Modified Media for Repeated In Vitro Cutting Cycles of <i>Cannabis sativa</i> Without the Use of Cytokinin

In vitro hedging; combined with the fed-batch liquid media process is an innovative system that generates multiple sterile plants without the use of exogenous cytokinin. This combined process was demonstrated with <i>Cannabis sativa</i> (‘Cherry1’, ‘BaOx’, ‘T1’, ‘Peach’) grown in vessels...

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Main Authors: Molly McKay, James E. Faust, Matthew Taylor, Jeffrey Adelberg
Format: Article
Language:English
Published: MDPI AG 2025-04-01
Series:Plants
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Online Access:https://www.mdpi.com/2223-7747/14/7/1138
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author Molly McKay
James E. Faust
Matthew Taylor
Jeffrey Adelberg
author_facet Molly McKay
James E. Faust
Matthew Taylor
Jeffrey Adelberg
author_sort Molly McKay
collection DOAJ
description In vitro hedging; combined with the fed-batch liquid media process is an innovative system that generates multiple sterile plants without the use of exogenous cytokinin. This combined process was demonstrated with <i>Cannabis sativa</i> (‘Cherry1’, ‘BaOx’, ‘T1’, ‘Peach’) grown in vessels of three different physical states—stationary agar (A); stationary Oasis<sup>®</sup> infused with liquid (OILs); and agitated Oasis<sup>®</sup> infused with liquid (OILa). Vessels were pre-selected as control or supplemented; where supplement vessels received 15 mL DKW liquid media each cycle harvest. The number of shoot tips harvested; shoot length; and dry shoot mass from repeated cutting cycles was recorded. In a single harvest; ‘BaOx’ and ‘Cherry 1’ produced one shoot per plant from the original 15 planted on all treatments. ‘Peach’ and ‘T1’ produced less shoots on average; but the most in OIL treatments. All shoots harvested were longer in OIL compared to A; regardless of genotype. Over multiple cycles; ‘Peach’ and ‘T1’ were unable to reliably produce shoots on a repeated schedule and were, therefore, eliminated from the experiment. By cycle 3; maximum number of plants were produced; regardless of supplementation (‘Cherry 1’; 30; ‘BaOx’; 22). Shoot length was above 10 mm (planting standard) for both genotypes until after the third cycle (10 weeks) where number and quality decreased (nodes and internodes easily discerned). By the end of the experiment; the only shoots that remained productive for over 16 weeks and multiple repeated harvest cycles were those in OIL treatments with supplements.
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spelling doaj-art-900f3a056b2d4d5486913fd4acf702c22025-08-20T03:03:25ZengMDPI AGPlants2223-77472025-04-01147113810.3390/plants14071138Modified Media for Repeated In Vitro Cutting Cycles of <i>Cannabis sativa</i> Without the Use of CytokininMolly McKay0James E. Faust1Matthew Taylor2Jeffrey Adelberg3Plant and Environmental Science Department, 171 Poole Agricultural Center, Clemson University, Clemson, SC 29630, USAPlant and Environmental Science Department, 171 Poole Agricultural Center, Clemson University, Clemson, SC 29630, USACurio Wellness, P.O. Box 5470, Towson, MD 21285, USAPlant and Environmental Science Department, 171 Poole Agricultural Center, Clemson University, Clemson, SC 29630, USAIn vitro hedging; combined with the fed-batch liquid media process is an innovative system that generates multiple sterile plants without the use of exogenous cytokinin. This combined process was demonstrated with <i>Cannabis sativa</i> (‘Cherry1’, ‘BaOx’, ‘T1’, ‘Peach’) grown in vessels of three different physical states—stationary agar (A); stationary Oasis<sup>®</sup> infused with liquid (OILs); and agitated Oasis<sup>®</sup> infused with liquid (OILa). Vessels were pre-selected as control or supplemented; where supplement vessels received 15 mL DKW liquid media each cycle harvest. The number of shoot tips harvested; shoot length; and dry shoot mass from repeated cutting cycles was recorded. In a single harvest; ‘BaOx’ and ‘Cherry 1’ produced one shoot per plant from the original 15 planted on all treatments. ‘Peach’ and ‘T1’ produced less shoots on average; but the most in OIL treatments. All shoots harvested were longer in OIL compared to A; regardless of genotype. Over multiple cycles; ‘Peach’ and ‘T1’ were unable to reliably produce shoots on a repeated schedule and were, therefore, eliminated from the experiment. By cycle 3; maximum number of plants were produced; regardless of supplementation (‘Cherry 1’; 30; ‘BaOx’; 22). Shoot length was above 10 mm (planting standard) for both genotypes until after the third cycle (10 weeks) where number and quality decreased (nodes and internodes easily discerned). By the end of the experiment; the only shoots that remained productive for over 16 weeks and multiple repeated harvest cycles were those in OIL treatments with supplements.https://www.mdpi.com/2223-7747/14/7/1138micropropagationhedgingfed-batchcannabistissue culturecytokinin
spellingShingle Molly McKay
James E. Faust
Matthew Taylor
Jeffrey Adelberg
Modified Media for Repeated In Vitro Cutting Cycles of <i>Cannabis sativa</i> Without the Use of Cytokinin
Plants
micropropagation
hedging
fed-batch
cannabis
tissue culture
cytokinin
title Modified Media for Repeated In Vitro Cutting Cycles of <i>Cannabis sativa</i> Without the Use of Cytokinin
title_full Modified Media for Repeated In Vitro Cutting Cycles of <i>Cannabis sativa</i> Without the Use of Cytokinin
title_fullStr Modified Media for Repeated In Vitro Cutting Cycles of <i>Cannabis sativa</i> Without the Use of Cytokinin
title_full_unstemmed Modified Media for Repeated In Vitro Cutting Cycles of <i>Cannabis sativa</i> Without the Use of Cytokinin
title_short Modified Media for Repeated In Vitro Cutting Cycles of <i>Cannabis sativa</i> Without the Use of Cytokinin
title_sort modified media for repeated in vitro cutting cycles of i cannabis sativa i without the use of cytokinin
topic micropropagation
hedging
fed-batch
cannabis
tissue culture
cytokinin
url https://www.mdpi.com/2223-7747/14/7/1138
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