New Technique to Create a Suspension Containing Bacteriophages and How It Can Be Used to Control Cabbage Leaf Spot Caused by Xanthomonas campestris PV. campestris
Affiliation(s)
Department of Basic Health Sciences, Rua Sarmento Leite, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil.
Department of Basic Health Sciences, Rua Sarmento Leite, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil.
ABSTRACT
The purpose of the present study was to develop a new method for producing and identifying a bacteriophage suspension and research its potential against plant bacteria. The suspension obtained with the new technique was effective in controlling Xanthomonas campestris PV. campestris both in vitro and in vivo. A difference was observed between the suspension with pathogen (PLP) and crude suspension with lesioned plant without the pathogen originated from it (PLN). Clear zones were observed at 48 h/28°C in both (PLS) suspension added saccharose and (PLP). Although PLP and PLS treatments showed no statistical difference in CFU values (5 × 105 and 1 × 105, respectively), they differed from the crude suspension (PLN) and the control. Then, in vivo assessment of suspensions with clear zones in the dishes was performed. In vivo (PLPS) and (PLPP) treatments consisting of inoculum were added to the crude suspensions (PLS and PLP) to increase bacteriophages. A decrease in the disease severity index compared to the control was observed, showing that treatment with the phage suspension protects the plant. The lowest disease indexes were reported with (PLP) and (PLS), with values of 1.46 and 1.40, respectively. The highest averages in dry matter (5.1 inboth treatments) were obtained with (PLP) and (PLS) treatments.
Cite this paper
Marroni, I. and Germani, J. (2014) New Technique to Create a Suspension Containing Bacteriophages and How It Can Be Used to Control Cabbage Leaf Spot Caused by Xanthomonas campestris PV. campestris. Agricultural Sciences, 5, 286-297. doi: 10.4236/as.2014.54031.
Marroni, I. and Germani, J. (2014) New Technique to Create a Suspension Containing Bacteriophages and How It Can Be Used to Control Cabbage Leaf Spot Caused by Xanthomonas campestris PV. campestris. Agricultural Sciences, 5, 286-297. doi: 10.4236/as.2014.54031.
References
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[1] Mahy, B.W.J. and van Regenmortel, M.H.V. (2010) Desk Encyclopedia of Plant and Fungal Virology. Academic Press, Oxford, 613. [2] Summers, W.C. (2005) Bacteriophage Research: Early History. In: Kutter, E. and Sulakvelidze, A., Eds., Bacteriophages: Biology and Applications, CRC Press, Boca Raton. [3] Boyd, R.J., Hildebrandt, A.C. and Allen, O.N. (1971) Retardation of Crown Gall Enlargement after Bacteriophages Treatment. Plant Disease, 55, 145-148. [4] Okabe, M. and Goto, M. (1963) Bacteriophages of Plant Pathogens. Annual Review of Phytopathology, 1, 397-418.
http://dx.doi.org/10.1146/annurev.py.01.090163.002145 [5] Goto, M. (1992) Fundamental of Bacterial Plant Pathology. Academic Press, San Diego. [6] Johnson, K.L., Minsavage, G.V., Le, T., Jones, J.B. and Walcott, R.R. (2011) Efficiency of a Nonpathogenic Acidovorax citrulli Strain as a Biocontrol Seed Treatment for Bacterial Fruit Blotch of Cucurbits. Plant Disease, 95, 697-704.
http://dx.doi.org/10.1094/PDIS-09-10-0660 [7] Jones, J.B., Jackson, L.E., Balogh, B., Obradovic, A., Iriarte, F.B. and Momol, M.T. (2007) Bacteriophages for Plant Disease Control. Phytopathology, 45, 245-262. http://dx.doi.org/10.1146/annurev.phyto.45.062806.094411 [8] Stockwell, V.O. and Stack, J.P. (2007) Using Pseudomonas spp. for Integrated Biological Control. Phytopathology, 37, 12-18. [9] Maketon, M., Apisitsantikul, J. and Siriraweekul, C. (2008) Greenhouse Evaluation of Bacillus subtilis AP-01 and Trichoderma harzianum AP-001 in Controlling Tobacco Diseases. Brazilian Journal of Microbiology, 39, 296-300.
http://dx.doi.org/10.1590/S1517-83822008000200018 [10] Fritze, D. (2004) Taxonomy of the Genus Bacillus and Related Genera: The Aerobic Endospore Forming Bacteria. Phytopathology, 94, 1245-1248. http://dx.doi.org/10.1094/PHYTO.2004.94.11.1245 [11] Hagens, S. and Offerhaus, M.L. (2008) Bacteriophages—New Weapons for Food Safety. Food Technology, 62, 46-54. [12] Nagy, J.K., Király, L. and Schwarczinger, I. (2012) Phage Therapy for Plant Disease Control with a Focus on Fire Blight. Central European Journal of Biology, 1, 1-12. http://dx.doi.org/10.2478/s11535-011-0093-x [13] Matsuzaki, S., Rashel, M., Uchiyama, J., Sakurai, S., Ujihara, T., Kuroda, M., Ikeuchi, M., Tani, T., Fujieda, M., Wakiguchi, H. and Imai, S. (2005) Bacteriophage Therapy: A Revitalized Therapy against Bacterial Infectious Diseases. Japanese Society of Chemotherapy and the Japanese Association for Infectious Diseases, 11, 211-219. [14] Shasha, S.M, Sharon, N. and Inbar, M. (2004) Bacteriophages as Antibacterial Agents. Harefuah, 143, 121. [15] Balogh, B., Jones, J.B., Momol, M.T., Olson, S.M., Obradovic, A., King, P. and Jackson, L.E. (2003) Improved Efficacy of Newly Formulated Bacteriophages for Management of Bacterial Spot on Tomato. Plant Disease, 87, 949-954.
http://dx.doi.org/10.1094/PDIS.2003.87.8.949 [16] Romeiro, R.S. (2001) Métodos em Bacteriologia de Plantas. Editora UFV, Viçosa. [17] Svensson, U. and Christiansson, A. (1991) Methods for Phage Monitoring. Bulletin of the International Dairy Federation, 263, 29-39. [18] Marroni, I.V., Ueno, B. and Moura, A.B. (2012) Chemical and Biological Treatments of Castor Bean Seeds: Effect on Germination, Emergence and Associated Microorganism. Revista Brasileira de Sementes, 34, 1-8.
http://dx.doi.org/10.1590/S0101-31222012000100003 [19] Carzaniga, R., Fiocco, D., Bowyer, P. and O’Connell, R.J. (2002) Localization of Melanin in Conidia of Alternaria alternata Using Phage Display Antibodies. Molecular Plant-Microbe Interactions, 15, 216-224.
http://dx.doi.org/10.1094/MPMI.2002.15.3.216 [20] Rademaker, J.L.W., Norman, D.J., Forster, R.L., Louws, F.J., Schultz, M.H. and de Bruijn, F.J. (2006) Classification and Identification of Xanthomonas translucens Isolates, Including Those Pathogenic to Ornamental Asparagus. Phytopathology, 96, 876-884. http://dx.doi.org/10.1094/PHYTO-96-0876 [21] Kohl, J., Postma, J., Nicot, P., Ruocco, M. and Blum, B. (2011) Stepwise Screening of Microorganisms for Commercial Use in Biological Control of Plant-Pathogenic Fungi and Bacteria. Biological Control, 57, 1-12.
http://dx.doi.org/10.1016/j.biocontrol.2010.12.004 [22] Henriques, A.P. de A.R. (2003) Uso de Bacteriófagos para Controlo de Salmonella em Avicultura. Dissertação de Mestrado, Universidade de Aveiro, Portugal. [23] Griep, R.A., Prins, M., van Twisk, C., Keller, H.J., Kerschbaumer, R.J., Kormelink, R., Goldbach, R.W. and Schots, A. (2000) Application of Phage Display in Selecting Tomato Spotted wilt Virus-Specific Single-Chain Antibodies (scFvs) for Sensitive Diagnosis in ELISA. Phytopathology, 90, 183-190. http://dx.doi.org/10.1094/PHYTO.2000.90.2.183 [24] Lang, J.M., Gent, D.H. and Schwartz, H.F. (2007) Management of Xanthomonas Leaf Blight of Onion with Bacteriophages and a Plant Activator. Plant Disease, 91, 871-878. http://dx.doi.org/10.1094/PDIS-91-7-0871 [25] Obradovic, A., Jones, J.B., Momol, M.T., Balogh, B. and Olson, S.M. (2004) Management of Tomato Bacterial Spot in the Field by Foliar Applications of Bacteriophages and SAR Inducers. Plant Disease, 88, 736-740.
http://dx.doi.org/10.1094/PDIS.2004.88.7.736 [26] Balogh, B., Canteros, B.I., Stall, R.E. and Jones, J.B. (2008) Control of Citrus Canker and Citrus Bacterial Spot with Bacteriophages. Plant Disease, 92, 1048-1052. http://dx.doi.org/10.1094/PDIS-92-7-1048