The spreading property of mannosylerythritol lipids (MELs) was investigated in connection with our search for new application in agriculture. The wetting ability of MEL solutions for hydrophobic surfaces was evaluated based on contact angle measurements for several surfactant solutions on abiotic and biotic surfaces. The contact angle of MEL-A solution on a hydrophobic plastic surface at 100 s after placement decreased to 8.4°, and those of other MEL solutions decreased more significantly compared to those of commonly-used nonionic surfactants. In addition, the contact angle of MEL solutions also dropped down to around 10° on various plant leaf surfaces. MEL solutions, in particular, efficiently spread even on poorly wettable Gramineae plant surfaces on which general nonionic surfactant solutions could not. Moreover, the wetting ability of MEL solutions was found to be greatly affected by the structural difference in their carbohydrate configuration. Furthermore, surface pretreatment with MEL solution led to more efficient spreading and fixing of microbial cells onto plant leaf surface compared to several conventional surfactants used in this study. These results suggested that MELs have a potential to use as a natural bio-based spreading agent, particularly as agrochemical spreader for biopesticides.