Biofilm formation by Xanthomonas campestris pv. viticola affected by abiotic surfaces and culture media
Biofilms are dense surface-associated communities formed by microorganisms. Formation of these structures by the plant pathogenic bacterium Xanthomonas campestris pv. viticola (bacterial canker of grapevine) had not previously been studied. The ability of seven strains of this bacterium to adhere to abiotic surfaces and to form biofilms in vitro was evaluated under different conditions. The surfaces tested were polystyrene and glass using microtiter plates and tubes, respectively. Four liquid culture media were used, nutrient-dextrose-yeast extract (NYD), yeast extract-dextrose-calcium carbonate (YDC), Kado 523 (KADO) and Luria-Bertani (LB). The biofilm architecture was examined by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). Seven strains adhered to polystyrene in the microtiter plates and formed biofilms in all culture media at weak, moderate, and strong levels. In glass tubes, only strains Xcv229 and Xcv158 formed biofilms. SEM of Xcv229 and Xcv158 revealed typical biofilm architectures. CLSM showed that only Xcv229 formed an initial matrix structure characteristic of biofilms. The X. campestris pv. viticola strains exhibited different levels of biofilm formation in different culture media, of which LB and KADO were the best. Therefore, bacterial growth in polystyrene microtiter plates using LB and KADO media is a good qualitative method for the detection of biofilms of this pathogen.
KeywordsVitis vinifera Bacterial canker disease CLSM SEM
The authors thank FACEPE (IBPG-0138-5.01/11), CNPq (Proc. 307348/2011-3, APQ 477521/2011-8), and Professor Carlos Termignoni for providing the Biotechnology Laboratory (UFRGS) facilities in support of this research.
- Araujo JSP (2001) Perfil epidemiológico e subsídios para o controle de Xanthomonas campestris pv. viticola (Nayudu) Dye, agente do cancro bacteriano da videira (Vitis vinifera L.) no Brasil. PhD Thesis, Universidade Federal Rural do Rio de Janeiro. Seropédica, BrazilGoogle Scholar
- Estela CRL, Alejandro PR (2012) Biofilms: a survival and resistance mechanism of microorganisms. In: Pana M (ed) Antibiotic resistant bacteria: A continuous challenge in the new millennium. In Tech Europe, Rijekam, pp 159–178Google Scholar
- Gama MAS (2014) Caracterização polifásica de Xanthomonas campestris pv. viticola e reposicionamento de espécies de Xanthomonas patogênicas à videira e cajueiro. PhD Thesis, Universidade Federal Rural de Pernambuco. Recife, BrazilGoogle Scholar
- Harrison JJ, Ceri H, Yerly J, Stremick CA, Hu Y, Martinuzzi R, Turner RJ (2006) The use of microscopy and three-dimensional visualization to evaluate the structure of microbial biofilms cultivated in the Calgary biofilm device. Biological Procedures Online 8:194–215CrossRefPubMedPubMedCentralGoogle Scholar
- Janissen R, Murillo DM, Niza B, Sahoo PK, Nobrega MM, Cesar CL, Temperini MLA, Carvalho HF, Souza AA, Cotta MA (2015) Spatiotemporal distribution of different extracellular polymeric substances and filamentation mediate Xylella fastidiosa adhesion and biofilm formation. Scientific Reports 5. https://doi.org/10.1038/srep09856
- Lazzarotto JJ, Fioravanço JC (2013) Tendências e sazonalidades nas exportações e importações brasileiras de uva de mesa. Informações Econômicas 43:43–58Google Scholar
- Lorite GS, Janissen R, Clerici JH, Rodrigues CM, Tomaz JP, Mizaikoff B, Kranz C, Souza AA, Cotta MA (2013) Surface physicochemical properties at the micro and nano length scales: role on bacterial adhesion and Xylella fastidiosa biofilm development. PLoS One 8:e75247CrossRefPubMedPubMedCentralGoogle Scholar
- Martínez LC, Vadyvaloo V (2014) Mechanisms of post-transcriptional gene regulation in bacterial biofilms. Frontiers in Cellular and Infection Microbiology. https://doi.org/10.3389/fcimb.2014.00038
- Naue CR, Costa VSO, Barbosa MAG, Batista DC, Souza EB, Mariano RLR (2014) Xanthomonas campestris pv. viticola on grapevine cutting tools and water: survival and disinfection. Journal of Plant Pathology 96:451–458Google Scholar
- Rigano LA, Siciliano F, Enrique R, Sendín L, Filippone P, Torres PS, Qüesta J, Dow JM, Castagnaro AP, Vojnov AA, Marano MR (2007) Biofilm formation, epiphytic fitness, and canker development in Xanthomonas axonopodis pv. citri. Molecular Plant-Microbe Interactions 20:1222–1230CrossRefPubMedGoogle Scholar
- Simões M, Simões LC, Vieira MJ (2010) A review of current and emergent biofilm control strategies. Food Science and Technology 43:573–583Google Scholar
- Trentin DS, Giordani RB, Macedo AJ (2013) Biofilmes bacterianos patogênicos: aspectos gerais, importância clínica e estratégias de combate. Revista Liberato 14:113–238Google Scholar