Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지
DOI QR코드

DOI QR Code

PCR-Based Assay for Rapid and Specific Detection of the New Xanthomonas oryzae pv. oryzae K3a Race Using an AFLP-Derived Marker

  • Song, Eun-Sung (National Academy of Agricultural Science, Rural Development Administration) ;
  • Kim, Song-Yi (National Academy of Agricultural Science, Rural Development Administration) ;
  • Noh, Tae-Hwan (National Institute of Crop Science, Rural Development Administration) ;
  • Cho, Heejung (National Academy of Agricultural Science, Rural Development Administration) ;
  • Chae, Soo-Cheon (Department of Horticulture, Kong-Ju National University) ;
  • Lee, Byoung-Moo (National Academy of Agricultural Science, Rural Development Administration)
  • Received : 2013.11.05
  • Accepted : 2014.02.27
  • Published : 2014.06.28
Download PDF
⟨ Previous Next ⟩

Abstract

We describe the development of a polymerase chain reaction method for the rapid, precise, and specific detection of the Xanthomonas oryzae pv. oryzae (Xoo) K3a race, the bacterial blight pathogen of rice. The specific primer set was designed to amplify a genomic locus derived from an amplified fragment length polymorphism specific for the K3a race. The 1,024 bp amplicon was generated from the DNA of 13 isolates of Xoo K3a races out of 119 isolates of other races, pathovars, and Xanthomonas species. The assay does not require isolated bacterial cells or DNA extraction. Moreover, the pathogen was quickly detected in rice leaf 2 days after inoculation with bacteria and at a distance of 8 cm from the rice leaf 5 days later. The results suggest that this PCR-based assay will be a useful and powerful tool for the detection and identification of the Xoo K3a race in rice plants as well as for early diagnosis of infection in paddy fields.

Keywords

References

  1. Adachi N, Oku T. 2000. PCR-mediated detection of Xanthomonas oryzae pv. oryzae by amplification of the 16S- 23S rDNA spacer region sequence. J. Gen. Plant Pathol. 33: 303-309.
  2. Adhikari TB, Mew TW, Leach JE. 1999. Genotypic and pathotypic diversity of Xanthomonas oryzae pv. oryzae in Nepal. Phytopathology 89: 687-694. https://doi.org/10.1094/PHYTO.1999.89.8.687
  3. Adhikari TB, Mew TW, Teng PS. 1994. Phenotypic diversity of Xanthomonas oryzae pv. oryzae in Nepal. Plant Dis. 78: 68-72. https://doi.org/10.1094/PD-78-0068
  4. Adhikari TB, Vera Cruz CM, Zhang Q, Nelson RJ, Skinner DZ, Mew TW, Leach JE. 1995. Genetic diversity of Xanthomonas oryzae pv. oryzae in Asia. Appl. Environ. Microbiol. 61: 966-971.
  5. Cho MS, Kang MJ, Kim CK, Seol YJ, Hhan JH, Park SC, et al. 2011. Sensitive and specific detection of Xanthomonas oryzae pv. oryzae by real-time bio-PCR using pathovarspecific primers based on an rhs family gene. Plant Dis. 95: 589-594. https://doi.org/10.1094/PDIS-06-10-0399
  6. George MLC, Bustamam M, Cruz WT, Leach JE, Nelson RJ. 1997. Movement of Xanthomonas oryzae pv. oryzae in southeast Asia detected using PCR-based DNA fingerprinting. Phytopathology 87: 302-309. https://doi.org/10.1094/PHYTO.1997.87.3.302
  7. Leach JE, Rhoads ML, Vera Cruz CM, White FF, Mew TW. 1992. Assessment of genetic diversity and population structure of Xanthomonas oryzae pv. oryzae with a repetitive DNA element. Appl. Environ. Microbiol. 52: 2188-2195.
  8. Liu H, Yang W, Hu B, Liu F. 2007. Virulence analysis and race classification of Xanthomonas oryzae pv. oryzae in China. J. Phytopathol. 155: 129-135. https://doi.org/10.1111/j.1439-0434.2007.01197.x
  9. Mew TW, Alvarez AM, Leach JE, Swing J. 1993. Focus on bacterial blight of rice. Plant Dis. 77: 5-12. https://doi.org/10.1094/PD-77-0005
  10. Mew TW, Vera Cruz CM, Medalla ES. 1992. Changes in race frequency of Xanthomonas oryzae pv. oryzae in response to rice cultivars planted in the Philippines. Plant Dis. 76: 1029-1032. https://doi.org/10.1094/PD-76-1029
  11. Noda T, Li C, Li J, Ochiai H, Ise K, Kaku H. 2001. Pathogenic diversity of Xanthomonas oryzae pv. oryzae strains from Yunnan Province, China. Jpn. Agric. Res. Q. 35: 97-103. https://doi.org/10.6090/jarq.35.2
  12. Noh TH, Lee DK, Kang MH, Shin MS, Shim HK, Na SY. 2003. Identification of new race of Xanthomonas oryzae pv. oryzae (Xoo) in Korea. Phytopathology 93: s66.
  13. Ochiai H, Horino O, Miyajima K, Kaku H. 2000. Genetic diversity of Xanthomonas oryzae pv. oryzae strains from Sri Lanka. Phytopathology 90: 415-421. https://doi.org/10.1094/PHYTO.2000.90.4.415
  14. Sakthivel N, Mortensen CN, Mathur SB. 2001. Detection of Xanthomonas oryzae pv. oryzae in artificially inoculated and naturally infected rice seeds and plants by molecular techniques. Appl. Microbiol. Biotechnol. 56: 435-441. https://doi.org/10.1007/s002530100641
  15. Song ES, Lee BM, Lee CS, Park YJ. 2012. PCR-based rapid assay for discriminative detection of latent infections of rice bacterial blight. J. Phytopathol. 160: 195-200. https://doi.org/10.1111/j.1439-0434.2012.01883.x
  16. Vos P, Hogers R, Bleeker M, Reijans M, van de Lee T, Hornes M, et al. 1995. AFLP: a new technique for DNA fingerprinting. Nucleic Acids Res. 23: 4407-4414. https://doi.org/10.1093/nar/23.21.4407
  17. Yun MS, Lee EJ, Cho YS. 1985. Pathogenic specialization of the rice bacterial leaf blight pathogen, Xanthomonas campestris pv. oryzae: race classification based on reaction of Korean differential varieties. Kor. J. Plant Protect. 24: 97-102.

Cited by

  1. Development of sandwich ELISA and immunochromatographic strip methods for the detection of Xanthomonas oryzae pv. oryzae vol.7, pp.15, 2014, https://doi.org/10.1039/c5ay01166c
  2. PCR-based Assay for the Specific Detection of Pseudomonas syringae pv. tagetis using an AFLP-derived Marker vol.21, pp.1, 2014, https://doi.org/10.5423/rpd.2015.21.1.001
  3. Transcriptome-Based Identification of Differently Expressed Genes from Xanthomonas oryzae pv. oryzae Strains Exhibiting Different Virulence in Rice Varieties vol.17, pp.2, 2014, https://doi.org/10.3390/ijms17020259
  4. Development of race-specific molecular marker for Xanthomonas campestris pv. campestris race 3, the causal agent of black rot of crucifers vol.98, pp.5, 2014, https://doi.org/10.1139/cjps-2018-0035
  5. Development of Molecular Marker through Genome Realignment for Specific Detection of Xanthomonas campestris pv. campestris Race 5, a Pathogen of Black Rot Disease vol.29, pp.5, 2014, https://doi.org/10.4014/jmb.1901.01050
  6. Development of a marker for detection of Xanthomonas campestris pv. campestris races 1 and 2 in Brassica oleracea vol.60, pp.4, 2014, https://doi.org/10.1007/s13580-019-00143-7
  7. Development of PCR-Based Molecular Marker for Detection of Xanthomonas campestris pv. campestris Race 6, the Causative Agent of Black Rot of Brassicas vol.36, pp.5, 2020, https://doi.org/10.5423/ppj.oa.06.2020.0103