Comparative analysis of hydroxyl radical production in fresh and desiccated fronds of Asplenium ceterach L. examined by fluorescence microscopy

Abstract

Group of higher plants, known as desiccation tolerant or resurrection plants, possess uniquely effective mechanism to withstand extreme dehydration and rapid rehydration of vegetative tissues without cell damage. Resurrection plants survive the loss of most of their tissues water content until a quiescent stage is achieved. Upon watering, the plants revive and are restored to their former state within 24h. In order to cope with desiccation, resurrection plants have to overcome a number of stresses, among them the most critical being oxidative stress. In our previous investigations, we have shown that during the rehydration process in A. ceterach, •OH radicals are produced. In vivo productions of free radicals remain unexplained and difficult for examination. By application of specific fluorescence probe for detection of hydroxyl radical APF (3’-(p-aminophenyl) fluorescein) it is possible to get insight into processes occurring in fronds during the dehydration process. In reaction of hydroxyl radical with APF, fluorescent product emerges. Examination under fluorescence microscope and image capturing in time intervals provides determination of hydroxyl radical production in different plant tissues. Through image analysis based on multivariate techniques, it is possible to simultaneously distinguish plant tissues according to its morphological and physiological properties.