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Abstract The numerous ephemeral glacial meltwater streams that flow during the summer in the McMurdo Dry Valleys of South Victoria Land, Antarctica, provide habitats for microbial mats. One of the common mat types is composed of Chlorophyta (colloquially known as a ‘green mat’ due to its colour). While the presence of these mats is regularly monitored, their taxonomic makeup is still under investigation. Using 18S rRNA gene sequencing, the composition of the chlorophyte-dense mats from between rocks and in the main channel from several streams across two valleys was examined. Samples were maintained in native stream water, and select samples from representative locations were transferred to Bristol Medium. The appearance of other eukaryotic species - diatoms and tardigrades - in these green mats completed this integrated study. The results show that the relative abundance of Chlorophyta was significantly increased with the introduction of inorganic nitrogen from Bristol Medium. Chlorophyte taxa in theHazeniaandPleurastrumgenera dominated the samples across both sample types (rock or exposed) and treatments (Antarctic water or Bristol Medium). Furthermore, a reduction in overall sample diversity was observed in samples in Bristol Medium, suggesting preferential nitrogen utilization by these chlorophytes.