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.     

Pollen morphology in the genus Veronica L. (Plantaginaceae) and its systematic significance

Pollen grains of 30, mainly annual, species from 134 populations of the genus Veronica (Plantaginaceae, formerly Scrophulariaceae) from the Mediterranean Region, have been studied with light and scanning electron microscopy. Three pollen types are defined based on pollen size and exine surface sculpture. In some cases within the study group pollen size can be useful in species determination. Hypothetical relationships of pollen grain size and aperture number with ploidy level, style length and corolla diameter are discussed. The ornamentation of the exine in Veronica, although generally a conservative character within the genus, gives some additional support to the most recent infrageneric classification of Veronica.     

Cold tolerance of New Zealand alpine insects

New Zealand has extensive alpine and subalpine habitats where, together with some lowland sites, insects are exposed to subzero temperatures. Studies of cold tolerance in New Zealand insects have centred on an alpine weta (Hemideina maori), which is the world's largest freezing tolerant insect, and an alpine cockroach (Celatoblatta quinquemaculata). Both of these insects are moderately freezing tolerant and have ice nucleating agents in their haemolymph and guts. There is some evidence for the survival of intracellular ice formation in the isolated gut tissue of C. quinquemaculata. Trehalose is a suggested cryoprotectant in both H. maori and C. quinquemaculata whilst proline also provides this role in H. maori. Cells and tissues of both insects maintain viability and physiological function during freezing to moderately low temperatures but viability declines at lower temperatures, the most vulnerable tissue presumably setting the limit to the survival of the animal. Antifreeze proteins are found in the gut tissue of C. quinquemaculata and may protect this tissue when freezing occurs in the gut. Several other New Zealand insects are also moderately freezing tolerant and the apparent dominance of this cold tolerance strategy in the New Zealand fauna may reflect the relatively mild climate but unpredictable exposure to subzero temperatures that is typical of many Southern Hemisphere environments.     

Red (anthocyanic) leaf margins do not correspond to increased phenolic content in New Zealand Veronica spp.

Background and Aims

Red or purple coloration of leaf margins is common in angiosperms, and is found in approx. 25 % of New Zealand Veronica species. However, the functional significance of margin coloration is unknown. We hypothesized that anthocyanins in leaf margins correspond with increased phenolic content in leaf margins and/or the leaf entire, signalling low palatability or leaf quality to edge-feeding insects.

Methods

Five species of Veronica with red leaf margins, and six species without, were examined in a common garden. Phenolic content in leaf margins and interior lamina regions of juvenile and fully expanded leaves was quantified using the Folin–Ciocalteu assay. Proportions of leaf margins eaten and average lengths of continuous bites were used as a proxy for palatability.

Key Results

Phenolic content was consistently higher in leaf margins compared with leaf interiors in all species; however, neither leaf margins nor more interior tissues differed significantly in phenolic content with respects to margin colour. Mean phenolic content was inversely correlated with the mean length of continuous bites, suggesting effective deterrence of grazing. However, there was no difference in herbivore consumption of red and green margins, and the plant species with the longest continuous grazing patterns were both red-margined.

Conclusions

Red margin coloration was not an accurate indicator of total phenolic content in leaf margins or interior lamina tissue in New Zealand Veronica. Red coloration was also ineffective in deterring herbivory on the leaf margin, though studies controlling for variations in leaf structure and biochemistry (e.g. intra-specific studies) are needed before more precise conclusions can be drawn. It is also recommended that future studies focus on the relationship between anthocyanin and specific defence compounds (rather than general phenolic pools), and evaluate possible alternative functions of red margins in leaves (e.g. antioxidants, osmotic adjustment).     

Evolutionary Rates in Veronica L. (Plantaginaceae): Disentangling the Influence of Life History and Breeding System

The evolutionary rate at which DNA sequences evolve is known to differ between different groups of organisms. However, the reasons for these different rates are seldom known. Among plants, the generation-time hypothesis, which states that organisms that reproduce faster also have more DNA substitutions per time, has gained most popularity. We evaluate the generation-time hypothesis using 131 DNA sequences from the plastid trnLF region and the nuclear ribosomal ITS region of the genus Veronica (Plantaginaceae). We also examine the alternative hypothesis that a higher substitution rate is correlated with selfing breeding system. Selfing is associated with annual life history in many organisms and may thus often be the underlying reason for observed correlations of annual life history with other characters. We provide evidence that annual life history is more likely to be the responsible factor for higher substitution rates in Veronica than a selfing breeding system. Nevertheless, the way in which annual life history may influence substitution rate in detail remains unknown, and some possibilities are discussed.     

Mating system variation in Veronica (Plantaginaceae): inferences from pollen/ovule ratios and other reproductive traits

The pollen–ovule ratio (P/O) is commonly used to estimate the mode of sexual reproduction in flowering plants. In previous studies, a clear correspondence has been detected between this character and the degree of autogamy. We here investigate variation in this character and its expected correlates in the genus Veronica (Plantaginaceae). Pollen–ovule ratios of 45 species representing eleven percent of all the species in the genus were investigated and compared with results from crossing experiments from previous studies. In addition, multiple populations of 17 of the 45 studied species were sampled and a controlled‐environment experiment was conducted to evaluate the extent of intraspecific variation. Moreover, relationships between P/O and other primary and secondary reproductive characters of the Veronica flower were investigated in relation to a phylogenetic hypothesis in order to determine the phylogenetic constraints on reproductive characters. The differences in P/O among species correspond well to the diversity of mating systems in Veronica and correlate well with other floral characters such as corolla size. These characters together seem to allow a powerful and fast tool to infer mating systems. However, causes for intraspecific variation of P/O, such as different cytotypes, ecotypes or different growth conditions, need to be considered.     

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.     

Notes on the capsule dehiscence inAcanthophyllum (Caryophyllaceae) and allied genera

The shape and the mode of dehiscence of the capsule had been regarded as good differential characters betweenAcanthophyllum and related genera.—Studies of these characters, including the shape of the ovary, in species ofAcanthophyllum, Diaphanoptera, Ochotonophila andScleranthopsis show, however, that they cannot be used as differential characters for the genusAcanthophyllum.Dedicated to Prof.K. H. Rechinger on the occasion of his 80th birthday.     

Differentiating between hypotheses of lineage sorting and introgression in New Zealand alpine cicadas (Maoricicada Dugdale)

Lineage sorting and introgression can lead to incongruence among gene phylogenies, complicating the inference of species trees for large groups of taxa that have recently and rapidly radiated. In addition, it can be difficult to determine which of these processes is responsible for this incongruence. We explore these issues with the radiation of New Zealand alpine cicadas of the genus Maoricicada Dugdale. Gene trees were estimated from four putative independent loci: mitochondrial DNA (2274 nucleotides), elongation factor 1-alpha (1275 nucleotides), period (1709 nucleotides), and calmodulin (678 nucleotides). We reconstructed phylogenies using maximum likelihood and Bayesian methods from 44 individuals representing the 19 species and subspecies of Maoricicada and two outgroups. Species-level relationships were reconstructed using a novel extension of gene tree parsimony, whereby gene trees were weighted by their Bayesian posterior probabilities. The inferred gene trees show marked incongruence in the placement of some taxa, especially the enigmatic forest and scrub dwelling species, M. iolanthe. Using the species tree estimated by gene tree parsimony, we simulated coalescent gene trees in order to test the null hypothesis that the nonrandom placement of M. iolanthe among gene trees has arisen by chance. Under the assumptions of constant population size, known generation time, and panmixia, we were able to reject this null hypothesis. Furthermore, because the two alternative placements of M. iolanthe are in each case with species that share a similar song structure, we conclude that it is more likely that an ancient introgression event rather than lineage sorting has caused this incongruence.     

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.     

Phylogeographic patterns in the Australasian genus Chionohebe (Veronica s.l., Plantaginaceae) based on AFLP and chloroplast DNA sequences

The alpine genus Chionohebe is one of seven genera in the southern hemisphere Hebe complex. The main aims of this study were to infer the evolutionary relationships and assess phylogeographic patterns among the six species of Chionohebe, determine the origin of the two species with trans-Tasman distributions, and test species delimitations and specimen identifications based on morphology. Analyses of AFLP data recovered five major lineages within Chionohebe, some of which corresponded to species and varieties as currently circumscribed. Although the cushion chionohebes were strongly supported as monophyletic, the sole non-cushion species, C. densifolia, was sister to Parahebe trifida, and thus the AFLP data do not support a monophyletic Chionohebe as usually circumscribed. Strong north/south and west/east phylogeographic patterns were found among and within the main AFLP lineages in New Zealand. Analyses of chloroplast DNA (cpDNA) revealed eight haplotypes in Chionohebe, but these did not correspond to current taxonomy or geography due to widespread interspecific haplotype sharing. Based on both AFLP and cpDNA results, the two trans-Tasman species are shown to have originated in New Zealand and dispersed to Australia independently.     

Bryophytes as invertebrate habitat in two New Zealand alpine streams

SUMMARY. 1. Quantitative samples of lotic benthic invertebrates and associated organic material were taken monthly for 18 months from bryophyte and gravel habitats in two New Zealand alpine streams in Arthur's Pass National Park, South Island; an unshaded site above the tree-line and a shaded stream flowing through mountain beech forest. This was to assess the ecological role of bryophytes in these habitats and to determine the relative effects on the fauna of algal and detrital resources. 2. Bryophytes were confined to stable bedrock substrata in shallow (mean depth of 4.5 and 10.5 cm), fast water (mean velocity of 0.21 ms^?1 and 0.30ms^?1) at the unshaded and shaded sites. Mean ash-free dry weight (AFDW) of organic material from bryophyte areas was similar at the unshaded and shaded sites (335.6 and 272.4 gm^?2). Low stream-bed stability of the gravel areas resulted in poor retention of organic matter, with average AFDW of 59.1 and 77.9 gm^?2 at the unshaded and shaded sites, respectively. Periphyton biomass, however, was much higher at the unshaded site. 3. Within bryophytes, water velocity is reduced and large quantities of periphyton and detritus accumulate. This explains the greatly enhanced invertebrate densities within bryophytes (218400 and 53500 individuals m^?2), compared with gravel areas (20900 and 7400 individuals m^?2) in the unshaded and shaded sites, respectively. The higher invertebrate densities at the unshaded site reflect greater periphyton biomass there. 4. Invertebrate densities followed clear seasonal patterns, attributable to changes in chironomid densities. These reflected larval chironomid hatching following adult oviposition. This illustrates the importance of these plants as oviposition sites, and as protective nurseries for small larvae where they can obtain shelter during floods and consume the abundant food sources available to them. 5. DECORANA ordination revealed taxonomically distinct invertebrate communities in each stream, and in bryophytes and gravels. Several taxa appeared restricted to either the unshaded site (e.g. Acroperla spiniger and Hydrobiosis silvicola) or the shaded site (e.g. Zelolessica     

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.     

Chemotaxonomic markers in Digitalideae (Plantaginaceae)

In a chemosystematic investigation of Digitalideae (Plantaginaceae), the water-soluble part of extracts of two species of Digitalis, two species of Isoplexis, as well as Erinus alpinus and Lafuentea rotundifolia were studied with regard to their content of main carbohydrates, iridoids and caffeoyl phenylethanoid glycosides (CPGs). Digitalis and Isoplexis contained sorbitol, cornoside and a number of other phenylethanoid glycosides including the new tyrosol beta-D-mannopyranoside, sceptroside but were found to lack iridoid glucosides. Erinus contained mainly glucose, the new 8,9-double bond iridoid, erinoside, and a number of known iridoid glucosides including two esters of 6-rhamnopyranosylcatalpol, as well as the CPG poliumoside. Finally, Lafuentea was characterized by the presence of glucose, aucubin and cryptamygin B but apparently lacked CPGs. The chemosystematic significance of the isolated compounds is discussed.     

Amplified fragment length polymorphisms and sequence data in the phylogenetic analysis of polyploids: multiple origins of Veronica cymbalaria (Plantaginaceae)

The origin of polyploid Veronica cymbalaria (Plantaginaceae) was investigated using DNA sequence data and amplified fragment length polymorphism (AFLP) fingerprints to reveal the parentage of this taxon. The use of AFLP fingerprints in phylogenetic analysis is problematic and various methods have therefore been compared. DNA sequence data (for the internal transcribed spacer (ITS) region and the plastid trnL-F region (trnL intron, 3'exon, and trnL-F spacer)) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the ITS region suggested a reliable hypothesis for the evolution of the V. cymbalaria complex. This hypothesis allowed evaluation of the effect of different distance measures (Jaccard and Nei-Li) in phenetic, character-state weighted parsimony, and Bayesian analyses of AFLP markers. The study establishes that tetraploid V. cymbalaria originated at least twice in the eastern Mediterranean, with one parent differing in the two separate origins. Hexaploid V. cymbalaria originated even more often. The results illustrate that even subtle differences in the analyses of AFLP markers can lead to drastically different conclusions. The study reveals multiple origins of a Mediterranean polyploid species. Furthermore, it demonstrates that the analysis of a complex marker system such as AFLP fingerprints using only one type of analysis can easily be misleading.     

Succession in a New Zealand alpine cushion community: a Markovian model

A permanent plot in a low-alpine cushion/tussock/shrub community was mapped at ca. decade intervals from 1953 to 1984. Transition probabilities between vegetation types were calculated, and a Markovian model was fitted.In spite of non-stationarity a Markov model was consistent with the observations, and produced reasonable predictions, especially for the major components of the vegetation. The model predictions were conservative in terms of future changes, in that future changes would be smaller than those observed, but such predictions could be supported by independent evidence, such as the nature of the non-stationarity, plant counts and seedling numbers.The general pattern was of a decrease in the cover of both Chionochloa rigida tussocks and mixed turf, and an increase in the cover of cushion bog species, especially Donatia novae-zelandiae. Shrubs, the suggested climax, showed no tendency to increase. Known climatic changes cannot explain the trend, which therefore seems to be a local retrogressive succession. There may be a cycle of alternating cushion and turf.     

Microclimate and variations in haemolymph composition in the freezing-tolerant New Zealand alpine weta Hemideina maori Hutton (Orthoptera: Stenopelmatidae)

The microclimate in the habitat of the New Zealand alpine weta Hemideina maori is very variable with winter temperatures down to −6 °C under the rocks where the insects are found. Subfreezing temperatures may in winter prevail for up to 17 days but diurnal cycles of freezing and thawing are common, as is also the case in summer. Rates of temperature change can be very high and up to −7.20 °C/h. During winter, humidity was high for extended periods ranging from 70% to 100% relative humidity (RH). In the summer, humidity ranged from 30% RH during the day to 100% RH at night. The supercooling point of the haemolymph was approximately −8 °C year round, caused by a heat labile substance. The supercooling point of the haemolymph of an insect of the same genus, Hemideina femorata not regularly exposed to subfreezing temperatures, was ca. −16.5 °C. Thermal hysteresis was not detected in the haemolymph of H. maori. Haemolymph osmolality varied from 380 mOsm (summer) to 700 mOsm (winter). Body water content was ca. 75% all year round. Total concentrations of sodium, potassium and chloride in haemolymph varied from 170 mM (winter) to 250 mM (summer). The total concentration of free amino acids varied from 58 mM (summer) to 263 mM (winter). This variation was mostly due to proline which varied from ca. 15 mM (summer) to ca. 100 mM (winter). The freeze-tolerant weta H. maori is exposed to a highly variable and cold environment all year round and several properties of its haemolymph composition can be attributed to these climatic conditions, e.g. the presence of ice-nucleating agents and an increase in the concentration of proline during cold hardening in the autumn. Accepted: 22 February 1999     

A wingless alpine stonefly from New Zealand,and further information on genus Holcoperla (Plecoptera: Gripopterygidae)

Abstract

Holcoperla magna n.sp. is described and figured from material collected on Headlong Peak, Mt Aspiring National Park. New information is given on H. angularis (Wisely) and H. jacksoni McLellan, and the 3 species are compared. Larvae designated as type specimens of H. angularis are shown to be Apteryoperla monticola Wisely.     

The New Zealand Hydroptilidae (Trichoptera)

Two new species of the genus Paroxyethira are described and figured. The females of the four other New Zealand Hydroptilidae are redescribed and illustrated, and a new key to the species is given. Taxonomic differences between the larvae and pupae are discussed.     

Ice nucleation and freezing tolerance in New Zealand alpine and lowland weta, Hemideina spp. (Orthoptera; Stenopelmatidae)

Abstract.The alpine tree weta Hemidiena maori Pictet et Saussure (Orthoptera: Stenopelmatidae) is a large, flightless insect found above the treeline on many of the mountain ranges of the South Island of New Zealand. The population found on the Rock and Pillar Range, Central Otago has been identified as freezing tolerant with a haemolymph ice nucleating agent. The ability of H. maori to survive freezing is compared to the lowland weta Hemideina thoracica Walker and H. crassidens Blanchard, both of which are able to survive the formation of some ice in their bodies. Mortality is associated with time spent frozen in H. thoracica , and it is hypothesized that this species is killed when a critical proportion of its body water is frozen. All five subalpine and alpine populations of H. maori surveyed were found to be freezing tolerant.
Comparison of temperatures of first nucleation and mean supercooling point of haemolymph droplets suggest that haemolymph ice nucleating activity varies between populations of H. maori. Hemideina maori collected from the Mt Cook region appear to lack a haemolymph ice nucleator. This population is nevertheless freezing tolerant, suggesting that the haemolymph ice nucleating agent described in H. maori is not essential for freezing tolerance. Hemideina crassidens and H. ricta Hutton, both of which are found in lowland habitats, also had high mean supercooling point and temperatures of first nucleation of haemolymph droplets, suggesting that these species also have a haemolymph ice nucleator.
Comparison of ice nucleation characteristics of haemolymph and faecal material (representing gut contents) suggests that gut nucleators in H. maori may be at least as efficient as the haemolymph nucleator. It is concluded that freezing tolerance is probably not an adaptation to the alpine environment. This highlights the need for inter- and intraspecific comparative studies if physiological data are to be used to draw evolutionary conclusions.