We are trying to analyze Pseudomonas syringae cells growing on the leaf surface, using MALDI mass spec. We need to determine a chemical marker in the bacteria to differentiate between the bacteria and the leaf surface.
Does any have ideas of chemicals unique to bacteria that would be absent on the leaf surface (as many as you can think of)?
It is now the latest method for the diagnosis of various fungal infections including the invasive fungal infections (IFI's) which result in lots of morbidity and mortality
Why not analyse both separate and look for differences - then go to complex sample of interaction.
Microbes have lots of unique lipids - may you can differentiate them by those? Also, cell membrane components are possibly quite different to plant material.
Dear Michael, I agree with Manuel's comment - compare the leaf surface with and without bacteria, and the compare spectra statistically without an a priori expectation of differences or the need to identify peaks. If you do identify consistent differences, then you can chase as many as you want with further work. We used this type of blind spectral comparison using raman spectroscopy of individual bacterial cells (Pseudomonas fluorescens SBW25) where we were able to demonstrated that cells grown on different carbon sources produced different spectra - including cells grown in the rhizopshere and on the leaves of sugar beet (Ude et al., Microbial Ecology 2007, 53:494–497). In a follow-up, we showed that P. syringae DC3000, P. putida KT2440 and P. aeruginosa PA01 showed similar differentiation on carbon sources - all suggesting that pseudomonas will produce slightly different metabolites in different environments. Regards, Andrew.
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