Molecular assessment of dietary niche partitioning in an endemic island radiation of tropical mammals.

Island radiations represent unique evolutionary histories in unique ecological contexts. These radiations provide opportunities to investigate ecological diversification in groups that typically exhibit niche partitioning among their constituents, including partitioning of food resources. DNA metabarcoding produces finer levels of diet identification than traditional methods, allowing us to examine dietary niche partitioning in communities or clades in which species share superficially similar diets. Here, we use DNA metabarcoding to investigate dietary niche partitioning in an endemic radiation of mammals in the Philippines. Our data reveal niche partitioning as well as phylogenetically-uncorrelated adaptive evolution in this small mammal community. Because 70% of the focal species belong to the tribe Chrotomyini, an endemic Philippine radiation of murid rodents that feed extensively on earthworms, this study sheds light on dietary adaptation and its role in the co-occurrence of closely related species. Our results reveal fine-scale resource partitioning within this community; our data provide compelling evidence for niche partitioning of diet that was masked by previous diet categories and will help in further dissecting the model adaptive radiation of endemic small mammals on Luzon. This study reinforces the notion that DNA metabarcoding can be a valuable tool for investigating both ecological relationships and evolutionary ecology at the community and phylogenetic level, respectively.

Interspecific diversity of testes mass and sperm morphology in the Philippine chrotomyine rodents: implications for differences in breeding systems across the species.

The high diversity of native Philippine murid rodents includes an old endemic group, the chrotomyines, which are the sister group of the Australasian hydromyines. Herein we detail their interspecific diversity of relative testes mass (RTM) and sperm morphology. We find that in chrotomyines, as in the Australasian hydromyines, testes mass relative to body mass differs by an order of magnitude across the species and ranges from a large RTM in Soricomys and Chrotomys species to a small RTM in Apomys. Sperm morphology is associated with these findings, with individuals in species of Soricomys and Chrotomys producing relatively larger spermatozoa with a prominent apical hook and long tail, whereas, by contrast, the Apomys species have a sperm head that either has a very short or no apical hook and a shorter tail. These findings indicate coevolution of RTM with sperm morphological traits across the species, with the marked interspecific differences in RTM suggesting differences in the intensity of intermale sperm competition and hence breeding system. Thus, we hypothesise that species of Soricomys and Chrotomys that produce more streamlined spermatozoa with longer tails have a polyandrous or promiscuous mating system, whereas the Apomys species, which produce smaller and less streamlined spermatozoa, may exhibit monogamy.

Colonization and diversification of the white-browed shortwing (Aves: Muscicapidae: Brachypteryx montana) in the Philippines.

Molecular phylogenetic approaches have greatly improved our knowledge of the pattern and process of biological diversification across the globe; however, many regions remain poorly documented, even for well-studied vertebrate taxa. The Philippine archipelago, one of the least-studied 'biodiversity hotspots', is an ideal natural laboratory for investigating the factors driving diversification in an insular and geologically dynamic setting. We investigated the history and geography of diversification of the Philippine populations of a widespread montane bird, the White-browed Shortwing (Brachypteryx montana). Leveraging dense archipelago-wide sampling, we generated a multi-locus genetic dataset (one nuclear and two mtDNA markers), which we analyzed using phylogenetic, population genetic, and coalescent-based methods. Our results demonstrate that Philippine shortwings (1) likely colonized the Philippines from the Sunda Shelf to Mindanao in the late Miocene or Pliocene, (2) diversified across inter-island barriers into three divergent lineages during the Pliocene and early Pleistocene, (3) have not diversified within the largest island, Luzon, contrary to patterns observed in other montane taxa, and (4) colonized Palawan from the oceanic Philippines rather than from Borneo, challenging the assumption of Palawan functioning exclusively as a biogeographic extension of the Sunda Shelf. Additionally, our finding that divergent (c. 4.0 mya) lineages are coexisting in secondary sympatry on Mindanao without apparent gene flow suggests that the speciation process is likely complete for these shortwing lineages. Overall, these investigations provide insight into how topography and island boundaries influence diversification within remote oceanic archipelagos and echo the results of many other studies in demonstrating that taxonomic diversity continues to be underestimated in the Philippines.

A re-assessment of diversity among Philippine gymnures (Mammalia: Erinaceidae: Podogymnura), with a new species from eastern Mindanao.

Podogymnura, a Philippine endemic genus of gymnures ("soft-furred hedgehogs"), has been known previously only from the highlands of central Mindanao (P. truei truei and P. t. minima) and from two small islands off the northeast tip of Mindanao (P. aureospinula). Based on a combination of mitochondrial genetic and qualitative and quantitative morphological data, we recognize P. minima, formerly considered a subspecies of P. truei, as a distinct species. We also recognize specimens from two little-studied montane regions of southeastern Mindanao, Mt. Hamiguitan and Mt. Kampalili, as a distinctive new species, P. intermedia n. sp. We estimate that dispersal of a common ancestor from Borneo to Mindanao ca. 1.9 to 3.5 Ma gave rise to Podogymnura, followed by speciation within Mindanao beginning 0.53 to 0.97 Ma. These results further increase the rich diversity of mammals on Mindanao, and underscore the importance of the eastern Mindanao highlands as a likely important but little-studied center of Philippine mammalian diversity.

Single-locus species delimitation: a test of the mixed Yule-coalescent model, with an empirical application to Philippine round-leaf bats.

Prospects for a comprehensive inventory of global biodiversity would be greatly improved by automating methods of species delimitation. The general mixed Yule-coalescent (GMYC) was recently proposed as a potential means of increasing the rate of biodiversity exploration. We tested this method with simulated data and applied it to a group of poorly known bats (Hipposideros) from the Philippines. We then used echolocation call characteristics to evaluate the plausibility of species boundaries suggested by GMYC. In our simulations, GMYC performed relatively well (errors in estimated species diversity less than 25%) when the product of the haploid effective population size (N(e)) and speciation rate (SR; per lineage per million years) was less than or equal to 10(5), while interspecific variation in N(e) was twofold or less. However, at higher but also biologically relevant values of N(e) × SR and when N(e) varied tenfold among species, performance was very poor. GMYC analyses of mitochondrial DNA sequences from Philippine Hipposideros suggest actual diversity may be approximately twice the current estimate, and available echolocation call data are mostly consistent with GMYC delimitations. In conclusion, we consider the GMYC model useful under some conditions, but additional information on N(e), SR and/or corroboration from independent character data are needed to allow meaningful interpretation of results.

A new genus and species of shrew-like mouse (Rodentia: Muridae) from a new center of endemism in eastern Mindanao, Philippines.

The Philippine archipelago hosts an exceptional diversity of murid rodents that have diversified following several independent colonization events. Here, we report the discovery of a new species of rodent from Mt. Kampalili on eastern Mindanao Island. Molecular and craniodental analyses reveal this species as a member of a Philippine "New Endemic" clade consisting of Tarsomys, Limnomys, and Rattus everetti (tribe Rattini). This new species of "shrew-mouse" is easily distinguished from its relatives in both craniodental and external characteristics including a long, narrow snout; small eyes and ears; short, dark, dense fur dorsally and ventrally; stout body with a tapering, visibly haired tail shorter than head and body length; stout forepaws; bulbous and nearly smooth braincase; narrow, tapering rostrum; short incisive foramina; slender mandible; and narrow, slightly opisthodont incisors. This new genus and species of murid rodent illustrates that murids of the tribe Rattini have exhibited greater species and morphological diversification within the Philippines than previously known and provides evidence that Mt. Kampalili represents a previously unrecognized center of mammalian endemism on Mindanao Island that is deserving of conservation action.

Wallace's line, Wallacea, and associated divides and areas: history of a tortuous tangle of ideas and labels.

Due to its position between the highly distinct Oriental and Australasian biogeographical realms, much effort has been spent demarcating associated separations and transitions in the faunal assemblages of the Indo-Australian Archipelago. Initially, sharp boundary lines were proposed, with the earliest dating from the mid-1800s. Notably, the one published by Alfred R. Wallace in 1863, based upon land-mammal and land-bird distributions, has since achieved iconic status and today its significance is recognized well beyond the confines of the biogeography community. Over the next four decades many such divides were engraved onto plates and inked onto charts of SE Asia using additional information, different organisms or other criteria. However, it became apparent that, as Wallace had noted, all such lines were to some degree permeable, and by the 1880s transition zones were being put forward instead; the label 'Wallacea' was introduced in 1924. Interestingly, the last decade has seen new divides and sub-regions being proposed, some departing markedly from earlier offerings. Although currently there is general agreement regarding much of the terminology associated with both the lines and the areas, the record of publication indicates that this consensus has emerged obliquely, and in some cases is weakly founded. This review does not present new data nor new analyses; rather it summarizes the development of ideas and reflects upon attendant issues that have emerged. After reviewing the key proposals, recommendations are presented that should in future alleviate perceived difficulties or inadequacies. Reference to specific divides must be true to their original definitions; there are many instances where the secondary literature has portrayed them incorrectly and with some this has rippled through into later publications. Moreover, Wallace's 1863 line is not the one that he finally settled upon (in 1910); its path around Sulawesi was transferred from the west to the east of the Island. Ideally, Huxley's divide (1868) should carry his name rather than Wallace's; the latter never accepted the proposition. Lydekker's Line (1896) ought to be labelled the Heilprin-Lydekker Line in recognition of Angelo Heilprin's 1887 contribution. Concerning transition zones, ideally Wallacea should correspond to its original 1924 description, which incorporated the Philippine islands bar the Palawan group. Notably, though, a smaller form (introduced by Darlington in 1957, used frequently from 1998 onwards) in which all of the Philippine islands are excluded is entrenched within the recent literature, but this is often without evident justification. It should also be recognized that the 'reduced' (=southern) Wallacea area was effectively defined by Heilprin in 1887, but was then labelled the 'Austro-Malaysian Transition Zone'. Finally, the application in recent years of modern analytical techniques has not led to a consensus view on where the lines/areas should run/be placed; with such a large, diverse set of organisms, each with differing histories, this is perhaps not surprising.

Philippine bats of the genus Kerivoula (Chiroptera: Vespertilionidae): Overview and assessment of variation in K. pellucida and K. whiteheadi.

Bats of the genus Kerivoula (Mammalia, Chiroptera) are widespread in the Philippines with four reported species, but have been poorly known due to a paucity of specimens. We provide the first molecular phylogeny for Philippine Kerivoula, which supports the existence of four distinct clades that we treat as species (K. hardwickii, K. papillosa, K. pellucida, and K. whiteheadi); these four overlap broadly geographically. Each of these may be recognized on the basis of cytochrome b sequences and external and craniodental morphology. Detailed examination of K. pellucida shows little geographic differentiation within the Philippines, but they differ subtly from those on the Sunda Shelf. We consider K. whiteheadi to be composed of four recognizable clades, each restricted to a geographic region within the Philippines. We consider K. bicolor, from peninsular Thailand, and K. pusilla, from Borneo, to be distinct from K. whiteheadi. Our data indicate the presence of two species within the Philippines currently lumped as K. hardwickii; further study of these is needed. A calibrated phylogeny suggests that Kerivoula began arriving in the Philippines about 10 MYA, with each of the four current lineages arriving independently.

Tempo and mode of mandibular shape and size evolution reveal mixed support for incumbency effects in two clades of island-endemic rodents (Muridae: Murinae).

Existing radiations in a spatially limited system such as an oceanic island may limit the ecological opportunity experienced by later colonists, resulting in lower macroevolutionary rates for secondary radiations. Additionally, potential colonists may be competitively excluded by these incumbent (resident) species, unless they are biologically distinct (biotic filtering). The extant phenotypic diversity of secondary colonists may thus be impacted by lower rates of phenotypic evolution, exclusion from certain phenotypes, and transitions to new morphotypes to escape competition from incumbent lineages. We used geometric morphometric methods to test whether the rates and patterns of mandibular evolution of the Luzon "old endemic" rodent clades, Phloeomyini and Chrotomyini, are consistent with these predictions. Each clade occupied nearly completely separate shape space and partially separate size space. We detected limited support for decelerating and clade-specific evolutionary rates for both shape and size, with strong evidence for a shift in evolutionary mode within Chrotomyini. Our results suggest that decelerating phenotypic evolutionary rates are not a necessary result of incumbency interactions; rather, incumbency effects may be more likely to determine which clades can become established in the system. Nonincumbent clades that pass a biotic filter can potentially exhibit relatively unfettered evolution.

Diversification rates of the "Old Endemic" murine rodents of Luzon Island, Philippines are inconsistent with incumbency effects and ecological opportunity.

Diversity-dependent cladogenesis occurs when a colonizing lineage exhibits increasing interspecific competition as it ecologically diversifies. Repeated colonization of a region by closely related taxa may cause similar effects as species within each lineage compete with one another. This may be particularly relevant for secondary colonists, which could experience limited diversification due to competition with earlier, incumbent colonists over evolutionary time. We tested the hypothesis that an incumbent lineage may diminish the diversification of secondary colonists in two speciose clades of Philippine "Old Endemic" murine rodents-Phloeomyini and Chrotomyini-on the relatively old oceanic island of Luzon. Although phylogenetic analyses confirm the independent, noncontemporaneous colonization of Luzon by the ancestors of these two clades, we found no support for arrested diversification in either. Rather, it appears that diversification of both clades resulted from constant-rate processes that were either uniform or favored the secondary colonists (Chrotomyini), depending on the method used. Our results suggest that ecological incumbency has not played an important role in determining lineage diversification among Luzon murines, despite sympatric occurrence by constituent species within each lineage, and a substantial head start for the primary colonists.

Convergent evolution of an extreme dietary specialisation, the olfactory system of worm-eating rodents.

Turbinal bones are key components of the mammalian rostrum that contribute to three critical functions: (1) homeothermy, (2) water conservation and (3) olfaction. With over 700 extant species, murine rodents (Murinae) are the most species-rich mammalian subfamily, with most of that diversity residing in the Indo-Australian Archipelago. Their evolutionary history includes several cases of putative, but untested ecomorphological convergence, especially with traits related to diet. Among the most spectacular rodent ecomorphs are the vermivores which independently evolved in several island systems. We used 3D CT-scans (N = 87) of murine turbinal bones to quantify olfactory capacities as well as heat or water conservation adaptations. We obtained similar results from an existing 2D complexity method and two new 3D methodologies that quantify bone complexity. Using comparative phylogenetic methods, we identified a significant convergent signal in the rostral morphology within the highly specialised vermivores. Vermivorous species have significantly larger and more complex olfactory turbinals than do carnivores and omnivores. Increased olfactory capacities may be a major adaptive feature facilitating rats' capacity to prey on elusive earthworms. The narrow snout that characterises vermivores exhibits significantly reduced respiratory turbinals, which may reduce their heat and water conservation capacities.

Oceanic island biogeography through the lens of the general dynamic model: assessment and prospect.

The general dynamic model of oceanic island biogeography (GDM) has added a new dimension to theoretical island biogeography in recognizing that geological processes are key drivers of the evolutionary processes of diversification and extinction within remote islands. It provides a dynamic and essentially non-equilibrium framework generating novel predictions for emergent diversity properties of oceanic islands and archipelagos. Its publication in 2008 coincided with, and spurred on, renewed attention to the dynamics of remote islands. We review progress, both in testing the GDM's predictions and in developing and enhancing ecological-evolutionary understanding of oceanic island systems through the lens of the GDM. In particular, we focus on four main themes: (i) macroecological tests using a space-for-time rationale; (ii) extensions of theory to islands following different patterns of ontogeny; (iii) the implications of GDM dynamics for lineage diversification and trait evolution; and (iv) the potential for downscaling GDM dynamics to local-scale ecological patterns and processes within islands. We also consider the implications of the GDM for understanding patterns of non-native species diversity. We demonstrate the vitality of the field of island biogeography by identifying a range of potentially productive lines for future research.

Mammalian species richness on islands on the Sunda Shelf, Southeast Asia.

A rich mammalian fauna is found on islands that lie on the Sunda Shelf, a continental shelf extending from Vietnam to Borneo and Java that was periodically exposed as dry land during the Pleistocene. The correlation between log of island area and number of species is high (r 2=0.94); the slope of the curve is moderate (z=0.235). Distance from small islands to "source areas" (=Borneo, Sumatra, and the Malay Peninsula) does not appear to affect species richness, nor does depth of water to source area (a measure of isolation time). A species-area curve for forest reserves of varying sizes on the Malay Peninsula has a low slope (z=0.104); comparison of the mainlaind and island curves indicates that decreasing island area is strongly correlated with increased extinction. Extinction has left reduced but ecologically balanced sets of species on all islands, except that carnivores are under-represented on all but the largest islands. Initial body size and rarity appear to play a significant role in determining the probability of extinction of individual species.

The pattern and timing of diversification of Philippine endemic rodents: evidence from mitochondrial and nuclear gene sequences.

The 22 genera and 64 species of rodents (Muridae: Murinae) distributed in the Philippine Islands provide a unique opportunity to study patterns and processes of diversification in island systems. Over 90% of these rodent species are endemic to the archipelago, but the relative importance of dispersal from the mainland, dispersal within the archipelago, and in situ differentiation as explanations of this diversity remains unclear, as no phylogenetic hypothesis for these species and relevant mainland forms is currently available. Here we report the results of phylogenetic analyses of the endemic Philippine murines and a wide sampling of murine diversity from outside the archipelago, based on the mitochondrial cytochrome b gene and the nuclear-encoded IRBP exon 1. Analysis of our combined gene data set consistently identified five clades comprising endemic Philippine genera, suggesting multiple invasions of the archipelago. Molecular dating analyses using parametric and semiparametric methods suggest that colonization occurred in at least two stages, one ca. 15 Mya, and another 8 to 12 million years later, consistent with the previous recognition of "Old" and "New" endemic rodent faunas. Ancestral area analysis suggests that the Old Endemics invaded landmasses that are now part of the island of Luzon, whereas the three New Endemic clades may have colonized through either Mindanao, Luzon, or both. Further, our results suggest that most of the diversification of Philippine murines took place within the archipelago. Despite heterogeneity between nuclear and mitochondrial genes in most model parameters, combined analysis of the two data sets using both parsimony and likelihood increased phylogenetic resolution; however, the effect of data combination on support for resolved nodes was method dependent. In contrast, our results suggest that combination of mitochondrial and nuclear data to estimate relatively ancient divergence times can severely compromise those estimates, even when specific methods that account for rate heterogeneity among genes are employed. [Biogeography; divergence date estimation; mitochondrial DNA; molecular systematics; Murinae; nuclear exon; Philippines; phylogeny.].