A Protocol for EndophyteFree Callus Tissue of the Grape Vitis aestivalis 'Norton/Cynthiana' (Vitaceae)

Abstract

Vitis aestivalis is used in commercial wine production. Propagation rates of V. aestivalis are low, therefore, plant tissue culture may be a viable method for propagation. However, a fungal endophyte exists within the plant which complicates tissue culture methods. This study reports a protocol for obtaining fungus-free callus from V. aestivalis. Explant tissue was chosen from healthy plants grown in a plant growth room and in a vineyard. Tissues were sterilized with isopropanol, bleach, and chlorine dioxide gas and plated onto media containing chlorothalonil. Successful results showed endophyte-free callus using explants taken from plants grown in a plant growth room, sterilized in alcohol, bleach, and chlorine dioxide, and plated on selection media containing an antifungal agent. This technique can be used with plants that have endophytes or other contamination problems to establish callus tissue for research and/or commercial propagation.