Ioannidis Kostas, Tomprou Ioanna, Mitsis Vangelis
Laboratory of Sylviculture, Forest Genetics and Biotechnology, Institute of Mediterranean and Forest Ecosystems, Hellenic Agricultural Organization "Demeter", 11528 Athens, Greece.
Ekati Alchemy Lab SL, Carretera Barcelona 11, 08180 Moia, Spain.
Plants (Basel). 2022 May 18;11(10):1333. doi: 10.3390/plants11101333.
An alternative in vitro propagation protocol for medical L. cultivars for pharmaceutical industrial use was established. The aim of the protocol was to reduce the culture time, offering healthy and aseptic propagating material, while making the whole process more economic for industrial use. The propagation procedure was performed using plastic autoclavable vented and non-vented vessels, containing porous rooting fine-milled sphagnum peat moss-based sponges, impregnated in ½ Murashige and Skoog liquid growth medium, supplemented with indole-3-butyric acid (IBA) at various concentrations (0, 2.46, 4.92, and 9.84 µM) or by dipping nodal cuttings into 15 mM IBA aqueous solution. The highest average root numbers per cutting, 9.47 and 7.79 for high cannabidiol (H_CBD) and high cannabigerol (H_CBG) varieties, respectively, were achieved by dipping the cuttings into IBA aqueous solution for 4 min and then placing them in non-vented vessels. The maximum average root length in H_CBD (1.54 cm) and H_CBG (0.88 cm) was ascertained using 2.46 μM filter sterilized IBA in non-vented vessels. Filter-sterilized IBA at concentrations of 2.46 μM in vented and 4.92 μM in non-vented vessels displayed the maximum average rooting percentages in H_CBD (100%) and H_CBG (95.83%), respectively. In both varieties, maximum growth was obtained in non-vented vessels, when the medium was supplemented with 4.92 μM filter-sterilized IBA. Significant interactions between variety and vessel type and variety and IBA treatments were observed in relation to rooting traits. Approximately 95% of plantlets were successfully established and acclimatized in field. This culture system can be used not only for propagating plant material at an industrial scale but also to enhance the preservation and conservation of genetic material.
建立了一种用于制药工业的药用大麻品种的体外繁殖方案。该方案的目的是缩短培养时间，提供健康无菌的繁殖材料，同时使整个过程在工业应用中更具经济性。繁殖过程使用可高压灭菌的塑料通气和不通气容器，容器中装有基于多孔生根细磨水苔泥炭藓的海绵，海绵浸渍在½ Murashige和Skoog液体生长培养基中，并添加了不同浓度（0、2.46、4.92和9.84 μM）的吲哚-3-丁酸（IBA），或者将带节插条浸入15 mM IBA水溶液中。对于高CBD（H_CBD）和高CBG（H_CBG）品种，通过将插条浸入IBA水溶液中4分钟，然后将其置于不通气容器中，分别获得了最高的平均每插条根数，即9.47和7.79。在不通气容器中使用2.46 μM过滤灭菌的IBA确定了H_CBD（1.54 cm）和H_CBG（0.88 cm）的最大平均根长。在通气容器中浓度为2.46 μM和不通气容器中浓度为4.92 μM的过滤灭菌IBA分别在H_CBD（100%）和H_CBG（95.83%）中显示出最高的平均生根率。在两个品种中，当培养基添加4.92 μM过滤灭菌的IBA时，在不通气容器中获得了最大生长量。在生根性状方面观察到品种与容器类型以及品种与IBA处理之间存在显著相互作用。大约95%的植株在田间成功定植并适应环境。这种培养系统不仅可用于工业规模繁殖植物材料，还可用于加强遗传材料的保存和保护。
