CN104131011A - Pathogen-induced rice promoter P71Z2 and application thereof - Google Patents

Abstract

The invention relates to a pathogen-induced rice promoter P71Z2 and application thereof and belongs to the field of rice organism breeding. The invention aims to provide a DNA sequence fragment which is separated from rice, can respond to intrusion of Xanthomonas oryzae and can serve as the pathogen-induced rice promoter. The DNA sequence fragment is a transcriptional control region of rice P450 monooxygenase gene Oscyp71Z2 (NCBI registry number of Os07g0217600), and is characterized by comprising a promoter region nucleotide sequence from the first locus to the 1098th site. The DNA sequence fragment can serve as the pathogen-induced promoter or is used for related regulatory sequences of other transgenic plants and is applied to breeding disease-resistant plants.

Description

A kind ofly be subject to pathogeny evoked rice starter P71Z2 and application thereof

Technical field

The present invention relates to a kind of be subject to pathogeny evoked rice starter P71Z2 and application thereof, belong to plant gene engineering technology field.Be specifically related to clone, functional verification and the applied analysis of a rice pathogen inducible promoter P71Z2.

Background technology

Paddy rice, in growth and development process, often can be subject to the infringement of (comprising Pyricularia oryzae, bacterial leaf spot pathogenic bacteria, bacterium Population of Xanthomonas Oryzae Pv and fractilinea oryzae etc.) of various cause of diseases.With the long-term interactional process of cause of disease in, paddy rice has been evolved into Various Complex and effective disease-resistant mechanism is resisted infecting of cause of disease.The clone of disease-resistant gene and the control that is applied as rice disease provide an economical and effective and eco-friendly method.

At present, by transgenic technology, disease-resistant gene being transferred to rice cultivar susceptible and that economical character is good is the important channel that utilizes resistance resource improvement rice disease resistance.Although large quantities of disease-resistant genes is cloned and applies, because the promotor majority that is applied to transgene carrier belongs to constitutive promoter.This has not only caused the burden of plant materials, make plant in the time not needing disease resistance response occurs, express extra albumen, cause waste, and seriously also cause the disorder of plant materials physiology and metabolism, cause it and produce variation or dead, make transgenic paddy rice need in life to consume a large amount of energy.This is to utilize transgenic technology to prevent and treat the most obvious defect of rice disease.In order to address this problem, applicant attempts utilization and is subject to pathogenic bacterium inducing promoter regulation destination gene expression.

Promotor is to be positioned at structure gene upstream can be combined and form transcription initiation complex with RNA polymerase, and the expression of gene is played to the section of DNA sequence of regulating and controlling effect.A typical promotor comprises the central role such as CAAT-box and TATA-box element.TATA frame, claims again Hogness frame, and its concensus sequence is: TATAAAA or TATATAT, and formed by the sequence that is rich in G and C base pair in its both sides, near be generally all positioned at-35bp.TATA frame has determined the selection of transcripting start point, and it is the position of RNA polymerase and DNA chain combination.CAAT box, its concensus sequence is GGC (T) CAATCT.Near be generally positioned at-75bp, once the base of this frame disappearance or sudden change will cause the sharply reduction of transcribing efficiency, CAAT box may controlled the frequency of transcription initiation.Enhanser (enhancer), is called again upstream sequence far away, generally all-more than 100bp, it all has to relying on and do not rely on transcribing of TATA frame the effect of transcribing of enhancing.In addition, in different promotors, also have other different cis (cis) controlling elements, the binding site of their (trans) regulatory factors that is trans or transcriptional regulation protein.Different trans regulatory factor/transcriptional regulation proteins is specific is combined with cis-regulating element, and to the expression time of gene, space or expression amount carry out specific regulation and control.Can roughly be divided into composition type expression promoter, tissue specificity expression promoter and inducible expression promotor three major types according to function and effect mode.Just as mentioned before, composition type expression promoter orders about goal gene and continues great expression in genetically modified crops, waste crop energy also brings loss, and tissue-specific promoter specifically expresses goal gene in a certain tissue, can solve preferably this problem on the one hand.At present, inducible promoter is the promotor that has better application prospect of generally acknowledging.The promotor of induction type Cucumber to external world, as pathogen, chemical substance or adverse circumstance are reacted, and start corresponding genetic expression in plant materials.Therefore, utilizing the expression of the promoters driven resistant gene of the pathogen abduction delivering in paddy rice source, is one of optimal selection of improvement paddy rice resistance.

Summary of the invention

Technical problem

The object of the invention is the above-mentioned deficiency for prior art, provide one come from paddy rice be subject to pathogenic bacterium inducing express promotor P71Z2.

Another object of the present invention is to provide the clone of pathogenic bacterium inducing type promotor and the method for Function Identification.

Another object of the present invention is to provide the application of pathogenic bacterium inducing type promotor.

The object of the invention is to clone the promotor P71Z2DNA sequence of disease-resistant related gene Oscyp71Z2 from paddy rice, and utilize transgenosis to carry out functional verification, analyze the tissue expression pattern of this gene in paddy rice, for utilizing resistant gene improvement rice disease resistance that resource is provided simultaneously.

Technical scheme

The present invention relates to a kind of pathogeny evoked rice starter P71Z2 that is subject to, it is characterized in that: this promotor comes from Oscyp71Z2 gene transcription regulation district in the fine genome of japonica rice variety Japan, be one section of DNA that is positioned at transcription initiation site upstream 1098bp, sequence is as SEQ.ID.NO.1.

Described rice starter P71Z2 can be applied by goal of regulation and control genetic expression in paddy rice Micobial Disease resistance.

Adopt PCR (Polymerase chain reaction) technology from genome, to increase to obtain promotor P71Z2 of the present invention and any section of DNA or the section of DNA with its homology.The DNA fragmentation being cloned into is built to the genetic transformation carrier that carries reporter gene gus, utilize agriculture bacillus mediated genetic transformation method, obtain transgenic paddy rice.Utilize laboratory Inoculation Method to carry out bacterial leaf-blight to transgenic paddy rice and connect bacterium, thereby by histochemical staining method and the active ability that detects qualification promotor P71Z2 response cause of disease of GUS.

Beneficial effect

Clone and the application thereof of the rice pathogen evoked promoter P71Z2 that this institute provides, tool has the following advantages:

Obtain rice pathogen inducible promoter P71Z2 by the present invention.

Clone's promotor P71Z2 is connected to binary vector regulation and control tolerant gene expression, utilize transgenic technology to proceed to paddy rice, can be subject to induce target gene to express after pathogen infection by render transgenic paddy rice, thus the damage of avoiding plant because of overexpression target gene, self to be caused.This has also made up traditional constitutive promoter and has caused because starting by force all the time destination gene expression a series of critical defects such as be obstructed of growing.

Brief description of the drawings

Fig. 1 P71Z2 promotor clone.1, promotor P71Z2PCR amplification; 3, the checking of promotor P71Z2 expression vector pBI121/Pro double digestion; 2 and 5, DNAmarkerDL2000; 4, DNAMarker λ-HindIII.

Fig. 2 genetic transformation carrier collection of illustrative plates.LB and RB are respectively borders, T-DNA left and right; NOS, rouge alkali synthetase encoding gene terminator; NOSP, rouge alkali synthetase encoding gene promotor; NPTII is kalamycin resistance gene (marker gene); Pro, promotor P71Z2; Gus is β-glucuronidase gene (GRD beta-glucuronidase, reporter gene).

Fig. 3 P71Z2 promotor is induced by bacterial leaf spot pathogenic bacteria.R is that resisting bacterial leaf-blight turns hrf1 trans-genetic hybrid rice strain NJH12; S is susceptible wild-type paddy rice R109.

Fig. 4 Oscyp71Z2 gene expression pattern.(A) blade; (C) stipes; (D) clever shell; (G) main root; (F) lemma; (B) be and (E) square section of blade and stipes.

The GUS activity that Fig. 5 turns P71Z2 promotor paddy rice is induced by bacterial leaf spot pathogenic bacteria.

Embodiment

In embodiment, method therefor is ordinary method if no special instructions below.

(1) experimental technique

1.1 vector construction

The Oscyp71Z2 gene promoter P71Z2 sequence of predicting according to promotor on-line analysis software PROSCAN (http://bimas.dcrt.nih.gov/molbio/proscan) is as SEQ.ID.NO.1, utilize primer-design software Oligo6 design promotor P71Z2 upstream primer Up:cccaagcttGTACAAGGGCTGACAGATGGG, upstream primer adds HindIII restriction enzyme site; Downstream primer Down:cgcggatccGCCCGTAGGATCGATGTGCTGTA, downstream primer adds BamHI restriction enzyme site.Oscyp71Z2 coding region 5 ' end 1098bp sequence fragment increases from the fine genome of pattern japonica rice variety Japan with HS high-fidelity enzyme.Amplification condition: 95 DEG C of denaturation 4min; 95 DEG C of sex change 40s, 58 DEG C of renaturation 45s, 72 DEG C of extension 45s, 32 circulations; Last 72 DEG C are extended 10min.Gel electrophoresis is presented at 1098bp place feature band (Fig. 1).With after PCR product test kit purify DNA, with HindIII and the BamHI promotor PCR product D NA that double digestion is purified respectively, pBI121 binary expression vector (Shao M, WangJ, Dean RA, Lin Y, GaoX, Hu S.Expression of a harpin-encoding gene in rice confers durable nonspecific resistance to Magnaporthe grisea.Plant BiotechnolJ, 2008, 6 (1): 73-81), gel electrophoresis, cut respectively glue and reclaim promoter DNA, pBI121 large fragment DNA, T4 ligase enzyme spends the night to connect and builds pBI121::P71Z2 plasmid, transform bacillus coli DH 5 alpha (Shao M, WangJ, Dean RA, Lin Y, Gao X, HuS.Expression of a harpin-encoding gene in rice confers durable nonspecific resistance to Magnaporthe grisea.Plant BiotechnolJ, 2008, 6 (1): 73-81), picking mono-clonal, in the LB liquid nutrient medium that inoculation contains kantlex, after 37 DEG C of incubated overnight, extract plasmid respectively with HindIII and the checking of BamHI double digestion, gel electrophoresis result is presented at 1098bp place and has feature band (Fig. 1).Send subsequently the handsome Bioisystech Co., Ltd order-checking to Shanghai of 2ml bacterium liquid, result shows does not have base mispairing, now successfully builds the binary expression vector pBI121/Pro (Fig. 2) that is driven gus genetic expression by promotor P71Z2.

1.2 plasmids transform

1.2.1 plasmid transformation escherichia coli (1) recipient bacterium DH5 α is at the flat lining out of LB, 37 DEG C of activation culture; Single colony inoculation of picking is in the LB liquid nutrient medium of 20ml, and 37 DEG C of shaking culture are spent the night;

(2) by 1% inoculum size, draw 0.5ml the mother liquor of incubated overnight is inoculated into 50mlLB liquid nutrient medium, cultivate 2-3h in 37 DEG C of thermal agitations;

(3) under aseptic condition, bacterium is transferred in a sterilizing 50ml centrifuge tube, placed 10min on ice, make culture be cooled to 0 DEG C; (5-9 step below must complete under condition of ice bath.)

(4) centrifugal collection somatic cells (4,000r/min, 4 DEG C, 10min).Pour out nutrient solution, pipe is inverted to 1min so that the trace nutrient solution of final residual flows to end.

(5) 0.1mol/LCaCl of use 5ml ice precooling ₂eddy diffusion bacterial sediment, is positioned over 10min on ice.

(6) centrifugal collection thalline (4,000r/min, 4 DEG C, 10min).To going out CaCl ₂, pipe is inverted to 1min so that residual CaCl ₂flow to end.

(7) 0.1mol/LCaCl of use 1ml ice precooling ₂resuspended thalline.Now, suspension is distributed into aliquot with eppendorf pipe, every pipe 200 μ l;

(8) in every pipe, add DNA (volume≤10 μ l, DNA≤50ng) with aseptic suction nozzle, rotate gently or flick tube wall to mix content, in ice, place 30min;

(9) pipe is put into heat shock 90s in 42 DEG C of water-baths, does not shake test tube;

(10) fast test tube is transferred in ice bath, made the cooling 1-2min of cell;

(11) in every pipe, add after 800 μ lLB liquid nutrient mediums, be put into 37 DEG C of shaking table shaking culture 45min;

(12) draw the competent cell that transformed of proper volume and be added on the LB solid plate containing Km (20 μ g/ml), lightly cell is coated onto to agar plate surface equably with L glass rod;

(13) flat board being placed in to room temperature to liquid is absorbed;

(14) be inverted culture dish in 37 DEG C of cultivation 12-16h, after single bacterium colony grows, extract plasmid enzyme restriction checking.

1.2.2 plasmid transforms Agrobacterium (1) recipient bacterium EHA105 (Shao M, Wang J, Dean RA, Lin Y, Gao X, HuS.Expression of a harpin-encoding gene in rice confers durable nonspecific resistance to Magnaporthe grisea.Plant BiotechnolJ, 2008,6 (1): 73-81) at the flat lining out of YEB, 28 DEG C of activation culture;

(2) single colony inoculation of picking is in the YEB liquid nutrient medium of 2ml, and 28 DEG C of shaking culture are spent the night;

(3) in the YEB of 200ml substratum, dilute 28 DEG C of shaking culture 3-4h.

(4) under aseptic condition, bacterium is transferred in a sterilizing 50ml centrifuge tube, 3000rpm4 DEG C of centrifugal 10min, removes supernatant.

(5) wash with the TE (pH7.5) of 10ml precooling.

(6) 3000rpm4 DEG C of centrifugal 10min, removes supernatant.

(7) resuspended with 20mlYEB.

(8) by every pipe 500 μ l packing, freezing in liquid nitrogen, be competent cell.

(9) competent cell is placed in to ice bath, adds 0.5-1.0 μ g recombinant plasmid dna, place 5min on ice.

(10) put 5min in liquid nitrogen.37 DEG C of insulation 5min.

(11) with 1mlYEB dilution, jolt 2-4h at 28 DEG C.

(12) get 200 μ l and coat on the YEB flat board that contains kantlex 50mg/L+ Rifampin 50mg/L, 28 DEG C of overnight incubation are extracted plasmid checking after single bacterium colony grows.

The induction of 1.3 Mature Embryos of Rice callus and subculture

(1) the embryo grain of rice that contains of surface sterilization is inoculated on N6D2 substratum, 28 DEG C of biochemical training casees are cultivated about 2 weeks, and evoked callus produces;

(2) callus newly growing is peeled from seed, proceed on new N6D2 substratum succeeding transfer culture.Once, subculture 2 times, gets fresh light yellow callus and infects for Agrobacterium every two weeks subcultures altogether.

The common cultivation of 1.4 callus and Agrobacterium

(1) callus is immersed in resuspended Agrobacterium bacterium liquid, shake up rear standing 30min;

(2) shift out callus, with sterilizing filter paper exhaustion raffinate, tiling is to the N6D2 solid culture primary surface that fills aseptic filter paper, and by the wetting callus of 2-3mlN6D2-AS nutrient solution, 23 DEG C of biochemical cultivation cases are secretly cultivated 3d.

1.5 acquisitions turn P71Z2 promotor paddy rice

(1) select the resistant calli of 1-2mm well-grown, compact structure, go to MS ₁on-CG substratum, 26 DEG C of intelligent illumination boxs, light intensity 30000lux, breaks up 2 weeks in advance.

(2) go to division culture medium MS ₂the upper cultivation of-CG, 26 DEG C of intelligent illumination boxs, light intensity 30000lux, differentiation.Every 2 weeks subcultures once, until obtain resistant plant.

(3) the short and small seedling of growing thickly of part goes to MS ₀on G substratum, make its elongation.

(4) seedling having broken up goes on root media 1/2MSNG.Screen, take root until grow up to whole plant.

(5) rice plant complete growth is moved in sterilizing soil, 26 DEG C of intelligent illumination boxs, light intensity 30000lux, illumination 14h, dark 10h cultivates or moves into incubation growth in greenhouse.

1.6 pathogenic bacteria inoculations

Resisting bacterial leaf-blight is turned to hrf1 trans-genetic hybrid rice strain NJH12 and susceptible wild-type paddy rice R109, and (material is public, see Li Wenqi, Shao Min, Zhong Weigong, Yang Jie, Kazunori Okada, Hisakazu Yamane, Zhang Lei, Wang Guang, Wang Dong, Xiao Shanshan, Chang Shanshan, Qian Guoliang, Liu Fengquan.Ectopic expression of hrf1 enhances bacterial resistance via regulation of diterpene phytoalexins, silicon and reactive oxygen species burst in rice.PLoS ONE, 2012, 7 (9): e43914.) and to turn P71Z2 promotor paddy rice potted plant in solarium of food crop institute of academy of agricultural sciences of Nanjing Jiangsu Province, with bacterial leaf spot pathogenic bacteria (Xanthomonas oryzaepv.oryzae, Xoo) strong pathogenic strains PXO99 ^a(public, see Li Wenqi, Shao Min, Zhong Weigong, Yang Jie, Kazunori Okada, Hisakazu Yamane, Zhang Lei, Wang Guang, Wang Dong, Xiao Shanshan, Chang Shanshan, Qian Guoliang, Liu Fengquan.Ectopic expression of hrf1 enhances bacterial resistance via regulation of diterpene phytoalexins, silicon and reactive oxygen species burst in rice.PLoS ONE, 2012, 7 (9): e43914.) respectively in tillering phase and inoculation in boot stage, concrete operation method is with reference to " planting disease research method ".

1.7GUS histochemical stain

(1) get 2 1.5ml centrifuge tubes, respectively add 0.5ml1mol/LGUS staining fluid;

(2) take appropriate big or small transgenosis and wild-type rice tissue, put into respectively above-mentioned centrifuge tube;

(3) 37 DEG C of soaked overnight (or 2-5h);

(4) outwell staining fluid, add 75% ethanol decolorization; Change 75% ethanol decolorization one time;

(5) observe dyeing situation, take pictures.If there is GUS expression product, there is the blue blueness that occurs.

The configuration of GUS staining fluid: in 450mlddH2O, add the 82mg Tripotassium iron hexacyanide, 105.6mg yellow prussiate of potash, 50ml1mol/L potassium phosphate buffer (pH7.0), adds water to 500ml, and 4 DEG C store for future use.

1mol/LGUS staining fluid: dissolve 100mgX-Gluc with DMSO, then add GUS working fluid, constant volume 100ml, is distributed into tubule, is placed in-20 DEG C and stores for future use.

(2) result shows

Utilize promoter Analysis software prediction promotor P71Z2 sequence, and utilize round pcr this promotor of clone (Fig. 1) from japonica rice type species Japan is fine.Successfully build the binary expression vector (Fig. 2) for pathogenic bacterium inducing type promotor P71Z2, obtained and turn P71Z2 promotor paddy rice by agriculture bacillus mediated rice transformation method.With the strong pathogenic strains PXO99 of bacterial leaf spot pathogenic bacteria ^ainoculation resisting bacterial leaf-blight turns after hrf1 trans-genetic hybrid rice strain NJH12 and susceptible wild-type paddy rice R109, and this promotor P71Z2 is subject to bacterial leaf spot pathogenic bacteria induction (Fig. 3).Turning in P71Z2 promotor paddy rice, GUS activity (Fig. 4) mainly in root, joint and blade, detected.Turn P71Z2 promotor paddy rice and be subject to bacterial leaf spot pathogenic bacteria PXO99 ^aafter infecting, gus protein is active significantly to raise, and is approximately doubly (Fig. 5) of 2-4 that does not connect bacterium.These results show, this promotor P71Z2 can respond infecting of bacterial leaf spot pathogenic bacteria.

Annex

1, LB substratum (cultivating intestinal bacteria)

LB liquid nutrient medium composition: take Tryptones 10g; Sodium-chlor 10g; Yeast powder 5g; 500ml adds water; After heating for dissolving, be settled to 1000ml, adjust pH is to 7.0-7.2, autoclaving after every bottle of 50ml packing.LB solid medium (LA) is for adding 15g agar in every 1000ml liquid nutrient medium, add pour into after corresponding microbiotic (ammonia benzyl mycin and Totomycin) dull and stereotyped for subsequent use.

2, YEB substratum (cultivating Agrobacterium)

Liquid nutrient medium composition: take beef extract 5g; Sucrose 5g; Polyprotein peptone 5g; Magnesium sulfate 2mmol/L; Yeast powder 1g; Add 500ml water, after heating for dissolving, be settled to 1000ml, adjust pH to 7.2, autoclaving after packing.When preparation YEB solid medium, in every 1000ml liquid nutrient medium, add 15g agar, pour flat board into after adding corresponding microbiotic.

3, the culture medium prescription that transgenosis is used

Involved substratum:

N6 Media Components:

Every liter of substratum is with sucrose 30g, agar 15g, pH5.8.For the medium pH 5.2 of cultivating altogether.

MS Media Components:

Every liter of substratum adds inositol 100mg, sucrose 30g, agar 15g, pH5.8.

4, P71Z2 promoter DNA sequence

Gtacaagggctgacagatgggtactcatgacggttgtgacagttatggcggtttctaggatggtttcacctgcatgtgggcctctctatccttgagcttgcttaagagtgagtttttccttttttttcttctccattaatttatgtgctaatctctgcaaacaaataatactccaagtacaagtggaaaaatattattctaaaataaatatgcattgcaagcatagctacttcttctttattttaagtatattgatggtctaaattagtctataatgaccgtcaacaaccatcttcacgtgctataggttagctgtcagatcctttttccaaatactacttttagatcttttagatctctcttattttctctttttccataaacttatttaagagagaagacgtataaaaatatagctagaaatgtaatatgctaaccatacatcaaaatattttcacaaaaaaatttggtagcacatccattcctcctcctttcactttgtagcaatcgcttgagctggaggagcaagaatacctataaaaaggatatgacctttaatactagttggtgttgaactttagtacaccagccaatacataattaatatcacgggggcctaaaaatgatttttagaactggctaaagattttaaagtgccgttgggtattttaaccagtaataaatatcatttttagcaccgattcgttttacatccgagatatttacgattttggccgatcgagcgaagatcggttctccaatagtgctcattgattccactaattaagaaccgctaactacgttgggtttagagcccagcatatatgatgagaggttgacggttaatttagaacattagcccctagtgtgaacatagccctagagttccgtggtgggcatgctagccaccatcggtcactgtcgtatatgccgtagaagtgacggcgtctcgccaatactgttatcacgccacaacaatcgaatctctcaaaaaatctcggcaaactctcacactagaaatgctgtggctttggctcagcatcgctatttatactagtggctaccagcctactagtacagcacatcgatcctacgggc。

SEQUENCE LISTING

<110> Jiangsu Province Agriculture Science Institute

Mono-kind of <120> is subject to pathogeny evoked rice starter P71Z2 and application thereof

<130> 0

<160> 3

<170> PatentIn version 3.1

<210> 1

<211> 1098

<212> DNA

<213> Oryza sativa

<220>

<221> P71Z2 promoter DNA sequence

<222> (1)..(1098)

<223>

<400> 1

gtacaagggc tgacagatgg gtactcatga cggttgtgac agttatggcg gtttctagga 60

tggtttcacc tgcatgtggg cctctctatc cttgagcttg cttaagagtg agtttttcct 120

tttttttctt ctccattaat ttatgtgcta atctctgcaa acaaataata ctccaagtac 180

aagtggaaaa atattattct aaaataaata tgcattgcaa gcatagctac ttcttcttta 240

ttttaagtat attgatggtc taaattagtc tataatgacc gtcaacaacc atcttcacgt 300

gctataggtt agctgtcaga tcctttttcc aaatactact tttagatctt ttagatctct 360

cttattttct ctttttccat aaacttattt aagagagaag acgtataaaa atatagctag 420

aaatgtaata tgctaaccat acatcaaaat attttcacaa aaaaatttgg tagcacatcc 480

attcctcctc ctttcacttt gtagcaatcg cttgagctgg aggagcaaga atacctataa 540

aaaggatatg acctttaata ctagttggtg ttgaacttta gtacaccagc caatacataa 600

ttaatatcac gggggcctaa aaatgatttt tagaactggc taaagatttt aaagtgccgt 660

tgggtatttt aaccagtaat aaatatcatt tttagcaccg attcgtttta catccgagat 720

atttacgatt ttggccgatc gagcgaagat cggttctcca atagtgctca ttgattccac 780

taattaagaa ccgctaacta cgttgggttt agagcccagc atatatgatg agaggttgac 840

ggttaattta gaacattagc ccctagtgtg aacatagccc tagagttccg tggtgggcat 900

gctagccacc atcggtcact gtcgtatatg ccgtagaagt gacggcgtct cgccaatact 960

gttatcacgc cacaacaatc gaatctctca aaaaatctcg gcaaactctc acactagaaa 1020

tgctgtggct ttggctcagc atcgctattt atactagtgg ctaccagcct actagtacag 1080

cacatcgatc ctacgggc 1098

<210> 2

<211> 30

<212> DNA

<213> is artificial

<220>

<221> promotor P71Z2 upstream primer

<222> (1)..(30)

<223>

<400> 2

cccaagcttg tacaagggct gacagatggg 30

<210> 3

<211> 32

<212> DNA

<213> is artificial

<220>

<221> promotor P71Z2 downstream primer

<222> (1)..(32)

<223>

<400> 3

cgcggatccg cccgtaggat cgatgtgctg ta 32

Claims (3)

1. be subject to a pathogeny evoked rice starter P71Z2, it is characterized in that: this promotor comes from the fine genome of japonica rice variety Japan oscyp71Z2gene transcription regulation district, is one section of DNA that is positioned at transcription initiation site upstream 1098 bp, and sequence is as SEQ.ID.NO.1.

2. a kind of application that is subject to pathogeny evoked rice starter P71Z2 described in claim 1.

3. application according to claim 1, rice starter P71Z2 is by the application of goal of regulation and control genetic expression in paddy rice Micobial Disease resistance.