CN102181455A - Rice bacterial leaf streak resistance relevant gene NRRB and coded protein and applications thereof - Google Patents
Rice bacterial leaf streak resistance relevant gene NRRB and coded protein and applications thereof Download PDFInfo
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Abstract
The invention discloses rice bacterial leaf streak resistance relevant gene NRRB and a coded protein and applications thereof, relates to a rice gene, and provides rice bacterial leaf streak resistance relevant gene NRRB and a coded protein and applications thereof. The rice bacterial leaf streak resistance relevant gene NRRB can be used for improving the resistance of rice on the bacterial leaf streak, and culturing the rice enhancing the bacterial leaf streak resistance; the average length of the bacterial leaf streak of a ribonucleic acid (RNA) interference transgene plant strain of the rice bacterial leaf streak resistance relevant gene NRRB is obviously less than control at T0 or T1 generation; and the RNA interference transgene plant strain expressing the gene can obviously improve the resistance of the rice on the bacterial leaf streak. The invention provides an important path for culturing the rice enhancing the bacterial leaf streak resistance, and has great significance in culturing the rice enhancing the bacterial leaf streak resistance on production, reducing the use of pesticides and increasing the grain output and the like.
Description
Technical field
The present invention relates to paddy gene, particularly the sick resistance related gene NRRB of paddy rice slice and proteins encoded and application.
Background technology
Paddy rice bacterium cecospora spot (Bacterial Leaf Streak abbreviates the slice disease as) is an important disease in the Rice Production, belongs to national agricultural plants quarantine sex object.This disease in 1918 in Philippines's reported first, and be found in the Delta of the Pearl River early than nineteen fifty-three in China.The slice disease mainly is distributed in south China Dao Qu at home, also has big area to take place in recent years on the late hybrid rice of the Yangtze valley.Hybridisation rice is very responsive to paddy rice slice disease, and along with the big area of hybridisation rice is promoted, paddy rice slice disease has risen to the predominantly bacteria disease in south China, Central-South rice district, and its hazard rating has surpassed bacterial blight of rice, and the underproduction can reach 5%～25%.In addition, paddy rice bacterium cecospora spot also has generation in Southeast Asian countries and middle part, Africa.This shows that paddy rice slice disease has the characteristics wide, that harm is serious that distribute.The pathogenic bacteria of paddy rice slice disease is the living Xanthomonas campestris of rice (Xanthomonas oryzae pv.Oryzicola), belongs to Gracilicutes Xanthomonas campestris section Xanthomonas.By strengthening quarantine, stop pathogen transmission, select disease-resistant variety for use, cultivate anosis strong seedling, strengthen rich water quality management and in time carry out measure such as medication control and can prevent and control the bacterial stripe generation to a certain extent and cause harm.The sick paddy rice resource of existing anti-slice only limit to some local ancient kinds and proterties relatively poor, this has limited their application on producing.And use chemical agent to prevent and treat, and it is not ideal enough to have effect, and pathogenic bacteria is easy to generate shortcomings such as resistance and contaminate environment.Utilize biotechnology to excavate the sick resistance related gene of slice, and use it for the rice varieties improvement, cultivate the sick rice varieties of anti-slice,, improve rice yield, opened up a new road for control paddy rice slice disease.
In plant, by strengthening or disturbing the expression of certain or some genes involveds (especially resistance related gene) can improve the resistance of plant to disease.Wherein disturb the method for genetic expression to have that overexpression justice gene, gene antisense are expressed, the RNA of double chain RNA mediate disturbs and artificial Microrna interference etc. multiple, and RNA disturbs (RNAinterference, RNAi), promptly the specific gene silence of double chain RNA mediate is one of interference gene expression method of comparatively effectively and easily row.Some studies show that, express hair clip formula double-stranded RNA (hairpin RNAi) than expressing general double-stranded RNA (non-hair clip formula) and have better interference effect.The RNAi technology notable feature of this expression hair clip formula double-stranded RNA is to add an intron between sense dna sequence and antisense sequences, and make double-stranded RNA be hairpin structure through transcribing processing and the generation of splicing back, so more can effectively cause target gene mRNA degraded (1, Smith, N.A., S.P.Singh, et al.Total silencing by intron-splicedhairpin RNAs[J] .Nature.2000.407 (6802): 319-320; 2, Wesley, S.V., C.A.Helliwell, et al.Construct design for efficient, effective and high-throughput gene silencing in plants[J] .PlantJ.2001.27 (6): 581-590).
The report of RNAi technology aspect the plant trait improved application is a lot, the research report is arranged, by the RNAi technology disturb in the cotton fatty acid desaturation enzyme expression of gene can improve the Oleum Gossypii semen composition (3, Liu, Q., S.P.Singh, et al.High-stearic andHigh-oleic cottonseed oils produced by hairpin RNA-mediated post-transcriptional genesilencing[J] .Plant Physiol.2002.129 (4): 1732-1743.); Other there are some researches show, the expression that suppresses xanthosine methyl transferase gene CaMXMT1 with the RNAi technology can reduce the content of Theobromine and caffeine in the transfer-gen plant, thereby be expected to obtain decaffeinated the cocoa kind (4, Ogita, S., H.Uefuji, et al. Application of RNAi to confirmtheobromine as the major intermediate for caffeine biosynthesis in co
Summary of the invention
First purpose of the present invention is to provide the paddy rice slice sick resistance related gene NRRB.
Second purpose of the present invention is to provide the sick resistance related gene NRRB of paddy rice slice encoded protein.
The 3rd purpose of the present invention is to provide the application of the sick resistance related gene NRRB of paddy rice slice in cultivating bacterium cecospora spot resistance enhanced paddy rice.
The nucleotide sequence of the sick resistance related gene NRRB of described paddy rice slice is shown in the SEQ ID No:1 in the sequence table.
The aminoacid sequence of the sick resistance related gene NRRB of described paddy rice slice encoded protein is shown in the SEQ ID No:2 in the sequence table.
The sick resistance related gene NRRB of described paddy rice slice can be used for improving the resistance of paddy rice to the bacterium cecospora spot, cultivates bacterium cecospora spot resistance enhanced paddy rice.
Described cultivation bacterium cecospora spot resistance enhanced paddy rice can be adopted following method:
Make up the rnai expression carrier of the sick genes involved NRRB of paddy rice slice, and it is transformed into paddy rice, screening obtains bacterium cecospora spot resistance enhanced paddy rice.
Described expression vector can be Ti class plasmid vector; Described conversion can be adopted agrobacterium mediation converted method, particle gun mediated transformation method, preferred agrobacterium mediation converted method.
The bacterium cecospora spot scab mean length of the RNA interference of transgene plant strain system of the sick genes involved NRRB of the paddy rice slice among the present invention is at T
0And T
1In generation, show that significantly less than contrast the RNA interference of transgene plant of this gene can significantly improve the resistance of paddy rice to the bacterium cecospora spot.The present invention provides an important channel for cultivating bacterium cecospora spot resistance enhanced paddy rice.And cultivation bacterium cecospora spot resistance enhanced paddy rice is gone up in production, and is significant to reducing agricultural chemicals use, increase grain yield etc.
Description of drawings
Fig. 1 is that the NRRB gene disturbs the pcr amplification electrophorogram of expressing transgenic rice plant in the embodiment of the invention.In Fig. 1, M is DL 5000DNAMarker, and 1 is the wild-type rice plant, 2 positive plasmid DNA, 3～13 positive transgenic rice plants.
Fig. 2 is that the NRRB gene is at T
0And T
1For the horizontal comparison diagram of the relative expression in the rnai expression transgenic rice plant.In Fig. 2, WT is the wild-type rice plant, and R1～R11 and Ra1～Ra7 are respectively 11 strain T
0Generation and 7 strain T
1Disturb the expression transgenic rice plant for the NRRB gene; Column diagram is represented NRRB gene relative expression level, and vertical line is represented 3 technology multiple standard errors on it.
Fig. 3 is T
0And T
1Disturb the mean length comparison diagram of the bacterium cecospora spot scab of expressing transgenic rice plant and corresponding wild-type rice plant for the NRRB gene.In Fig. 3, WT is the wild-type rice plant, and R1～R11 and Ra1～Ra7 are respectively 11 strain T
0Generation and 7 strain T
1Disturb the expression transgenic rice plant for the NRRB gene, column diagram is represented the scab mean length, vertical line is represented standard error on it, and " * * " expression NRRB gene disturbs the scab mean length of expressing between transgenic rice plant and the wild-type plant to have utmost point significant difference (P＜0.01).
Fig. 4 is for meeting bacterium whole 11 strain T after 14 days
0Generation and whole 7 strain T
1In generation, disturbed the scab length comparison diagram of expressing transgenic rice plant and corresponding wild-type plant.In Fig. 4, WT is the wild-type rice plant, and RNAi expresses transfer-gen plant for the NRRB gene disturbs, and column diagram is represented the scab mean length, and vertical line is represented standard error on it; " * * " expression NRRB gene disturbs the scab mean length of expressing between transfer-gen plant and the wild-type plant to have utmost point significant difference (P＜0.01).
Fig. 5 is respectively the NRRB gene at T
0And T
1For the horizontal comparison diagram of the relative expression in the rnai expression transgenic rice plant.In Fig. 5, WT is the wild-type rice plant, and R1～R11 and Ra1～Ra7 are respectively 11 strain T
0Generation and 7 strain T
1Disturb the expression transgenic rice plant for the NRRB gene; Column diagram is represented NRRB gene relative expression level, and vertical line is represented 3 technology multiple standard errors on it.
Fig. 6 is T
0And T
1Disturb the mean length comparison diagram of the bacterium cecospora spot scab of expressing transgenic rice plant and corresponding wild-type rice plant for the NRRB gene.In Fig. 6, WT is the wild-type rice plant, and R1～R11 and Ra1～Ra7 are respectively 11 strain T
0Generation and 7 strain T
1Disturb the expression transgenic rice plant for the NRRB gene, column diagram is represented the scab mean length, vertical line is represented standard error on it, and " * * " expression NRRB gene disturbs the scab mean length of expressing between transgenic rice plant and the wild-type plant to have utmost point significant difference (P＜0.01).
Fig. 7 is for meeting bacterium whole 11 strain T after 14 days
0Generation and whole 7 strain T
1In generation, disturbed the scab length comparison diagram of expressing transgenic rice plant and corresponding wild-type plant.In Fig. 7, WT is the wild-type rice plant, and RNAi expresses transfer-gen plant for the NRRB gene disturbs, and column diagram is represented the scab mean length, and vertical line is represented standard error on it; " * * " expression NRRB gene disturbs the scab mean length of expressing between transfer-gen plant and the wild-type plant to have utmost point significant difference (P＜0.01).
Embodiment
Following examples will the present invention is further illustrated:
The acquisition of the sick resistance related gene NRRB of embodiment 1 paddy rice slice
The sick resistance related gene NRRB of paddy rice slice is the protein coding gene of a differential expression in the paddy rice of bacterium streak infection process.Concrete preparation method is, the technology of employing albumen two-dimensional electrophoresis (2-DE) and substance assistant laser desorpted ionized flight time tandem mass spectrum (Matrix assisted laser desorption ionization-time of flight-mass spectrometry) is found out the protein of differential expression from the rice protein group of bacterium streak infection process, according to the relevant rice protein database of peptide fingerprint search of these differential expression proteins, obtain to comprise the protein information of these peptide fingerprints then.By search GenBank nucleic acid sequence data storehouse, find the nucleotide sequence and the gene numbering thereof of this differentially expressed protein of coding in the Japanese fine rice genome again.
The structure of the sick resistance related gene NRRB of embodiment 2 paddy rice slices gene RNA interference expression vector
Select 5 ' end 346bp nucleotide sequence of NRRB gene cDNA, synthetic a pair of primer NRRB-f and NRRB-r:
NRRB-f：
caccttagtt?aatcagttgg?gaac；
NRRB-r：atgctcatcg?gaatcatctg；
Wherein introduce TOPO clone recognition site among the forward primer NRRB-f, so that the gene DNA segment is cloned on the pENTR/D-TOPO carrier with the TOPO cloning process.
Use above-mentioned primer, and be template, carry out PCR and expanded once, to obtain 5 ' end 346bp dna segment of NRRB gene cDNA with the genomic dna.
The amplification condition of PCR is 94 ℃ of pre-sex change 5min; 94 ℃ of sex change 30s, 60 ℃ of annealing 30s, 72 ℃ of 1min, 35 circulations; 72 ℃ are extended 5min.
Reclaim target 346bp dna segment, according to pENTR
TMDirectional TOPO
Cloning Kits (Invitrogen, USA) operation instructions of test kit is cloned this dna segment on the pENTR/D-TOPO carrier, selects positive recombinant clone and checks order, the positive colony called after pENTR34RNAi that sequencing result is correct; Extract pENTR34RNAi and rnai expression carrier pH7GWIWG2 (II) plasmid DNA, get the pENTR34RNAi of 50ng and the pH7GWIWG2 of 100ng (II) plasmid DNA, with reference to Gateway LR clonase II Enzyme Mix (Invitrogen, USA) process specifications preparation reaction system is connected in the rnai expression box by 5 ' the end 346bp dna segment of Gateway LR recombining reaction with the NRRB gene cDNA.The rnai expression box of reorganization has comprised 5 ' end 346bp sense dna segment of a NRRB gene cDNA and 5 ' end 346bp antisense DNA segment of this gene cDNA.PH7GWIWG2 (II) expression vector data is referring to document (Kafimi, M., Inz é, D., Depicker, A., Gateway vectors forAgrobacterium-mediated plant transformation.Trends Plant Sci.2002May; 7 (5): 193-195.).
Carry out the positive recombinant clone of bacterium colony PCR evaluation and screening with NRRB-f and NRRB-r primer, and with positive recombinant clone called after pH34i; Get 1 μ g pH34i plasmid DNA and transform Agrobacterium EHA105; Bacterium colony PCR identifies the Agrobacterium clone on the resistant panel, can obtain positive Agrobacterium clone, adds 20% glycerine in its bacterium liquid, be stored in-80 ℃ standby.
The sick resistance related gene NRRB of embodiment 3 paddy rice slices gene RNA disturbs expression to make up rice transformation and the PCR detection is identified
The preparation of agrobacterium suspension: get the Agrobacterium glycerine preservation liquid that 20 μ L contain the pH34i plasmid, be inoculated among the 10mL LB (containing the 50mg/L kantlex), 28 ℃ of shaking culture are spent the night, centrifugal collection thalline, resuspended with proper volume AAM nutrient solution, make agrobacterium suspension OD
600Value adds Syringylethanone at last and makes it ultimate density and reach 50mg/L between 0.3～1.
Get Japan fine or in spend mature seed No. 11, after the shelling, be chlorine bleach liquor's (reactive chlorine is about 3%) of 1/3 with volume ratio, surface sterilization 30min, usefulness sterile water wash 2～5 times; After drying on the aseptic filter paper, be inoculated in the inducing culture of N6D, evoked callus under the condition of 28 ℃ of dark, after 2 weeks with callus succeeding transfer culture on fresh inducing culture.
Get fresh eugonic callus, be immersed in the agrobacterium suspension, leave standstill 30min, remove agrobacterium suspension, callus placed on the aseptic filter paper dries, transfer to then 2N6-AS altogether on the culture medium (add 50mg/L Syringylethanone and aseptic filter paper of pad on culture medium altogether) in advance then place under 19～23 ℃ of dark conditions and cultivated altogether 3 days.After 3 days, to be total to cultured calli takes out, clean Agrobacterium with sterilized water, and place on the aseptic filter paper and dry, transfer to then on the N6D-S screening culture medium (adding 50mg/L Totomycin and 400mg/L carboxylic Bian penicillin), cultivate some weeks down in 28 ℃ of dark conditions, during most of callus brownization gradually, the small part callus can grow the more vigorous fresh resistant calli of faint yellow, loose growth.With this resistant calli at succeeding transfer culture on the screening culture medium after 1 week, be transferred on the MS-NK division culture medium, carry out differentiation culture, resistant calli can begin to differentiate green regrowth about 2 weeks, the regrowth of differentiation continues to cultivate on the MS-HF root media, treat that foundation originally grows up to, can be transplanted in the nutrition soil, transplant to the field subsequently.But wherein said AAM nutrient solution, 2N6-AS be culture medium, N6D-S screening culture medium, MS-NK division culture medium and MS-HF root culture based formulas reference (Nishimura altogether, A., I.Aichi, et al. (2006). " A protocolforAgrobacterium-mediated transformation in rice. " Nature Protocols 1 (6): 2796-2802).
Get the blade of above-mentioned conversion seedling respectively, extract genomic dna, and be template, carry out the PCR detection with following 1 couple of special primer P35s-f and NRRB-idr with the genomic dna:
P35s-f：gacgtaaggg?atgacgcaca?a；
NRRB-idr：atgctcatcg?gaacactg。
Since primer P35s-f target cauliflower mosaic virus (CaMV) 35S promoter DNA, and NRRB-idr target NRRB gene DNA, thereby can be used to detect positive transfer-gen plant, and expected results is negative when detecting the wild-type plant.After 11 strain transfer-gen plants being carried out PCR detection evaluation, the agarose gel electrophoresis result shows, the transfer-gen plant detected result of numbering 3～13 is positive, can specific amplified to the target dna band of about 420bp, and be that the detected result of PCR of template is all negative with wild-type plant (numbering 1) genomic dna, promptly can not specific amplified to the target dna band (referring to Fig. 1) of about 420bp.
Embodiment 4T
0Generation and T
1For the NRRB gene RNA disturb to express in the transgenic rice plant NRRB genetic expression and to the resistance analysis of bacterium cecospora spot
Respectively 11 strains are in together the T in boot stage
0Carry out quantitative analysis and the evaluation of bacterium cecospora spot resistance for the NRRB genetic expression of rnai expression transfer-gen plant and corresponding wild-type plant.Obtaining T
0Behind transgenic seed, get T
0Disturb the seed and the corresponding wild-type rice paddy seed of expressing transfer-gen plant for the NRRB gene RNA, respectively after planting, cultivating method is cultivated routinely, treats that plant is long to boot stage, to wild-type plant and T
1Disturb the NRRB genetic expression of expressing transfer-gen plant to carry out quantitative analysis for the NRRB gene RNA, and test the resistance of these transfer-gen plants bacterial stripe.Concrete operation method is as follows:
Before rice leaf inoculation paddy rice slice germ Xanthomonas oryzae pv.oryzicola (Xoc) RS105 bacterial strain, disturb the expression transfer-gen plant respectively to get the about 5cm blade of 3 segment lengths from wild-type plant and NRRB gene RNA respectively, and therefrom extract total RNA, with reverse transcription reagent its reverse transcription is become cDNA then.Be template with these cDNA respectively, Actin-rqF/Actin-rqR and 34real-f/34real-r carried out quantitative analysis to Actin gene in the sample and NRRB gene respectively with gene specific primer.During the analyzing gene expression level, with the Actin gene is internal standard gene, with the relative expression quantity method of calculation (Pfaffl MW.A new mathematical model for relative quantification in real-time RT-PCR[J] .Nucleic Acids Res.2001,29 (9): 2002-2007.) analyze the relative expression level of NRRB gene in different transfer-gen plants.In the quantitative analysis of genetic expression, each gene repeats fluorescent quantitative PCR experiment at least 2 times, and each experiment is provided with 3 technology and repeats.
Analyze the NRRB gene at 11 strain T by aforesaid method
0Generation and 7 strain T
1Disturb relative expression's level of expressing in transfer-gen plant and the corresponding wild-type plant for the NRRB gene RNA, the result shows that the NRRB gene is at 11 strain T
0Disturb relative expression's level of expressing in the transfer-gen plant to be about wild-type plant 0.18～0.90 times (referring to Fig. 2) for the NRRB gene RNA.The NRRB gene is at 7 strain T
1Disturb relative expression's level of expressing in the transfer-gen plant to be about wild-type plant 0.21～1.20 times (referring to Fig. 5) for the NRRB gene RNA.This shows, at 11 strain T
0Generation and 7 strain T
1Disturb in the expression transfer-gen plant for the NRRB gene RNA, the NRRB expression of gene is suppressed in various degree.
The primer sequence is:
34real-F：cttattttgc?tgcatggggt；
34real-R：ctgttttgga?cttgggcaat；
Actin-rqF：tgtatgccag?tggtcgtacc?a；
Actin-rqR：ccagcaaggt?cgagacgaa。
Before the resistance test experiments that carries out paddy rice bacterium cecospora spot, be that NA substratum (+100mg/L Rifampin) was gone up streak culture 2～3 days with bacterial stripe bacterium X.oryzae pv.oryzicola (Xoc) RS105 virulent strain at nutrient agar earlier, wash thalline with sterilized water, preparation germ suspension makes it OD
600Value reaches about 1.0, last in the suspension adding Tween-20 make it final concentration and reach 0.1%.Dip in bacterium liquid with the safety pin needle point, connect bacterium launching on the blade acupuncture treatment fully, specifically connecing the bacterium method is to connect bacterium in blade vein both sides and by leaf sheath end, intermediate ends and the acupuncture treatment of leaf end, and every blade meets bacterium 6～8 places altogether, and every strain connects bacterium altogether and handles 3～5 blades.Use sterilized water (containing 0.1%Tween-20) to replace bacterium liquid to carry out simulation process simultaneously, to carry out disease symptom relatively.Connect bacterium after 14 days, connecing the scab length of bacterium blade with the ruler measurement.Scab is the boundary with the water stain edge that can see, and the scab length data is measured and write down to all scabs that connect the bacterium blade.
The statistical study of scab length data averages the different significance of value difference with the incidental statistics program of origin 8.0 softwares or extremely significantly analyzes.To 11 strain T
0Disturb the scab length data of expressing transfer-gen plant and corresponding wild-type plant to carry out statistical study for the NRRB gene RNA, the result shows, 11 strain T
0Disturbing expression transfer-gen plant scab mean length for the NRRB gene RNA is 4.8～7.0mm, and wild-type plant scab mean length is about 7.6mm, wherein 4 strains (being numbered R5, R6, R7 and R9) T
0Scab mean length that disturb to express transfer-gen plant for the NRRB gene RNA significantly (" * ", P＜0.05) or extremely significantly (" * * ", P＜0.01) be shorter than the scab mean length (referring to Fig. 3) of wild-type plant.And whole 11 strain NRRB gene RNAs disturb the scab mean length of expressing transfer-gen plant to be about 6.3 mm, and extremely significantly (" * * ", P＜0.01) is shorter than the mean length 7.6mm (referring to Fig. 4) of wild-type plant scab.The result shows, compares T with the wild-type rice plant
0Disturb the expression transfer-gen plant that the resistance of bacterium cecospora spot is significantly strengthened for the NRRB gene RNA.
Same method statistic is analyzed wild-type plant and 7 strain T
1Disturb the scab length of expressing transfer-gen plant, 7 strain T for the NRRB gene RNA
1Disturbing expression transfer-gen plant scab mean length for the NRRB gene RNA is 5.1～8.9mm, and wild-type plant scab mean length is about 8.2mm, wherein 5 strains (being numbered Ra1-Ra5) NRRB gene RNA disturbs the scab mean length remarkable (" * " of expressing transfer-gen plant, P＜0.05) or extremely significantly (" * * ", P＜0.01) is shorter than the scab mean length (referring to Fig. 6) of wild-type plant.And whole 7 strain T
1Disturb the scab mean length of expressing transfer-gen plant to be about 6.2mm for the NRRB gene RNA, extremely significantly (" * * ", P＜0.01) is shorter than the mean length 8.2mm (referring to Fig. 7) of wild-type plant scab.The result shows, compares 7 strain T with the wild-type rice plant
1Disturb the expression transfer-gen plant that the resistance of bacterium cecospora spot is significantly strengthened for the NRRB gene RNA.
Claims (6)
1. the sick resistance related gene NRRB of paddy rice slice is characterized in that its nucleotides sequence classifies the SEQ ID No:1 in the sequence table as.
2. the sick resistance related gene NRRB of paddy rice slice encoded protein according to claim 1 is characterized in that its aminoacid sequence is the SEQ ID No:2 in the sequence table.
3. the application of the sick resistance related gene NRRB of paddy rice slice in cultivating bacterium cecospora spot resistance enhanced paddy rice according to claim 1.
4. as the application of the sick resistance related gene NRRB of paddy rice slice as described in the claim 3 in cultivating bacterium cecospora spot resistance enhanced paddy rice, it is characterized in that adopting following method:
Make up the rnai expression carrier of the sick genes involved NRRB of described paddy rice slice, and it is transformed into paddy rice, screening obtains described bacterium cecospora spot resistance enhanced paddy rice.
5. as the application of the sick resistance related gene NRRB of paddy rice slice as described in the claim 4 in cultivating bacterium cecospora spot resistance enhanced paddy rice, it is characterized in that described expression vector is a Ti class plasmid vector.
6. as the application of the sick resistance related gene NRRB of paddy rice slice as described in the claim 4 in cultivating bacterium cecospora spot resistance enhanced paddy rice, it is characterized in that described conversion adopts agrobacterium mediation converted method or particle gun mediated transformation method.
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