Evaluating hypotheses on the origin and evolution of the New Zealand alpine cicadas (Maoricicada) using multiple-comparison tests of tree topology

The statistical testing of alternative phylogenetic trees is central to evaluating competing evolutionary hypotheses. Fleming proposed that the New Zealand cicada species Maoricicada iolanthe is the sister species to the major radiation of both low-altitude and montane Maoricicada species. However, using 1,520 bp of mitochondrial DNA sequence data from the cytochrome oxidase subunit I, tRNA aspartic acid, and the ATPase subunit 6 and 8 genes, we inferred that both M. iolanthe and another low-altitude species, Maoricicada campbelli, are nested within the montane Maoricicada radiation. Therefore, we examined the stability of the inferred phylogenetic placement of these two species using the newly developed Shimodaira-Hasegawa test (SH test) implemented in a maximum-likelihood framework. The SH test has two advantages over the more commonly used Kishino-Hasegawa (KH) and Templeton tests. First, the SH test simultaneously compares multiple topologies and corrects the corresponding P: values to accommodate the multiplicity of testing. Second, the SH test is correct when applied to a posteriori hypotheses, unlike the KH test, because it readjusts the expectation of the null hypothesis (that two trees are not different) accordingly. The comparison of P: values estimated under the assumptions of both the KH test and the SH test clearly demonstrate that the KH test has the potential to be misleading when the issue of comparing of a posteriori hypotheses is ignored and when multiple comparisons are not taken into account. The SH test, in combination with a variety of character-weighting schemes applied to our data, reveals a surprising amount of ambiguity in the phylogenetic placement of M. iolanthe and M. campbelli.     

New Zealand cicadas of the genus Maoricicada (Homoptera: Tibicinidae)

Abstract

The genus Maoricicada Dugdale, 1972 includes 19 taxa, of which 15 are described here. Ten from the South Island mountains are described as new: M. alticola, M. clamitans, M. mangu gourlayi, M. m. multicostata, M. m. celer, M. nigra frigida, M. otagoensis otagoensis, M. o. maceweni, M. phaeoptera, and M. tenuis. The distributions of most taxa are mapped, their songs are represented by sonograms, and their known periods of emergence are recorded. Evidence of hybridisation is unknown. Most species are characteristic of open, unforested sites; M. nigra nigra and M. oromelaena occur close to the summer snowline, a zone not known to be inhabited by cicadas elsewhere in the world.     

Evolutionary radiation of the cicada genus Maoricicada Dugdale (Hemiptera: Cicadoidea) and the origins of the New Zealand alpine biota

We have used ancestral character state reconstruction and molecular dating to test hypotheses on the evolution of New Zealand alpine cicadas of the genus Maoricicada Dugdale. Gene trees were estimated from mitochondrial DNA and the nuclear loci elongation factor 1-α, period, and calmodulin and species-level relationships were reconstructed using gene tree parsimony. These analyses suggest that the alpine habitat character state had a single origin and that the ancestral Maoricicada lived in low to mid-elevation habitats. Our reconstructions also strongly support the hypothesis that this ancestor was darkly coloured and had increased pubescence, classic adaptations of alpine insects. Using relaxed-clock Bayesian dating methods, we estimated that the radiation of the alpine Maoricicada species was coincident with the late Miocene acceleration in the rate of uplift of the Southern Alps rather than uplift in the early Miocene. These dates are very similar to those of other alpine taxa, indicating that the New Zealand alpine biota is very young. Our reconstructions suggest that the ancestral Maoricicada may have been preadapted to the alpine environment because it existed before the origin of high mountainous habitats, dwelt at mid-to-low altitudes, and yet possessed the classic alpine insect adaptations of heavy pubescence and dark coloration.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 91 , 419–435.     

Hygrochastic capsule dehiscence in New Zealand alpine Veronica (Plantaginaceae)

? Premise of the study: Plant movement is more widespread than often recognized, involving different organs and mechanisms. Hygrochasy (opening in response to moisture) is a capsule-opening movement that is widely believed to be predominantly a feature of plants of desert and arid zones, where it may protect against seed predators and harsh climatic conditions and restrict dispersal to favorable germination times and sites. However, recently it has been reported from a wider range of environments. This study demonstrates that hygrochasy is much more common than previously realized, extending the phenomenon to plants of alpine habitats. ? Methods: Capsules of 23 species of New Zealand Veronica were collected, and we used light microscopy to investigate the anatomy and biomechanics responsible for the opening mechanism. Additionally, we collected morphological data to identify common traits of hygrochastic species. ? Key results: Hygrochastic capsule dehiscence was found in 10 alpine Veronica species. The opening mechanism is based on an antagonistic reaction between a nonlignified swelling tissue and a lignified resistance tissue. In Veronica, hygrochasy is associated with erect, narrowly angustiseptate capsules on short peduncles of creeping subshrubs or cushion plants. ? Conclusion: Hygrochasy is a common dehiscence type in New Zealand alpine Veronica, and for the first time, this mechanism is described in detail for plants in alpine habitats. We propose that hygrochasy provides an effective seed dispersal mechanism in solitary capsules embedded in cushion plants and may restrict dispersal within habitat patches.     

Phylogeny and phylogeography of the Liolaemus darwinii complex (Squamata: Liolaemidae): evidence for introgression and incomplete lineage sorting

Although mitochondrial DNA markers have several properties that make them suitable for phylogeographic studies, they are not free of difficulties. Phylogeographic inferences within and between closely related species can be mislead by introgression and retention of ancestral polymorphism. Here we combine different phylogenetic, phylogeographic, and population genetic methods to extract the maximum information from the Liolaemus darwinii complex. We estimate the phylogeographic structure of L. darwinii across most of its distributional range, and we then estimate relationships between L. darwinii and the syntopic species L. laurenti and L. grosseorum. Our results suggest that range expansion of these lineages brought them into secondary contact in areas where they are presently in syntopy. Here we present the first evidence for introgression in lizards from temperate South America (of L. danwinii mitochondrial DNA into L. laurenti and L. grosseorum), and for incomplete lineage sorting (between L. darwinii and L. laurenti). We show that a combination of methods can provide additional support for inferences derived from any single method and thus provide more robust interpretations and narrow the range of plausible hypotheses about mechanisms and processes of divergence. Additional studies are needed in this group of lizards and in other codistributed groups to determine if Pleistocene climatic changes could be a general factor influencing the evolutionary history of a regional biota.     

Mitochondrial introgression and incomplete lineage sorting through space and time: phylogenetics of crotaphytid lizards

We investigate the roles of mitochondrial introgression and incomplete lineage sorting during the phylogenetic history of crotaphytid lizards. Our Bayesian phylogenetic estimate for Crotaphytidae is based on analysis of mitochondrial DNA sequence data for 408 individuals representing the 12 extant species of Crotaphytus and Gambelia. The mitochondrial phylogeny disagrees in several respects with a previously published morphological tree, as well as with conventional species designations, and we conclude that some of this disagreement stems from hybridization-mediated mitochondrial introgression, as well as from incomplete lineage sorting. Unidirectional introgression of Crotaphytus collaris (western collared lizard) mitochondria into C. reticulatus (reticulate collared lizard) populations in the Rio Grande Valley of Texas has resulted in the replacement of ancestral C. reticulatus mitochondria over approximately two-thirds of the total range of the species, a linear distance of approximately 270 km. Introgression of C. collaris mitochondria into C. bicinctores (Great Basin collared lizard) populations in southwestern Arizona requires a more complex scenario because at least three temporally separated and superimposed introgression events appear to have occurred in this region. We propose an "introgression conveyor" model to explain this unique pattern of mitochondrial variation in this region. We show with ecological niche modeling that the predicted geographical ranges of C. collaris, C. bicinctores, and C. reticulatus during glacial maxima could have provided enhanced opportunities for past hybridization. Our analyses suggest that incomplete lineage sorting and/or introgression has further confounded the phylogenetic placements of additional species including C. nebrius, C. vestigium, C. insularis, C. grismeri, and perhaps G. copei. Despite many independent instances of interspecific hybridization among crotaphytid lizards, the species continue to maintain morphological and geographic cohesiveness throughout their ranges.     

Genetic introgression, incomplete lineage sorting and faulty taxonomy create multiple cases of polyphyly in a montane clade of tyrant-flycatchers (Elaenia, Tyrannidae)

Species-level paraphyly and polyphyly are pervasive phenomena in modern phylogenetic research and can be due to a number of factors. We explore a complicated pattern of nuclear and mitochondrial polyphyly in montane Neotropical Elaenia flycatchers. Using a combination of phylogenetic and population genetic methods, we demonstrate that no single factor is sufficient to account for this pattern of polyphyly, and that it is likely based on an interplay of three different factors: (i) faulty taxonomy which has led to the recognition of two polyphyletic species that are better classified as four biological species; (ii) a late Pleistocene hybridization event that resulted in two morphologically and ecologically distinct species sharing extremely similar mitochondrial DNA but distinct nuclear DNA profiles; and (iii) incomplete lineage sorting in a nuclear marker that results in a polyphyletic placement of species that are otherwise well-differentiated in mitochondrial DNA, morphology and ecology. Additionally, we demonstrate that the two clades of montane Elaenia exhibit a reverse pattern of mitochondrial and nuclear diversity, with high mitochondrial and low nuclear genetic diversity in one clade and vice versa in the other clade. A possible cause for this pattern is differences in population histories, with large panmictic population structures being conducive to the retention of ancient nuclear polymorphisms in Elaenia albiceps chilensis .     

Influence of landscape structure on endemic cicadas in New Zealand kiwifruit orchards

• 1 The endemic cicada species Amphipsalta cingulata (Fabricius) and Amphipsalta zelandica (Boisduval) are pests of New Zealand kiwifruit.
• 2 We determined the abundance of A. cingulata and A. zelandica by counting final‐instar exuviae in a block of ‘Hayward’ kiwifruit, the dominant cultivar, on each of 70 blocks on separate orchards in the Bay of Plenty, New Zealand.
• 3 We used a geographic information system and fragstats to generate predictive variables describing landscape structure in four nested landscapes ranging in size between 6.25 and 400 ha for each site. Other variables described the physical characteristics of the site and management practices. Data were analyzed by boosted regression trees, a method that combines the advantages of regression trees and machine learning.
• 4 The most influential variables differed for each species. Modified coastal landscapes with high densities of ‘Hayward’ kiwifruit were most favourable for A. cingulata. For A. zelandica, favourable landscapes contained significant areas of native forest. The 12 most influential variables accounted for 51% and 46% of the total influence of all variables measured for A. cingulata and A. zelandica, respectively.
• 5 Landscape structure was more influential than insecticide use and local site factors. Despite the apparent low vagility of cicadas, landscape structure at relatively large scales of ≥25 ha was influential for both A. cingulata and A. zelandica. The ability to use a wide range of hosts within the production landscape may account for this pattern. Key variables need to be confirmed by identifying the same patterns in other landscapes.

     

Hygrochastic capsule dehiscence supports safe site strategies in New Zealand alpine Veronica (Plantaginaceae)

Background and Aims

Hygrochasy is a capsule-opening mechanism predominantly associated with plants in arid habitats, where it facilitates spatially and temporally restricted dispersal. Recently, hygrochastic capsules were described in detail for the first time in alpine Veronica in New Zealand. The aim of the present study was to investigate whether hygrochastic capsules are an adaptation of alpine Veronica to achieve directed dispersal to safe sites. We expect that by limiting dispersal to rainfall events, distances travelled by seeds are short and confine them to small habitat patches where both seedlings and adults have a greater chance of survival.

Methods

Dispersal distances of five hygrochastic Veronica were measured under laboratory and field conditions and the seed shadow was analysed. Habitat patch size of hygrochastic Veronica and related non-hygrochastic species were estimated and compared.

Key Results

Dispersal distances achieved by dispersal with raindrops did not exceed 1 m but weather conditions could influence the even distribution of seeds around the parent plant. Compared with related Veronica species, hygrochastic Veronica mostly grow in small, restricted habitat patches surrounded by distinctly different habitats. These habitat patches provide safe sites for seeds due to their microtopography and occurrence of adult cushion plants. Non-hygrochastic Veronica can be predominantly found in large habitats without clearly defined borders and can be spread over long distances along rivers.

Conclusions

The results suggest that hygrochasy is a very effective mechanism of restricting seed dispersal to rainfall events and ensuring short-distance dispersal within a small habitat patch. It appears that it is an adaptation for directed dispersal to safe sites that only exist within the parent habitat.     

Analysis of DNA sequences of six chloroplast and nuclear genes suggests incongruence, introgression, and incomplete lineage sorting in the evolution of Lespedeza (Fabaceae)

The genus Lespedeza (Fabaceae) consists of 40 species disjunctively distributed in East Asia and eastern North America. Phylogenetic relationships of all Lespedeza species and closely related genera were reconstructed using maximum parsimony, maximum likelihood, and Bayesian analyses of sequence data from five chloroplast (rpl16, rpl32-trnL, rps16-trnQ, trnL-F, and trnK/matK) and one nuclear (ITS) DNA regions. All analyses yielded consistent relationships among major lineages. Our results suggested that Campylotropis, Kummerowia, and Lespedeza are monophyletic, respectively. Lespedeza is resolved as sister to Kummerowia and these two together are further sister to Campylotropis. Neither of the two subgenera, subgen. Lespedeza and subgen. Macrolespedeza, in Lespedeza based on morphological characters, is recovered as monophyletic. Within Lespedeza, the North American clade is retrieved as sister to the Asian clade. The nuclear and chloroplast markers showed incongruent phylogenetic signals at shallow-level phylogeny, which may point to either introgression or incomplete lineage sorting in Lespedeza. The divergence times within Lespedeza and among related genera were estimated using Bayesian approach with BEAST. It is assumed that following the divergence between Kummerowia and Lespedeza in Asia in the late Miocene, the ancestor of Lespedeza diverged into the North American and the Asian lineages. The North American ancestor quickly migrated to North America through the Bering land bridge in the late Miocene. The North American and Asian lineages started to diversify almost simultaneously in the late Miocene but resulted in biased numbers of species in two continents.     

Micro-pattern in an area of New Zealand alpine vegetation

An area of New Zealand alpine herbfield, 22×22 m, was sampled on a 3×3 m grid.Two community types could be distinguished, dominated respectively by the cushion epacrid Dracophyllum muscoides and the subshrub asterad Celmisia viscosa. Each type was subdivided into three communities. The distribution of these communities was correlated with the environmental factors measured, with a predictability of up to 81%. Some of the factors, such as soil organic matter, could have been influenced by the vegetation, but others were microtopographic ones, unlikely to have been so influenced. There was no evidence of cyclic succession. Possibly the distribution of the two major types reflects previous patch burns, but distributions of individual communities are related to the current environment.Association between the more common species revealed two very clear groups, which characterise, but are not exclusive to, the two community types.     

New Zealand geckos (Diplodactylidae): Cryptic diversity in a post-Gondwanan lineage with trans-Tasman affinities

We used a multi-gene approach to assess the phylogenetic relationships of New Zealand diplodactylid geckos to their Australian and New Caledonian relatives and to one another. Data from nuclear (RAG-1, PDC) and mitochondrial (ND2, 16S) genes from >180 specimens representing all 19 recognized New Zealand taxa and all but two of 20 putatively new species suggested by previous studies were analyzed using Maximum Parsimony, Maximum Likelihood and Bayesian inference. All analyses retrieved a monophyletic New Zealand clade, most closely related to the Australian Diplodactylidae exclusive of Pseudothecadactylus. Hoplodactylus is paraphyletic and composed of two morphological groups: a broad-toed clade, consisting of the island-restricted, largest extant species, Hoplodactylus duvaucelii, and the species-rich, wide-ranging Hoplodactylus maculatus clade; and a narrow-toed clade, comprising five monophyletic subgroups: Naultinus, the Hoplodactylus pacificus and Hoplodactylus granulatus clades, and the distinctive species Hoplodactylus rakiurae and Hoplodactylus stephensi. Each of these lineages is here recognized at the generic level. Our data support recognition of 16 new species (36 total), and five new or resurrected genera (seven total). The New Zealand diplodactylid radiation split from its Australian relatives 40.2mya (95% highest posterior density estimate 28.9-53.5), after the opening of the Tasman Sea. Their distribution cannot, therefore, be regarded as derived as a result of Gondwanana vicariance. The age of the New Zealand crown group, 24.4mya (95% highest posterior density estimate 15.5-33.8), encompasses the period of the 'Oligocene drowning' of New Zealand and is consistent with the hypothesis that New Zealand was not completely inundated during this period. Major lineages within New Zealand geckos diverged chiefly during the mid- to late Miocene, probably in association with a suite of geological and climatological factors that have characterized the region's complex history.     

Using patterns of genetic structure based on microsatellite loci to test hypotheses of current hybridization,ancient hybridization and incomplete lineage sorting in Conradina (Lamiaceae)

Hybridization and/or incomplete sorting of ancestral polymorphism are commonly implicated to explain discordant phylogenetic analyses of closely related species complexes. One genus in which these phenomena have been suggested to have played major roles based on phylogenetic data is Conradina, a genus of mints (Lamiaceae) endemic to the southeastern USA containing several endangered species. The goals of this study were to use microsatellite data to better understand patterns of genetic structure in Conradina, to test hypotheses of recent or ancient hybridization and incomplete lineage sorting, and to clarify species boundaries. Individuals from 55 populations representing all Conradina species were genotyped using 10 microsatellite loci. Analyses of the patterns of genetic structure in Conradina revealed a clear differentiation of populations following recognized species boundaries, indicating that species have diverged from one another genetically and interspecific hybridization has not occurred recently. Neither ancient hybridization nor incomplete lineage sorting is supported as the sole cause of species nonmonophyly, suggesting that both may have contributed to patterns found in phylogenetic trees; however, analyses of other types of data may be more appropriate to distinguish between these two hypotheses. Because all described species appear to be valid entities, the current listing status of most endangered species of Conradina is appropriate; however, populations of Conradina canescens are genetically differentiated into three groups, each of which may merit species status, and several recently discovered populations of Conradina in Dunn's Creek State Park in Florida are highly differentiated genetically and also appear to represent a new species.     

Water relations of the freeze-tolerant New Zealand alpine cockroach Celatoblatta quinquemaculata (Dictyoptera: Blattidae)

Celatoblatta quinquemaculata is a freeze-tolerant alpine cockroach found on the Rock and Pillar Range, Central Otago, New Zealand. This study investigated seasonal changes in water content, as well as desiccation tolerance, and the relationship between desiccation and cold tolerance. Whole body water contents from field-fresh cockroaches collected over a 20 month period ranged from 69.9+/-1.0% fresh weight (FW) in February 1998 to 60.3+/-1.1% FW in July 1998. Water contents were significantly lower in winter than summer, and were positively correlated to microhabitat temperatures over the week preceding collection. Cockroaches survived the loss of up to 82% (mean: 56.7%+/-10.2) of their initial body water content, and the amount of water loss sustained was not dependent on the rate of water loss. Cockroaches did not suffer further mortality due to desiccation after removal to 99% relative humidity, but only regained lost water if given access to liquid water. Experimental dehydration did not enhance freeze-tolerance, but did slightly lower the supercooling point. It is concluded that reduction of body water content in winter may be a consequence of cold hardening responses, but desiccation does not constitute the cold hardening mechanism itself.     

Cold tolerance of a New Zealand alpine cockroach, Celatoblatta quinquemaculata (Dictyoptera, Blattidae)

Abstract. Ecophysiological features, including survival and recovery from freezing and determination of the freezable water content, are reported for a cold-adapted cockroach Celatoblatta quinquemaculata Johns 1966 (Dictyoptera, Blattidae) inhabiting alpine communities at altitudes greater than 1300 m a.s.l. in mountains of Central Otago, New Zealand. Nymphs ranged from 15 to 51 mg live weight of which 67% was water. Cockroaches had a mean supercooling point temperature of ?5.4 ± 0.1°C; with recovery from freezing close to this temperature being rapid, but no recovery was observed when frozen at ?9 to ?10°C. The duration of exposure to freezing conditions and the time allowed for recovery (24–96 h) both influenced individual recovery and subsequent survival. Comparison of supercooling point data and survival shows that this species possesses a few degrees of freeze tolerance, and individuals have been found frozen in the field when subzero temperatures occur. Differential scanning calorimetry showed ≈ 74% of body water froze during cooling and between 24 and 27% of total body water was osmotically inactive (unfreezable under the experimental conditions). Carbohydrates, other than glucose at 7.5μg/mg fresh weight, were in low concentrations in the body fluids, suggesting little cryoprotection. No thermal hysteresis from antifreeze protein activity was detected in haemolymph samples using calorimetric techniques. It is suggested that slow environmental cooling rates, together with high individual supercooling points, confer a small amount of freezing tolerance on this species enabling it to survive low winter temperatures. This has allowed it to colonize and maintain populations in alpine habitats > 1300 m a.s.1. in New Zealand.     

Seed dispersal effectiveness increases with body size in New Zealand alpine scree weta (Deinacrida connectens)

Weta are giant, flightless orthopterans that are endemic to New Zealand. Although they are known to consume fleshy fruits and disperse seeds after gut passage, which is unusual among insects, their effectiveness as seed dispersal mutualists is debated. We conducted a series of laboratory experiments on alpine scree weta (Deinacrida connectens) and mountain snowberries (Gaultheria depressa) to investigate how fruit consumption rates, the proportion of ingested seeds dispersed intact and weta movement patterns vary with weta body sizes. On average weta dispersed 252 snowberry seeds nightly and travelled at a rate of 4 m min^?1. However, seed dispersal effectiveness varied over three orders of magnitude and was strongly associated with body sizes. Smaller weta consumed few snowberry seeds and acted primarily as seed predators. On the other hand, the largest weta consumed and dispersed thousands of seeds each night and appear to be capable of transporting seeds over large distances. Overall results indicate that scree weta shift from being weakly interacting seed predators to strongly interacting, effective seed dispersers as they increase in size.     

Electrochemical characteristics of ion secretion in malpighian tubules of the New Zealand alpine weta (Hemideina maori)

Characteristics of ion and fluid secretion were investigated in isolated Malpighian tubules of the New Zealand Alpine Weta (Hemideina maori). Fluid secretion by tubules in iso-osmotic saline (500mOsm) occurred at a rate of 15+/-3nlh(-1) and was enriched in K(+) (approx. 125mmoll(-1)) relative to the saline (10mmoll(-1)). Maximal fluid secretion (112nlh(-1)) during simultaneous exposure to hypo-osmolality and dibutyryl cAMP resulted in an 8.8x increase in the quantity of K(+) secreted, compared to only a 2.4x increase in Na(+) secretion. Measurements of intracellular ion activities and membrane potentials indicated that Na(+) and K(+) were transported against a strong electrochemical gradient across the apical surface, regardless of saline osmolality. On the basolateral surface, there was a large driving force for Na(+) entry, while K(+) was distributed near its equilibrium potential. Neither bumetanide nor ouabain in the bathing saline had a significant effect on fluid secretion, but Ba(2+) and amiloride decreased fluid secretion by 79 and 57%, respectively. The effect of Ba(2+) on fluid secretion was consistent with a high basolateral permeability to K(+), relative to Na(+) and Cl(-). These results indicate that the characteristics of fluid secretion in this primitive insect are largely conserved with characteristics reported for other insects.