Isopentenyladenosine and cytokinin-like activity of different humic substances

Increasing attention has been devoted in the last few years to the mechanisms by which humic substances (HS) influence plant growth. Nevertheless, among the biologically active compounds that might be present in HS, only indoleacetic acid has been identified. An enzyme-linked immunosorbent assay (ELISA) was applied to detect the presence of isopentenyladenosine (IPA), a cytokinin, in two lignosulphonate-humates, leonardite humic acid and in a high molecular weight humic fraction extracted from earthworm faeces. The IPA concentration ranged from 34 to 145 pmol mg-1 humic C and was estimated by using a linear dose-response curves between the carbon (C) concentration and percentage of binding of an anti-IPA monoclonal antibody. The cytokinin-like activity of HS was evaluated by weighing small radish (Raphanus sativus L.) cotyledon leaves and measuring chlorophyll and protein contents, the activities of ATP sulfurylase and O-acetylserine sulfhydrylase on maize plants. It is noteworthy that, among the four HS, those from earthworm faeces showed the highest amount of IPA and the strongest cytokinin-like activity in both plants. The presence of IPA confirmed that HS influence plant metabolism at different levels. Our results open new perspectives in the plant-HS relationship and shed new light on the active components of HS.