Application of AFLP for the detection of sex-specific markers in hemp

Two dioecious hemp accessions (Can18 and Can17) were tested by bulked segregant analysis for polymorphisms between male and female bulks with amplified fragment length polymorphisms. Thirty‐nine primer combinations were tested and 20 of these yielded one to three male‐specific bands. In contrast, no female‐specific band was detected. Eight of these primer combinations were used for testing 80 progeny plants from a cross between two plants from Can18 and 30 plants from Can17. A total of 16 and 17 male‐specific fragments were obtained for Can 18 and Can 17, respectively. Eleven fragments exhibited the same fragment size in both accessions. All male plants, but not one female plant, showed the respective polymorphic band with each of the eight primer combinations. Problems regarding sex determination under field conditions were successfully overcome by testing plants that had been grown in small pots in a greenhouse. The abundant number of potential markers for the male sex, their complete cosegregation with male plants and the absence of markers for the female sex support the presence of a male sex chromosome in hemp.