KR101444244B1 - Primer Set for Detecting Xanthomonas oryzae pv. oryzae and Method for Detecting by Using the Same - Google Patents

Abstract

The present invention relates to a method for treating rice blast fungus such as Xanthomonas oryzae pv. oryzae ), and the use of the primer set for detecting a pathogenic bacterium ( Xanthomonas oryzae pv. oryzae , a composition for detecting Xanthomonas oryzae pv. oryzae , and a PCR kit containing the same.
When the primer set according to the present invention is used, the pathogenic bacteria such as Xanthomonas oryzae pv. oryzae ) can be detected quickly and accurately. Thus, it is possible to effectively diagnose the infection of rice blast fungus, and thus it can contribute to prevention and control of rice blast fungus blight.

Description

TECHNICAL FIELD [0001] The present invention relates to a primer set for detecting pathogenic bacilli of rice blossom, and a method for detecting the same. oryzae and Method for Detecting by Using the Same}

The present invention relates to a method for producing rice bran blight pathogen ( Xanthomonas oryzae pv. oryzae ), a set of primers for detecting rice blast pathogens ( Xanthomonas oryzae pv. oryzae ), a method of detecting the pathogenic fungus such as Xanthomonas oryzae pv. oryzae ) and a PCR kit.

Blight of rice blight is caused by a pathogenic bacterium such as Xanthomonas oryzae pathovar oryzae ), and the damage pattern is reduced in yield and quality due to loss of photosynthesis and fruit ripening rate. Blight of rice blight occurs in most rice cultivation areas including Asia, Africa, Latin America and Australia as well as Korea, and is known to be a serious disease particularly in Southeast Asia.

Until now, there is no germicide specific to rice blast fungus, and it does not help seed quality control (QC). It is very time consuming to propagate bacterial growth rate during incubation. Previous methods for identifying and classifying pathogenic fungi of rice blight are based on the morphological, biochemical, and growth characteristics of the fungus. These methods are troublesome, complicated, and take a long time.

In recent years, rapid and simpler methods of targeting genes have been increasingly used and being newly developed. In this method, specific genes (genus-specific or pathovar-specific) A primer or a nucleic acid probe is used.

In this background, Xanthomonas oryzae pv. Systematic analysis of oryzae is made through comparison of 16S rRNA or its nucleic acid sequence. This is based on the fact that the 16S rRNA gene is highly conserved and diverse in species (Moore, ERB, M. Mau, A. Arnscheidt, EC Boottger, RA Huston, MD Collins, Y. van de Peer, R de Wachter, and KN Timmis 1996. Determination and comparison of the 16S rRNA gene sequences of species of the genus Pseudomonas (sensu stricto) and estimation of the natural intrageneric relationships. Syst. Appl. Microbiol. 19: 478-492) . However, the 16S rRNA gene has some limitations in the identification of the species because it has some similarity to some species (Fox, GE, JD Wisotzkey, and PJ Jurtshumk 1992). .. not be sufficient to guarantee species identity Int J. Syst Bacteriol 42:... 166-170; Yamamoto, S., H. Kasai, DL Arnold, RW Jackson, A. Viavian, and S. Harayama 2000. phylogeny of the genus Pseudomonas: reconstructed from the nucleotide sequences of gyrB and rpoD genes. Microbiology 146: 2385-2394).

Currently, Xanthomonas oryzae pv. oryzae is increasingly likely to occur along with global warming, and a new identification and diagnosis method that shows specific genetic differences in order to accurately identify the pathovar that is required for their prediction and prevention through the proliferation of free trade between countries Is required to be developed.

Therefore, the present invention relates to a method for producing Xanthomonas oryzae pv. a primer set capable of effectively detecting oryzae at a gene level, Xanthomonas oryzae pv. oryzae detection method, and Xanthomonas oryzae pv. oryzae, and a PCR kit.

To address the problem, the present invention has been made to a primer having the nucleotide sequence of the primers and SEQ ID NO: 2 with a nucleic acid sequence of SEQ ID NO: 1 Rice huinip diamond pathogen (Xanthomonas oryzae pv. oryzae ) detection primer set.

The present invention also includes a step of PCR amplifying a DNA extracted from a plant sample using a primer set consisting of a primer having a nucleic acid sequence of SEQ ID NO: 1 and a primer having a nucleic acid sequence of SEQ ID NO: 2 to confirm the presence or absence of an amplification product Of Xanthomonas oryzae pv. oryzae ).

The present invention also relates to a method for screening for a pathogenic bacterium belonging to the genus Xanthomonas , which comprises a primer having a nucleic acid sequence of SEQ ID NO: 1 and a primer set of a primer having a nucleic acid sequence of SEQ ID NO: oryzae pv. oryzae ).

In another aspect, the present invention rice plant comprising a primer set consisting of primers having the nucleic acid sequence of the primers and SEQ ID NO: 2 with a nucleic acid sequence of SEQ ID NO: 1 huinip diamond pathogen (Xanthomonas oryzae pv. oryzae ) detection kit.

Using the primer set according to the present invention, Xanthomonas oryzae pv. oryzae can be detected quickly and accurately with high efficiency, and it is possible to effectively diagnose the infection of rice blast fungus, thereby contributing to prevention and control of rice blight blight.

1 is a photograph showing a result of a conventional PCR reaction using a primer set (a) according to the present invention and a primer set (b) used as a comparative example. (M: size marker, 1-36: strain of the number shown in Table 1)
Fig. 2 is a photograph showing an appropriate sample position that can be taken from the transferred rice.
FIG. 3 shows the results of a real-time PCR reaction using a primer set according to the present invention.
FIG. 4 shows the result of real-time PCR to determine the detection of a specific DNA fragment from water immersed in sterilized water after immersing a portion of the leaves of the leaves infected with the pathogens of rice blast fungus using the primer set according to the present invention (1: Xanthomonas oryzae pv. or yzae genomic DNA, 2 to 12: Xanthomonas oryzae pv. oryzae , 13-15: untreated rice control, 16: no template control)

The present invention also relates to a method for screening a rice blotch pathogen ( Xanthomonas spp.) Comprising a primer having a nucleic acid sequence of SEQ ID NO: 1 and a primer having a nucleic acid sequence of SEQ ID NO: oryzae pv. oryzae ) detection primer set.

The primers having the nucleic acid sequences of SEQ ID NO: 1 or SEQ ID NO: 2 amplify a 114 bp fragment of sequence 661 to 774 of the rhs family sequence (SEQ ID NO: 3) of Xanthomonas oryzae pv. Oryzae, respectively (XOO114F; SEQ ID NO: 1) and reverse (XOO114R; SEQ ID NO: 2) primers.

The inventors of the present invention have found that Xanthomonas oryzae pv. In the course of research on effective detection methods of oryzae , we tried to quickly and accurately diagnose rice blast fungus pathogen using the method of identifying the species at the level of DNA (DNA). The NCBI (National Center for Biotechnology Information) Using the GenBank and blastn programs, Xanthomonas oryzae pv. Nucleic acid sequence information capable of specifically detecting oryzae was searched and a primer capable of detecting the specific nucleotide sequence was prepared. As a result of performing general PCR and real-time PCR amplification reaction using the prepared primer set, the primer set according to the present invention was found to be Xanthomonas oryzae pv. It was confirmed that Oryzae can be detected specifically with high sensitivity.

Xanthomonas oryzae pv. oryzae is a causative agent of rice blight blight. Using the primer set according to the present invention, other Xanthomonas No band was detected in the genus and other pathovars in the genus, and Xanthomonas oryzae pv. It is possible to detect bands specifically for Oryzae only for about 2 hours, so that it is possible to quickly and accurately determine the authenticity of the pathogenic bacterium ( Xanthomonas oryzae pv. Or yzae) at the DNA level.

Accordingly, the present invention includes a step of PCR amplifying a DNA extracted from a plant sample using a primer set consisting of a primer having a nucleic acid sequence of SEQ ID NO: 1 and a primer having a nucleic acid sequence of SEQ ID NO: 2 to confirm the presence or absence of an amplification product ( Xanthomonas oryzae pv. oryzae ).

PCR amplification of DNA extracted from a plant sample using the primer set according to the present invention revealed that Xanthomonas oryzae pv. If there is a nucleic acid amplification product specific for oryzae , the plant sample can be diagnosed as infected with rice blast fungus.

In the present invention, the term "plant sample" means a target plant to be inspected for infection with rice blast, for example, rice, but is not limited thereto.

The method of extracting DNA from a plant sample can be carried out by a conventional method known in the art, which is appropriately selected by a person skilled in the art.

In one embodiment of the present invention, the DNA extracted by a simple procedure of immersing a 1 cm * 1 cm piece of a plant specimen in sterilized water is amplified through the primer of the present invention, thereby clearly and rapidly diagnosing whether or not it is infected with rice blast fungus .

In one embodiment of the invention, the PCR amplification can utilize conventional PCR amplification reactions, preferably using conventional PCR or real-time PCR amplification , Real-time quantitative analysis is possible using real-time PCR amplification reactions. Conventional PCR amplification reaction refers to a conventional PCR amplification reaction in which a step of amplifying a specific DNA and then confirming whether or not a specific DNA is amplified is further performed, and the step of confirming amplification of the specific DNA is performed by electrophoresis The size of the amplified DNA product can be confirmed. Specific PCR amplification methods can be suitably selected and carried out by those skilled in the art.

The present invention also relates to a method for screening for a pathogenic bacterium belonging to the genus Xanthomonas , which comprises a primer having a nucleic acid sequence of SEQ ID NO: 1 and a primer set of a primer having a nucleic acid sequence of SEQ ID NO: oryzae pv. oryzae ).

In the composition of the present invention, Xanthomonas oryzae pv. and various reagents necessary for confirming the results of the experiment and the results required for detecting oryzae , such as PCR reaction mixture, restriction enzyme, agarose, buffer solution necessary for hybridization or electrophoresis, and the like.

More specifically, the present invention relates to a method for screening a rice blotch pathogen ( Xanthomonas spp.) Comprising a primer having a nucleic acid sequence of SEQ ID NO: 1 and a primer set of a primer having a sequence of SEQ ID NO: oryzae pv. oryzae ) detection kit.

The PCR kit for detection according to the present invention, in addition to the primer set described above, also includes Xanthomonas oryzae pv. a reaction buffer solution, a polymerase, a dNTP, a stabilizer and any biological or chemical reagents necessary for detecting oryzae , instructions for use, and the like. Other configurations of such kits may be suitably selected by those skilled in the art.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

< Example  1> Experimental strain

The Xanthomonas oryzae pv. or yzae were deposited in the Korean Agricultural Culture Collection (KACC) of the National Institute of Agricultural Science and Technology, the Belgian Co-ordinated Collections of Micro-organisms (BCCM ^™ ) and the National Collection of Plant Pathogenic Bacteria (NCPPB) And is summarized in Table 1.

Xanthomonas strains were grown on NA media (D - (+) - glucose 2.0%, CaCO ₃ 2.0%, Yeast extract 1.0%: 1L), Fusarium spp., Potato dextrose agar (PDA, Difco), Escherichia coli LB agar Maniatis 1989), and other bacteria were cultured on nutrient agar (NA, Difco).

No. Bacterial and Fungal Isolates Source Geographical origin rhs family gene XOO114 One Xanthomonas oryzae pv . oryzae KACC 10331 Republic of Korea + 2 Xanthomonas oryzae pv . oryzae KACC 10878 Republic of Korea + 3 Xanthomonas oryzae pv . oryzae KACC 10381 Republic of Korea + 4 Xanthomonas oryzae pv . oryzae KACC 10384 Republic of Korea + 5 Xanthomonas oryzae pv . oryzae KACC 10385 Republic of Korea + 6 Xanthomonas oryzae pv . oryzae KACC 10883 Philippines + 7 Xanthomonas oryzae pv . oryzae KACC 10885 Philippines + 8 Xanthomonas oryzae pv . oryzae MAFF 311018 Japan + 9 Xanthomonas oryzae pv . oryzae MAFF 311019 Japan + 10 Xanthomonas oryzae pv. oryzicola LMG 797 Malaysia - 11 Xanthomonas oryzae pv. oryzicola LMG 657 India - 12 Xanthomonas campestris pv. campestris ATCC 33913 United Kingdom - 13 Xanthomonas campestris pv. carotae ATCC 10547 USA - 14 Xanthomonas campestris pv. glycines LMG 7403 Zambia - 15 Xanthomonas campestris pv. pelargoni DSMZ 50857 Germany - 16 Xanthomonas campestris pv. juglandis DSMZ 1049 United Kingdom - 17 Xanthomonas campestris pv. malvacearum DSMZ 1220 Germany - 18 Xanthomonas axonopodis pv. citri KACC10443 Republic of Korea - 19 Xanthomonas axonopodis pv. dieffenbachiae LMG 695 Brazil - 20 Xanthomonas axonopodis pv. vasculorum LMG 901 Mauritius - 21 Xanthomonas axonopodis pv. begoniae LMG 551 United Kingdom - 22 Xanthomonas axonopodis pv. phaseoli LMG 7455 Belgium - 23 Xanthomonas axonopodis pv. phyllanthi LMG 844 Sudan - 24 Xanthomonas axonopodis pv. aurantifolii KACC 10161 Not known - 25 Xanthomonas axonopodis pv. vesicatoria LMG 905 Tonga - 26 Xanthomonas translucence pv. phleipratensis LMG 843 USA - 27 Xanthomonas translucence pv. cerealis LMG 679 USA - 28 Xanthomonas translucence pv. hordei LMG 882 Canada - 29 Xanthomonas arboricola pv. poinsettiicola LMG 5403 New Zealand - 30 Xanthomonas cassavae LMG 673 Malawi - 31 Xanthomonas cucurbitae LMG 8662 New Zealand - 32 Xanthomonas theicola LMG 8684 Japan - 33 Pectobacterium carotovorum subsp. carotovorum LMG 2435 Italy - 34 Pseudomonas syringae pv. tomato LMG 5093 United Kingdom - 35 Escherichia coli  ( O157 : H7 ) ATCC 35150 Not known - 36 Fusarium oxysporum f. sp . dianthia ATCC 11939 Not known -

LMG, The Belgian Co-ordinated Collections of Microorganisms (BCCM ^TM ), Belgium; NCPPB, National Collection of Plant Pathogenic Bacteria; KACC, Korean Agricultural Culture Collection, Republic of Korea, ATCC, American Type Culture Collection.

< Example  2> primer  Set production

Example 2 -One: DNA  extraction

Total DNA from Xanthomonas strains cultured on NA medium was extracted by CTAB method and total DNA from other microorganisms was extracted using Qiagen (Hilden, Germany) genomic DNA extraction kit (Genomic-tips).

Example 2 -2: Specific Nucleic acid sequence  Explore Information

Xanthomonas oryzae pv. or yzae Specifically Xanthomonas &lt; / RTI &gt; (registered at Genbank (www.ncbi.nlm.nih.gov/) of National Center for Biotechnology Information (NCBI) oryzae pv. or yzae genome information (registration number: GenBank Accession No. NC_006834, gi | 58579623: 124816-125631) In genome information, rhs family (SEQ ID NO: 3: 816 bp) was confirmed by blastn and blastx programs.

Example  2-3: primer  Set production

(5'-gatcc gctcg gtcgc aatgt g -3 ') (SEQ ID NO: 1: 21mer) and XOO114R (5'-ggctc gcccc agtcc gtcgt a-3') SEQ ID NO: 2: 21mer) was prepared.

 As a comparative example, a primer consisting of XOO290F (5'-GCGCACCGAGTATTCCTA-3 ') (SEQ ID NO: 4 18mer) and XOO290R (5'-CTTCGCCGGTCCAGATGA-3') (SEQ ID NO: 5 18mer) amplifying a 290 bp- .

The sequence of the primers is shown in Table 2 below.

SEQ ID NO: designation order One XOO114F 5'-GATCCGCTCGGTCGCAATGTG -3 ' 2 XOO114R 5'-GGCTCGCCCCAGTCCGTCGTA -3 ' 4 XOO290F 5'-GCGCACCGAGTATTCCTA-3 ' 5 XOO290R 5'-CTTCGCCGGTCCAGATGA-3 '

The primers were prepared by confirming specificity of the sequence information of GenBank YD repeat protein through blastn program.

< Example  3> primer  Identify the specificity of the set

Example 3 -1: General PCR  Amplification reaction

The primer set of the present invention consisting of SEQ ID NO: 1 (XOO114F) and SEQ ID NO: 2 (XOO114R) and the primer set of SEQ ID NO: 4 (XOO290F) and SEQ ID NO: 5 (XOO290R) Were subjected to PCR amplification.

The PCR reaction was carried out using a PTC-200 ^TM thermocycler (MJ research, Watertown, Mass.). Each reaction was performed with Tris-HCl 10 mM, KCl 50 mM, MgCl ₂ 1.5 mM, gelatin 0.01%, dNTP 0.2 mM, was carried out with an amount of total 50㎕ PCR mixture containing each of ^{(. Promega TM, Madison, Wis} ) 10pM, Taq polymerase 2unit. The amount of genomic DNA of some microorganisms added to the PCR mixture was about 50 ng. The reaction was denatured at 95 ° C for 3 minutes, and the reaction was repeated 45 times at 95 ° C for 10 seconds, 60 ° C for 20 seconds, and 72 ° C for 60 seconds, followed by reaction at 72 ° C for 7 minutes. After amplification reaction, 10 μl of the PCR product was electrophoresed on 1.5% agarose gel, stained with ethidium bromide, and confirmed on a UV transilluminator.

The results are shown in Fig.

1A is a primer set amplifying 114 bp of the present invention, and FIG. 1B is a test result of a primer set amplifying 290 bp of a comparative example.

In FIG. 1, M is a DNA size measurement marker (1 kb ladder, Gibco BRL ^TM ), and numbers 1 to 36 mean strains of the numbers shown in Table 1 above. As can be seen from Fig. 1, the band representing the PCR amplification product is Xanthomonas oryzae pv. oryzae 1 to 9, a primer set amplifying a part of the rhs family according to the present invention was identified as Xanthomonas oryzae pv. oryzae strains can be specifically detected. In particular, when the primer set of the present invention is used, a clearer band has been obtained. As a result, Xanthomonas oryzae pv. oryzae strains could be detected.

Example 3 -2: Real - time PCR  Amplification reaction

In order to measure the real-time PCR diagnostic sensitivity from the leaves, a part of the rice leaves (about 1 cm x 1 cm) was soaked in 1.0 ml of sterilized water for 30-40 minutes and then taken 2 μl of the sample immersion liquid. Real-time PCR was performed using XOO114F (SEQ ID NO: 1) and XOO114R (SEQ ID NO: 2). Appropriate sample locations that can be taken from the transferred rice are shown in FIG.

Real-time PCR amplification was performed using a CFX96 real-time PCR system (Bio-Rad laboratories, Inc., USA). Each reaction was performed with iQ ^™ SYBR® Green Supermix and SYBR green PCR Mixed solution in an amount of 20 쨉 l in total. The amount of genomic DNA of some microorganisms added to the PCR mixture was about 5 ng. The reaction was performed at 95 ° C for 3 minutes, followed by repeating the reaction at 95 ° C for 10 seconds and 60 ° C for 20 seconds for 45 times. The reaction was continued at 65 ° C from 0.5 ° C for 10 seconds and then heated to 95 ° C to perform a melting curve and a melting peak . As a result, as shown in FIG. 3, specific curves and peaks of each of the individual leaves were confirmed.

Fig. 4 shows the results obtained by using XOO114F (SEQ ID NO: 1) and XOO114R (SEQ ID NO: 2) as the primer set of the present invention, Xanthomonas oryzae pv. or yzae after real-time PCR, and 1 lane is Xanthomonas oryzae pv. or yzae genomic DNA , lanes 13-15 are Xanthomonas oryzae pv. or yzae, and lanes 2 to 12 are Xanthomonas oryzae pv. or yzae, and lane 16 is a negative control containing only distilled water. Here, in lanes 1 to 12, each Xanthomonas oryzae pv. or yzae, and amplification does not occur in lanes 13-16.

Accordingly, the primer set consisting of primers having the nucleic acid sequence of SEQ ID NO: 1 (XOO114F) primers and SEQ ID NO: 2 (XOO114R) having a nucleic acid sequence of the Xanthomonas oryzae pv. or yzae, it can be used as a PCR primer for the diagnosis of blight of rice blast. The conventional PCR amplification degree is superior to that of existing primers, but the amplification target nucleic acid sequence is 114 bp It can be seen that amplification of the target product is possible even with a real-time PCR method. Also, Xanthomonas oryzae pv. or yzae can be detected specifically and quantitatively even in a plant specimen of 1 cm x 1 cm which is infected with yzae. Therefore, it is very useful for diagnosis of rice blast fungus in future.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that such detail is solved by the person skilled in the art without departing from the scope of the invention. will be. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

<110> Rural Development Administration <120> Primer Set for Detecting Xanthomonas oryzae pv. oryzae and Method          Detecting by Using the Same <130> P110367 <160> 5 <170> Kopatentin 1.71 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> primer for detecting Xanthomonas oryzae pv. oryzae <400> 1 gatccgctcg gtcgcaatgt g 21 <210> 2 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> primer for detecting Xanthomonas oryzae pv. oryzae <400> 2 ggctcgcccc agtccgtcgt a 21 <210> 3 <211> 816 <212> DNA <213> Xanthomonas oryzae <400> 3 ttgatgattc cagcccaggt gcgtttgcaa ggaatctttg atggcaagaa tcatcgcgtg 60 ttatggaatg agtttgaaga cgacgtcgga ctcggatcat ggaccggtcg cgtcgtcaag 120 cagacgcagg cagatggtgg tgtgttgacc atcgactacg cccatgtcga aggaaacgtg 180 cttggtagat tgatcaccaa gcctgacggc agcaagcgac gcgtgacgtt tgatccggct 240 accctttatc cgaagaccga cactgctgcc tatggtacga acctcgctca gaccatcacc 300 tacgaacgca gtgccggcgg gcagatcact gcgcagatcg ccccactggg gcggcgcacc 360 gagtattcct acgatggcag cggtcgcaag acgcagatca cgatgatggc ggggaccact 420 caagcacgta ctacaatcct ggtctataaa gctaatggcg atcttgaatc cattactgac 480 ccactaaaac acaagacgac ctttggttac accagtcgtt gtctgacgtc tgtcaccaat 540 cccctgggca acgtgaccac cgcaacctgc aatgctgcag gtcaacgtct cacgctgact 600 gatgccttga atcacactac cagcttcatc tggaccggcg aagacctgac ccagatcagc 660 gatccgctcg gtcgcaatgt gcacttccgc tatgacgtgc tggggcggat gattgccgcc 720 caggacgtgc agggtaacct cacacgaatg gagtacgacg gactggggcg agccgtaaag 780 tccatcgatc cgttagttcc actgtgttac aaataa 816 <210> 4 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> primer for detecting Xanthomonas oryzae pv. oryzae <400> 4 gcgcaccgag tattccta 18 <210> 5 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> primer for detecting Xanthomonas oryzae pv. oryzae <400> 5 cttcgccggt ccagatga 18

Claims (6)

A primer set for detection of a 114 bp fragment consisting of a rhs family gene 661 to 774 of a rice bloom pathogen ( Xanthomonas oryzae pv. Oryzae ) consisting of a primer having a nucleic acid sequence of SEQ ID NO: 1 and a primer having a nucleic acid sequence of SEQ ID NO: .
1) immersing the plant specimen in sterilized water; And
2) Real-time PCR was carried out after adding the primer set of the above 1 to the sterilized water containing the sample of the step 1), and the rhs family gene 661 of the rice blotch pathogen ( Xanthomonas oryzae pv. Oryzae ) A method for detecting a 114 bp fragment consisting of the rhs family gene 661 to 774 of a rice blast fungus pathogen ( Xanthomonas oryzae pv. Oryzae ) comprising amplifying a 114 bp fragment consisting of SEQ ID NO: 774 and confirming the presence of an amplification product .
3. The method of claim 2,
A method for detecting a 114 bp fragment comprising the rhs family gene 661 to 774 of a rice blast fungus pathogen ( Xanthomonas oryzae pv. Oryzae ), wherein the plant sample is rice.
delete ( Xanthomonas oryzae pv. Oryzae ) comprising a primer set consisting of a primer having a nucleic acid sequence of SEQ ID NO: 1 and a primer set having a nucleic acid sequence of SEQ ID NO: 2. The 114 bp fragment consisting of the rhs family genes 661 to 774 .
( Xanthomonas oryzae pv. Oryzae ) comprising a primer set consisting of a primer having a nucleic acid sequence of SEQ ID NO: 1 and a primer set having a nucleic acid sequence of SEQ ID NO: 2. The 114 bp fragment consisting of the rhs family genes 661 to 774 PCR kit for detection.

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