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type of cannabis by seed id

Cannabis seed identification by chloroplast and nuclear DNA

The Control Substances Hazard Prevention Act classifies Cannabis sativa as a controlled substance under schedule II of the Act in Taiwan. The punishment for trafficking may be a sentence of up to life in prison. Huo Ma Jen is usually imported as a Chinese medicinal remedy and also as bird food. Importers of cannabis material make use of loopholes in national legislation and claim that the botanical material is from the plant known as Huo Ma Jen and as such avoids normal regulations. The term Huo Ma Jen is therefore frequently used as a cover for the importation of cannabis seeds for germination.

The identification of C. sativa plant material is conducted using morphological observation based upon the cystolithic hairs on the leaves [1], chemical tests to identify of the presence of cannabinoids [2] and more recently using DNA analysis [3,4].

When seeds suspected of contravening national legislation are seized, the seeds are germinated into a viable plant so that morphological and chemical tests can be conducted. There is a delay in the process while the seeds germinate. In the case outlined it was possible to obtain species identification from the seeds directly without the need for propagation. The methods used analyzed species conserved loci and were used in conjunction with the 2,3,5-triphenyltetrazolium chloride (TTC) method [5] to detect viability. The gene loci used were the intergenic spacer (IGS) of trnL (UAA)-trnF (GAA) in the chloroplast genome and the internal transcribed spacer I (ITS1) of nuclear rDNA.

From 2001 to 2004, a total of eleven containers of suspect seeds labeled as ‘Huo Ma Jen’ were seized by custom officials at Keelung Harbor. The seeds were suspected to be C. sativa but the owners claimed that the seeds were from a Chinese medicinal plant called ‘Huo Ma Jen’. Random samples were collected from the 11 containers for identification. Approximately 1 kg of suspect seeds from each container was collected. Leaves of C. sativa and the closest related species Humulus japonicus and Humulus lupulus were also collected and used as reference samples for species identification. Ten seed samples from each container were germinated in culture medium to observe seedling growth and 100 seed samples from each container were used to test the rate of viability by TTC. Ten seed samples from each container and leaves of seedlings.