Opuntioideae, composed of roughly 370 species, occur in almost every biome in the Americas, from seasonally dry tropical forests (SDTF) to high-elevation Andean grasslands, American deserts and temperate zones. The interrelationships among the three major clades of Opuntioideae (Cylindropuntieae, Opuntieae and Tephrocacteae) are not well resolved, and thus, the ancestral habitat, biogeographic history and evolution of morphological characters, such as large photosynthetic leaves and flattened stems, are poorly understood. To test their geographic origin and evolution of key morphological characters, we built the largest phylogenomic dataset for Cactaceae to date using 103 plastid genes of 107 taxa of Opuntioideae. The subfamily Opuntioideae likely evolved in South America in a combination of seasonally dry tropical forest (SDTF)/desert habitats. Opuntieae most likely evolved in South America in SDTF and, from there, moved into desert regions, Chaco and temperate/subtropical zones, while Tephrocacteae and Cylindropuntieae evolved in South America in desert regions and moved into SDTF, Chaco and temperate/subtropical zones. Analyses of morphological evolution suggest that, although large leaves are plesiomorphic in Opuntioideae, long-lived, photosynthetically active leaves in Cylindropuntieae and Tephrocacteae are homoplasious and do not represent retained plesiomorphy, as is often assumed. Flattened stems are synapomorphic for Opuntieae, possibly representing adaptation to competition for light resources in SDTF, their most likely ancestral area.

• Premise of the study: Genetic studies of Cactaceae can at times be impeded by difficult sampling logistics and/or high mucilage content in tissues. Simplifying sampling and DNA isolation through the use of cactus spines has not previously been investigated.• Methods and Results: Several protocols for extracting DNA from spines were tested and modified to maximize yield, amplification, and sequencing. Sampling of and extraction from spines resulted in a simplified protocol overall and complete avoidance of mucilage as compared to typical tissue extractions. Sequences from one nuclear and three plastid regions were obtained across eight genera and 20 species of cacti using DNA extracted from spines.• Conclusions: Genomic DNA useful for amplification and sequencing can be obtained from cactus spines. The protocols described here are valuable for any cactus species, but are particularly useful for investigators interested in sampling living collections, extensive field sampling, and/or conservation genetic studies.

Cirsium (i.e., “thistles”) are one of the most taxonomically challenging groups of Compositae in North America. This study represents the first attempt to infer a broadly sampled phylogeny of Cirsium in North America. The two main objectives are to: (i) test whether currently hypothesized species variety complexes (C. arizonicum, C. clavatum, C. eatonii, and C. scariosum) constitute monophyletic lineages, and (ii) recircumscribe any taxa that are identified as problematic. Phylogeny reconstructions based on DNA sequence data from two nuclear ribosomal regions and four plastid markers were used to infer evolutionary lineages and test species’ delimitations. Eight species varietal complexes were resolved as polyphyletic. We recircumscribed these complexes and in doing so found evidence to support the recognition of six new taxa. We hypothesize that the extensive taxonomic difficulty within Cirsium is the result of several factors: (i) previously undescribed taxa, (ii) inadequate representation of taxa from herbarium specimens, (iii) phenotypic convergence, (iv) hybridization, and (v) incipient speciation. While we can provide evidence to support the recircumscription of some taxa, others remain unresolved. Therefore, we are working on a phylogenomic study of North American Cirsium to answer remaining taxonomic questions as well as provide a robust framework for biogeographic studies.