A simple procedure for the manual isolation and identification of plant heterokaryons

Abstract

A simple procedure is described for the aseptic manual isolation of individual heterokaryons. Heterokaryons were identified with bright field illumination using an inverted microscope and isolated by means of a micro-manipulator and capillary pipette coupled to a specially constructed syringe. When cell suspension protoplasts were labelled with fluorescein isothiocyanate and fused with mesophyll protoplasts, the heterokaryons exhibited an apple green cytoplasmic fluorescence (from cell suspension protoplasts) and a red chloroplast fluorescence (from mesophyll protoplasts). By this double fluorescence procedure, these manually isolated heterokaryons, identified initially using brightfield could be confirmed as heterokaryons.