Edophytic plant probiotic bacteria produce appreciable amount of IAA in vitro. We wish to quantify IAA production by the endophytic bacteria in vivo.
A lot of literature exists for IAA profiling/ quantification using (HP)TLC, HPLC and even LC-MS unless you have access to immuno-screens. Depends what you have access to. Thanks,
there are a wealth of published work on methods of determination of all plant hormones.Please, read my published work on plant hormones quantification procedures .
To Tofazzel: Quantifying IAA in production by the endophytic bacteria.
Do you want to quantify the level, the rate of biosynthesis or the expression of auxin response genes (which do not directly correlate to IAA levels)? For a review of auxin determination methods, see my article Tivendale and Cohen (2015) J Plant Plant Growth Regul DOI: 10.1007/s00344-015-9519-4.
Hi there,
you may need a knockout plant not able to produce IAA, so the IAA you quantify comes from the bacteria only. Other way would be if the IAA biosynthetic pathways are different in bacteria and plant (they use diff. precursors; no idea about this) you could feed a labeled precursor that the bacteria uses it and you can track you labeled IAA afterwards.
Agreed, you need to disentangle the competing IAA sources, but I do not known of any mutants that produce no IAA. A complete lack of IAA would be lethal.
The bacteria and the plant probably use different pathway to make IAA, so you could feed differently-labelled forms of the precursors (e.g. if the bacteria uses IAM and the plant uses IPyA, you could feed a 13C4 version of one and 13C6 version of the other), then you could quantify using isotope dilution (add a fully labelled internal standard after extraction) on LC-MS. This would enable you to tell the level of IAA in the bacteria vs that in the plant. Although you would have to rule out transfer of IAA between the two.
Thanks to Nathan Tivendale and others for sharing your thoughts. Do anyone have IAA knockout mutant plant or protocol for making IAA knockout bacteria?
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