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The role of motility in the in vitro attachment of Pseudomonas putida PaW8 to wheat roots
Authors:Turnbull G A  Morgan J AW  Whipps J M  Saunders J R
Institution:Department of Plant Pathology and Microbiology, Horticulture Research International, Wellesbourne, Warwickshire CV35 9EF, UK;School of Biological Sciences, Life Sciences Building, Crown Street, Liverpool L69 7ZB, UK
Abstract:The attachment of motile and non-motile strains of Pseudomonas putida PaW8 to sterile wheat roots was assessed in both non-competitive and intra-specific competitive assays. The motile strain showed significantly greater attachment to wheat roots than non-motile strains in phosphate buffer. Overall, the motile strain attached better than the non-motile strain at 10(6), 10(7) and 10(8) cfu ml(-1) in competitive assays and at 10(6) and 10(7) cfu ml(-1) in non-competitive assays. When attachment was studied in Luria broth no significant difference between motile and non-motile strains was detected. P. putida PaW8 cells marked with the luxAB genes were used to compare direct detection of attached cells by luminometry with indirect detection by dilution plate counts following extraction from root material. Although direct detection permitted a rapid assessment (60 s) of attachment to surfaces, dilution plate counts provided a more sensitive method for quantification of bacteria. The detection limits were approximately 10 cfu root(-1) using dilution plate counts compared with 1000 cfu root(-1) using luminometry. All results highlighted the importance of motility for the attachment of P. putida to plant roots in simple model systems. To take this work further, studies to assess the role of motility using complex non-sterile systems are needed.
Keywords:Bacterial motility              Pseudomonas putida PaW8  Wheat root              luxAB            Attachment
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