Isolation of intact mesophyll cells from the leaves of higher plants

Abstract

A total of 146 species of angiosperms belonging to 35 taxonomically diverse families were screened for the isolation of living mesophyll cells from the leaves. Seventy-three species belonging to 22 families, on mild maceration in mortar with the isolation medium (pH 5.8) containing 0.7 M mannitoJ, 2 mM EDTA, 5 mM MgCl₂, 5 mM K₂HPO₄ and 1 mM NaNO₃, followed by fractional centrifugation, yielded intact mesophyll cells as seen under a research microscope. The high frequency of cell release, associated with the high percentage recovery of chlorophyll in cells was a common feature of most of the plant species examined by us. Nearly 87% of the chlorophyll present in the leaf could be recovered from the isolated cells in Dolichos lablab. The isolated cells retained active photosynthetic carbon metabolism as evidenced by high rates of ferricyanide reduction as well as carbon assimilation.