is seeded in the aperture (cutting bottom) of 96 hole PCR, and every kind of strain continuously sows two rows；After planting prior to being urged in running water
Bud, seedling cultivates 20d in illumination cultivation room (28 DEG C, 12h illumination/12h is dark), treats young rice seedlings growth to the heart stage of two leaf one
After move to 45 DEG C of light temperature incubator (humidity：85%RH, illumination：10000Lx) high-temperature process 36h, respectively extract 0h, 1h, 3h,
Rice leaf total serum IgE after 5h, 12h heat treatment, the cDNA of reverse transcription synthesis analyzes OsTBZ1 genes as quantitative PCR template
Expression, as a result as figure 5 illustrates, under 0-36h hot shock conditions, compared with WT strain, the expression of Os TBZ1 turns in RNAi interference
Substantially lowered in gene plant, the signal induction and expression of wild type Os TBZ1 is heated is more sensitive to heat stress, with heat
The expression quantity of the increases Os TBZ1 of the time of swashing is in first rising the fluctuation tendency that declines afterwards, and the expression quantity highest in heat shock 1h.
Rice seedling after heat treatment is placed in 28 DEG C of room temperature condition restoration ecosystem 5d, Phenotypic Observation and analysis is carried out.Through
After crossing 45 DEG C of high-temperature process 36h, wild type Kitaake, the phenotype of transgenic line OsTBZ1-RNAi-2, OsTBZ1-RNAi-4
Obvious difference (Fig. 6) is presented, with wild type ratio, OsTBZ1-RNAi-2, OsTBZ1-RNAi-4 strain are impacted serious, blade
Wither here dehydration, it is withered turn yellow, be placed in room temperature 5d major part strains and be unable to restoration ecosystem；And though WT strain has dry in various degree
Point, blade is withered to turn yellow, but dead without plant.
SEQUENCE LISTING
<110>Agricultural University Of Hunan
<120>A kind of adjusting and controlling rice seedling stage heat resistance gene TBZ1 and application
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 1727
<212> DNA
<213>OsTBZ1 DNA sequence dnas
<400> 1
ccgcggcgcg cccagcgcct cgtggatgag tcgtggtggt aggtaggtag ctagatcgat 60
gaaggtgctg tgctccgcgt gcgaggcggc ggaggcgcgg gtgctctgct gcgccgacga 120
cgccgccctc tgcgcgcgct gcgacctcca cgtccacgcc gccaaccgcc tcgccggcaa 180
gcaccaccgc ctccccctcc tctcctcctc ctcttcttcc tcctctccct cccccccgac 240
ctgcgacatc tgccaggacg cccacgccta cttcttctgc gtcgaggacc gcgccctcct 300
ctgccgcgcc tgcgacgtcg ccgtccacac cgccaacgcc ctcgtctccg cccaccgccg 360
cttcctcctc accggcgtcc acgtcggcct tgacgccgcc gccgacgacg acgacaaaca 420
ccccccacac cccttgtcgt cgtcgctgcc gcgcaacacg gcaccgcccc cgcagccgcc 480
gccgaagcgc agcccctcgc cgatctacag cgatgacgac gtcatcgact gggccaccgg 540
tggccacgac atcggcatca ccggcaacct gcccgactgg tcgctcgtcg acgagcagtt 600
caacacccct gcgctgccgc cggtggtgac caagaccccg ccgaagcggg cctcccgtgg 660
ccccgtcacg gccggcaccg ccgcggcggt gttcggcaac ctcgccggcg gatcgccgga 720
ctggccgctc aacgagttct tcggcttcgc cgacttcagc tccggcttcg gcttcgccga 780
gaacggcacg tccaaggtca gcatccaaat tctcgtcaga tttttctttt cttttcaata 840
tctcatcaaa ttggtactac gaataatcaa aatactacta aaaaatcgat ctatttatga 900
cctcattatt cgctcatgct gatctgagcg atcgatggca tgagatattt tgcgagacac 960
agcttgagct aacgtggcgc gttccaatgg catgcaggcg gacagcggca agatcgggag 1020
catggacggc tcgccgaacg gcggcaggtc gtcgtcgtcg tcctcctcct cctccgccgc 1080
cgccgccggc ggcggcggcg gcggccagga cttcttcggc caggtgccgg aagttcactg 1140
ggccgtgccg gagctcccct cgccgcccac ggcgtcaggg ctccactggc aacgggaccc 1200
gcgctacggt ggcggcgcca ccgacgccag cgcggtgttc gtgccggaca tctcctcgcc 1260
ggagaacccc ttccgttgct tcgccgccgc cgccgccggt gaccatacca tgaaacgccg 1320
gaggagatgc taattaagaa gcgcggcaac aaaatgagag aaaaaaaatg aaagagagag 1380
aaaatgttcg agttggtaca tgtagtagga gtatttgagt gtgagattaa ttagtggtaa 1440
tccgttgaag taaattgatg tacttggcta gctagcctgc tactaggtga tcgcttactc 1500
tgtattagtg gatataatag ctttggatct gattaattag gggctgcaat gctgcatttg 1560
tgtaacgttt tgtgttagtt actactctcc tccgtcccta aatataagtg attttggctg 1620
tctaattaaa atcccttata tttagagacg gagggaatag aaattaagga cagtgaaaag 1680
aaaacgaaat aaagtggaga aaattgcaaa tcaatttgtt tcataca 1727
<210> 2
<211> 1074
<212> DNA
<213>OsTBZ1 coded sequences
<400> 2
atgaaggtgc tgtgctccgc gtgcgaggcg gcggaggcgc gggtgctctg ctgcgccgac 60
gacgccgccc tctgcgcgcg ctgcgacctc cacgtccacg ccgccaaccg cctcgccggc 120
aagcaccacc gcctccccct cctctcctcc tcctcttctt cctcctctcc ctcccccccg 180
acctgcgaca tctgccagga cgcccacgcc tacttcttct gcgtcgagga ccgcgccctc 240
ctctgccgcg cctgcgacgt cgccgtccac accgccaacg ccctcgtctc cgcccaccgc 300
cgcttcctcc tcaccggcgt ccacgtcggc cttgacgccg ccgccgacga cgacgacaaa 360
caccccccac accccttgtc gtcgtcgctg ccgcgcaaca cggcaccgcc cccgcagccg 420
ccgccgaagc gcagcccctc gccgatctac agcgatgacg acgtcatcga ctgggccacc 480
ggtggccacg acatcggcat caccggcaac ctgcccgact ggtcgctcgt cgacgagcag 540
ttcaacaccc ctgcgctgcc gccggtggtg accaagaccc cgccgaagcg ggcctcccgt 600
ggccccgtca cggccggcac cgccgcggcg gtgttcggca acctcgccgg cggatcgccg 660
gactggccgc tcaacgagtt cttcggcttc gccgacttca gctccggctt cggcttcgcc 720
gagaacggca cgtccaaggc ggacagcggc aagatcggga gcatggacgg ctcgccgaac 780
ggcggcaggt cgtcgtcgtc gtcctcctcc tcctccgccg ccgccgccgg cggcggcggc 840
ggcggccagg acttcttcgg ccaggtgccg gaagttcact gggccgtgcc ggagctcccc 900
tcgccgccca cggcgtcagg gctccactgg caacgggacc cgcgctacgg tggcggcgcc 960
accgacgcca gcgcggtgtt cgtgccggac atctcctcgc cggagaaccc cttccgttgc 1020
ttcgccgccg ccgccgccgg tgaccatacc atgaaacgcc ggaggagatg ctaa 1074
<210> 3
<211> 357
<212>Protein
<213>OsTBZ1 amino acid sequences
<400> 3
MKVLCSACEA AEARVLCCAD DAALCARCDL HVHAANRLAG KHHRLPLLSS SSSSSSPSPP 60
TCDICQDAHA YFFCVEDRAL LCRACDVAVH TANALVSAHR RFLLTGVHVG LDAAADDDDK 120
HPPHPLSSSL PRNTAPPPQP PPKRSPSPIY SDDDVIDWAT GGHDIGITGN LPDWSLVDEQ 180
FNTPALPPVV TKTPPKRASR GPVTAGTAAA VFGNLAGGSP DWPLNEFFGF ADFSSGFGFA 240
ENGTSKADSG KIGSMDGSPN GGRSSSSSSS SSAAAAGGGG GGQDFFGQVP EVHWAVPELP 300
SPPTASGLHW QRDPRYGGGA TDASAVFVPD ISSPENPFRC FAAAAAGDHT MKRRRRC 357
<210> 4
<211> 848
<212> DNA
<213>OsTBZ1-RNAi nucleotide sequences
<400> 4
actagtgaga gagaaaatgt tcgagttggt acatgtagta ggagtatttg agtgtgagat 60
taattagtgg taatccgttg aagtaaattg atgtacttgg ctagctagcc tgctactagg 120
tgatcgctta ctctgtatta gtggatataa tagctttgga tctgattaat taggggctgc 180
aatgctgcat ttgtgtaacg ttttgtgtta gttactactc tcctccgtcc ctaaatataa 240
gtgattttgg ctgtctaatt aaaatccctt atatttagag acggagggaa tagaaattaa 300
ggacagtgaa aagaaaacga aataaagtgg agaaaattgc aaatcaattt gtttcataca 360
aggtacgtag tactctacta cccatcactt tctgtgaaga cttttgctga agaaacacat 420
tagaattttg agatatttat gtgtgatcga ttgatcactt acctacttat aacatgcatc 480
atgcagggtg tatgaaacaa attgatttgc aattttctcc actttatttc gttttctttt 540
cactgtcctt aatttctatt ccctccgtct ctaaatataa gggattttaa ttagacagcc 600
aaaatcactt atatttaggg acggaggaga gtagtaacta acacaaaacg ttacacaaat 660
gcagcattgc agcccctaat taatcagatc caaagctatt atatccacta atacagagta 720
agcgatcacc tagtagcagg ctagctagcc aagtacatca atttacttca acggattacc 780
actaattaat ctcacactca aatactccta ctacatgtac caactcgaac attttctctc 840
tcctgcag 848
Claims (10)
1. a kind of adjusting and controlling rice seedling stage heat resistance gene TBZ1, it is characterised in that open reading frame sequence is：SEQ ID NO.1,
The sequence of its code area is：SEQ ID NO.2.
2. a kind of B-box zinc finger proteins of adjusting and controlling rice seedling stage Holstein Cattle, it is characterised in that with SEQ ID No：Shown in 3
Amino acid sequence.
3. a kind of carrier of the gene containing described in claim 1.
4. carrier according to claim 3, it is characterised in that it is plant expression vector, is suitable to be expressed in paddy rice.
5. carrier according to claim 4, it is characterised in that with SEQ ID No：PUbi-OsTBZ1-RNAi shown in 4
Expression vector.
6. a kind of host cell, it is characterised in that the cell contains the gene described in claim 1, or contains claim
Carrier described in 3~5 any one.
7. host cell according to claim 6, it is characterised in that it is thin that the cell is selected from Bacillus coli cells, Agrobacterium
Born of the same parents or plant cell.
8. it is a kind of influence plant seedling stage heat resistance method, it is characterised in that methods described include use any one of claim 3~5
Described carrier converts the cell or tissue of plant, and the plant cell that will convert or tissue cultivating are into plant.
9. it is according to claim 8 influence plant seedling stage heat resistance method, it is characterised in that the conversion pass through agriculture bar
Bacterium mediated method or particle bombardment are carried out.
10. the method for influence plant seedling stage heat resistance according to claim 8 or claim 9, it is characterised in that the plant is standing grain
Graminaceous plant.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102747099A (en) * | 2011-04-21 | 2012-10-24 | 华中农业大学 | Application of rice gene OsbZIP46 in heat resistance and cold resistance regulation |
| CN103848906A (en) * | 2012-12-05 | 2014-06-11 | 浙江大学 | Rice high-temperature-resistant related gene OsZFP, selection marker and separating method of related gene |
| CN103981196A (en) * | 2014-06-11 | 2014-08-13 | 湖南农业大学 | Application of PFP (Peroxidase Family Protein) gene for controlling seedling-stage heat resistance of rice |
| CN104073491A (en) * | 2014-07-08 | 2014-10-01 | 安徽省农业科学院水稻研究所 | High-temperature-induced expressed plant promoter Posheat2 and application thereof |
-
2017
- 2017-03-02 CN CN201710121075.9A patent/CN106868017B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102747099A (en) * | 2011-04-21 | 2012-10-24 | 华中农业大学 | Application of rice gene OsbZIP46 in heat resistance and cold resistance regulation |
| CN103848906A (en) * | 2012-12-05 | 2014-06-11 | 浙江大学 | Rice high-temperature-resistant related gene OsZFP, selection marker and separating method of related gene |
| CN103981196A (en) * | 2014-06-11 | 2014-08-13 | 湖南农业大学 | Application of PFP (Peroxidase Family Protein) gene for controlling seedling-stage heat resistance of rice |
| CN104073491A (en) * | 2014-07-08 | 2014-10-01 | 安徽省农业科学院水稻研究所 | High-temperature-induced expressed plant promoter Posheat2 and application thereof |
Non-Patent Citations (5)
| Title |
|---|
| GENBANK: "登录号：CAW36830 unnamed protein product", 《GENBANK》 * |
| GENBANK: "登录号：XP_015630321 B-box zinc finger protein 22", 《GENBANK》 * |
| PUZIO,P.等人: "登录号：FB790027 Sequence 9300 from Patent WO2008034648", 《GENBANK》 * |
| REUBER,T.L等: "登录号：GY458330 Sequence 6105 from patent US 8030546", 《GENBANK》 * |
| 王冬等: "水稻耐热生长的保护者——RNA解旋酶TOGR1", 《遗传》 * |
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