Video registration as a method for studying kinetic parameters of chlorophyll fluorescence in Ficus benjamina leaves

Abstract

A method of imaging and analysis of fluorescence kinetic parameters distributed over leaf area is described in details. Video data recorded by the CCD-camera were processed using free programs VirtualDub and ImageJ. The method allows not only to image the distribution of fluorescence decrease ratio (Rfd), but also to obtain fluorescence induction curves, which are corresponded to any region of interest selected within the leaf image. A considerable mosaicism of shade leaves of Ficus benjamina L. was shown on the basis of Rfd values, whose functional significance is discussed from the point of view of adaptation to different light intensities. A pronounced non-uniformity of F. benjamina leaves was revealed in relation to the retention time of the secondary fluorescence maximum (peak M). A method of differential imaging was proposed to allow visualization of this non-uniformity and to demonstrate the existence of the previously unknown phenomenon of secondary chlorophyll fluorescence “wave” in leaves. It was also shown the absence of M peak in the chlorophyll-deficient, lacking the mesophyll leaf sectors, where the residual chlorophyll is originated from guard cell chloroplasts.