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A Protocol for Endophyte-­Free Callus Tissue of the Grape Vitis aestivalis “Norton/Cynthiana” (Vitaceae)
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作者 Aimee Wilson Matthew Fuller +2 位作者 shannon smith Tony V. Johnston John D. Du Bois 《Agricultural Sciences》 2016年第10期680-692,共14页
Vitis aestivalis is used in commercial wine production. Vegetative propagation of V. aestivalis has shown a low success rate. Although plant tissue culture has been a successful method to propagate many species, V. ae... Vitis aestivalis is used in commercial wine production. Vegetative propagation of V. aestivalis has shown a low success rate. Although plant tissue culture has been a successful method to propagate many species, V. aestivalis has not yet been reliably grown as pure callus culture due to a fungal endophyte that exists within the plant. This study reports a viable protocol for obtaining fungus-free tissue culture callus from V. aestivalis. Explant tissue was chosen from healthy, actively growing plants grown in a growth room and in a vineyard. Tissues were sterilized with a combination of isopropanol, bleach, and chlorine dioxide gas and plated onto media containing chlorothalonil. The results from this study suggest that in order to obtain endophyte-free callus tissue, vine explants are to be taken from plants grown in a growth chamber simulating springtime conditions, sterilized in a combination of alcohol, bleach, and chlorine dioxide, and plated on selection media containing an antifungal agent, such as chlorothalonil. This technique could potentially be used with plants that have associated endophytes or other contamination problems to establish callus tissue for research and/or commercial propagation efforts. 展开更多
关键词 CHLOROTHALONIL ENDOPHYTE Vitis aestivalis NORTON Cynthiana
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Determination of Optimal Hormone and Mineral Salts Levels in Tissue Culture Media for Callus Induction and Growth of Industrial Hemp (<i>Cannabis sativa</i>L.)
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作者 Xoe Thacker Kayla Thomas +2 位作者 Matthew Fuller shannon smith John DuBois 《Agricultural Sciences》 2018年第10期1250-1268,共19页
Historically, industrial hemp (Cannabis sativa L.) has been a valuable source of metabolites and compounds, such as cannabidiols. There is a need for large amounts of plant tissue to be grown under controlled environm... Historically, industrial hemp (Cannabis sativa L.) has been a valuable source of metabolites and compounds, such as cannabidiols. There is a need for large amounts of plant tissue to be grown under controlled environments, and plant tissue culture is one unique way to yield this tissue. The purposes of this study were to determine: 1) the optimal concentrations (μM)/ratios of auxin:cytokinin in media and;2) the optimal mineral salts formulation for callus induction and callus growth in select hemp cultivars. To find the optimal concentration/ratios, 16 different combinations of auxin:cytokinin and three different mineral salts formulations were evaluated. The three mineral salts formulations tested were MS salts, MB5D1K and an MTSU formulation. The top performing hormone formulations were determined to be equal concentrations (1:1, 2:2, 3:3 μM) of auxin and cytokinin. The top performing media hormone formulations for callus induction were determined to be 2:1, 2:2, 2:3, and 3:2 μM (auxin:cytokinin). The optimal mineral salts formulation was determined to be MD5D1K. Therefore, the overall optimal media formulation for hemp callus production would be MB5D1K salts with the concentration/ratio of 2:2 μM (auxin:cytokinin). 展开更多
关键词 Industrial Hemp CANNABIS Sativa TISSUE Culture Hormones MINERAL SALTS Callus
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