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Applied and Environmental Microbiology, January 2005, p. 417-422, Vol. 71, No. 1
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.1.417-422.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Contribution of Folate Biosynthesis to Ralstonia solanacearum Proliferation in Intercellular Spaces

Rena Shinohara,¹ Ayami Kanda,¹ Kouhei Ohnishi,² Akinori Kiba,¹ and Yasufumi Hikichi^1*

Laboratory of Plant Pathology and Biotechnology,¹ Research Institute of Molecular Genetics, Kochi University, Nankoku, Kochi, Japan²

Received 24 April 2004/ Accepted 19 August 2004

The vigorous proliferation of Ralstonia solanacearum OE1-1 in host intercellular spaces after the invasion of host plants is necessary for the virulence of this bacterium. A folate auxotroph, RM, in which a mini-Tn5 transposon was inserted into pabB encoding para-aminobenzoate synthase component I, lost its ability to vigorously proliferate in intercellular spaces along with its systemic infectivity and virulence after inoculation into roots and infiltration into leaves of tobacco plants. Complementation of RM with the pabB gene allowed the mutant to multiply in intercellular spaces and to cause disease. In tobacco plants that were pretreated with folate, RM was able to vigorously proliferate in the intercellular spaces and cause disease. Interestingly, when it was inoculated through cut stems, the mutant multiplied in the plants and was virulent. Moreover, the mutant multiplied well in stem fluids but not in intercellular fluids, suggesting that the folate concentration within intercellular spaces may be a limiting factor for bacterial proliferation. Therefore, folate biosynthesis contributes to the vigorous proliferation of bacteria in intercellular spaces and leads to systemic infectivity resulting in virulence.

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* Corresponding author. Mailing address: Laboratory of Plant Pathology and Biotechnology, Kochi University, 200 Monobe, Nankoku, Kochi 783-8502, Japan. Phone: 81-88-864-5218. Fax: 81-88-864-5219. E-mail: yhikichi{at}cc.kochi-u.ac.jp.

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Applied and Environmental Microbiology, January 2005, p. 417-422, Vol. 71, No. 1
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.1.417-422.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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