Chloroplast simple sequence repeats amplicons in 5 subspecies of Brassica oleracea were sequenced, and one chloroplast SNP was detected in amplicon ACP43. Through the introduction of an RsaI recognition site by adding one mismatch in the forward primer, combined with the increased primer length and raised annealing temperature, the dCAPS (derived cleaved amplified polymorphic sequences) marker ACP43-93 RsaI was successfully developed. By using the dCAPS marker, the subspecies-specific SNP was assayed in 206 materials representing the wide distribution of B. oleracea. This is the first report of chloroplast DNA (cpDNA) variation in cultivated subspecies of B. oleracea, which showed that chloroplast diversity existed at the intersubspecies level. Unlike other subspecies, most of the broccoli and all of the cauliflower materials sharing the same haplotype showed closer relationships in cpDNA level. Furthermore, the dCAPS haplotype of the offspring from 7 male sterile backcross populations was the same as the female parents, indicating maternal inheritance.
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