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Iris series Hexagonae is a small, monophyletic complex of five species and associated hybrid populations, popularly known as the ‘Louisiana irises’. Series Hexagonae has been recognized as a textbook case of introgressive hybridization based on numerous studies in Louisiana. We previously explored patterns of genetic structure and diversity in populations of the complex in Florida. Populations that occupy high, dry habitats have plants with a distinctive floral ‘highlands’ phenotype. Aquatic populations of I. savannarum also have a consistent floral phenotype (‘coastal’). Jacks Branch slough in Glades County harbors the largest population of series Hexagonae sampled in our previous studies. It is an ecologically heterogeneous site, punctuated by sandy uplands that remain above high water and harbor Iris with the highlands floral phenotype as well as coastal types in the wetter areas of the slough. We hypothesized that mesic, but non-inundated, sites in the slough would host hybrids of the two phenotypic groups, and tested these hypotheses with 19 microsatellite loci. All data analyses support our hypothesis of hybridization between the upland and hydric subpopulations. Although two methods of introgression analysis identify some of the individuals from the admixture zone as first-generation hybrids, subsequent admixture was asymmetric with introgression from the ‘highland’ parent. This is the first report of such assortative processes occurring in the confines of a single population of Iris. We suggest that the evolutionary processes described interspecifically for series Hexagonae commence early at the population level.

Ten microsatellite loci isolated from Iris hexagona are described. All these loci are polymorphic, with three to 13 alleles across 24 individuals from a single natural population. Heterozygosity ranged from 0.125 to 0.870. Three loci depart significantly from Hardy–Weinberg equilibrium in our test population. The test population shows significant heterozygote deficiency in these and two other loci. Three loci exhibit significant linkage disequilibrium. These loci will be utilized to investigate patterns of genetic variation in the species throughout the Florida peninsula.