RAPD and SCAR molecular markers of sexuality in the dioecious species Cannabis sativa L.

In this study the random amplified polymorphic DNA (RAPD) technque was employed with the objective of finding markers linked to sex determination in hemp. Ten male individual DNA samples or ten female individual DNA samples or ten female individual DNA sampes were prepared and contributed to two DNA pools: one was male DNA pool and another female DNA pool, with the intention of providing a common genetic background for each pool and leaving the sex-linked materials as the primary difference between the two pools. A total of 30 10-mer primers was tested with at least three different PCR cycling procedures. A male-associated fragment with a length of about 2.5 kb was generated with S401 primer. The fragment was cloned and sequenced. In order to convert the RAPD marker into SCAR (sequence characterized amplified regions) marker, one 20-mer and another 22-mer specific primers were constructed and used for PCR amplifying. The male-linked dominant SCAR marker was obtained, which would favor the screening of all-gynoecious hemp lines.