Study of phenotypic characteristics and genetic diversity of Pseudomonas syringae pv. Syringae, the causal agent of almond bacterial canker in Khorasan Razavi province using BOX-PCR

Document Type : Research Paper

Authors

Abstract

Bacterial canker caused by pathovars of Pseudomonas syringae Van Hall is one of the serious problems in stone fruit productions. In this research samples with canker symptoms were collected from Khorasane Razavi province during 2011–2012 and bacterial isolation was performed. The bacterial strains were identified as Pseudomonas syringae pv. syringae (Pss) on basis of LOPAT and GATTa tests and were confirmed as the causal agent of the almond bacterial canker based on the pathogenicity tests. A total of 16 isolates were characterized based on physiological and biochemical tests. The isolates showed a few differences in phenotypic characteristics. Genomic DNA from strains was used in PCR with D21 and D22 primers. To assess genetic diversity among the strains, rep-PCR analysis was performed using BOXA1R primer. Similarity matrix of the strains was calculated using the Jaccard's coefficient and cluster analysis was performed by UPGMA method using NTSYS-pc. The strains were divided into 3 clusters with BOXA1R at 68% similarity level. The results demonstrated considerable genetic diversity among strains causing canker in almond trees.
 

Keywords

References

  1. Abbasi V, Rahimian H and Tajick-Ghanbari A. 2013. Genetic variability of Iranian strains of Pseudomonas syringae pv. syringae causing bacterial canker disease of stone fruits. European Journal Plant Pathology 135: 225–235
  2. Aldaghi M, Mohammadi M, Rahimian H and Sharifi-Tehrani A. 2000. Serological and molecular comparison of Pseudomonas syringae pv. syringae strains isolated from stone fruit trees, wheat and barley. Paper presented at: 14th Iranian Plant Protection Congress; 5–8 September; Isfahan, Iran.
  3. Alvarez A M. 2004. Integrated approaches for detection of plant pathogenic bacteria and diagnosis of bacterial diseases. Annual Review of Phytopathology 42: 339–366.
  4. Arabi F, Nikravesh Z, Babaeezad V, Rezaeean V and Rahimian H. 2006. Occurrence of bacterial leaf spot of garden beet caused by Pseudomonas syringae pv. aptata in Iran. Iranian Journal of Plant Pathology 42: 655–671.
  5. Bahar M, Mochtahedi H and Akhiani A. 1982. Bacterial canker of apricot in Esfahan. Iranian Journal of Plant Pathology 18: 58–68.
  6. Banapour A, Zakiee Z and Amani G. 1990. Isolation of Pseudomonas syringae from sweet cherry in Tehran Province. Iranian Journal of Plant Pathology. 26:67–72.
  7. Bradbury J F. 1986. Guide to Plant Pathogenic Bacteria. CAB International Mycological Institute, Kew, England. 332p.
  8. Bultreys A and Kaluzna M. 2010. Bacterial cankers caused by Pseudomonas syringae on stone fruit species with special emphasis on the pathovars syringae and morsprunorum race1 and race2. Journal of Plant Pathology 92: S1.21–S1.33.
  9. Dawson S L, Fry J C and Dancer B N. 2002. A comparative evaluation of five typing techniques for determining the diversity of fluorescent Psudomonas. Journal of Microbiological Methods50: 9–22.
  10. Garden L, Shafik H, Belouin S, Broch R, Grimont F and Grimont P A. 1999. DNA relatedness among the pathovars of Pseudomonas syringae and description of Pseudomonas tremae sp. nov. and Pseudomonas cannabina sp. nov. (ex Sutic and Dowson 1959). International Journal of Systematic Bacteriology 49: 469-478.
  11. Gilbert V, Legros F, Maraite H and Bultreys A. 2009. Genetic analyses of Pseudomonas syringae isolates from Belgian fruit orchards reveal genetic variability and isolate-host relationships within the pathovar syringae, and help identify both races of the pathovar morsprunorum. European Journal of Plant Pathology124: 199–218.
  12. Hildebrand D C, Schorth M N and Huisman O C. 1982. The DNA homology matrix and non-random variation concepts as the basis for the taxonomic treatment of plant pathogenic and other bacteria. Annual Review of Phytopatholog. 20: 235–256.
  13. 13.  Kaku H. 2003. Genetic diversity and infection mechanism of plant pathogenic bacteria. Journal of General Plant Pathology 69: 412–414.
  14. 14.  Kang H P and Dunne W M. 2003. Stability of repetitive-sequence PCR patterns with respect to culture age and subculture frequency. Journal of Clinical Microbiology41: 2694–2696.
  15. Kennelly M M, Cazorla F M, Vicente A, Ramos C and Sundin G W. 2007. Pseudomonas syringae diseases of fruit trees progress toward understanding and control. Plant Disease 91: 4–17.
  16. Little E L, Bostock R M and Kirkpatric B C. 1998. Genetic characterization of Pseudomonas syringae pv. syringae from stone fruits in California. Applied and Environmental Microbiology 64: 3818–3823.
  17. Lopez M M, Llop P, Olmos A, Mcro-Noales E, Cambra M and Bertolini E. 2009. Are the molecular tools solving the challenges posed by detection of plant pathogenic bacteria and viruses?. Current Issues in Molecular Biology 11: 13–46.
  18. Lopez M M, Rosello M and Palacio-Bielsa A. 2010. Diagnosis and detection of the main bacterial pathogens of stone fruit and almond. Journal of plant pathology 92: S1.57–S1.66
  19. Manceau C and Horvais, A. 1997. Assessment of genetic diversity among strains of Pseudomonas syringae by PCR-Restriction Fragment Length Polymorphism analysis of rRNA operons with special emphasis on P. syringae pv. tomato. Applied and Environmental Microbiology 63: 491–505.
  20. Menard M, Sutra L, Luisetti J, Prunier J P and Gardan L. 2003. Pseudomonas syringae pv. avii (pv. nov.) the causal agent of bacterial canker of wild cherries (Prunus avium) in France. European Journal of Plant Pathology109: 565–576.
  21. Mohammadi M, Ghasemi A and Rahimian H. 2001. Phenotypic characterization of Iranian of Pseudomonas syringae pv. syringae Van Hall, the causal agent of bacterial canker disease of stone fruit trees. Journal Agricultural Science and Technology3: 51–65.
  22. Mousivand M, Rahimian H and Shamsbakhsh M. 2009. Phenotypic and genotypic relatedness among Pseudomonas syringae pv. syringae isolates from sugarcane, stone fruits and wheat. Iranian Journal of Plant Pathology 45: 75–85.
  23. Najafi Pour G and Taghavi S M. 2011. Comparison of P. syringae pv. syringae from different hosts based on pathogenicity and BOX-PCR in Iran. Journal of Agricultural Science and Technology 13: 431–442.
  24. Palacio-Bielsa A, Cambra M A and López M M. 2009. PCR detection and identification of plant-pathogenic bacteria: updated review of protocols (1989-2007). Journal of Plant Pathology 91: 249–297.
  25. Palleroni N J. 2005. Genus Ӏ. Pseudomonas Migula 1894. pp. 323–379. In D J Brenner, N R Krieg and J T Staley (eds). Bergey᾽s Manual of Systematic Bacteriology (2nd edition). The Proteobacteria, Part B, The Gammaproteobacteria, Vol. 2. New York: Springer.
  26. Radmarker J W, Hoste B, Louws F J, Kersters K, Swing J, Vauterine L, Vauterin P and de Brujin F J. 2000. Comparison of AFLP and rep-PCR genomic fingerprinting with DNA-DNA homology studies: Xanthomonas as a model system. International Journal of Systematic and Evolutionary Microbiology50: 665–677.
  27. 27.  Samavatian H. 2006. Identification and distribution of bacterial disease agent of almond tree canker in Isfahan province. Acta Horticulture726: 667–671.
  28. Schaad N W, Jones J B and Chun W. 2001. Laboratory guide for identification of plant pathogenic bacteria, Third edition. St Paul: APS Press. USA. 373p.
  29. Thomidis T, Tsipouridis C, Exadaktylou E and Drogoudi P. 2005. Comparison of three laboratory methods to evaluate the pathogenicity and virulence of ten Pseudomonas syringae pv. syringae strains on apple, pear, cherry and peach trees. Phytopathology 33: 137–140.
  30. Versalovic J, Koeuth T and Lupski J R. 1991. Distribution of repetitive DNA-sequences in eubacteria and application to fingerprinting of bacterial genomes. Nucleic Acids Research 19: 6823–6831.
  31. Vicent J G and Roberts S J. 2007. Discrimination of Pseudomonas syringae isolated from sweet and wild cherry using rep-PCR. European Journal of Plant Pathology 117: 383–392.
  32. Vicent J G, Alves J P, Russell K and Roberts S J. 2004. Identification and discrimination of Pseudomonas syringae isolates from wild cherry in England. European Journal of Plant Pathology 110: 337–351.

Files

Share

How to cite

Statistics