, 2004). Rhizobium leguminosarum swarm cells are also characterized by an increase in flagellation in 3841 and hyperflagellation in VF39SM. The hyperflagellation observed in VF39SM swarm cells is coupled with an increased expression of flagellin genes. Hyperflagellation of swarmer cells has been demonstrated in a number of bacteria including Vibrio parahaemolyticus (McCarter, 1999), P. mirabilis (Allison et al., 1993), R. etli (Braeken et al., 2008), E. coli, and Salmonella typhimurium (Harshey & Matsuyama, 1994). We also looked at the expression of the transcriptional activators VisN and Rem under swarming conditions. We have shown in a previous study that VisN is a transcriptional activator of rem, while

Rem regulates the expression of a subset of flagellin genes in R. leguminosarum (Tambalo et al., 2010). It appears that the upregulation of flagellin synthesis for R. leguminosarum swarmer PFT�� mouse EPZ015666 cells occurs at the level of the transcriptional activator VisN because increased expression was also observed for visN under swarming conditions. This type of regulation is similar to what has been reported

in P. mirabilis, where the expression of the master regulator FlhDC increased 30-fold in swarmer cells (Fraser & Hughes, 1999). Although slightly higher, the expression of rem under swarming conditions was very similar to cells grown in liquid media. It is possible that Rem is involved in the activation of motility-related genes under both swimming and swarming conditions. There might also be additional transcriptional activators of flagellar genes under swarming conditions, aside from Rem, thus Urocanase the observed upregulation of flagellin genes in swarmer cells. We demonstrated

that a nutrient-rich medium is essential for surface migration in R. leguminosarum. Without supplementation of a carbon source to the basal swarm medium, swarming motility was significantly reduced. We have shown that differentiation into swarm cells involves increased flagellation. Because flagellar synthesis and function is energetically costly (Wei & Bauer, 1998; Soutourina & Bertin, 2003), we speculate that a significant amount of energy is needed for differentiation, thus the need for an energy-rich medium. In addition, the supplemented sugar might be metabolized by the bacteria to produce the extracellular matrix. Plasmid-cured strains that are unable to metabolize the sugar did not swarm and they formed dry colonies, which could indicate the absence of the extracellular matrix that is needed for surface translocation. Although swarming motility is not dependent on the type of carbon source used, VF39SM exhibited slightly different swarming patterns using different types of carbon sources. The differences in the swarming patterns could be attributed to the different types and amounts of extracellular slime produced using these carbon sources. Rhizobium leguminosarum swarmed faster in mannitol compared with glycerol (data not shown).