PHYLOGEOGRAPHY AND WITHIN-STREAM GENETIC VARIATION OF BATRACHOSPERMUM HELMENTOSUM (RHODOPHYTA) USING INTER-SIMPLE SEQUENCE REPEATS

A relatively new molecular marker technique, inter-simple sequence repeats (ISSR), was used to assess the within-stream reach genetic variation and phylogeography of the freshwater red alga, Batrachospermum helmentosum. The within-stream reach study was undertaken in Southeastern Ohio, where 58 individuals were collected from an approximately 200 m stream length. Individuals appeared to be congregated into three distinct areas, with no plants between areas. Using three primers, 100 polymorphic bands were scored. Although it was hypothesized that genetic variation would reflect physical distance, the results showed six distinct genetic groupings, some of which consisted of physically distant individuals. Overall, genetic diversity was greater among individuals than between geographic and genetic groups. To investigate phylogeography, 11 stream reaches (3 from Ohio and one each from Connecticut, Massachusetts, Rhode Island, North Carolina, Tennessee, Indiana, Michigan and Louisiana) were sampled throughout the alga's known distribution in North America. Approximately 15 individuals were collected per stream for ISSR analysis. Five primers, yielding 243 polymorphic bands, were used to examine variation among all individuals. Neighbor-joining trees generated from the Dice, Jaccard and Excoffier coefficients showed that individuals from the same stream formed a clade. Among streams there was no distinct correlation between geographic distance and genetic similarity. Genetic groupings varied slightly among distance coefficients employed. Michigan and Tennessee, Connecticut and Rhode Island, and Indiana and Massachusetts streams consistently showed the same relationship, but the relationships among the three Ohio streams and Louisiana stream were variably related to each other.