School of Pharmacy, University of Mississippi, University, MS, USA.
Planta Med. 2010 Oct;76(14):1629-33. doi: 10.1055/s-0030-1249773. Epub 2010 Mar 30.
An efficient in vitro propagation protocol for rapidly producing Cannabis sativa plantlets from young leaf tissue was developed. Using gas chromatography-flame ionization detection (GC-FID), high THC yielding elite female clone of a drug-type CANNABIS variety (MX) was screened and its vegetatively propagated clones were used for micropropagation. Calli were induced from leaf explant on Murashige and Skoog medium supplemented with different concentrations (0.5, 1.0, 1.5, and 2.0 µM) of indole- 3-acetic acid (IAA), indole- 3- butyric acid (IBA), naphthalene acetic acid (NAA), and 2,4-dichlorophenoxy-acetic acid (2,4-D) in combination with 1.0 µM of thidiazuron (TDZ) for the production of callus. The optimum callus growth and maintenance was in 0.5 µM NAA plus 1.0 µM TDZ. The two-month-old calli were subcultured to MS media containing different concentrations of cytokinins (BAP, KN, TDZ). The rate of shoot induction and proliferation was highest in 0.5 µM TDZ. Of the various auxins (IAA, IBA, and NAA) tested, regenerated shoots rooted best on half strength MS medium (1/2 - MS) supplemented with 2.5 µM IBA. The rooted plantlets were successfully established in soil and grown to maturity with no gross variations in morphology and cannabinoids content at a survival rate of 95 % in the indoor growroom.
从年轻的叶片组织快速生产大麻植物的高效体外繁殖方案。利用气相色谱-火焰离子化检测（GC-FID），对一种药用大麻品种（MX）的高 THC 产生的优良雌性无性系进行筛选，并将其营养繁殖无性系用于微繁殖。将叶片外植体在添加不同浓度（0.5、1.0、1.5 和 2.0 μM）吲哚-3-乙酸（IAA）、吲哚-3-丁酸（IBA）、萘乙酸（NAA）和 2,4-二氯苯氧乙酸（2,4-D）的Murashige 和 Skoog 培养基上诱导愈伤组织，同时添加 1.0 μM 的噻二唑（TDZ）用于产生愈伤组织。最佳的愈伤组织生长和维持是在 0.5 μM NAA 加 1.0 μM TDZ。两个月大的愈伤组织在含有不同浓度细胞分裂素（BAP、KN、TDZ）的 MS 培养基中进行继代培养。在 0.5 μM TDZ 中，诱导和增殖芽的速率最高。在所测试的各种生长素（IAA、IBA 和 NAA）中，再生芽在补充有 2.5 μM IBA 的半强度 MS 培养基（1/2 - MS）上生根最好。生根的幼苗成功地在土壤中建立，并在室内生长室中以 95%的存活率生长成熟，形态和大麻素含量没有明显变化。
