KR100489010B1 - Ascorbate peroxidase and peroxidase coding genes from Capsicum annuum L. cv. Hanbyul induced by plant pathogen and method for detecting plant defense response using the genes - Google Patents

Abstract

The present invention relates to a defense gene derived from red pepper induced by a plant pathogenic bacterium or plant disease resistance inducer, and a method for detecting a plant defense reaction using the gene. There is an excellent effect of providing genes encoding ascorbate peroxidase and peroxidase derived from pepper, which are induced defense genes, respectively. In addition, the present invention has an excellent effect of providing a method for searching for plant defense reactions using genes encoding ascorbate peroxidase and peroxidase derived from pepper, which is a defense gene. Therefore, the gene encoding the ascorbate peroxidase and the peroxidase derived from red pepper of the present invention can be used as a genetic material for labeling plant defense reactions and pepper molecular breeding.

Description

Ascorbate peroxidase and peroxidase genes derived from red pepper induced by phytopathogenic bacteria and methods for detecting plant defense reactions using the genes {Capsule peroxidase and peroxidase coding genes from Capsicum annuum L. cv. Hanbyul induced by plant pathogen and method for detecting plant defense response using the genes}

The present invention relates to a defense gene derived from pepper induced by plant pathogenic bacteria and a method for searching for plant defense reaction using the gene. More specifically, the present invention relates to a gene encoding ascorbate peroxidase and peroxidase obtained from red pepper induced by a plant pathogenic bacterium or plant disease resistance inducer, and a method for detecting a plant defense reaction using the genes. will be.

As part of efforts to increase yields by minimizing the decrease in yield caused by pest outbreaks of crops, research and commercialization of transgenic plants using plant disease resistant genes and molecular breeding of crops are underway. Red pepper is one of the major crops in Korea, and diseases such as late blight, anthrax, and bacterial spot pattern diseases are known during cultivation. Studies on plant defense mechanisms against the invasion of these disease-causing pathogens have revealed several enzymes and genes involved in pathogenic response. In particular, ascorbate peroxidase is a kind of antioxidant enzyme that removes hydrogen peroxide, and is known to be involved in plant defense mechanisms in response to attack of pathogens, wounds, and other environmental stresses. By understanding and using the biochemical mechanisms related to plant disease resistance, it may be possible to develop or commercially apply transformed plants to impart new disease resistance or enhance resistance to plants.

Accordingly, an object of the present invention is to isolate the asperate peroxidase-encoding CAPOA1 gene and the CAPO1 gene encoding peroxidase, and their base sequences, which are expressed in pepper by plant pathogenic bacteria or plant disease resistance inducers. Investigate the analysis and expression patterns to identify plant defense reactions.

In addition, another object of the present invention to provide a method for detecting a plant defense reaction using the above-described defense genes expressed in pepper by plant pathogenic bacteria or plant disease resistance inducer.

The object of the present invention is to isolate the protective gene from peppers infected with red pepper bacilli bacterium bacterium using differential hybridization, determine the nucleotide sequence of the selected gene, and then investigate the expression patterns of the organs in the whole red pepper plants. To investigate the total peroxidase activity and the hydrogen peroxide content in pepper leaves infected with red pepper bacterium bacterium, and to investigate the expression of the protective gene in the pepper leaves infected with red pepper bacterium bacterium or red pepper anthracnose bacterium. Treatment with hydrogen peroxide was achieved by examining the expression of the protective gene.

Hereinafter, the specific configuration of the invention will be described in detail.

The present invention is the isolation of the protective gene in peppers infected with the bacterium bacillus bacillus pathogen, the base sequence determination step; Investigation of total peroxidase activity and hydrogen peroxide content in pepper leaves infected with red pepper bacterium fungal pathogens in healthy red pepper plants; Investigating the expression of the protective gene in the pepper leaf infected with pepper blight bacteria or pepper berry anthrax; And investigating whether the defense gene is expressed by salicylic acid and hydrogen peroxide, which are plant disease resistance inducers.

Hereinafter, the specific configuration of the present invention will be described through Examples, but the scope of the present invention is not limited to these Examples.

EXAMPLE

Example 1 Isolation of Protective Genes from Peppers Infected with Red Pepper

In order to isolate the red pepper-derived protective gene induced by the red pepper bacterium bacterium of the present invention, Capsicum annuum L.cv.Hanbyul, a variety showing a resistance to the red pepper bacterium, was used. To this end, Xanthomonas campestris pv.vesicatoria strain BV5-4a, which is a non-pathogenic strain of red pepper bacilli, is inoculated into red pepper and treated with hypersensitivity. At this time, the plants were removed from the plants and the leaves were collected. MRNA was extracted from the leaves to prepare a cDNA library. To find genes related to disease resistance from the cDNA library, probes were labeled with dioxygenin (digoxygenin) from the leaves of pepper plants inoculated with non-pathogenic strains of pepper bacillus bacteria and the leaves of healthy pepper plants not inoculated. And differential hybridization was performed. As a result, clones of genes presumed to be involved in pathogenicity were obtained to determine sequencing and compared to gene databases registered in GenBank using a blast program. The gene selected above was confirmed to be a gene encoding ascorbate peroxidase, and the gene was named CAPOA1 . In addition, by determining the nucleotide sequence of another selected gene, it was confirmed that the selected gene was a gene encoding a peroxidase of the plant and named the gene CAPO1 .

As shown in FIG. 1, the CAPOA1 gene of red pepper induced by the red pepper bacilli bacterium shows a homology of about 66 to 67% with the amino acid sequence of ascorbate peroxidase of other crops, such as tobacco, cucumber, and tomato. It was.

The predicted amino acid sequence of the CAPO1 gene of peppers induced by the infection of the bacterium bacillus bacterium of FIG. 2 showed a high homology of 89% with that of tomato disease resistance-related peroxidase, 73% with peanut, and baby 49% with pole and 43% with alfalfa.

Example 2: Expression of CAPOA1 Gene and CAPO1 Gene in Healthy Pepper Plants

This study was conducted to investigate the expression patterns of organs in healthy pepper plants of CAPOA1 gene and CAPO1 gene derived from red pepper induced by red pepper bacterial infection. For this purpose, Northern blotting was performed, and the probe used was labeled with ³² P of the CAPOA1 gene. When Northern blotting was performed, it was confirmed that the same amount of RNA sample was loaded by monitoring the amount of ribosomal RNA.

As shown in FIG. 3, as a result of examining organ expression patterns of CAPOA1 gene and CAPO1 gene in healthy red pepper plants, CAPOA1 gene was strongly expressed in red berries of red pepper, but not in leaves, stems, roots, flowers, and blue berries. The CAPO1 gene was not expressed in all healthy organs such as leaves, stems, roots, flowers, green and red fruits.

Example 3 Investigation of Total Peroxidase Activity and Hydrogen Peroxide Content in Red Pepper Leaves Infected with Red Pepper Bacterial Spot Bacteria

Changes in the activity of peroxidase are known to be involved in the plant's pathogenic response. Therefore, in order to examine the activities of total peroxidase during the resistance reaction against red pepper bacterial bacterial pathogens, pathogenic and non-pathogenic strains of red pepper bacterial fungal pathogens were treated on pepper leaves to measure total peroxidase activity over time. It was.

First, to inoculate the bacteriophage bacillus bacterium, incubate the pathogenic strains Ds1 and the non-pathogenic strain Bv5-4a and vacuum infiltrate (vacuum-infiltration) the bacterium with a concentration of 1 X 10 ⁸ cfu / mL on the back of the six-leaf pepper. After the inoculation by the method was subjected to a wet treatment for 18 hours under conditions of 28 ℃, 100% relative humidity. After inoculation of the pathogen, the leaves were obtained at 2, 6, 12, 18, 24 and 30 hours to extract the total protein, and reacted for 5 minutes at 25 ° C using guaiacol, followed by 470 nm. Absorbance was measured at. The activity of peroxidase was calculated using the extinction coefficient of tetraguaiacol (26.6 X 10 ³ / mol / cm).

In addition, Northern blotting was performed by separating total RNA from the leaves obtained after inoculation of the pathogen.

As shown in Fig. 4 (a), the activity of peroxidase in pepper leaves infected with non-pathogenic strains (resistance reaction) was observed 6 hours after bacterial inoculation and peaked at 18 hours when hypersensitivity was observed in pepper leaf tissue. After that, it showed a tendency to decrease rapidly. On the other hand, pepper leaves infected with pathogenic strains (sensitivity reactions) showed a tendency to increase continuously from 18 hours to 30 hours after bacterial inoculation.

In addition, the accumulation of oxides such as hydrogen peroxide is a typical characteristic in plant tissues during hypersensitivity reactions that are plant disease resistant. Therefore, the accumulation of hydrogen peroxide over time was measured in pepper leaf tissues inoculated with non-pathogenic strains or pathogenic strains of pepper bacterial pathogens in order to examine the correlation between the bacterial disease resistance response of peppers and the hydrogen peroxide content accumulated in peppers. .

According to the method of Example 2, after inoculating the red pepper bacterium bacterium bacterium on the leaves of pepper, liquid nitrogen and 5 mL of 0.2 M HClO ₄ were added to the leaves to be pulverized, followed by ultracentrifugation to obtain a supernatant. The supernatant was neutralized to pH 7.5 using 4 M KOH, and then 200 μl of the supernatant obtained by centrifugation was added to the anion-exchange column filled with AG1 X 8 resin to add 800 μl of sterilized water. I was. The resulting 1 mL sample contained 400 μl of 12.5 mM 3-dimethylaminobenzoic acid and 80 μl of 1.3 mM 3-methyl-2-benzothiazoline hydrazone (3-methyl-2-benzothiazoline hydrazone ), 20 μl of peroxidase (0.25 unit) was added, and the absorbance of 590 nm was measured at 25 ° C. to calculate the amount of hydrogen peroxide.

As shown in Fig. 4 (b), after inoculation of the pathogenic strain and the non-pathogenic strain, respectively, a slight accumulation of hydrogen peroxide was observed at 2 hours after inoculation, and then gradually decreased after 6 hours and 12 hours, respectively. On the other hand, 12 hours after bacterial inoculation, hydrogen peroxide accumulated rapidly in incompatible reactions (non-pathogenic strains), and about 1200 nmol / g of hydrogen peroxide accumulated in 24 and 30 hours, and gradually increased in friendly reactions (pathogenic strains). About 30 nmol / g of hydrogen peroxide accumulated after 30 hours of inoculation.

On the other hand, red pepper leaves were infected with red pepper bacillus bacteria and observed the expression of CAPOA1 gene and CAPO1 gene.

As shown in FIG. 4 ( c), the expression of strong CAPOA1 gene was observed from 2 hours after bacterial inoculation in both friendly and incompatible reactions. However, after 18 hours after inoculation, the CAPOA1 gene was not expressed in an incompatible reaction, but continued to be expressed in a friendly response.

The CAPO1 gene was strongly expressed at 6 hours after bacterial inoculation in both friendly and incompatible reactions. However, after 18 hours after inoculation, the CAPO1 gene was not expressed in an incompatible reaction, but continued to be expressed in a friendly response.

Example 4 Expression of Protective Genes in Pepper Leaves Infected with Pepper Blight

In order to determine whether the protective genes induced during the infection of the bacterium bacillus bacillus were also expressed in pepper leaves infected with the pepper bacterium, the pathogenic strain S197 and non-pathogenic strain CBS178.26 of the pepper bacterium were incubated at 28 ° C. for 7 days, followed by jujube sac. After forming the pasteurized water and low-temperature treatment at 4 ℃ and incubated at room temperature so that the strainer flows out of the sachet. A 1 × 10 ⁵ strainer / mL concentration of the strainer was inoculated by immersing it in a sterile cotton pad and wound it on a stem of a wounded 6-leafed pepper. After inoculation of pepper blight, 12, 24, 36 and 48 hours later, the inoculated pepper stalks were taken to extract total RNA and subjected to northern blotting.

Figure 5 (a) shows the expression of the CAPOA1 gene and CAPO1 gene in the friendly reaction appearing in the pepper stem inoculated with the pathogenic strains of pepper blight and non-pathogenic strains. In the friendly response, the CAPOA1 gene was continuously expressed from 12 hours after inoculation, and the incompatible response was strongly expressed at 24 and 36 hours and then decreased. The expression of CAPO1 gene was not observed in the friendly response, but in the incompatible reaction, expression was observed from 12 hours after inoculation and strongly expressed at 36 and 48 hours.

Example 5 Expression of Protective Genes in Pepper Leaves Infected with Red Pepper Anthrax

In the pepper leaves infected with the pepper anthrax aneurysm, it was examined whether the protective genes induced during the infection of the bacterium bacilli.

To inoculate red pepper anthracnose bacterium, strain C.C.C.C.2, which causes anthrax, was incubated at 28 ° C. for 5 days. After collecting conidia, 40 μl each of 1 × 10 ⁶ conidia / mL suspension was used. It was inoculated by dropping it onto the blue and red fruits of red pepper. After incubation in a humid condition at 28 ℃, 12, 24, 48, 72 hours after the inoculated fruit parts were extracted, total RNA was extracted and Northern blotting was performed.

5 (b) shows the expression patterns of the CAPOA1 gene and the CAPO1 gene after inoculating the strain CHC2 causing anthrax to the green fruit of red pepper to the blue fruit and red fruit of red pepper. In immature blue fruit, strong expression of CAPOA1 gene was continuously induced after 12 hours, but in mature red fruit, the expression of CAPOA1 gene in healthy fruit decreased rather after anthrax inoculation and showed a strong expression at 48 hours. Seemed. CAPO1 gene was not expressed in the immature green fruit is the CAPO1 genes weakly expressed strongly expressed after 12 hours is induced at 24 hours and 48 hours showed me a red fruit maturity decreased second to 72 hours.

Example 6: Expression of CAPOA1 and CAPO1 Genes of Red Pepper by Salicylic Acid and Hydrogen Peroxide Treatment

To be involved in plant disease resistance responses induced by treatment with hydrogen peroxide, salicylic acid, known in pepper leaves was investigated whether expression of the genes and patterns of CAPOA1 CAPO1 gene. First, 5 mM of salicylic acid was sprayed onto 6 leaf pepper leaves and treated leaves were obtained after 1, 2, 6, 12, 18, 24 hours, and total RNA was extracted.

In addition, 0.001, 0,01, 0.1, 1, 5, 10 mM hydrogen peroxide was treated by vacuum infiltration method on 6-leaf pepper leaves after 18 hours to obtain the treated leaves, and 10 mM The pepper leaves treated with hydrogen peroxide were harvested after 0.25, 0.5, 1, 2, 6, 12, 18, 24 hours, and total RNA was extracted from each sample. Northern blotting was performed using the RNA obtained above.

As shown in Fig. 6, since a high concentration of oxides in the plant is harmful to the plant, the plant has its own antioxidant mechanism. Ascorbate peroxidase, encoded by the CAPOA1 gene, is known to play an important role in the removal of oxides such as hydrogen peroxide. Northern blotting was performed to observe the expression pattern of CAPOA1 gene when the pepper leaves were treated with hydrogen peroxide by time or concentration. As a result, strong expression of CAPOA1 gene was observed 1 hour after hydrogen peroxide treatment, and continuous expression was observed up to 24 hours thereafter. It also showed strong expression when treated with hydrogen peroxide of 10 M or more. The strong expression of CAPO1 gene was observed after 12 hours of hydrogen peroxide treatment, and strong expression was observed at 10 M or more hydrogen peroxide treatment.

As shown in Figure 7, salicylic acid was treated to pepper leaves, the expression of the CAPOA1 gene was induced 6 hours after the treatment was observed to be strongly expressed at 18 hours and 24 hours. CAPO1 gene was observed weak expression of CAPO1 gene 18 hours and 24 hours after treatment.

As described above, the present invention provides an excellent effect of providing genes encoding ascorbate peroxidase and peroxidase, respectively, derived from red pepper, which are defense genes induced by plant pathogenic bacteria or plant disease resistance inducers. There is. In addition, the present invention has an excellent effect of providing a method for searching for plant defense reactions using genes encoding ascorbate peroxidase and peroxidase derived from pepper, which is a defense gene. Therefore, the present invention can be used as a genetic material for the labeling of plant defense reaction and pepper molecular breeding using genes encoding ascorbate peroxidase and peroxidase derived from red pepper, which is a very useful invention in molecular breeding industry.

Figure 1 shows the nucleotide sequence of the cloned CAPOA1 gene obtained from the Capsicum annuum L. cv. Hanbyul varieties infected with red pepper bacilli, and its expected amino acid sequence.

Figure 2 shows the nucleotide sequence of the cloned CAPO1 gene obtained from the red pepper varieties infected with red pepper fungal bacillus bacteria and its expected amino acid sequence.

Figure 3 shows the expression pattern of CAPOA1 gene and CAPO1 gene in each organ of healthy pepper plants.

Figure 4 shows the activity of total peroxidase, accumulation of hydrogen peroxide (H ₂ O ₂ ) and mRNA expression patterns of CAPOA1 gene and CAPO1 gene according to the treatment of pathogenic and non-pathogenic strains of red pepper bacterial spot pattern bacteria on the leaves .

Figure 5 shows the expression pattern of the CAPOA1 gene and CAPO1 gene by treating the pathogenic strain and non-pathogenic strains of the causal pepper bacterium on the stems of the Korean pepper, respectively.

Figure 6 shows the hydrogen peroxide treatment concentration and time- dependent expression of CAPOA1 gene and CAPO1 gene according to the treatment of hydrogen peroxide on the leaves of Hanseol pepper.

Figure 7 shows the time- dependent expression of the CAPOA1 gene and CAPO1 gene according to the treatment of salicylic acid on the leaves of Hanseol pepper.

<110> Korea University Foundation <120> The defense genes from Capsicum annuum L.cv.Hanbyul induced by plant pathogen and method for detection plant defense response using the gene <160> 4 <170> KopatentIn 1.71 <210> 1 <211> 1138 <212> DNA <213> Capsicum annuum <220> <221> CDS (222) (24) .. (884) <400> 1 gcacgaggtc ggttctctct cca atg gcg aag cca att gtc gat act 47 Met Ala Lys Pro Ile Val Asp Thr 1 5 gaa tac atc aaa gaa att gag aaa gct cgt cgc gac ctc cgc gct ctc 95 Glu Tyr Ile Lys Glu Ile Glu Lys Ala Arg Arg Asp Leu Arg Ala Leu 10 15 20 atc tcc aac aaa aac tgt gct cct atc atg ctt cgc tta gca tgg cac 143 Ile Ser Asn Lys Asn Cys Ala Pro Ile Met Leu Arg Leu Ala Trp His 25 30 35 40 gat gca gga acg tat gat gct aag tca aag act ggt gga ccg aat ggt 191 Asp Ala Gly Thr Tyr Asp Ala Lys Ser Lys Thr Gly Gly Pro Asn Gly 45 50 55 tct atc aga aat gag gaa gaa ttc act cac ggt gct aat aat ggc ttg 239 Ser Ile Arg Asn Glu Glu Glu Phe Thr His Gly Ala Asn Asn Gly Leu 60 65 70 aaa atc gcc ctt gat ttc tgt gaa gca gtg aag tcc aaa cat cca aag 287 Lys Ile Ala Leu Asp Phe Cys Glu Ala Val Lys Ser Lys His Pro Lys 75 80 85 att aca tat gca gat ctg tac cag ctt gca cgt gtt gtt gct gtt gag 335 Ile Thr Tyr Ala Asp Leu Tyr Gln Leu Ala Arg Val Val Ala Val Glu 90 95 100 gtc act ggt ggt cct act att gat ttt gtc cct ggt agg aag gat tcc 383 Val Thr Gly Gly Pro Thr Ile Asp Phe Val Pro Gly Arg Lys Asp Ser 105 110 115 120 agt att tct cca aag gaa ggg cgg tta cca gat gct aaa caa ggt gtg 431 Ser Ile Ser Pro Lys Glu Gly Arg Leu Pro Asp Ala Lys Gln Gly Val 125 130 135 cca cat ctt aaa gat gtc ttt tat agg atg ggt ttg tct gac aaa gat 479 Pro His Leu Lys Asp Val Phe Tyr Arg Met Gly Leu Ser Asp Lys Asp 140 145 150 ata gtg gcg cta tca ggg ggt cat aca ctg gga agg gca cat cca gag 527 Ile Val Ala Leu Ser Gly Gly His Thr Leu Gly Arg Ala His Pro Glu 155 160 165 aga tca ggc ttt gat ggt cca tgg aca aag gag cct ctg aaa ttt gac 575 Arg Ser Gly Phe Asp Gly Pro Trp Thr Lys Glu Pro Leu Lys Phe Asp 170 175 180 aat tca tat ttt gtg gag ctg ctt aag ggg gaa agt gag ggc cta ctg 623 Asn Ser Tyr Phe Val Glu Leu Leu Lys Gly Glu Ser Glu Gly Leu Leu 185 190 195 200 aaa cta ccc aca gac att gct tta ttg gat gat cct gag ttc aga cac 671 Lys Leu Pro Thr Asp Ile Ala Leu Leu Asp Asp Pro Glu Phe Arg His 205 210 215 tat gtc gag ctg tat gct aag gat gaa gat gcc ttc ttt agg gat tac 719 Tyr Val Glu Leu Tyr Ala Lys Asp Glu Asp Ala Phe Phe Arg Asp Tyr 220 225 230 gcc ata tca cac aag aaa ctg tct gag cta ggg ttt acc cca agt tct 767 Ala Ile Ser His Lys Lys Leu Ser Glu Leu Gly Phe Thr Pro Ser Ser 235 240 245 ggt tcc aag gcc aca gtg agg gat ggt act att tta gca caa agt gct 815 Gly Ser Lys Ala Thr Val Arg Asp Gly Thr Ile Leu Ala Gln Ser Ala 250 255 260 gta gga gtt gta gtg gct gct gca gtt gtg gca ctg agt tac tgg tat 863 Val Gly Val Val Val Ala Ala Ala Val Val Ala Leu Ser Tyr Trp Tyr 265 270 275 280 gaa gtc cgg aaa aag atg aag taaatc gtacgaccac attcttatct 910 Glu Val Arg Lys Lys Met Lys 285 ccatatttac tactacgtac tcttcaatga taaaagagga ccttattttt atggttataa 970 cacactttcc tcaaataata gcacgtagtg tcgactaaat ggaattagta caaaaatata 1030 gtaagaaact ggttgtaccc ttggaatata tgtttatgtt tgatacccat actttggagg 1090 gtttcttaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaa 1138 <210> 2 <211> 287 <212> PRT <213> Capsicum annuum <400> 2 Met Ala Lys Pro Ile Val Asp Thr Glu Tyr Ile Lys Glu Ile Glu Lys 1 5 10 15 Ala Arg Arg Asp Leu Arg Ala Leu Ile Ser Asn Lys Asn Cys Ala Pro 20 25 30 Ile Met Leu Arg Leu Ala Trp His Asp Ala Gly Thr Tyr Asp Ala Lys 35 40 45 Ser Lys Thr Gly Gly Pro Asn Gly Ser Ile Arg Asn Glu Glu Glu Phe 50 55 60 Thr His Gly Ala Asn Asn Gly Leu Lys Ile Ala Leu Asp Phe Cys Glu 65 70 75 80 Ala Val Lys Ser Lys His Pro Lys Ile Thr Tyr Ala Asp Leu Tyr Gln 85 90 95 Leu Ala Arg Val Val Ala Val Glu Val Thr Gly Gly Pro Thr Ile Asp 100 105 110 Phe Val Pro Gly Arg Lys Asp Ser Ser Ile Ser Pro Lys Glu Gly Arg 115 120 125 Leu Pro Asp Ala Lys Gln Gly Val Pro His Leu Lys Asp Val Phe Tyr 130 135 140 Arg Met Gly Leu Ser Asp Lys Asp Ile Val Ala Leu Ser Gly Gly His 145 150 155 160 Thr Leu Gly Arg Ala His Pro Glu Arg Ser Gly Phe Asp Gly Pro Trp 165 170 175 Thr Lys Glu Pro Leu Lys Phe Asp Asn Ser Tyr Phe Val Glu Leu Leu 180 185 190 Lys Gly Glu Ser Glu Gly Leu Leu Lys Leu Pro Thr Asp Ile Ala Leu 195 200 205 Leu Asp Asp Pro Glu Phe Arg His Tyr Val Glu Leu Tyr Ala Lys Asp 210 215 220 Glu Asp Ala Phe Phe Arg Asp Tyr Ala Ile Ser His Lys Lys Leu Ser 225 230 235 240 Glu Leu Gly Phe Thr Pro Ser Ser Gly Ser Lys Ala Thr Val Arg Asp 245 250 255 Gly Thr Ile Leu Ala Gln Ser Ala Val Gly Val Val Val Ala Ala Ala 260 265 270 Val Val Ala Leu Ser Tyr Trp Tyr Glu Val Arg Lys Lys Met Lys 275 280 285 <210> 3 <211> 1222 <212> DNA <213> Capsicum annuum <220> <221> CDS (222) (62) .. (1057) <400> 3 gcacgaggaa agagtataga attgtattat cataaatagt agcagtacat aatcaccatt 60 t atg gag tac tac tat aat tac aat tca att aac aaa atg gtt 103 Met Glu Tyr Tyr Tyr Asn Tyr Asn Ser Ile Asn Lys Met Val 1 5 10 tcg att att ttt att ctc gtg cta gca att gat ttg act atg gta tta 151 Ser Ile Ile Phe Ile Leu Val Leu Ala Ile Asp Leu Thr Met Val Leu 15 20 25 30 ggc caa ggg act cgt gtt gga ttt tat tcc agt acg tgc cca aga gct 199 Gly Gln Gly Thr Arg Val Gly Phe Tyr Ser Ser Thr Cys Pro Arg Ala 35 40 45 gaa tcc att gtt caa tca acg gtg agg tct cat ttt cag tct gat cca 247 Glu Ser Ile Val Gln Ser Thr Val Arg Ser His Phe Gln Ser Asp Pro 50 55 60 acg gtg gcg cca gga ttg ctg aca atg cac ttc cat gat tgt ttc gta 295 Thr Val Ala Pro Gly Leu Leu Thr Met His Phe His Asp Cys Phe Val 65 70 75 caa ggt tgt gat gct tcc atc ctc att tct ggt tct ggc act gag aga 343 Gln Gly Cys Asp Ala Ser Ile Leu Ile Ser Gly Ser Gly Thr Glu Arg 80 85 90 aca gct ccc cca aat tca ttg ttg aga gga tac gag gtt atc gat gat 391 Thr Ala Pro Pro Asn Ser Leu Leu Arg Gly Tyr Glu Val Ile Asp Asp 95 100 105 110 gct aag cag caa att gaa gct att tgt cca gga gtt gtc tcc tgt gct 439 Ala Lys Gln Gln Ile Glu Ala Ile Cys Pro Gly Val Val Ser Cys Ala 115 120 125 gac att ctt gct ctt gct gcc cgt gat tcc gtt ctt gtg act aaa gga 487 Asp Ile Leu Ala Leu Ala Ala Arg Asp Ser Val Leu Val Thr Lys Gly 130 135 140 ttg acc tgg tct gtg ccc acg ggg cgc agg gat gga cta gtt tca aga 535 Leu Thr Trp Ser Val Pro Thr Gly Arg Arg Asp Gly Leu Val Ser Arg 145 150 155 gca tca gac acg tct gat ttg cca ggc ttt act gag tct gtt gat tct 583 Ala Ser Asp Thr Ser Asp Leu Pro Gly Phe Thr Glu Ser Val Asp Ser 160 165 170 caa aag caa aag ttt tct gca aag ggt ctc aac act caa gat ctc gtc 631 Gln Lys Gln Lys Phe Ser Ala Lys Gly Leu Asn Thr Gln Asp Leu Val 175 180 185 190 acc ctt gtt ggt ggg cac aca att ggg acc tca gca tgc caa ttc ttc 679 Thr Leu Val Gly Gly His Thr Ile Gly Thr Ser Ala Cys Gln Phe Phe 195 200 205 agc tac agg cta tac aat ttc aac tcc act ggt ggc cct gac cct tca 727 Ser Tyr Arg Leu Tyr Asn Phe Asn Ser Thr Gly Gly Pro Asp Pro Ser 210 215 220 atc gac gca tcc ttt ctt cct acg ctt cga gga tta tgt cca caa aac 775 Ile Asp Ala Ser Phe Leu Pro Thr Leu Arg Gly Leu Cys Pro Gln Asn 225 230 235 gga gat ggc tca aag cgt gtg gca ctg gac act gga agc gtg aac aat 823 Gly Asp Gly Ser Lys Arg Val Ala Leu Asp Thr Gly Ser Val Asn Asn 240 245 250 ttt gac acc tcg tac ttc tct aac ttg agg aat ggt cgc gga att ctg 871 Phe Asp Thr Ser Tyr Phe Ser Asn Leu Arg Asn Gly Arg Gly Ile Leu 255 260 265 270 gaa tca gac cag aaa ttg tgg act gat gat tcc acc aag gtg ttt atc 919 Glu Ser Asp Gln Lys Leu Trp Thr Asp Asp Ser Thr Lys Val Phe Ile 275 280 285 caa agg tat tta ggg ctc agg gga ttt ctt gga ttg aga ttt ggt gtc 967 Gln Arg Tyr Leu Gly Leu Arg Gly Phe Leu Gly Leu Arg Phe Gly Val 290 295 300 gaa ttt gga agg tcg atg gtt aaa atg agc aat att gaa gtt aag act 1015 Glu Phe Gly Arg Ser Met Val Lys Met Ser Asn Ile Glu Val Lys Thr 305 310 315 ggg act aat ggt gaa att cgt aaa gtt tgc tca gca atc aac tga 1060 Gly Thr Asn Gly Glu Ile Arg Lys Val Cys Ser Ala Ile Asn 320 325 330 ttcataatat catcctctta tttatgtact taattacact ctttttacat aggttcgttc 1120 atgttagcgc aagctgcaaa gtcgaattgt tatagtggta tccaataata tcaataaata 1180 aaagcattta ccgagtgaac ttgcaaaaaa aaaaaaaaaa aa 1222 <210> 4 <211> 332 <212> PRT <213> Capsicum annuum <400> 4 Met Glu Tyr Tyr Tyr Asn Tyr Asn Ser Ile Asn Lys Met Val Ser Ile 1 5 10 15 Ile Phe Ile Leu Val Leu Ala Ile Asp Leu Thr Met Val Leu Gly Gln 20 25 30 Gly Thr Arg Val Gly Phe Tyr Ser Ser Thr Cys Pro Arg Ala Glu Ser 35 40 45 Ile Val Gln Ser Thr Val Arg Ser His Phe Gln Ser Asp Pro Thr Val 50 55 60 Ala Pro Gly Leu Leu Thr Met His Phe His Asp Cys Phe Val Gln Gly 65 70 75 80 Cys Asp Ala Ser Ile Leu Ile Ser Gly Ser Gly Thr Glu Arg Thr Ala 85 90 95 Pro Pro Asn Ser Leu Leu Arg Gly Tyr Glu Val Ile Asp Asp Ala Lys 100 105 110 Gln Gln Ile Glu Ala Ile Cys Pro Gly Val Val Ser Cys Ala Asp Ile 115 120 125 Leu Ala Leu Ala Ala Arg Asp Ser Val Leu Val Thr Lys Gly Leu Thr 130 135 140 Trp Ser Val Pro Thr Gly Arg Arg Asp Gly Leu Val Ser Arg Ala Ser 145 150 155 160 Asp Thr Ser Asp Leu Pro Gly Phe Thr Glu Ser Val Asp Ser Gln Lys 165 170 175 Gln Lys Phe Ser Ala Lys Gly Leu Asn Thr Gln Asp Leu Val Thr Leu 180 185 190 Val Gly Gly His Thr Ile Gly Thr Ser Ala Cys Gln Phe Phe Ser Tyr 195 200 205 Arg Leu Tyr Asn Phe Asn Ser Thr Gly Gly Pro Asp Pro Ser Ile Asp 210 215 220 Ala Ser Phe Leu Pro Thr Leu Arg Gly Leu Cys Pro Gln Asn Gly Asp 225 230 235 240 Gly Ser Lys Arg Val Ala Leu Asp Thr Gly Ser Val Asn Asn Phe Asp 245 250 255 Thr Ser Tyr Phe Ser Asn Leu Arg Asn Gly Arg Gly Ile Leu Glu Ser 260 265 270 Asp Gln Lys Leu Trp Thr Asp Asp Ser Thr Lys Val Phe Ile Gln Arg 275 280 285 Tyr Leu Gly Leu Arg Gly Phe Leu Gly Leu Arg Phe Gly Val Glu Phe 290 295 300 Gly Arg Ser Met Val Lys Met Ser Asn Ile Glu Val Lys Thr Gly Thr 305 310 315 320 Asn Gly Glu Ile Arg Lys Val Cys Ser Ala Ile Asn 325 330

Claims (3)

A gene encoding the peroxidase derived from red pepper, which is represented by the nucleotide sequence set forth in SEQ ID NO: 3 of the Sequence Listing and expressed by a plant pathogenic bacterium or a plant disease resistance inducer. [Claim 2] The gene encoding peroxidase derived from red pepper according to claim 1, wherein the phytopathogenic bacterium is any one of red pepper bacilli, red pepper germ disease, and red pepper anthrax. [Claim 2] The gene encoding peroxidase derived from red pepper according to claim 1, wherein the plant disease resistance inducer is either salicylic acid or hydrogen peroxide.