National Center for Natural Products Research, School of Pharmacy, The University of Mississippi, University, MS 38677, USA.
Planta Med. 2010 Jan;76(1):97-100. doi: 10.1055/s-0029-1185945. Epub 2009 Jul 27.
Inter-simple sequence repeat (ISSR) markers were used to evaluate the genetic stability of the micropropagated plants of Cannabis sativa over 30 passages in culture and hardening in soil for 8 months. A total of 15 ISSR primers resulted in 115 distinct and reproducible bands. All the ISSR profiles from micropropagated plants were monomorphic and comparable to mother plants, confirming the genetic stability among clones and mother plants. Chemical analysis of cannabinoids, using gas chromatography/flame ionization detection (GC/FID), was done to further confirm whether the qualitative and quantitative differences in the major secondary metabolites exist between the mother plant and micropropagated plants. Six major cannabinoids - Delta(9)-THC, THCV, CBD, CBC, CBG, and CBN - were identified and compared with the mother plant. Our results clearly showed a similar cannabinoid profile and insignificant differences in THC content between the two types of plants. These results suggest that the micropropagation protocol developed by us for rapid IN VITRO multiplication is appropriate and applicable for clonal mass propagation of C. SATIVA.
利用 ISSR 标记对大麻的微繁殖植株在培养中经过 30 代和在土壤中硬化 8 个月的遗传稳定性进行了评估。总共使用了 15 个 ISSR 引物，产生了 115 个独特且可重复的条带。微繁殖植株的所有 ISSR 图谱均为单态，与母本相似，证实了克隆和母本之间的遗传稳定性。使用气相色谱/火焰离子化检测（GC/FID）对大麻素进行化学分析，以进一步确认母本植株和微繁殖植株之间主要次生代谢产物的定性和定量差异是否存在。鉴定并比较了 6 种主要大麻素 - Delta(9)-THC、THCV、CBD、CBC、CBG 和 CBN - 。我们的结果清楚地显示了两种类型的植物具有相似的大麻素图谱和 THC 含量无显著差异。这些结果表明，我们开发的用于快速体外繁殖的微繁殖方案适合并适用于大麻的无性系大规模繁殖。
