A Protocol for EndophyteFree Callus Tissue of the Grape Vitis aestivalis 'Norton/Cynthiana' (Vitaceae)
A Protocol for EndophyteFree Callus Tissue of the Grape Vitis aestivalis 'Norton/Cynthiana' (Vitaceae)
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Date
2016-12
Authors
Wilson, Aimee
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Publisher
University Honors College, Middle Tennessee State University
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.