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Mesophyll cells are desirable targets for studying responses to pathogens or pathogen-induced toxins. Based on host-pathogen or host-toxin interaction studies at the cellular level it can be determined whether a toxin can be used as a selective agent. Suspension cells are suitable selection units for in vitro selection of potentially useful somaclonal variants. Protocols for the isolation of muskmelon mesophyll and suspension cells were developed in order to study the effects of roridin E, a toxin produced by  Myrothecium roridum , on leaf spot tolerant and sensitive muskmelon cultivars. Viable mesophyll cells were obtained by exposing leaf tissue to 1% cellulysin and 5% macerase in B 5  medium with 0.4M sucrose for one hour. Viable suspension cells were maintained a medium consisting of MS salts, 3% sucrose, 3 (μM thiamine·HCl, 555 μM myo-Inositol, 28 μM kinetin and 9 μM IAA. Fluorescein diacetate was used to determine viability over time. Membrane stability was monitored by measuring changes in the fluorescence of cells stained with Merocyanine 540 (MC 540), an optical probe for changes in transmembrane electrical potential (PD).

ISOLATION OF CUCUMIS MELO L. MESOPHYLL AND SUSPENSION CELLS FOR HOST-PATHOGEN INTERACTION STUDIES.