Development and Characterization of Microsatellite Markers for Three Pollination Morphs of Cimicifuga simplex (Ranunculaceae)

Cimicifugasimplex Wormsk. (Ranunculaceae) is a perennial herbdistributed in eastern and northeastern Asia for which at least three differentpollination morphs have been reported. It is classified as endangered or nearthreatened in some Japanese regions, and its rhizome is commercially used as acrude drug. To examine genetic differentiation and gene flow among the threemorphs, we developed eight microsatellite markers by using next-generationsequencing and estimated the genetic structure of C. simplex. We tested eight primer pairs on 93 individuals from sixpopulations of C. simplex in Nagano,central Japan, and found that heterozygosity in morphs I and III was lowcompared to expected heterozygosity. Bayesian clustering performed with theSTRUCTURE program clearly distinguished the three morphs of C. simplex, and only a little gene flowwas detected among the morphs. These eight microsatellite markers are expectedto be useful in conservation genetic studies of this species and for futureconservation planning.