Distinguishing the distributions of two cryptic frogs (Anura: Discoglossidae) using molecular data and environmental modeling

Currently, the identification of two cryptic Iberian amphibians, Discoglossus galganoi Capula, Nascetti, Lanza, Bullini and Crespo, 1985 and Discoglossus jeanneae Busack, 1986, relies on molecular characterization. To provide a means to discern the distributions of these species, we used 385-base-pair sequences of the cytochrome b gene to identify 54 Spanish populations of Discoglossus. These data and a series of environmental variables were used to build up a logistic regression model capable of probabilistically designating a specimen of Discoglossus found in any Universal Transverse Mercator (UTM) grid cell of 10 km × 10 km to one of the two species. Western longitudes, wide river basins, and semipermeable (mainly siliceous) and sandstone substrates favored the presence of D. galganoi, while eastern longitudes, mountainous areas, severe floodings, and impermeable (mainly clay) or basic (limestone and gypsum) substrates favored D. jeanneae. Fifteen percent of the UTM cells were predicted to be shared by both species, whereas 51% were clearly in favor of D. galganoi and 34% were in favor of D. jeanneae, considering odds of 4:1. These results suggest that these two species have parapatric distributions and allow for preliminary identification of potential secondary contact areas. The method applied here can be generalized and used for other geographic problems posed by cryptic species.

Protein markers of B. xylophilus Steiner & Buhrer, 1934 (Nickle, 1970) populations using quantitative proteomics and character compatibility.

The Pine Wood Nematode (PWN) Bursaphelenchus xylophilus is a severe forest pathogen in countries where it has been introduced and is considered a worldwide quarantine organism. In this study, protein markers for differentiating populations of this nematode were identified by studying differences among four selected Iberian and one American population. These populations were compared by quantitative proteomics (iTRAQ). From a total of 2860 proteins identified using the public database from the B. xylophilus genome project, 216 were unambiguous and significantly differentially regulated in the studied populations. Comparisons of their pairwise ratio were statistically treated and supported in order to convert them into discrete character states, suggesting that 141 proteins were not informative as population specific markers. Application of the Character Compatibility methodology on the remaining 75 proteins (belonging to families with different biological functions) excludes 27 which are incompatible among them. Considering only the compatible proteins, the method selects a subset of 30 specific unique protein markers which allowed the compared classification of the Iberian isolates. This approach makes it easier search for diagnostic tools and phylogenetic inference within species and populations of a pathogen exhibiting a high level of genetic diversity.