Phenotypic and molecular characterization of the bacterial causal agents of sugar beet soft rot and vascular necrosis disease in Southeastern Iran
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Abstract
The bacterial soft rot and vascular necrosis disease in sugar beet (Beta vulgaris var. saccharifera) can be caused by different bacteria such as Pectobacterium betavasculorum and P. carotovorum subsp. carotovorum. During the 2018-2019 growing season, tissue samples from symptomatic sugar beet plants were collected from sugar beet growing areas in Kerman province, southeastern Iran. Totally, thirty-one bacterial isolates were isolated from the symptomatic tissues, of which nineteen isolates were identified as Pectobacterium spp., nine isolates as Bacillus spp. and three isolates as Flavobacterium spp. based on the phenotypic and biochemical tests. Pectobacterium isolates were further characterized by PCR amplification (using the EXPCCF/EXPCCR primer set) and genomic fingerprinting by rep-PCR. Only 27% of the Pectobacterium isolates were identified as P. carotovorum, while 73% of the isolates were identified as P. betavasculorum. Artificial inoculation of isolates into the wounded sugar beet petioles led to black streaking, root rot and vascular necrosis from which the same bacterial isolates were re-isolated and identified. PCR assays using BOX and ERIC primers, and combined assays clearly differentiated the bacterial isolates, indicating high variability among the isolates. The results shed light on the bacterial causal agents of the sugar beet soft rot and vascular necrosis disease in southeastren Iran, identifying Pectobacterium isolates as the main causal agents (61% of all isolates), and uncovered the phenotypic/genetic variability among the bacterial isolates
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References
Anonymous (2016). Agriculture statically year book of Iran. Ministry of Agriculture Jihad. Tehran, Iran.
Ayers, S. H., Rupp, P. and Johnson, W. T. (1919). A study of the alkali-forming bacteria in milk. USDA Bulletin, 787:1-39.
Baghaee-Ravari, S., Shams-Bakhsh, M., Rahimian, H., Lopez-Solanilla, E., Antúnez-Lamas, M. and Rodríguez-Palenzuela, P. (2011). Characterization of Pectobacterium species from Iran using biochemical and molecular methods. European Journal of Plant Patholology, 129:413-425.
Campos, E., Maher, E. A. and Kelman, A. (1982). Relationship of pectolytic Clostridia and Erwinia carotovora strains to the decay of potato tubers in storage. Plant Diseases, 66:543-546.
Charkowski, A. O. (2006). The soft rot Erwinia. In: S. S. Gnanamanickam (ed.). Plant-Associated Bacteria. Springer-Verlag, Berlin, Germany, pp.423-507.
Cuppels, D. A. and Kelman, A. (1974). Evaluation of selective media for isolation of soft rot bacteria from soil and plant tissue. Phytopathology, 64:468-475.
Dana, H., Khodakaramian, G. and Rouhrazi, K. (2015). Characterization of Pectobacterium carotovorum subsp. carotovorum causing watermelon soft rot disease in Iran. Journal of Phytopathology, 163:703-710.
De Boer, S. H. and Kelman, A. (2001). Erwinia soft rot group. In: Laboratory Guide for Identification of Plant Pathogenic Bacteria in: N. W. Schaad, J. B. Jones & W. Chun (Eds.). American Phytopathological Society, St. Paul, MN, USA. pp.56-72.
Dowson, W. J. (1943). Spore-forming bacteria in potatoes. Nature, 152:331.
Elbanna, K., Elnaggar, S. and Bakeer, A. (2014). Characterization of Bacillus altitudinis as a new causative agent of bacterial soft rot. Journal of Phytopathology, 162:712-722.
Fahy, P. C. and Persley, G. J. (1983). Plant Bacterial Diseases: A Diagnostic Guide. Academic Press, Sydney, Australia.
Fassihiani, A. and Nedaienia, R. (2008). Characterization of Iranian Pectobacterium carotovorum strains from sugar beet by phenotypic tests and whole-cell proteins profile. Journal of Phytopathology, 156:281-286.
Firoz, R., Bahar, M. and Sharif-Nabi, B. (2007). Detection of casual agents of potato soft rot and blackleg in Esfehan province. Iranian Journal of Plant Pathology, 43:145-162.
Gardan, L., Gouy, C., Christen, R. and Samson, R. (2003). Elevation of three subspecies of Pectobacterium carotovorum to species level: Pectobacterium atrosepticum sp. nov., Pectobacterium betavasculorum sp. nov. and Pectobacterium wasabiae sp. nov. International Journal of Systematic and Evolutionary Microbiology, 53:381-391.
Hauben, L., Moore, E. R. B., Vauterin, L., Steenackers, M., Mergaert, J., Verdonck, L. and Swings, J. (1998). Phylogenetic position of phytopathogens within the Enterobacteriaceae. Systematic and Applied Microbiology, 21:384-397.
Jacobsen, B. J. (2006). Root rot diseases of sugar beet. Matica Srpska Journal of Natural Sciences, 110:9-19.
Janse, J. D. (2006). Phytobacteriology, Principle and Practice. CABI Publishing.
Kang, H. W., Kwon, S. W. and Go, S. J. (2003). PCR-based specific and sensitive detection of Pectobacterium carotovora subsp. carotovora by primers generated from a URP-PCR fingerprinting-derived polymorphic band. Plant Pathology, 52:127-133.
Kelement, Z., Farkas, G. L. and Lovrekovich, L. (1964). Hypersensitive reaction induced by phytopathogenic bacteria in the tobacco leaf. Phytopathology, 54:474-477.
Lee, D. H, Kim, J. B., Lim, J. A., Han, S. W. and Heu, S. (2014). Genetic diversity of Pectobacterium carotovorum subsp. brasiliensis isolated in Korea. Plant Pathology Journal, 30:117-124.
Liao, C. H. and Wells, J. M. (1986). Properties of Cytophaga johnsonae strains causes spoilage of fresh produce at food markets. Applied Environmental Microbiology, 52:1261-1265.
Liao, C. H. and Wells, J. M. (1987a). Association of a pectolytic strain of Xanthomonas campestris with soft rots of fruits and vegetables at retail markets. Phytopathology, 77:418-422.
Liao, C. H. and Wells, J. M. (1987b). The diversity of pectolytic fluorescent Pseudomonas causing soft rots of fresh vegetables at produce markets. Phytopathology, 77:673-677.
Lund, B. M. (1969). Properties of some pectolytic, yellow-pigmented, gram-negative bacteria isolated from fresh cauliflower. Journal of Applied Bacteriology, 32:60-67.
Mahmoudi, E., Soleimani, M. J. and Taghavi, S. M. (2007). Detection of bacterial soft-rot of crown imperial caused by Pectobacterium carotovorum subsp. carotovorum using specific PCR primers. Phytopathologia Mediterranea, 46:1-8.
Mikiciński, A., Puławska, J., Sobiczewski, P. and Orlikowski, L. B. (2010). Pectolytic bacteria associated with soft rot of dieffenbachia (Dieffenbachia maculata). Phytopathologia, 58:21-32.
Pérombelon, M. C. M. (2002). Potato diseases caused by soft rot Erwinias: an overview of pathogenesis. Plant Pathology, 51:1-12.
Pérombelon, M. C. M. and Van Der Wolf, J. M. (2002). Methods for the detection and quantification of Erwinia carotovorum subsp. atrpsepticum (Pectobacterium carotovorum subsp. atrosepticum) on potatoes: a laboratory manual. Scottish Crop Research Institute Annual Report, 10.
Rezaei, R. and Taghavi, M. (2010). Phenotypic and genotypic characteristics of Iranian soft rot bacterial isolates from different hosts. Phytopathologia Mediterranea, 49:194-204.
Rohlf, F. J. (2000). NTSYS-PC, Numerical taxonomy and Multivariate analysis system, Version 2.11j. Exeter Software, Setauket, NY, USA.
Saleh, O. I., Huang, P. Y. and Huang, J. S. (1997). Bacillus pumilus, the cause of bacterial blotch of immature Balady peach in Egypt. Journal of Phytopathology, 145:447-453.
Saleh, O. I., Huang, P. Y. and Huang, J. S. (1996). Bacterial vascular necrosis and root rot diseases of sugar beet in Egypt. Journal of Phytopathology, 144:225-230.
Samavatian, H. (2006). Sugar beet bacterial diseases situation in Isfahan province. Proceeding of 17th Irananian Plant Protection Congress. Karaj, Iran, pp.108.
Schaad, N., Jones, J. B. and Chun, W. (2001). Laboratory Guide for Identification of Plant Pathogenic Bacteria. APS Press, St. Paul, MN.
Seo, S. T., Furuya, N., Lim, C. K., Takanam, Y. K. and Tsuchiya, K. (2003). Phenotypic and genetic characterization of Erwinia carotovora from mulberry (Morus spp.). Plant Pathology, 52:140-146.
Sheikholeslami, R. (2006). Evaluation of the Iranian sugar industry in the last four years. Journal of Sugar Beet, 21:16-17.
Soltani-Nejad, S., Taghavi, M., Hayati, J. and Mostofi, Z-G, R. (2005). Study of phenotypic and pathogenicity characteristics of Pectobacterium causing soft rot in Khozestan province. Iranian Journal of Plant Pathology, 41:585-611.
Thomson, S. V., Hildbrand, D. C. and Schorth, M. N. (1981). Identification and nutritional differentiation of the Erwinia sugarbeet pathogen from members of Erwinia carotovora and Erwinia chrysanthemi. Phytopathology, 71:1037-1042.
Thomson, S. V., Schroth, M. N., Hills, F. J., Whitney, E. D. and Hildebrand, D. C. (1977). Bacterial vascular necrosis and rot of sugar beet: general description and etiology. Phytopathology, 67:1183-1189.
Toth, I. K., Bell, K. S., Holeva, M. C. and Birch, P. R. J. (2003) Soft-rot Erwinia: from genes to genomes. Molecular Plant Pathology, 4:17-30.
Versalovic, J., Schneider, M., De Bruijn, F. J. and Lupski, J. R. (1994). Genome Fingerprinting of bacteria using repetitive sequence-based polymerase chain reaction. Methods in Molecular Cell Biology, 5:25-40.