Department of Biological Sciences, University of Lethbridge, 4401 University Drive, Lethbridge, Alberta T1K 3M4, Canada.
Plant Sci. 2025 Jan;350:112296. doi: 10.1016/j.plantsci.2024.112296. Epub 2024 Oct 18.
Large scale production of uniform disease-free plants is crucial for Cannabis sativa biotechnology. Existing micropropagation protocols rely heavily on shoot multiplication from existing meristems via direct organogenesis. Such protocols do not allow multiplication of plant material through continuous sub-culturing. Protocols that use indirect regeneration are usually not efficient enough and have very low multiplication rates. In the present study, an efficient protocol that uses a combination of direct organogenesis and callogenesis to induce multiple shoot development cultures is developed. Callogenesis was induced from various explants cultured on the media having various combinations of thidiazuron (TDZ) and naphthaleneacetic acid (NAA); best callogenesis and shoot regeneration was achieved from hypocotyl explants cultured on TDZ 0.4 mg l NAA 0.2 mg l. Hypocotyls with cotyledonary node and shoot apical meristem were significantly better for shoot regeneration than explants without it. Shoots obtained from multiple shoot cultures were successfully rooted and then acclimatized under greenhouse conditions to develop into adult cannabis plants.
大规模生产均匀无病的植物对于大麻生物技术至关重要。现有的微繁殖方案主要依赖于通过直接器官发生从现有分生组织中进行芽的增殖。这样的方案不允许通过连续的继代培养来繁殖植物材料。使用间接再生的方案通常效率不够高，增殖率很低。在本研究中，开发了一种使用直接器官发生和愈伤组织发生相结合的有效方案来诱导多个芽发育培养物。从在含有不同浓度噻苯隆（TDZ）和萘乙酸（NAA）的培养基上培养的各种外植体中诱导愈伤组织发生；从在 TDZ 0.4 mg l 和 NAA 0.2 mg l 的培养基上培养的下胚轴外植体中获得了最佳的愈伤组织发生和芽再生。具有子叶节点和茎尖分生组织的下胚轴明显比没有这些组织的外植体更有利于芽再生。从丛生芽培养物中获得的芽成功生根，然后在温室条件下适应，发育成成年大麻植物。
