ASYMMETRIC MATING PREFERENCES ACCOMMODATED THE RAPID EVOLUTIONARY LOSS OF A SEXUAL SIGNAL

Rapid evolution has been well documented in naturally selected traits, but few examples exist for sexually selected traits, particularly sexual signals. This may in part be due to the complex set of behaviors associated with sexual signals. For a sexual signal to change, the change must be favorable for the signaler, but must also be accommodated by the receiver's perception and preferences. We investigated female accommodation of an extreme change in the sexual signal of Polynesian field crickets, Teleogryllus oceanicus . The cricket is native to Australia, widely distributed on Pacific Islands, and was recently introduced to Hawaii. Selective pressure by a deadly parasitoid fly favored a wing mutation in Hawaii (flatwing) that eliminates males' singing ability altogether. Despite conventional wisdom that females require males to produce a courtship song before mating, we show that females from ancestral, unparasitized Australian and Pacific Island populations as well as parasitized Hawaiian populations, will mate with silent flatwing males, suggesting this behavioral option predates the change in sexual signal. Furthermore, ancestral Australian females discriminate against flatwing males more severely than island females. We suggest island colonization favored females with relaxed mating requirements (Kaneshiro's effect) facilitating the rapid evolutionary loss of song in Hawaii.     

Bottlenecks and multiple introductions: population genetics of the vector of avian malaria in Hawaii

Avian malaria has had a profound impact on the demographics and behaviour of Hawaiian forest birds since its vector, Culex quinquefasciatus the southern house mosquito, was first introduced to Hawaii around 1830. In order to understand the dynamics of the disease in Hawaii and gain insights into the evolution of vector-mediated parasite-host interactions in general we studied the population genetics of Cx. quinquefasciatus in the Hawaiian Islands. We used both microsatellite and mitochondrial loci. Not surprisingly we found that mosquitoes in Midway, a small island in the Western group, are quite distinct from the populations in the main Hawaiian Islands. However, we also found that in general mosquito populations are relatively isolated even among the main islands, in particular between Hawaii (the Big Island) and the remaining Hawaiian Islands. We found evidence of bottlenecks among populations within the Big Island and an excess of alleles in Maui, the site of the original introduction. The mitochondrial diversity was typically low but higher than expected. The current distribution of mitochondrial haplotypes combined with the microsatellite information lead us to conclude that there have been several introductions and to speculate on some processes that may be responsible for the current population genetics of vectors of avian malaria in Hawaii.     

Influence of invasion history on rapid morphological divergence across island populations of an exotic bird

There is increasing evidence that exotic populations may rapidly differentiate from those in their native range and that differences also arise among populations within the exotic range. Using morphological and DNA‐based analyses, we document the extent of trait divergence among native North American and exotic Hawaiian populations of northern cardinal (Cardinalis cardinalis). Furthermore, using a combination of historical records and DNA‐based analyses, we evaluate the role of founder effects in producing observed trait differences. We measured and compared key morphological traits across northern cardinal populations in the native and exotic ranges to assess whether trait divergence across the Hawaiian Islands, where this species was introduced between 1929 and 1931, reflected observed variation across native phylogeographic clades in its native North America. We used and added to prior phylogenetic analyses based on a mitochondrial locus to identify the most likely native source clade(s) for the Hawaiian cardinal populations. We then used Approximate Bayesian Computation (ABC) to evaluate the role of founder effects in producing the observed differences in body size and bill morphology across native and exotic populations. We found cardinal populations on the Hawaiian Islands had morphological traits that diverged substantially across islands and overlapped the trait space of all measured native North American clades. The phylogeographic analysis identified the eastern North American clade (C. cardinalis cardinalis) as the most likely and sole native source for all the Hawaiian cardinal populations. The ABC analyses supported written accounts of the cardinal's introduction that indicate the original 300 cardinals shipped to Hawaii were simultaneously and evenly released across Hawaii, Kauai, and Oahu. Populations on each island likely experienced bottlenecks followed by expansion, with cardinals from the island of Hawaii eventually colonizing Maui unaided. Overall, our results suggest that founder effects had limited impact on morphological trait divergence of exotic cardinal populations in the Hawaiian archipelago, which instead reflect postintroduction events.     

Strong founder effects and low genetic diversity in introduced populations of Coqui frogs

The success of non-native species may depend on the genetic resources maintained through the invasion process. The Coqui ( Eleutherodactylus coqui ), a frog endemic to Puerto Rico, was introduced to Hawaii in the late 1980s via the horticulture trade, and has become an aggressive invader. To explore whether genetic diversity and population structure changed with the introduction, we assessed individuals from 15 populations across the Hawaiian Islands and 13 populations across Puerto Rico using six to nine polymorphic microsatellite loci and five dorsolateral colour patterns. Allelic richness ( R _T) and gene diversity were significantly higher in Puerto Rico than in Hawaii populations. Hawaii also had fewer colour patterns (two versus three to five per population) than Puerto Rico. We found no isolation by distance in the introduced range, even though it exists in the native range. Results suggest extensive mixing among frog populations across Hawaii, and that their spread has been facilitated by humans. Like previous research, our results suggest that Hawaiian Coquis were founded by individuals from sites around San Juan, but unlike previous research the colour pattern and molecular genetic data (nuclear and mtDNA) support two separate introductions, one on the island of Hawaii and one on Maui. Coquis are successful invaders in Hawaii despite the loss of genetic variation. Future introductions may increase genetic variation and potentially its range.     

Genetic differentiation in endemic Silene (Caryophyllaceae) on the Hawaiian Islands

Endemic Hawaiian Silene colonize new lava flows and are outcompeted as the ground ages. They illustrate the genetic processes operating in the evolution of pioneering island plants. The volcanic history of the Hawaiian Islands allows an estimation of the age of these plant populations. In this study, populations of S. struthioloides from Maui and the older part of the island of Hawaii, and S. hawaiiensis from the youngest volcanoes of the island of Hawaii were analyzed by enzyme electrophoresis. The genetic structures of these populations were placed in a geographic and geologic context. Silene is much more polymorphic on the older island, Maui. Genetic variation appears to have been lost in the colonization of the youngest island, Hawaii. Interestingly, some loci for populations on the younger volcanoes of the island of Hawaii are monomorphic for alleles not found in populations on the older part of Hawaii. Recurrent colonizations are accompanied with founder effects, and restricted gene flow among populations has led to isolation and genetic drift. This has resulted in genetic and morphological differentiâtion of Silene populations on the youngest volcanoes.     

Genetic variation of introduced Hawaiian and native Costa Rican populations of an invasive tropical shrub, Clidemia hirta (Melastomataceae)

Clidemia hirta is one of the most common woody invasive plants in mesic to wet forests in Hawaii, where it was introduced around 1940. The species is relatively uncommon by comparison in its native range of Central and South America and some Caribbean Islands. We examined genetic variation in allozymes of 20 C. hirta populations on four Hawaiian Islands to determine the introduction history. For comparison, we measured genetic variation in 20 native populations across Costa Rica. Mean levels of genetic variation in Hawaiian and Costa Rican populations were low compared to other woody or introduced plants (11.5-12.5% polymorphic loci, 2.05-2.50 alleles per polymorphic locus, and 0.045-0.063 expected heterozygosity). Most genetic diversity was held within rather than among populations in both areas (G(ST) = 0.120 and 0.271 in Hawaii and Costa Rica, respectively). Hawaiian populations had a high degree of genetic similarity, and no genetic differentiation was found among the four Hawaiian Islands sampled. These patterns of genetic variation in Hawaii suggest that no intraspecific hybridization of genotypes from different parts of the native range has occurred and that introductions to the different islands came from the same or similar source populations. The low levels of genetic diversity in parts of both the native and introduced ranges suggest that genetic variation is unrelated to invasiveness in C. hirta.     

Microevolutionary patterns and processes of the Native Hawaiian colonizing fern Odontosoria chinensis (Lindsaeaceae)

Abstract.— The vascular‐plant flora of the Hawaiian Islands is characterized by one of the highest rates of species endemism in the world. Among flowering plants, approximately 89% of species are endemic, and among pteridophytes, about 76% are endemic. At the single‐island level, however, rates of species endemism vary dramatically between these two groups with 80% of angiosperms and only 6% of pteridophytes being single‐island endemics. Thus, in many groups of Hawaiian angiosperms, it is possible to link studies of phylogeny, evolution, and biogeographic history at the interspecific and interisland levels. In contrast, the low level of single‐island species endemism among Hawaiian pteridophytes makes similar interspecific and interisland studies nearly impossible. Higher levels of interisland gene flow may account for the different levels of single‐island endemism in Hawaiian pteridophytes relative to angiosperms. The primary question we addressed in the present study was: Can we infer microevolutionary patterns and processes among populations within widespread species of Hawaiian pteridophytes wherein gene flow is probably common? To address this broad question, we conducted a population genetic study of the native Hawaiian colonizing species Odontosoria chinensis. Data from allozyme analyses allowed us to infer: (1) significant genetic differentiation among populations from different islands; (2) historical patterns of dispersal between particular pairs of islands; (3) archipelago‐level patterns of dispersal and colonization; (4) founder effects among populations on the youngest island of Hawaii; and, (5) that this species primarily reproduces via outcrossing, but may possess a mixed‐mating system.     

Out of Hawaii: the origin and biogeography of the genus Scaptomyza (Diptera: Drosophilidae)

The Hawaiian Archipelago is the most isolated island system on the planet and has been the subject of evolutionary research for over a century. The largest radiation of species in Hawaii is the Hawaiian Drosophilidae, a group of approximately 1000 species. Dispersal to isolated island systems like Hawaii is rare and the resultant flora and fauna shows high disharmony with mainland communities. The possibility that some lineages may have originated in Hawaii and subsequently 'escaped' to diversify on continental landmasses is expected to be rarer still. We present phylogenetic analysis of 134 partially sequenced mitochondrial genomes of Drosophilidae (approx. 1.3 Mb of sequence total) to address major aspects of adaptive radiation and dispersal in Hawaii. We show that the genus Scaptomyza, a group that accounts for approximately one-third of the species-level diversity of Drosophilidae in the Hawaiian Islands, originated in Hawaii, diversified there, and subsequently colonized a number of island and continental landmasses elsewhere on the globe. We propose that a combination of small body size, rapid generation time and unique ecological and physiological adaptations have allowed this genus to effectively disperse and diversify.     

On the origin of the invasive olives (Olea europaea L., Oleaceae)

The olive tree (Olea europaea) has successfully invaded several regions in Australia and Pacific islands. Two olive subspecies (subspp. europaea and cuspidata) were first introduced in these areas during the nineteenth century. In the present study, we determine the origin of invasive olives and investigate the importance of historical effects on the genetic diversity of populations. Four invasive populations from Australia and Hawaii were characterized using eight nuclear DNA microsatellites, plastid DNA markers as well as ITS-1 sequences. Based on these data, their genetic similarity with native populations was investigated, and it was determined that East Australian and Hawaiian populations (subsp. cuspidata) have originated from southern Africa while South Australian populations (subsp. europaea) have mostly derived from western or central Mediterranean cultivars. Invasive populations of subsp. cuspidata showed significant loss of genetic diversity in comparison to a putative source population, and a recent bottleneck was evidenced in Hawaii. Conversely, invasive populations of subsp. europaea did not display significant loss of genetic diversity in comparison to a native Mediterranean population. Different histories of invasion were inferred for these two taxa with multiple cultivars introduced restoring gene diversity for europaea and a single successful founder event and sequential introductions to East Australia and then Hawaii for cuspidata. Furthermore, one hybrid (cuspidata x europaea) was identified in East Australia. The importance of hybridizations in the future evolution of the olive invasiveness remains to be investigated.     

Phylogeographic patterns of Hawaiian Megalagrion damselflies (Odonata: Coenagrionidae) correlate with Pleistocene island boundaries

The Pleistocene geological history of the Hawaiian Islands is becoming well understood. Numerous predictions about the influence of this history on the genetic diversity of Hawaiian organisms have been made, including the idea that changing sea levels would lead to the genetic differentiation of populations isolated on individual volcanoes during high sea stands. Here, we analyse DNA sequence data from two closely related, endemic Hawaiian damselfly species in order to test these predictions, and generate novel insights into the effects of Pleistocene glaciation and climate change on island organisms. Megalagrion xanthomelas and Megalagrion pacificum are currently restricted to five islands, including three islands of the Maui Nui super-island complex (Molokai, Lanai, and Maui) that were connected during periods of Pleistocene glaciation, and Hawaii island, which has never been subdivided. Maui Nui and Hawaii are effectively a controlled, natural experiment on the genetic effects of Pleistocene sea level change. We confirm well-defined morphological species boundaries using data from the nuclear EF-1alpha gene and show that the species are reciprocally monophyletic. We perform phylogeographic analyses of 663 base pairs (bp) of cytochrome oxidase subunit II (COII) gene sequence data from 157 individuals representing 25 populations. Our results point to the importance of Pleistocene land bridges and historical island habitat availability in maintaining inter-island gene flow. We also propose that repeated bottlenecks on Maui Nui caused by sea level change and restricted habitat availability are likely responsible for low genetic diversity there. An island analogue to northern genetic purity and southern diversity is proposed, whereby islands with little suitable habitat exhibit genetic purity while islands with more exhibit genetic diversity.     

Mating systems and genetic load in pioneer and non-pioneer Hawaiian Pteridophyta *

Sporophytes from natural populations of ferns occupying pioneer lava and mature rainforest habitats on the island of Hawaii, Hawaiian Islands, were investigated to determine their mating system and frequency of recessive lethal genes (genetic load). Species dominant in pioneer lava habitats were found to have intragametophytic mating systems and to be devoid of lethal genotypes. Species from intermediate and mature rain-forest habitats exhibited complex intergametophytic mating systems and higher levels of genetic load. It is suggested that natural selection has favoured intragametophytic mating and homozygosity in species of less diverse and less competitive pioneer habitats and intergametophytic mating and heterozygosity in species of more mature habitats.     

Origins,ages and population histories: comparative phylogeography of endemic Hawaiian butterflyfishes (genus Chaetodon)

Aim To investigate phylogeographic relationships, genetic connectivity and potential colonization routes for Hawaii’s endemic butterflyfishes (family Chaetodontidae). Location The Hawaiian Archipelago (central Pacific Ocean). Methods Molecular genetic analysis of mitochondrial DNA from three species of endemic Hawaiian butterflyfishes (Chaetodon multicinctus, n = 280; Chaetodon miliaris, n = 408; Chaetodon fremblii, n = 358) sampled from across the Hawaiian Archipelago was used in a suite of population genetic analyses to examine population histories and calculate coalescence times. We review a recent phylogenetic hypothesis for the Chaetodontidae and optimize ancestral distributions to nodes as a means of inferring colonization pathways to Hawaii. Results We found no evidence for population subdivisions across their ranges for any of the three endemic Hawaiian butterflyfish species (Φ_ST ≈ 0; P > 0.05 in each case). Coalescence analyses revealed that C. multicinctus, C. miliaris and C. fremblii date to genetic bottlenecks of c. 12 ka (95% confidence interval of τ, 0–46,732 years ago), c. 74 ka (95% confidence interval of τ, 62,918–105,699 years ago) and c. 301 ka (95% confidence interval of τ, 88,981–478,495 years ago), respectively. We found that the West Pacific appears to be the ancestral source for two of three independent colonization events to Hawaii (leading to the speciation of C. miliaris and C. fremblii). The third colonization of Hawaii (leading to the divergence of C. multicinctus) is ambiguous, but may have involved island‐hopping from the South Pacific via the Line Islands. Main conclusions Our results are consistent with the growing body of data indicating that ecological specialists may be more susceptible to severe population bottlenecks during large‐scale climatic variation on evolutionary time‐scales. The isolation of the Hawaiian Archipelago presents challenges for colonization by even the most highly dispersive marine organisms, and routes of colonization by endemic butterflyfishes show a variable pattern, indicating that there may be several pathways, both spatially and temporally, for marine fauna to colonize Hawaii. Hawaiian endemic butterflyfishes appear to represent genetically homogeneous populations across the archipelago.     

Fine-scale population genetic structure of a wildlife disease vector: the southern house mosquito on the island of Hawaii

The southern house mosquito, Culex quinquefasciatus, is a widespread tropical and subtropical disease vector. In the Hawaiian Islands, where it was introduced accidentally almost two centuries ago, it is considered the primary vector of avian malaria and pox. Avian malaria in particular has contributed to the extinction and endangerment of Hawaii's native avifauna, and has altered the altitudinal distribution of native bird populations. We examined the population genetic structure of Cx. quinquefasciatus on the island of Hawaii at a smaller spatial scale than has previously been attempted, with particular emphasis on the effects of elevation on population genetic structure. We found significant genetic differentiation among populations and patterns of isolation by distance within the island. Elevation per se did not have a limiting effect on gene flow; however, there was significantly lower genetic diversity among populations at mid elevations compared to those at low elevations. A recent sample taken from just above the predicted upper altitudinal distribution of Cx. quinquefasciatus on the island of Hawaii was confirmed as being a temporary summer population and appeared to consist of individuals from more than one source population. Our results indicate effects of elevation gradients on genetic structure that are consistent with known effects of elevation on population dynamics of this disease vector.     

Genetical variation in three Pacific House mouse (Mus musculus) populations

Studies of allozymic variation at 36 loci in samples of House mouse ( Mus musculus ) populations collected from the island of Hawaii and two islets in the Enewetak Atoll, Marshall Islands are reported. All three populations carried a great deal of inherited variation: the Hawaiian sample had a mean heterozygosity of 16.6% per locus, which is the highest value so far found for a mouse population. The mean heterozygosities for the two atoll populations were 11.4% and 10.9%.
There were no changes in either heterozygosity or allozyme frequencies with age. In this respect, these populations from the tropical Pacific differed from populations living in cold temperate regions where natural selection has been shown to affect allele frequencies in different ways at different stages of the life cycle.     

Seasonal surveillance confirms the range expansion of Aedes japonicus japonicus (Theobald) (Diptera: Culicidae) to the Hawaiian Islands of Oahu and Kauai

The Asian bush mosquito, Aedes japonicus japonicus (Theobald) was not known to occur in the Hawaii archipelago until it was identified on the island of Hawaii in 2003. This mosquito species remained undetected on the neighboring islands for 8?years before it was discovered at the Honolulu International Airport on Oahu in 2012. By 2015, four Ae. j. japonicus mosquitoes were collected in the western mountains of Oahu and one was collected in the central mountains of Kauai. The collection of this invasive mosquito species across the neighboring Hawaiian Islands of Oahu and Kauai indicated the need for increased seasonal surveillance on these islands. Following nearly four years of surveillance, Ae. j. japonicus was also confirmed to occur in the eastern mountains of Oahu and in the central mountainous region of Kauai. To expand the knowledge of the spread of invasive mosquitoes species further surveillance is necessary to identify all possible areas where populations of Ae. j. japonicus and other invasive mosquito species occur in Hawaiian archipelago.     

Hawaiian biogeography and the islands' freshwater fish fauna

Aim This paper describes known patterns in the distributions and relationships of Hawaiian freshwater fishes, and compares these patterns with those exhibited by Hawaii's terrestrial biota. Location The study is based in Hawaii, and seeks patterns across the tropical and subtropical Indo‐west Pacific. Methods The study is based primarily on literature analysis. Results The Hawaiian freshwater fish fauna comprises five species of goby in five different genera (Gobiidae). Four species are Hawaiian endemics, the fifth shared with islands in the western tropical Pacific Ocean. All genera are represented widely across the Indo‐west Pacific. All five species are present on all of the major Hawaiian islands. All five species are amphidromous – their larval and early juvenile life being spent in the sea. Although there has been some local phyletic evolution to produce Hawaiian endemics, there has been no local radiation to produce single‐island endemics across the archipelago. Nor is there evidence for genetic structuring among populations in the various islands. Main conclusions In this regard, the freshwater fish fauna of Hawaii differs from the well‐known patterns of local evolution and radiation in Hawaiian Island terrestrial taxa. Amphidromy probably explains the biogeographical idiosyncrasies of the fish fauna – dispersal through the sea initially brought the fish species to Hawaii, and gene flow among populations, across the archipelago, has hitherto inhibited the evolution of local island endemics, apparently even retarding genetic structuring on individual islands.     

Mating asymmetry and the direction of evolution in the Hawaiian cricket genus Laupala

Based on studies from native Hawaiian Drosophila, a model was proposed to explain sexual isolation and mating asymmetry, from which one could potentially infer the 'direction of evolution'. We examined sexual isolation between allopatric cricket species of the genus Laupala, another endemic Hawaiian insect with an elaborate mating system, to begin to explore the nature of sexual isolation and mating asymmetry in closely related Hawaiian organisms. We studied sexual isolation and mating asymmetry in two contrasts. First, an inter-island comparison, including L. makaio from the older island of Maui and L. paranigra from the younger island of Hawaii, and second, an intra-island (Hawaii) comparison, including L. nigra from the older volcano of Mauna Kea and L. paranigra with a primary distribution on the younger volcanoes of Mauna Loa and Kilauea. We used a 'no-choice' experimental design, pairing individual males and females in homospecific or heterospecific combinations. Several behavioural aspects of courtship (proportion of male singing, latency to male singing, production of spermatophores and courtship initiation speed) were quantified as well as the success or failure of matings. We demonstrate asymmetry in sexual isolation between reciprocal combinations of L. makaio and L. paranigra. This result is examined in light of the differences in courtship behaviour manifest in the experiments with these two species. We did not find evidence of asymmetry in sexual isolation between L. nigra and L. paranigra, although differences in courtship initiation speed were evident between reciprocal combinations of these two species. In addition to the geological argument that species on older islands and older volcanoes give rise to species on younger islands and younger volcanoes, we discuss phylogenetic evidence consistent with these biogeographic hypotheses of relationships among the focal taxa. The patterns of asymmetrical sexual isolation and mating asymmetry are consistent with those found in the native Hawaiian Drosophila.     

Biogeography and the evolution of flightlessness in a radiation of Hawaiian moths (Xyloryctidae: Thyrocopa)

Aim Although the ability to fly confers benefits to most insects, some taxa have become secondarily flightless. Insect flightlessness may be more likely to evolve in environments such as islands and other windswept and alpine areas, but this prediction has rarely been tested while controlling for phylogenetic effects. Here we present a phylogeny for the endemic Hawaiian Lepidoptera genus Thyrocopa, which has two flightless species that occur in alpine areas on Maui and Hawaii islands, in order to determine whether the flightless species are sister to each other or represent separate losses of flight. We also explore divergence times and biogeographic patterns of inter‐island colonization in Thyrocopa, and present the first Hawaiian study to sample a genus from nine islands. Location The Hawaiian Islands. Methods The phylogeny is composed of 70 individuals (including 23 Thyrocopa species and 7 outgroup species) sequenced for portions of cytochrome c oxidase subunit I, elongation factor 1α and wingless genes, for a total of 1964 base pairs, and was estimated using both parsimony (paup *) and Bayesian inference (Mr Bayes ). Divergence times were estimated using the beast software package. Results Our results indicate that two independent invasions of alpine habitats with concomitant loss of flight have occurred in Thyrocopa. Based on current taxon sampling, Thyrocopa colonized the Hawaiian Islands slightly before the formation of Kauai. In terms of overall patterns of diversification, subclades generally follow a progression from older to younger islands. The genus has the greatest number of species on Kauai, with species numbers generally decreasing with decreasing island age. Main conclusions Loss of flight ability has evolved twice in a short period of geological time in Thyrocopa, perhaps as a result of low temperatures, high winds and/or a lack of predation pressure. However, several other Thyrocopa species that live on small islands with consistently high winds, such as Necker and Nihoa islands, retain the ability to fly.     

Speciation and phylogeography of Hawaiian terrestrial arthropods

The Hawaiian archipelago is arguably the world's finest natural laboratory for the study of evolution and patterns of speciation. Arthropods comprise over 75% of the endemic biota of the Hawaiian Islands and a large proportion belongs to species radiations. We classify patterns of speciation within Hawaiian arthropod lineages into three categories: (i) single representatives of a lineage throughout the islands; (ii) species radiations with either (a) single endemic species on different volcanoes or islands, or (b) multiple species on each volcano or island; and (iii) single widespread species within a radiation of species that exhibits local endemism. A common pattern of phylogeography is that of repeated colonization of new island groups, such that lineages progress down the island chain, with the most ancestral groups (populations or species) on the oldest islands. While great dispersal ability and its subsequent loss are features of many of these taxa, there are a number of mechanisms that underlie diversification. These mechanisms may be genetic, including repeated founder events, hybridization, and sexual selection, or ecological, including shifts in habitat and/or host affiliation. The majority of studies reviewed suggest that natural selection is a primary force of change during the initial diversification of taxa.     

The diversity and biogeography of koa-finches (Drepanidini: Rhodacanthis), with descriptions of two new species

All known populations of koa-finches, genus Rhodacanthis , became extinct in the Holocene epoch. Two new species are described here from Quaternary fossil sites in the Hawaiian Islands. One new species, from Kauai and Maui, is roughly the size of the historically known greater koa-finch ( R. palmeri ) but differs in having a more robust skull and in bill morphology. The second new species, from Oahu and Maui, is similar in size to the lesser koa-finch ( R. flaviceps ) but closer to R. palmeri in qualitative osteological traits. The two species of koa-finches known historically from the island of Hawaii are distinct in osteology from the fossil koa-finches on the older Hawaiian islands, indicating that at least two of the four known speciation events in the genus took place within approximately the past 500 kyr. However, the similarity of maxillae from Pleistocene and Holocene sites on Oahu suggests that the Oahu population maintained morphological stasis through the climate changes of the late Quaternary. The evidence that speciation occurred on the youngest island in the archipelago suggests that the process of community assembly on newly emergent Hawaiian landscapes was a stimulus to evolutionary diversification in Rhodacanthis .  © 2005 The Linnean Society of London, Zoological Journal of the Linnean Society , 2005, 144 , 527–541.