Amphibia-Reptilia 28 (2007): 1-6 Genetic variation within African spiny-tailed lizards (Agamidae: Uromastyx) estimated using mitochondrial DNA sequences D. James Harris * , Raquel Vaconcelos, José C. Brito Abstract. African spiny-tailed lizards (Uromastyx) are large, herbivorous lizards extensively traded locally for food and internationally as pets. Several species have recently been described, although some remain controversial. To determine relationships within North African forms, twenty individuals were analysed for over 1000 bases of mitochondrial DNA sequences. Phylogenetic analyses indicate four deeply divergent lineages that correspond to sampling areas, but not to current species designations. These results indicate that present taxonomy does not reflect the evolutionary history of these species. Introduction Spiny-tailed lizards of the genus Uromastyx are large generalist herbivores distributed in arid re- gions from India to the Arabian Peninsula and across the Saharan region of Africa. Uromastyx is registered in appendix II of CITES due to their being collected for food as well as exten- sively for the pet trade – based on CITES data over 215 000 Uromastyx were legally traded be- tween 1977-2001, almost all wild-caught and primarily from Egypt and Mali (Knapp, 2004). Because of this, there are many publications regarding captive breeding and husbandry, but few related to ecology, behaviour in the wild, or phylogenetic relationships. Currently 16 species are recognized (Wilms, 2001), with several in- cluding Uromastyx geyri (Joger and Lambert, 1986), Uromastyx flavifasciata, Uromastyx oc- cidentalis (Mateo et al., 1998), Uromastyx al- fredschmidtii (Wilms and Böhme, 2001) and Uromastyx leptiens (Wilms and Böhme, 2000) being recently described. The acceptance of the specific status of some forms is not univer- sal (e.g. U. flavifasciata, reviewed in Geniez et al., 2004). Amer and Kumazawa (2005) presented the first phylogeny based on mito- chondrial DNA (mtDNA) sequences that in- Centro de Investigação em Biodiversidade e Recur- sos Genéticos (CIBIO\UP), ICETA, Campus Agrario de Vairão, 4485-661 Vila do Conde, Portugal * Corresponding author; e-mail: james@mail.icav.up.pt cluded several African and Arabian taxa. They clearly showed that the African forms were a clade, but included only four species within this clade and none of the more controversial recently described forms. Further, many were captive bred individuals, so geographic varia- tion within species could not be assessed. To determine relationships within the North–West African species, 20 individuals were analysed, all sampled in the field, from five species for approximately 1050 base pairs of mtDNA from three genes (cytochrome b, 12S rRNA and 16S rRNA). Genetic variation was then compared to the current taxonomic status. Methods Specimens were identified in the field using discriminant characters published in available guides (Geniez et al., 2004), digital photographs taken (available though the au- thors by request), and a toe or piece of tail was removed and stored in 100% ethanol (fig. 1 and table 1). Some specimens were roadkills. All live specimens were released immedi- ately after sampling. Total genomic DNA was extracted from these small pieces of tissue using standard methods, following Harris et al. (1998). Polymerase Chain Reaction primers used in both amplification and sequencing were 12Sa and 12Sb, 16SL and 16SH, and cytochrome b1 and cy- tochrome b3 from Kocher et al. (1989), Simon et al. (1990) and Palumbi et al. (1991). Amplified fragments were se- quenced on a 310 Applied Biosystem DNA Sequencing Ap- paratus. Mitochondrial DNA sequences were aligned using Clustal W (Thompson et al., 1994). Since North African Uromastyx are known to form a clade (Amer and Kumaza- wa, 2005), the Uromastyx sp. from Yemen was designated as an outgroup. A total of 20 taxa were analysed. Aligned © Koninklijke Brill NV, Leiden, 2007. Also available online - www.brill.nl/amre