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Premise of the study:  Polymorphic microsatellite loci were developed and used to genotype individuals of  Herbertia zebrina  (Iridaceae) as a first step for assessment of intraspecific genetic diversity.    Methods and Results:  Primer pairs for 47 markers were developed: 20 from a microsatellite‐enriched library and 27 from a next‐generation sequencing run using the Illumina MiSeq platform. Of those, 15 loci were considered successful, of which 12 were polymorphic and three were monomorphic. The primers were tested in 50 individuals from three populations of  H. zebrina . Two to 14 alleles per locus were identified, and observed and expected heterozygosity were 0.00–0.95 and 0.18–0.89, respectively. Tests of cross‐amplification to evaluate the applicability of these markers showed positive results in one congeneric species,  H. darwinii , and in a phylogenetically closely related species,  Calydorea crocoides .    Conclusions:  These microsatellite markers can be used for studies of genetic variation and genetic population structure, as well as to support conservation efforts.

Fifteen microsatellite markers for Herbertia zebrina (Iridaceae): An endangered species from South American grasslands