Genotypic and Phenotypic Diversity of the Casual agent of Bacterial Blight of Walnut in Fars and Lorestan Provinces

Document Type : Research Paper

Authors

Phd student of Plant protection department, Bu Ali Sina university

Abstract

During 2010 and 2011, 24 Xanthomonas strains were isolated from infected walnut trees from different regions of Fars and Lorestan provinces. For investigation of phenotypic and genotypic features, physiological, biochemical tests and rep-PCR with BOX, ERIC and REP primers. Isolates were rod-shaped, motile with polar flagella, gram negative, obligate parasite, oxidative negative, catalase positive and hydrolyzed esculin. All isolates produced H2S from cystein and acid from glucose, sucrose, D-mannose and D-sorbitol. Isolates showed differences in some characteristics such as tween 80 hydrolysis, starch hydrolysis, tolerance to 3% Nacl and utilization of L-alanin. Also pathogenicity tests and antibiogram were carried out. Based on phenotypic features, pathogenicity tests and amplification of 413 bp fragment with rev_seq R and rev_seq f primers in PCR assay, the isolates were identified as Xanthomonas arboricola pv. juglandis. Based on numerical analysis of phenotypic features most of the isolates were homogeneous and had about 87% similarity. The isolates divided into two groups on the basis of rep-PCR with BOX, ERIC and REP primers. Rep-PCR with above mentioned primers did not confirm clustering of theses isolates based on geographic origin. This is the first report of bacterial blight disease of walnut in Fars province of Iran.

Keywords

Main Subjects


Ark, P. A. 1944. Pollen as a source of walnut bacterial blight infection. Journal of Phytopathology, 34: 330-334.
Belisario, A. Palangio, C. and Zoina, A. 1997. Suscettibilit`a e fattori predisponenti a Xanthomonas campestris pv. juglandis in vivaio di noce comune. Informatore Fitopatologico, 4:60-64
Bergey, D. H. 1930. Bergey's Manual of Determinative Bacteriology. A key for the identification of organism of the class schizomycetes. Bergey’s manual of determinative Bacteriology. A Key for the Identification of Organisms of the class Schizomycetes. (Edn 3).
Dowson, D. 1939. On the systematic position and generic names of the Gram negative bacterial plant pathogens. Zentralblatt fur Bakteriologie, Parasitenkunde und Infektionskrankheiten, 100: 177-193.
Fahy, P. C. and Persley, G. J. 1983. Plant Bacterial Disease: A Diagnostic Guid. Academic Press. Sydney, Australia. 393p.
Garcin, A. and Duchesne, D. 2001. Walnut blight and apical necrosis. Acta Horticulturae, 544: 379-387.
Golmohammadi, M. Alizade, A and Rahimian, H. 2002. Homogenecity of the strains inciting bacterial blight of walnut in the central and northen provinces of Iran. Iranian Journal of Plant Pathology, 38: 11-20.
Hayward, A. C. 1964. Characteristics of Pseudomonas solanacearum. Journal of Appllied Bacteriology, 27: 265- 277.
Hugh, R. and Leifson, E. 1953. The taxonomic significance of fermentative versus oxidative metabolism of carbohydrates by varios gram negative bacteria. Journal of Bacteriology, 66: 24- 26.
Klement, Z. Farkas, G. L. and Lovrekovich, L. 1964. Hypersensitive reaction induced by phytopathogenic bacteria in the tobacco leaf. Phytopathology, 54: 474-477.
Kovacs, N. 1956. Identification of Pseudomonas pyocyanea by the oxidase reaction. Nature, 178:703.
Lelliot, R. A. Billing, E. and Hayward, A. C. 1966. A determinative scheme for the fluorescent plant pathogenic pseudomonads. Journal of Appllied  Bacteriology, 29: 470–489.
Louws, F. J. Fulbright, D. W. Stephens, C. T. and de Bruijn, F. J. 1994. Specific genomic fingerprints of phytopathogenic Xanthomonas and Pseudomonas pathovars and strains generated with repetitive sequences and PCR. Applied and Environmental Microbiology, 60: 2286-2295.
Macfadin, J. F. 1980. Biochemial Tests for Identification of Medical Bacteria. 2nd ed. The Williams and Wilkins, Baltimore, London, 527p.
Ngoc, L. B. T. Vernière, C. Jouen, E. Ah-You, N. Lefeuvre, P. Chiroleu, F. Gagnevin, L. and Pruvost, O. 2010. Amplified fragment length polymorphism and multilocus sequence analysis-based genotypic relatedness among pathogenic variants of Xanthomonas citri pv. citri and Xanthomonas campestris pv. bilvae. International journal of systematic and evolutionary microbiology, 60 (3):515-525.
Pierce, N. B. 1901. Walnut bacteriosis. Journal Botany, 31: 272-2.
Rohlf. F. J. 2000. NTSYS-PC, Numerical taxonomy and Multivariate analysis system, Version 2.11J. Exeter Software, Setauket, NY, USA.
Schaad, N. W. Jones, J. B. and Chun, W. 2001. Laboratory Guide for Identification of Plant Pathogenic Bacteria. American Phtopathology Society Minnesota, 373pp.
Scortichini, M. Marchesi, U. and Diprospero, P. 2001. Genetic Diversity of Xanthomonas arboricola pv. juglandis (synonyms: X. campestris pv. juglandis; X. juglandis pv. juglandis) strains from different geographical areas shown by repetitive polymerase chain reaction genomic fingerprinting. Journal of Phytopathology, 149: 325-332.
Smith, E.F. 1905. Bacteria in Relation to Plant Diseases. Carnegie Institute, Washington DC, USA.
Suslow, T. V. Schroth, M. V. and Isaka, M. 1982. Application of a rapid method for gram differentiation of plant pathogenic and saprophytic bacteria without staining. Phytopathology, 72: 917-918.
Tamponi, G. and Donati, G. P .1990. Walnut cultivars susceptible to Xanthomonas juglandis. Acta Horticolturae, 284: 301.
Thornley, M. J. 1960. The differentiation of Pseudomonas from other gram negative bacteria on the basis of argentine metabolism. Journal of Appllied Bacteriology, 3: 37-52.
Versalovic, J. Koeuth, T. and Lupski, J. 1991. Distribution of rerepetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes.  Journal of Nucleic Acids research,  19 (24): 6823-6831.
Zadeh- Mohammad, A and Marefat, A. 2011. PCR-based method for identifying and detection Xanthomonas strains blight of walnut. Thesis of MS. Zanjan University. 35-50.