Australian myobatrachid frogs of the genus Pseudophryne have only two classes of alkaloids in skin extracts, pseudophrynamines (PSs) and pumiliotoxins (PTXs). The former are unique to such Australian frogs, while the PTXs occur worldwide in all other genera of frogs/toads that contain lipophilic alkaloids. The major alkaloid of wild-caught frogs from one population of Pseudophryne semimarmorata was PTX 267C, while PSs were only minor or trace alkaloids. Captive-raised frogs from the same parental stock had no PTXs, but had larger amounts of PSs. A PTX fed to captive-raised frogs accumulated into skin along with dihydro and hydroxy metabolites. Thus, Pseudophryne frogs appear to biosynthesize PSs, but to sequester into skin dietary PTXs. In addition, biosynthesis of PSs appears reduced when high levels of dietary PTXs have accumulated into skin. This is the first evidence indicating that certain frogs are capable of synthesizing rather than merely sequestering alkaloids. A wide range of PSs, including many with molecular weights >500, were detected using both GC-mass spectral and LC-mass spectral analysis.
Arrenurus (Arrenuridae) is the most species-rich genus of mites with about 950 named species that inhabit standing, and to a lesser extent, running water habitats around the world. To date, distinguishing species of Arrenurus has been based on male reproductive morphology. Here, we use morphological and molecular approaches to examine species boundaries among 42 named species of Arrenurus, including four named species that have colour variants (red and green A. americanus, and red and blue A. intermedius, A. manubriator and A. apetiolatus), and two unnamed morphospecies. In this study, we examine male genital structures with the use of SEM techniques, and apply mitochondrial (COI barcode region) and nuclear (28S rRNA) gene fragments to test whether male morphology reflects species boundaries in Arrenurus assessed by molecular analyses. Our results reveal that male reproductive morphology parallels species boundaries as judged by molecular data. We discuss the cases of genetically poorly diversified, yet morphologically clearly defined named species. Moreover, we show that based on the species we examined, colour morphs within otherwise morphologically similar specimens represent within-species variation and, in the absence of other diagnostic traits, colour itself can be misleading in distinguishing species. Our outcomes on molecular taxonomy of Arrenurus provide a background for testing hypotheses about speciation rate in water mites.
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