Expediting cannabis seed examination by combining color reaction and DNA testing

Abstract

In Japan, possession of germinable cannabis seeds for cultivation purposes is subject to prosecution. Cannabis seeds are marketed after being treated to prevent germination ( heated or crushed ) Currently, forensic examination of cannabis seeds is conducted by cultivating the seeds after germination tests for several weeks and then performing morphological observation and chemical analysis on the true leaves. In this study, we attempted to construct a rapid and simple method for the identiˆcation of cannabis seeds by combining the color reaction using 2,3,5  Triphenyl  2 H  tetrazolium Chloride ( TTC ) , a reagent that discriminates between living and dead cells, and DNA testing using a commercially available simple kit. The color reaction using TTC can determine the viability of peeled embryos within 20 min at 45 ° C as previously reported. This method is eŠective for quickly determining whether a seed has been heat-treated or not. However, in the color reaction, a commercial health food seed that claimed to be unheated showed some colo-ration. This sample had been crushed to prevent germination and was easily identi-ˆed as non-germinable by morphological examination. After the color reaction, the embryos could be directly used for