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Myrtle rust is a plant disease caused through infection by the fungus Austropuccinia psidii and was first detected in Australia in 2010. The disease has spread through New South Wales, Victoria, Queensland, the Northern Territory, and Tasmania. In this short timeframe, myrtle rust has had a devastating impact on many native species in the family Myrtaceae, including several rainforest species that are now at risk of extinction. In 2022, myrtle rust was first detected in the northern part of Western Australia (WA)-the largest state in Australia. WA is home to ca. 2000 Myrtaceae taxa ( ca. 60% of Australia's Myrtaceae diversity), many of which form the dominant component of the vegetation across several ecosystems (e.g., Eucalyptus, Corymbia, Melaleuca, Agonis, Verticordia etc.). While modelling suggests that the environmental conditions in WA's north are less conducive to myrtle rust in comparison to the wet, temperate rainforests of the east coast, WA's temperate, Myrtaceae-rich south coast may be climatically suitable. Coupled with the sheer abundance of Myrtaceae species in WA, their high degree of endemism, high proportion of threatened species, and little available information on their susceptibility to myrtle rust, a pre-emptive strategy to conserve germplasm of at-risk species is warranted. This paper highlights the role of ex situ germplasm conservation in responding to biosecurity threats such as myrtle rust. With early intervention critical to sourcing healthy and genetically diverse germplasm, we present a prioritized list of genera and species of Myrtaceae in WA to inform strategic, coordinated, and timely ex situ conservation actions, along with case studies to illustrate the complementary approaches of seed banking, cryobiotechnology, and tissue culture necessary to conserve germplasm of WA's myrtaceous flora.

Larvae of the genus Megalopyge (Lepidoptera: Zygaenoidea: Megalopygidae), known as asp or puss caterpillars, produce defensive venoms that cause severe pain. Here, we present the anatomy, chemistry, and mode of action of the venom systems of caterpillars of two megalopygid species, the Southern flannel moth Megalopyge opercularis and the black-waved flannel moth Megalopyge crispata . We show that megalopygid venom is produced in secretory cells that lie beneath the cuticle and are connected to the venom spines by canals. Megalopygid venoms consist of large aerolysin-like pore-forming toxins, which we have named megalysins, and a small number of peptides. The venom system differs markedly from those of previously studied venomous zygaenoids of the family Limacodidae, suggestive of an independent origin. Megalopygid venom potently activates mammalian sensory neurons via membrane permeabilization and induces sustained spontaneous pain behavior and paw swelling in mice. These bioactivities are ablated by treatment with heat, organic solvents, or proteases, indicating that they are mediated by larger proteins such as the megalysins. We show that the megalysins were recruited as venom toxins in the Megalopygidae following horizontal transfer of genes from bacteria to the ancestors of ditrysian Lepidoptera. Megalopygids have recruited aerolysin-like proteins as venom toxins convergently with centipedes, cnidarians, and fish. This study highlights the role of horizontal gene transfer in venom evolution.

Environmentally cued germination may play an important role in promoting coexistence in Mediterranean annual plant systems if it causes niche differentiation across heterogeneous microsite conditions. In this study, we tested how microsite conditions experienced by seeds in the field and light conditions in the laboratory influenced germination in 12 common annual plant species occurring in the understorey of the York gum-jam woodlands in southwest Western Australia. Specifically, we hypothesized that if germination promotes spatial niche differentiation, then we should observe species-specific germination responses to light. In addition, we hypothesized that species' laboratory germination response may depend on the microsite conditions experienced by seeds while buried. We tested the laboratory germination response of seeds under diurnally fluctuating light and complete darkness, which were collected from microsites spanning local-scale environmental gradients known to influence community structure in this system. We found that seeds of 6 out of the 12 focal species exhibited significant positive germination responses to light, but that the magnitude of these responses varied greatly with the relative light requirement for germination ranging from 0.51 to 0.86 for these species. In addition, germination increased significantly across a gradient of canopy cover for two species, but we found little evidence to suggest that species' relative light requirement for germination varied depending on seed bank microsite conditions. Our results suggest that variability in light availability may promote coexistence in this system and that the microsite conditions seeds experience in the intra-growing season period can further nuance species germination behaviour., (© 2021. The Author(s).)

Changes in fire regimes due to climate change and fire management practices are affecting the timing, length, and distribution of vegetation fires throughout the year. Plant species responses and tolerances to fire differ from season to season and are influenced by species-specific phenological processes. The ability of seeds to tolerate extreme temperatures associated with fire is one of these processes, with survival linked to seed moisture content at the time of exposure. As fire is more often occurring outside historic dry fire seasons, the probability of fire occurring when seeds are hydrated may also be increasing. In this study, we set out to understand the seasonal dynamics of seed hydration for seeds of Banksia woodland species, and how certain seed traits interact with environmental conditions to influence survival of high temperatures associated with fire. We measured the moisture content of seeds buried to 2 cm in the soil seed bank for four common native species and one invasive species on a weekly basis throughout 2017, along with soil moisture content and environmental correlates. We determined water sorption isotherms at 20°C for seeds of each species and used these functions to model weekly variation in seed water activity and predict when seeds are most sensitive to soil heating. Using Generalised additive models (GAMs), we were able to describe approximately 67% of the weekly variance in seed water activity and explored differences in seed hydration dynamics between species. Seed water activity was sufficiently high (i.e., ≥ 0.85 a w ) so as to have created an increased risk of mortality if a fire had occurred during an almost continuous period between May and November in the study period (i.e., 2017). There were brief windows when seeds may have been in a dry state during early winter and late spring, and also when they may have been in a wet state during summer and late autumn. These data, and the associated analyses, provide an opportunity to develop approaches to minimize seed mortality during fire and maximize the seed bank response., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Tangney, Merritt and Miller.)

Seed-based restoration often experiences poor success due to a range of edaphic and biotic issues. Seed enhancement technologies (SETs) are a novel approach that can alleviate these pressures and improve restoration success. Broadly, SETs have been reviewed for agricultural and horticultural purposes, for specific types of SETs such as coating or priming, or for focal ecosystems. However, information is lacking for SETs within a restoration focused context, and how they are being used to alleviate certain barriers. This review aimed to synthesise the current literature on SETs to understand what SETs are being tested, in which sectors and locations they are being tested, what issues are faced within restoration using SETs, and how SETs are being used to approach these issues. Priming was highlighted as the main SET investigated. Inoculation, pesticide application and magnetic fields were also commonly tested (SETs we termed 'prospective techniques'). SET research mainly occurred in the agricultural sector. More recently, other sectors, such as restoration and rangeland management, have increased efforts into SET research. The restoration sector has focused on extruded pelleting and coating (with activated carbon), in combination with herbicide application, to overcome invasive species, and coating with certain additives to alleviate edaphic issues. Other sectors outside restoration were largely focused on evaluating priming for overcoming these barriers. The majority of priming research has been completed on crop species and differences between these species and ecosystems must be considered in future restoration efforts that focus on native seed use. Generally, SETs require further refinement, including identifying ideal additives and their optimum concentrations to target certain issues, refining formulations for coating and extruded pelleting and developing flash flaming. A bet-hedging approach using multiple SETs and/or combinations of SETs may be advantageous in overcoming a wide range of barriers in seed-based restoration., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Unseasonal fire occurrence is increasing globally, driven by climate change and other human activity. Changed timing of fire can inhibit postfire seedling recruitment through interactions with plant phenology (the timing of key processes, e.g., flower initiation, seed production, dispersal, germination), and therefore threaten the persistence of many plant species. Although empirical evidence from winter-rainfall ecosystems shows that optimal seedling recruitment is expected following summer and autumn (dry season) fires, we sought experimental evidence isolating the mechanisms of poor recruitment following unseasonal (wet season) fire. We implemented a seed-sowing experiment using nine species native to fire-prone, Mediterranean-climate woodlands in southwestern Australia to emulate the timing of postfire recruitment and test key mechanisms of fire seasonality effects. For seeds sown during months when fire is unseasonal (i.e., August-September: end of the wet winter season), seedling recruitment was reduced by up to 99% relative to seeds sown during seasonal fire months (i.e., May-June: end of the dry summer season) because of varying seed persistence, seedling emergence, and seedling survival. We found that up to 70 times more seedlings emerged when seeds were sown during seasonal fire months compared to when seeds were sown during unseasonal fire months. The few seedlings that emerged from unseasonal sowings all died with the onset of the dry season. Of the seeds that failed to germinate from unseasonal sowings, only 2% survived exposure on the soil surface over the ensuing hot and dry summer. Our experimental results demonstrate the potential for unseasonal fire to inhibit seedling recruitment via impacts on pregermination seed persistence and seedling establishment. As ongoing climate change lengthens fire seasons (i.e., unseasonal wildfires become more common) and managed fires are implemented further outside historically typical fire seasons, postfire seedling recruitment may become more vulnerable to failure, causing shifts in plant community composition towards those with fewer species solely dependent on seeds for regeneration., (© 2021 by the Ecological Society of America.)

Insects can become lethally infected by the oral intake of a number of insect-specific viruses. Virus infection commonly occurs in larvae, given their active feeding behaviour; however, older larvae often become resistant to oral viral infections. To investigate mechanisms that contribute to resistance throughout the larval development, we orally challenged Drosophila larvae at different stages of their development with Drosophila C virus (DCV, Dicistroviridae ). Here, we showed that DCV-induced mortality is highest when infection initiates early in larval development and decreases the later in development the infection occurs. We then evaluated the peritrophic matrix as an antiviral barrier within the gut using a Crystallin-deficient fly line ( Crys -/- ), whose PM is weakened and becomes more permeable to DCV-sized particles as the larva ages. This phenotype correlated with increasing mortality the later in development oral challenge occurred. Lastly, we tested in vitro the infectivity of DCV after incubation at pH conditions that may occur in the midgut. DCV virions were stable in a pH range between 3.0 and 10.5, but their infectivity decreased at least 100-fold below (1.0) and above (12.0) this range. We did not observe such acidic conditions in recently hatched larvae. We hypothesise that, in Drosophila larvae, the PM is essential for containing ingested virions separated from the gut epithelium, while highly acidic conditions inactivate the majority of the virions as they transit.

Venoms have evolved independently several times in Lepidoptera. Limacodidae is a family with worldwide distribution, many of which are venomous in the larval stage, but the composition and mode of action of their venom is unknown. Here, we use imaging technologies, transcriptomics, proteomics, and functional assays to provide a holistic picture of the venom system of a limacodid caterpillar, Doratifera vulnerans Contrary to dogma that defensive venoms are simple in composition, D. vulnerans produces a complex venom containing 151 proteinaceous toxins spanning 59 families, most of which are peptides <10 kDa. Three of the most abundant families of venom peptides (vulnericins) are 1) analogs of the adipokinetic hormone/corazonin-related neuropeptide, some of which are picomolar agonists of the endogenous insect receptor; 2) linear cationic peptides derived from cecropin, an insect innate immune peptide that kills bacteria and parasites by disrupting cell membranes; and 3) disulfide-rich knottins similar to those that dominate spider venoms. Using venom fractionation and a suite of synthetic venom peptides, we demonstrate that the cecropin-like peptides are responsible for the dominant pain effect observed in mammalian in vitro and in vivo nociception assays and therefore are likely to cause pain after natural envenomations by D. vulnerans Our data reveal convergent molecular evolution between limacodids, hymenopterans, and arachnids and demonstrate that lepidopteran venoms are an untapped source of novel bioactive peptides., Competing Interests: The authors declare no competing interest.

Gut microbes play an important role in the biology and evolution of insects. Australian native dung beetles (Scarabaeinae) present an opportunity to study gut microbiota in an evolutionary context as they come from two distinct phylogenetic lineages and some species in each lineage have secondarily adapted to alternative or broader diets. In this study, we characterised the hindgut bacterial communities found in 21 species of dung beetles across two lineages using 16S rRNA sequencing. We found that gut microbial diversity was more dependent on host phylogeny and gut morphology than specific dietary preferences or environment. In particular, gut microbial diversity was highest in the endemic, flightless genus Cephalodesmius that feeds on a broad range of composted organic matter. The hindgut of Cephalodesmius harbours a highly conserved core set of bacteria suggesting that the bacteria are symbiotic. Symbiosis is supported by the persistence of the core microbiota across isolated beetle populations and between species in the genus. A co-evolutionary relationship is supported by the expansion of the hindgut to form a fermentation chamber and the fermentative nature of the core microbes. In contrast, Australian species of the widespread dung beetle genus Onthophagus, specialise on a single food resource such as dung or fungus, exhibit minimal food processing behaviour, have a short, narrow hindgut and a variable gut microbiota with relatively few core bacterial taxa. A conserved, complex gut microbiota is hypothesised to be unnecessary for this highly mobile genus. IMPORTANCE Dung beetles are a very important part of an ecosystem because of their role in the removal and decomposition of vertebrate dung. It has been suspected that symbiotic gut bacteria facilitate this role, a hypothesis that we have explored with high throughput barcoding. We found that differences in hindgut morphology had the greatest effect on the bacterial community composition. Species with a hindgut fermentation chamber harboured a distinctly different hindgut community compared to those species with a narrow, undifferentiated hindgut. Diet and phylogeny were also associated with differences in gut community. Further understanding of the relationships between dung beetles and their gut microbes will provide insights into the evolution of their behaviours and how gut communities contribute to their fitness., (Copyright © 2020 Ebert et al.)

Altered fire regimes resulting from climate change and human activity threaten many terrestrial ecosystems. However, we lack a holistic and detailed understanding of the effects of altering one key fire regime component - season of fire. Altered fire seasonality can strongly affect post-fire recovery of plant populations through interactions with plant phenology. We identify seven key mechanisms of fire seasonality effects under a conceptual demographic framework and review evidence for these. We reveal negative impacts of altered fire seasonality and identify research gaps for mechanisms and climate types for future analyses of fire seasonality effects within the identified demographic framework. This framework and these mechanisms can inform critical decisions for conservation, land management, and fire management policy development globally., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Rehabilitation of degraded drylands is challenged by environmental and anthropogenic constraints, such as limited availability of locally-sourced topsoil and poor quality alternative soil substrates. Current rehabilitation practices, at times, utilise inorganic soil amendments to improve the physicochemical and biological characteristics of reconstructed soil profiles. These approaches may be appropriate for dryland rehabilitation, but there is limited research available regarding the benefits of using these amendments. Here, we present a study in the Pilbara region of Western Australia, an arid landscape subject to intensive mining that currently uses inorganic soil amendments (gypsum and urea) in post-mining rehabilitation. The aim of this study was to assess the effectiveness of these amendments to (1) promote seed germination, seedling emergence and seedling growth across five plant species and, (2) re-instate soil quality in mine waste substrates. A series of glasshouse experiments assessed eight application combinations of these amendments in two alternative substrates and compared these to unamended substrates and topsoil. Soil amendments had a limited influence on seed germination, were detrimental to seedling emergence and resulted in increased seedling mortality. Mortality in the waste ranged from 2 to 61% but increased to 7-92% in amended waste. Seedling growth improved with high doses of amendments in waste, with a 1.3-5.6-fold increase across all plant species. Soil quality was relatively unaffected by amendments with soil nitrogen ranging from 0.01 to 0.08%, organic carbon from 0.01 to 0.12% and soil microbial activity from 2.3 to 2.4 ppm-CO 2 in the amended and unamended waste. The use of soil amendments in mine rehabilitation requires consideration of the trade-off between initial reductions in seedling recruitment and enhanced seedling development at later stages. Future rehabilitation should consider the timing of amendment application to avoid detrimental impacts on seedling recruitment and maximise the benefits to seedling growth., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Eighteen native species of Nymphaea (waterlilies) inhabit a range of freshwater wetlands in northern Australia, which are threatened by increased development and the potential impacts of climate change. To investigate conservation seed banking of these vulnerable species, we aimed to characterize their seed storage physiology by determining (i) seed desiccation tolerance and (ii) the effects of moisture content and storage temperature on seed germination and viability. Seeds of N. immutabilis , N. lukei , N. macrosperma and N. violacea (including multiple collections of three species) were placed in experimental storage at a range of temperatures (25°C, 5°C, -20°C and -190°C) following pre-equilibration at different RHs (15%, 30%, 50%, 70% or 95%). Seeds were also experimentally aged at 60% RH and 45°C to assess comparative longevity. We found seeds of all species to be desiccation tolerant. However, the responses of seeds to experimental storage conditions were complex and variable between species and collections of the same species, and seeds of many species/collections were short-lived across many of the storage treatments. In many cases decreasing storage temperature did not increase longevity. Additional protocol development is necessary before we can have confidence that ex situ seed banking is a viable long-term germplasm conservation strategy for Nymphaea .

Wildfires are a natural component in many terrestrial ecosystems and often play a crucial role in maintaining biodiversity, particularly in the fire-prone regions of Australia. A prime example of plants that are able to persist in these regions is the genus Banksia . Most Banksia species that occur in fire-prone regions produce woody seed pods (follicles), which open during or soon after fire to release seeds into the post-fire environment. For population persistence, many Banksia species depend on recruitment from these canopy-stored seeds. Therefore, it is critical that their seeds are protected from heat and rapid oxidation during fire. Here, we show how different species of Banksia protect their seeds inside follicles while simultaneously opening up when experiencing fire. The ability of the follicles to protect seeds from heat is demonstrated by intense 180 s experimental burns, in which the maximum temperatures near the seeds ranged from ∼75°C for B. serrata to ∼90°C for B. prionotes and ∼95°C for B. candolleana , contrasting with the mean surface temperature of ∼450°C. Many seeds of native Australian plants, including those of Banksia , are able to survive these temperatures. Structural analysis of individual follicles from these three Banksia species demonstrates that all of them rely on a multicomponent system, consisting of two valves, a porous separator and a thin layer of air surrounding the seeds. The particular geometric arrangement of these components determines the rate of heat transfer more than the tissue properties alone, revealing that a strong embedment into the central rachis can compensate for thin follicle valves. Furthermore, we highlight the role of the separator as an important thermal insulator. Our study suggests that the genus Banksia employs a variety of combinations in terms of follicle size, valve thickness, composition and geometric arrangement to effectively protect canopy-stored seeds during fire.

Trait-based approaches have improved our understanding of plant evolution, community assembly and ecosystem functioning. A major challenge for the upcoming decades is to understand the functions and evolution of early life-history traits, across levels of organization and ecological strategies. Although a variety of seed traits are critical for dispersal, persistence, germination timing and seedling establishment, only seed mass has been considered systematically. Here we suggest broadening the range of morphological, physiological and biochemical seed traits to add new understanding on plant niches, population dynamics and community assembly. The diversity of seed traits and functions provides an important challenge that will require international collaboration in three areas of research. First, we present a conceptual framework for a seed ecological spectrum that builds upon current understanding of plant niches. We then lay the foundation for a seed-trait functional network, the establishment of which will underpin and facilitate trait-based inferences. Finally, we anticipate novel insights and challenges associated with incorporating diverse seed traits into predictive evolutionary ecology, community ecology and applied ecology. If the community invests in standardized seed-trait collection and the implementation of rigorous databases, major strides can be made at this exciting frontier of functional ecology., (© 2018 The Authors. New Phytologist © 2018 New Phytologist Trust.)

Background and Aims: Substantial evidence supports the hypothesis that morphophysiological dormancy (MPD) is the basal kind of seed dormancy in the angiosperms. However, only physiological dormancy (PD) is reported in seeds of the ANA-grade genus Nymphaea. The primary aim of this study was to determine the kind of dormancy in seeds of six species of Nymphaea from the wet-dry tropics of Australia., Methods: The effects of temperature, light and germination stimulants on germination were tested on multiple collections of seeds of N. immutabilis, N. lukei, N. macrosperma, N. ondinea, N. pubescens and N. violacea. Embryo growth prior to hypocotyl emergence was monitored., Key Results: Germination was generally <10 % after 28 d in control treatments. Germination percentage was highest at 30 or 35 °C for seeds exposed to light and treated with ethylene or in anoxic conditions in sealed vials of water, and it differed significantly between collections of N. lukei, N. macrosperma and N. violacea. Seeds of N. pubescens did not germinate under any of the conditions. Embryo growth (8-37 % in length) occurred before hypocotyl emergence (germination) in seeds of the five species that germinated., Conclusions: Fresh seeds were dormant, and the amount of pregermination embryo growth in seeds of N. lukei and N. immutabilis was relatively small, while in seeds of N. macrosperma, N. ondinea and N. violacea it was relatively large. Thus, seeds of N. lukei and N. immutabilis had PD and those of N. macrosperma, N. ondinea and N. violacea had MPD. Overall, we found that seeds in the most phylogenetically derived clades within Nymphaea have MPD, suggesting that PD is the most likely basal trait within the Nymphaeales. This study also highlights the broad range of dormancy types and germination strategies in the ANA-grade angiosperms.

Seed dormancy status regulates the response of seeds to environmental cues that can trigger germination. Anigozanthos flavidus (Haemodoraceae) produces seeds with morphophysiological dormancy (MPD) that are known to germinate in response to smoke, but embryo growth dynamics and germination traits in response to temperatures and after-ripening have not been well characterized. Seeds of A. flavidus , after-ripened for 28 months at 15 °C/15 % relative humidity, were incubated on water agar, water agar containing 1 μM karrikinolide (KAR 1 ) or 50 μM glyceronitrile at 5, 10, 15, 20, 25, 20/10 and 25/15 °C for 28 days. After incubation at 5, 10 and 25 °C for 28 days, seeds were transferred to 15 °C for another 28 days. Embryo growth dynamics were tested at 5, 10, 15 and 25 °C. Results demonstrated that fresh seeds of A. flavidus had MPD and the physiological dormancy (PD) component could be broken by either glyceronitrile or dry after-ripening. After-ripened seeds germinated to ≥80 % at 15-20 °C while no additional benefit of germination was observed in the presence of the KAR 1 or glyceronitrile. Embryo length significantly increased at 10 °C, and only slightly increased at 5 °C, while growth did not occur at 25 °C. When un-germinated seeds were moved from 5-10 °C to 15 °C for a further 28 days, germination increased from 0 to >80 % in significantly less time indicating that cold stratification may play a key role in the germination process during winter and early spring in A. flavidus . The lower germination (<50 %) of seeds moved from 25 to 15 °C was produced by the induction of secondary dormancy. Induction of secondary dormancy in seeds exposed to warm stratification, a first report for Anigozanthos species, suggests that cycling of PD may be an important mechanism of controlling germination timing in the field.

Many plants in fire-prone regions retain their seeds in woody fruits in the plant canopy until the passage of a fire causes the fruit to open and release the seeds. To enable this function, suitable tissues are required that effectively store and protect seeds until they are released. Here, we show that three different species of the Australian genus Banksia incorporate waxes at the interface of the two valves of the follicle enclosing the seeds, which melt between 45°C and 55°C. Since the melting temperature of the waxes is lower than the opening temperatures of the follicles in all investigated species ( B. candolleana , B. serrata , B. attenuata ), we propose that melting of these waxes allows the sealing of micro-fissures at the interface of the two valves while they are still closed. Such a self-sealing mechanism likely contributes to the structural integrity of the seed pods, and benefits seed viability and persistence during storage on the plants. Furthermore, we show in a simplified, bioinspired model system that temperature treatments seal artificially applied surface cuts and restore the barrier properties., (© 2018 The Author(s).)

We explore a recent, innovative variation of closed-system respirometry for terrestrial organisms, whereby oxygen partial pressure ( P O 2 ) is repeatedly measured fluorometrically in a constant-volume chamber over multiple time points. We outline a protocol that aligns this technology with the broader literature on aerial respirometry, including the calculations required to accurately convert O 2 depletion to metabolic rate (MR). We identify a series of assumptions, and sources of error associated with this technique, including thresholds where O 2 depletion becomes limiting, that impart errors to the calculation and interpretation of MR. Using these adjusted calculations, we found that the resting MR of five species of angiosperm seeds ranged from 0.011 to 0.640 ml g -1  h -1 , consistent with published seed MR values. This innovative methodology greatly expands the lower size limit of terrestrial organisms that can be measured, and offers the potential for measuring MR changes over time as a result of physiological processes of the organism., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2018. Published by The Company of Biologists Ltd.)

Heat-triggered fruit opening and delayed release of mature seeds are widespread among plants in fire-prone ecosystems. Here, the material characteristics of the seed-containing follicles of Banksia attenuata (Proteaceae), which open in response to heat frequently caused by fire, are investigated. Material analysis reveals that long-term dimensional stability and opening temperatures of follicles collected across an environmental gradient increase as habitats become drier, hotter, and more fire prone. A gradual increase in the biaxial curvature of the hygroscopic valves provides the follicles in the driest region with the highest flexural rigidity. The irreversible deformation of the valves for opening is enabled via a temperature-dependent reduction of the elastic modulus of the innermost tissue layer, which then allows releasing the stresses previously generated by shrinkage of the fiber bundles in the adjacent layer during follicle drying. These findings illustrate the level of sophistication by which this species optimizes its fruit opening mechanism over a large distribution range with varying environmental conditions, and may not only have great relevance for developing biomimetic actuators, but also for elucidating the species' capacity to cope with climatic changes.

Background: The Mantispidae are a distinctive group of Neuroptera known for the adults' possession of raptorial forelegs. There are four recognised, extant subfamilies of Mantispidae: the Mantispinae, Symphrasinae, Calomantispinae and Drepanicinae. The life history and larval behaviour of the subfamily Mantispinae is best known: the immatures are spider egg predators. Among the three remaining subfamilies, larval Symphrasinae and Calomantispinae most likely predate on other small arthropods, while the immature life history of Drepanicinae, until now, remained completely unknown., New Information: Here we provide observations of annual, near-synchronised, mass emergences of adults of the drepanicine, Ditaxis biseriata (Westwood), within a well-established Macadamia orchard in northern New South Wales, Australia. A female deposited fertile eggs, allowing this first report of egg batch and first instar morphology. The mass emergence of mobile pharate adults from the ground was observed in the same month in two consecutive years. The pharates climbed tree-trunks for a distance before undergoing eclosion. The newly-hatched first instar larvae are campodeiform and prognathous; a typical morphology among Mantispidae. After hatching, they drop to the ground and burrow into soil. They are unpigmented and appear to lack stemmata. Together, the observations infer that the immature component of the life cycle takes place underground in forested habitats. If this feature is common among the Drepanicinae, it might explain why so little is known of the biology of the immature stages.

Glowworms, members of the keroplatid fly genus, Arachnocampa, glow to attract prey. Here we describe substantial differences in the bioluminescence regulatory systems of two species; one is a troglophile with populations both in caves and outside of caves in wet forest (Arachnocampa tasmaniensis) and the other has no known cave populations (Arachnocampa flava). We find that A. tasmaniensis is ready to initiate bioluminescence at any time darkness is encountered. In contrast, A. flava shows a homeostatic control of bioluminescence; it is unlikely to initiate bioluminescence when exposed to dark pulses during the photophase and it does so with a long latency. Another difference between the two species is that A. tasmaniensis individuals synchronize their bioluminescence in the dark zone of caves under the control of the circadian system and A. flava individuals do not synchronize to each other, rather their circadian control system entrains to the light:dark cycle to promote nocturnal bioluminescence. Consequently, we produced a phase-response curve in response to photic entrainment under constant darkness for both species. The shape of the phase-response curves differs between the two species as does the overall sensitivity to the identical entrainment conditions. The phase-response curve of A. tasmaniensis facilitates synchronization whereas that of A. flava facilitates nocturnal glowing. The two-species comparison highlights possible pathways of divergence of circadian control of physiological functions that could be associated with the extreme ecological differences experienced in cave and surface habitats., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Fire is a topical issue in the management of many ecosystems globally that face a drying climate. Understanding the role of fire in such ecosystems is critical to inform appropriate management practices, particularly in the case of rare and ecologically specialized species. The Mediterranean heathlands are highly fire-prone and occur in a biodiversity hotspot increasingly threatened by human activities, and determining the reproductive thresholds of at-risk heathland species is critical to ensuring the success of future conservation initiatives. This study examined the germination biology of the threatened carnivorous subshrub Drosophyllum lusitanicum , with specific focus on the role of fire-related cues (heat and smoke) in combination with seasonal temperatures and moisture conditions to determine how these factors regulate seed dormancy and germination. We found that D. lusitanicum produces water-permeable, physiologically dormant seeds with a fully developed, capitate embryo that when fresh (~1 month old) and without treatment germinate to 20-40 % within 4-8 weeks. Seeds possess a restricted thermal window (15-20 °C) for germination and a neutral photoblastic response. Seed dormancy was overcome through precision nicking of the seed coat (>90 % germination) or by short exposure to dry heat (80 or 100 °C) for 5-30 min (60-100 % germination). We propose seedling emergence from the soil seed bank may be cued by the passage of fire, or by soil disturbance from the movement and browsing of animals. Long-term population viability is likely to be contingent upon appropriate management of the persistent soil seed bank, as well as the adequate management of key ecological disturbances such as fire. Drosophyllum lusitanicum faces an increasingly bleak future in the absence of conservation and management initiatives aimed at reducing habitat fragmentation in heathlands and aligning fire management and livestock practices with biodiversity outcomes.

Seed coating is the practice of covering seeds with external materials to improve handling, protection, and, to a lesser extent, germination enhancement and plant establishment. With an annual value exceeding US$1 billion dollars, this technology is mostly the preserve of the private research sector, with few links to the scientific community. Here, we analyse the science and industry of seed coating and its contribution to seed establishment and plant performance. We posit that a closer collaboration between academia and industry is critical to realising the potential of seed coating both as a tool for enhancing plant establishment in the face of the challenges posed to agricultural systems and to propel the multibillion-dollar global push for ecological restoration of degraded ecosystems., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Animals use adhesive secretions in a plethora of ways, either for attachment, egg anchorage, mating or as either active or passive defence. The most interesting function, however, is the use of adhesive threads to capture prey, as the bonding must be performed within milliseconds and under unsuitable conditions (movement of prey, variable environmental conditions, unfavourable attack angle, etc.) to be nonetheless successful. In the following study a detailed characterization of the prey capture system of the world-renowned glowworm group Arachnocampa from the macroscopic to the ultrastructural level is performed. The data reveal that the adhesive droplets consist mostly of water and display hygroscopic properties at varying humidity levels. The droplet core of Arachnocampa luminosa includes a certain amount of the elements sodium, sulphur and potassium (beside carbon, oxygen and nitrogen), while a different element composition is found in the two related species A. richardsae and A. tasmaniensis. Evidence for lipids, carbohydrates and proteins was negative on the histochemical level, however X-ray photoelectron spectroscopy confirm the presence of peptides within the droplet content. Different to earlier assumptions, the present study indicates that rather than oxalic acid, urea or uric acid are present in the adhesive droplets, presumably originating from the gut. Comparing the capture system in Arachnocampa with those of orb-spiders, large differences appear not only regarding the silky threads, but also, in the composition, hygroscopic properties and size of the mucous droplets., Competing Interests: The authors have declared that no competing interests exist.

Soil respiration (Rs) is the second largest carbon flux in terrestrial ecosystems and therefore plays a crucial role in global carbon (C) cycling. This biogeochemical process is closely related to ecosystem productivity and soil fertility and is considered as a key indicator of soil health and quality reflecting the level of microbial activity. Wildfires can have a significant effect on Rs rates and the magnitude of the impacts will depend on environmental factors such as climate and vegetation, fire severity and meteorological conditions post-fire. In this research, we aimed to assess the impacts of a wildfire on the soil CO 2 fluxes and soil respiration in a semi-arid ecosystem of Western Australia, and to understand the main edaphic and environmental drivers controlling these fluxes for different vegetation types. Our results demonstrated increased rates of Rs in the burnt areas compared to the unburnt control sites, although these differences were highly dependent on the type of vegetation cover and time since fire. The sensitivity of Rs to temperature (Q10) was also larger in the burnt site compared to the control. Both Rs and soil organic C were consistently higher under Eucalyptus trees, followed by Acacia shrubs. Triodia grasses had the lowest Rs rates and C contents, which were similar to those found under bare soil patches. Regardless of the site condition (unburnt or burnt), Rs was triggered during periods of higher temperatures and water availability and environmental factors (temperature and moisture) could explain a large fraction of Rs variability, improving the relationship of moisture or temperature as single factors with Rs. This study demonstrates the importance of assessing CO 2 fluxes considering both abiotic factors and vegetation types after disturbances such as fire which is particularly important in heterogeneous semi-arid areas with patchy vegetation distribution where CO 2 fluxes can be largely underestimated., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Wild-collected seed can no longer meet global demand in restoration. Dedicated Seed Production Areas (SPA) for restoration are needed and these require application of ecological, economic, and population-genetic science. SPA design and construction must embrace the ecological sustainability principles of restoration.

Glow-worms (larvae of dipteran genus Arachnocampa) are restricted to moist habitats where they capture flying prey using snares composed of highly extensible silk fibres and sticky mucus droplets. Little is known about the composition or structure of glow-worm snares, or the extent of possible convergence between glow-worm and arachnid capture silks. We characterised Arachnocampa richardsae silk and mucus using X-ray scattering, Fourier transform infrared spectroscopy and amino acid analysis. Silk but not mucus contained crystallites of the cross-β-sheet type, which occur in unrelated insect silks but have not been reported previously in fibres used for prey capture. Mucus proteins were rich in Gly (28.5%) and existed in predominantly a random coil structure, typical of many adhesive proteins. In contrast, the silk fibres were unusually rich in charged and polar residues, particularly Lys (18.1%), which we propose is related to their use in a highly hydrated state. Comparison of X-ray scattering, infrared spectroscopy and amino acid analysis data suggests that silk fibres contain a high fraction of disordered protein. We suggest that in the native hydrated state, silk fibres are capable of extension via deformation of both disordered regions and cross-β-sheet crystallites, and that high extensibility is an adaptation promoting successful prey capture. This study illustrates the rich variety of protein motifs that are available for recruitment into biopolymers, and how convergently evolved materials can nevertheless be based on fundamentally different protein structures., (Crown Copyright © 2015. Published by Elsevier Inc. All rights reserved.)

Background and Aims: Rock pools are small, geologically stable freshwater ecosystems that are both hydrologically and biologically isolated. They harbour high levels of plant endemism and experience environmental unpredictability driven by the presence of water over variable temporal scales. This study examined the hypothesis that the sediment seed bank in monsoon tropical freshwater rock pools would persist through one or more periods of desiccation, with seed dormancy regulating germination timing in response to rock pool inundation and drying events., Methods: Seeds were collected from seven dominant rock pool species, and germination biology and seed dormancy were assessed under laboratory conditions in response to light, temperature and germination stimulators (gibberellic acid, karrikinolide and ethylene). Field surveys of seedling emergence from freshwater rock pools in the Kimberley region of Western Australia were undertaken, and sediment samples were collected from 41 vegetated rock pools. Seedling emergence and seed bank persistence in response to multiple wetting and drying cycles were determined., Key Results: The sediment seed bank of individual rock pools was large (13 824 ± 307 to 218 320 ± 42 412 seeds m(-2) for the five species investigated) and spatially variable. Seedling density for these same species in the field ranged from 13 696 to 87 232 seedlings m(-2). Seeds of rock pool taxa were physiologically dormant, with germination promoted by after-ripening and exposure to ethylene or karrikinolide. Patterns of seedling emergence varied between species and were finely tuned to seasonal temperature and moisture conditions, with the proportions of emergent seedlings differing between species through multiple inundation events. A viable seed bank persisted after ten consecutive laboratory inundation events, and seeds retained viability in dry sediments for at least 3 years., Conclusions: The persistent seed bank in freshwater rock pools is likely to provide resilience to plant communities against environmental stochasticity. Since rock pool communities are often comprised of highly specialized endemic and range-restricted species, sediment seed banks may represent significant drivers of species persistence and diversification in these ecosystems., (© The Author 2015. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Venom represents one of the most extreme manifestations of a chemical arms race. Venoms are complex biochemical arsenals, often containing hundreds to thousands of unique protein toxins. Despite their utility for prey capture, venoms are energetically expensive commodities, and consequently it is hypothesized that venom complexity is inversely related to the capacity of a venomous animal to physically subdue prey. Centipedes, one of the oldest yet least-studied venomous lineages, appear to defy this rule. Although scutigeromorph centipedes produce less complex venom than those secreted by scolopendrid centipedes, they appear to rely heavily on venom for prey capture. We show that the venom glands are large and well developed in both scutigerid and scolopendrid species, but that scutigerid forcipules lack the adaptations that allow scolopendrids to inflict physical damage on prey and predators. Moreover, we reveal that scolopendrid venom glands have evolved to accommodate a much larger number of secretory cells and, by using imaging mass spectrometry, we demonstrate that toxin production is heterogeneous across these secretory units. We propose that the differences in venom complexity between centipede orders are largely a result of morphological restrictions of the venom gland, and consequently there is a strong correlation between the morphological and biochemical complexity of this unique venom system. The current data add to the growing body of evidence that toxins are not expressed in a spatially homogenous manner within venom glands, and they suggest that the link between ecology and toxin evolution is more complex than previously thought.

The Heteroptera show a diversity of glands associated with the epidermis. They have multiple roles including the production of noxious scents. Here, we examine the cellular arrangement and cytoskeletal components of the scent glands of pentatomoid Heteroptera in three families, Pentatomidae (stink bugs), Tessaratomidae, and Scutelleridae (shield-backed bugs or jewel bugs). The glands are; (1) the dorsal abdominal glands, (2) the tubular glands of the composite metathoracic gland, and (3) the accessory gland component of the composite metathoracic gland. The dorsal abdominal glands are at their largest in nymphs and decrease in size in adults. The metathoracic gland is an adult-specific gland unit with a reservoir and multiple types of gland cells. The accessory gland is composed of many unicellular glands concentrated in a sinuous line across the reservoir wall. The lateral tubular gland is composed of two-cell units. The dorsal abdominal glands of nymphs are composed of three-cell units with a prominent cuticular component derived from the saccule cell sitting between the duct and receiving canal. The cuticular components that channel secretion from the microvilli of the secretory cell to the exterior differ in the three gland types. The significance of the numbers of cells comprising gland units is related to the role of cells in regenerating the cuticular components of the glands at moulting in nymphs., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Flies of the genus Arachnocampa are sit-and-lure predators that use bioluminescence to attract flying prey to their silk webs. Some species are most common in rainforest habitat and others inhabit both caves and rainforest. We have studied the circadian regulation of bioluminescence in two species: one found in subtropical rainforest with no known cave populations and the other found in temperate rainforest with large populations in limestone caves. The rainforest species is typical of most nocturnal animals in that individuals are entrained by the light:dark (LD) cycle to be active at night; in this case, their propensity to bioluminesce is greatest at night. The dual-habitat species shows an opposite phase response to the same entrainment; its bioluminescence propensity rhythm is entrained by LD exposure to peak during the day. Nevertheless, in LD environments, individuals do not bioluminesce during the day because ambient light inhibits their bioluminescence (negative masking), pushing bioluminescence into the dark period. This unusual and unexpected phenomenon could be related to their association with caves and has been suggested to be an adaptation of the circadian system that promotes synchronization of a colony's output of bioluminescence. Here, we use controlled laboratory experiments to show that individuals do synchronize their bioluminescence rhythms when in visual contact with each other. Entrainment of the bioluminescence rhythm to the biological photophase causes colony-wide synchronization, creating a daily sinusoidal rhythm of the intensity of bioluminescence in the many thousands of individuals making up a colony. This synchronization could provide a group-foraging advantage, allowing the colony to glow most brightly when the prey are most likely to be active.

Background and Aims: Many terrestrial orchids have an obligate requirement for mycorrhizal associations to provide nutritional support from germination to establishment. This study will investigate the ability of orchid mycorrhizal fungi (OMF) to utilize a variety of nutrient sources in the nutrient-impoverished (low organic) soils of the Southwest Australian Floristic Region (SWAFR) in order to effectively compete, survive and sustain the orchid host., Methods: Mycorrhizal fungi representing key OMF genera were isolated from three common and widespread species: Pterostylis recurva, Caladenia flava and Diuris corymbosa, and one rare and restricted species: Drakaea elastica. The accessibility of specific nutrients was assessed by comparing growth including dry biomass of OMF in vitro on basal CN MMN liquid media., Key Results: Each of the OMF accessed and effectively utilized a wide variety of nutrient compounds, including carbon (C) sources, inorganic and organic nitrogen (N) and inorganic and organic phosphorus (P). The nutrient compounds utilized varied between the genera of OMF, most notably sources of N., Conclusions: These results suggest that OMF can differentiate between niches (micro-niche specialization) in a constrained, highly resource-limited environment such as the SWAFR. Phosphorus is the most limited macronutrient in SWAFR soils and the ability to access phytate by OMF indicates a characterizing functional capacity of OMF from the SWAFR. Furthermore, compared with OMF isolated from the rare D. elastica, OMF associating with the common P. recurva produced far greater biomass over a wider variety of nutritional sources. This suggests a broader tolerance for habitat variation providing more opportunities for the common orchid for recruitment and establishment at a site.

We provide insights into the secretory pathway of arthropod gland systems by comparing the royal jelly-producing hypopharyngeal glands and the venom-producing glands of the honeybee, Apis mellifera. These glands have different functions and different product release characteristics, but both belong to the class 3 types of insect glands, each being composed of two cells, a secretory cell and a microduct-forming cell. The hypopharyngeal secretory cells possess an extremely elongate tubular invagination that is filled with a cuticular structure, the end-apparatus, anchored against the cell membrane by a conspicuous series of actin rings. In contrast, venom glands have no actin rings, but instead have an actin-rich brush border surrounding the comparatively short and narrow end-apparatus. We relate these cytoskeletal differences to the production system and utilisation of secretions; venom is stored in a reservoir whereas royal jelly and enzymes are produced on demand. Fluorescence-based characterisation of the actin cytoskeleton combined with scanning electron microscopy of the end-apparatus allows for detailed characterisation of the point of secretion release in insect class 3 glands., (2012 Elsevier Ltd. All rights reserved)

Two new types of signaling compounds have been discovered in wildfire smoke due to their ability to stimulate seed germination. The first discovered were karrikins, which share some structural similarity with the strigolactone class of plant hormones, and both signal through a common F-box protein. However, karrikins and strigolactones operate through otherwise distinct signaling pathways, each distinguished by a specific α/β hydrolase protein. Genetic analysis suggests that plants contain endogenous compounds that signal specifically through the karrikin pathway. The other active compounds discovered in smoke are cyanohydrins that release germination-stimulating cyanide upon hydrolysis. Cyanohydrins occur widely in plants and have a role in defense against other organisms, but an additional role in endogenous cyanide signaling should also now be considered.

Arachnocampa species, commonly called glowworms, are flies whose larvae use light to attract prey. Here we compare rhythmicity in two of the nine described species: the Tasmanian species, Arachnocampa tasmaniensis, which inhabits caves and wet forest, and the eastern Australian mainland species, A. flava, primarily found in subtropical rainforest. Both species show the same nocturnal glowing pattern in external (epigean) environments and the same inhibition of bioluminescence by light and both species show circadian regulation of bioluminescence. We find that the underlying circadian bioluminescence propensity rhythm (BPR) of the two species peaks at opposite phases of the day:night cycle. Larvae of A. flava, placed in constant darkness in the laboratory, bioluminesce during the subjective scotophase, typical of nocturnal animals, whereas A. tasmaniensis shows the opposite tendency, bioluminescing most intensely during the subjective photophase. In A. tasmaniensis, which are exposed to natural day:night cycles, light exposure during the day overrides the high bioluminescence propensity through negative masking and leads to a release of bioluminescence after dusk when the BPR is on the wane. A consequence is that A. tasmaniensis is able to start glowing at any phase of the light:dark cycle as soon as masking by light is released, whereas A. flava is locked into nocturnal bioluminescence. We suggest that the paradoxical BPR of A. tasmaniensis is an adaptation for living in the cave environment. Observations of bioluminescence in colonies of A. tasmaniensis located in the transition from a cave mouth to the dark zone show that glowing is inhibited by light exposure but a peak bioluminescence follows immediately after "dusk" at their location. The substantial difference in the circadian regulation of bioluminescence between the two species probably reflects adaptation to the cave (hypogean) habitat in A. tasmaniensis and the forest (epigean) habitat in A. flava.

We describe a novel cytoskeletal element within secretory cells of an arthropod gland system, the hypopharyngeal gland of the honeybee, Apis mellifera. The hypopharyngeal secretory cells are the source of royal jelly in nurse bees and enzymes in foragers. Each cell possesses an elongate invagination that is occupied by a tubular cuticular structure, the end-apparatus, that accumulates secretion and transfers it into a cuticular microtube and then into a collecting duct. Within the secretory cell, a conspicuous series of actin rings, about 3 μm in diameter, follows the same path as the end-apparatus, surrounding it at spaced intervals. Transmission electron microscopy confirmed that the actin rings lie within septa of the secretory cell that are closely juxtaposed to the end-apparatus at regularly spaced intervals. We speculate that the function of the actin rings is to hold the end apparatus in place as secretion swells the extracellular compartments between the end apparatus and the cell membrane. To our knowledge, no such cytoskeletal component has been described in animal cells. © 2012 Wiley Periodicals, Inc., (Copyright © 2012 Wiley Periodicals, Inc.)

Background and Aims: Several ecologically important plant families in Mediterranean biomes have seeds with morphophysiological dormancy (MPD) but have been poorly studied. The aim of this study was to understand the seed ecology of these species by focusing on the prominent, yet intractably dormant Australian genus Hibbertia. It was hypothesized that the slow germination in species of this genus is caused by a requirement for embryo growth inside the seed before germination, and that initiation of embryo growth is reliant upon a complex sequence of environmental cues including seasonal fluctuations in temperature and moisture, and an interplay with light and smoke. Using the results, the classification of the MPD level in species of Hibbertia is considered., Methods: Four species of Hibbertia in winter rainfall south-western Australia were selected. These species, whilst differing in geographic distributions, are variously sympatric, and all are important understorey components of plant communities. The following aspects related to dormancy break, embryo growth and germination were investigated: temperature and moisture requirements; effects of karrikinolide, gibberellic acid and aerosol smoke; and phenology., Key Results: Following exposure to wet/dry cycles at low or high temperatures, embryo growth and germination occurred, albeit slowly in all species at low temperatures when moisture was unlimited, corresponding to winter in south-west Australia. Photo regime influenced germination only in H. racemosa. Aerosol smoke triggered substantial germination during the 1st germination season in H. huegelii and H. hypericoides., Conclusions: Although the study species are con-generic, sympatric and produce seeds of identical morphology, they possessed different dormancy-break and germination requirements. The physiological component of MPD was non-deep in H. racemosa but varied in the other three species where more deeply dormant seeds required >1 summer to overcome dormancy and, thus, germination was spread over time. Embryos grew during winter, but future studies need to resolve the role of cold versus warm stratification by using constant temperature regimes. To include Mediterranean species with MPD, some modifications to the current seed-dormancy classification system may need consideration: (a) wet/dry conditions for warm stratification and (b) a relatively long period for warm stratification. These outcomes have important implications for improving experimental approaches to resolve the effective use of broadcast seed for ecological restoration.

Background: The natural elastomeric protein, insect resilin, is the most efficient elastic material known, used to store energy for jumping and flight in a variety of insects. Here, an antibody to recombinant Drosophila melanogaster pro-resilin is used to examine resilin expression in Drosophila and a wider range of insects., Results: Immunostaining of Drosophila embryos reveals anti-resilin reactivity in epidermal patches that exhibit a dynamic spatial and temporal expression through late embryogenesis. Resilin is also detected in stretch receptors in the embryo. In developing adult Drosophila, resilin pads are described at the base of wings and at the base of flexible sensory hairs in pupae. Resilin is also detected in embryos of the tephritid fruitfly, Bactrocera tryoni, and two well-known concentrations of insect resilin: the flight muscle tendon of the dragonfly and the pleural arch of the flea., Conclusions: The anti-Rec1 antibody antibody developed using Drosophila pro-resilin as antigen is cross-reactive and is useful for detection of resilin in diverse insects. For the first time, resilin expression has been detected during embryogenesis, revealing segmental patches of resilin in the developing epidermis of Drosophila., (Copyright © 2011 Wiley Periodicals, Inc.)

Raspy crickets (Orthoptera: Gryllacrididae) are unique among the orthopterans in producing silk, which is used to build shelters. This work studied the material composition and the fabrication of cricket silk for the first time. We examined silk-webs produced in captivity, which comprised cylindrical fibers and flat films. Spectra obtained from micro-Raman experiments indicated that the silk is composed of protein, primarily in a beta-sheet conformation, and that fibers and films are almost identical in terms of amino acid composition and secondary structure. The primary sequences of four silk proteins were identified through a mass spectrometry/cDNA library approach. The most abundant silk protein was large in size (300 and 220 kDa variants), rich in alanine, glycine and serine, and contained repetitive sequence motifs; these are features which are shared with several known beta-sheet forming silk proteins. Convergent evolution at the molecular level contrasts with development by crickets of a novel mechanism for silk fabrication. After secretion of cricket silk proteins by the labial glands they are fabricated into mature silk by the labium-hypopharynx, which is modified to allow the controlled formation of either fibers or films. Protein folding into beta-sheet structure during silk fabrication is not driven by shear forces, as is reported for other silks.

Resilin is an important elastomeric protein of insects, with roles in the storage and release of energy during a variety of different functional categories including flight and jumping. To date, resilin genes and protein function have been characterised only in a small number of flying insects, despite their importance in fleas and other jumping insects. Microscopy and immunostaining studies of resilin in flea demonstrate the presence of resilin pads in the pleural arch at the top of the hind legs, a region responsible for the flea's jumping ability. A degenerate primer approach was used to amplify resilin gene transcripts from total RNA isolated from flea (Ctenocephalides felis), buffalo fly (Haematobia irritans exigua) and dragonfly (Aeshna sp.) pharate adults, and full-length transcripts were successfully isolated. Two isoforms (A and B) were amplified from each of flea and buffalo fly, and isoform B only in dragonfly. Flea and buffalo fly isoform B transcripts were expressed in an Escherichia coli expression system, yielding soluble recombinant proteins Cf-resB and Hi-resB respectively. Protein structure and mechanical properties of each protein before and after crosslinking were assessed. This study shows that resilin gene and protein sequences are broadly conserved and that crosslinked recombinant resilin proteins share similar mechanical properties from flying to jumping insects. A combined use of degenerate primers and polyclonal sera will likely facilitate characterisation of resilin genes from other insect and invertebrate orders., (Crown Copyright © 2011. Published by Elsevier Ltd. All rights reserved.)

The glowworm Arachnocampa flava is a carnivorous fly larva (Diptera) that uses light to attract prey into its web. The light organ is derived from cells of the Malpighian tubules, representing a bioluminescence system that is unique to the genus. Bioluminescence is modulated through the night although light levels change quite slowly compared with the flashing of the better-known fireflies (Coleoptera). The existing model for the neural regulation of bioluminescence in Arachnocampa, based on use of anaesthetics and ligations, is that bioluminescence is actively repressed during the non-glowing phase and the repression is partially released during the bioluminescence phase. The effect of the anaesthetic, carbon dioxide, on the isolated light organ from the present study indicates that the repression is at least partially mediated at the light organ itself rather than less directly through the central nervous system. Blocking of neural signals from the central nervous system through ligation leads to uncontrolled release of bioluminescence but light is emitted at relatively low levels compared with under anaesthesia. Candidate biogenic amines were introduced by several methods: feeding prey items injected with test solution, injecting the whole larva, injecting a ligated section containing the light organ or bathing the isolated light organ in test solution. Using these methods, dopamine, serotonin and tyramine do not affect bioluminescence output. Exposure to elevated levels of octopamine via feeding, injection or bathing of the isolated light organ indicates that it is involved in the regulation of repression. Administration of the octopamine antagonists phentolamine or mianserin results in very high bioluminescence output levels, similar to the effect of anaesthetics, but only mianserin acts directly on the light organ.

Background and Aims: Karrikinolide (KAR(1)) is a smoke-derived chemical that can trigger seeds to germinate. A potential application for KAR(1) is for synchronizing the germination of weed seeds, thereby enhancing the efficiency of weed control efforts. Yet not all species germinate readily with KAR(1), and it is not known whether seemingly non-responsive species can be induced to respond. Here a major agronomic weed family, the Brassicaceae, is used to test the hypothesis that a stimulatory response to KAR(1) may be present in physiologically dormant seeds but may not be expressed under all circumstances., Methods: Seeds of eight Brassicaceae weed species (Brassica tournefortii, Raphanus raphanistrum, Sisymbrium orientale, S. erysimoides, Rapistrum rugosum, Lepidium africanum, Heliophila pusilla and Carrichtera annua) were tested for their response to 1 µm KAR(1) when freshly collected and following simulated and natural dormancy alleviation, which included wet-dry cycling, dry after-ripening, cold and warm stratification and a 2 year seed burial trial., Key Results: Seven of the eight Brassicaceae species tested were stimulated to germinate with KAR(1) when the seeds were fresh, and the remaining species became responsive to KAR(1) following wet-dry cycling and dry after-ripening. Light influenced the germination response of seeds to KAR(1), with the majority of species germinating better in darkness. Germination with and without KAR(1) fluctuated seasonally throughout the seed burial trial., Conclusions: KAR(1) responses are more complex than simply stating whether a species is responsive or non-responsive; light and temperature conditions, dormancy state and seed lot all influence the sensitivity of seeds to KAR(1), and a response to KAR(1) can be induced. Three response types for generalizing KAR(1) responses are proposed, namely inherent, inducible and undetected. Given that responses to KAR(1) were either inherent or inducible in all 15 seed lots included in this study, the Brassicaceae may be an ideal target for future application of KAR(1) in weed management.

Cyanide is well known for its toxicity towards living organisms. Many plants use cyanide as a defensive agent against herbivores, releasing it through the enzymatic hydrolysis of endogenous cyanogenic compounds. At low concentrations, cyanide has been proposed to have a regulatory role in many plant processes including stimulation of seed germination. However, no ecological role for cyanide in seed germination has been established. In the present study, we show that burning plant material produces the cyanohydrin, glyceronitrile. We also show that, in the presence of water, glyceronitrile is slowly hydrolysed to release cyanide that stimulates seed germination of a diverse range of fire-responsive species from different continents. We propose that glyceronitrile serves as an ecological store for cyanide and is an important cue for stimulating seed germination and landscape regeneration after fires.

Trap-building, sit-and-wait predators such as spiders, flies and antlions tend to have low standard metabolic rates (SMRs) but potentially high metabolic costs of trap construction. Members of the genus Arachnocampa (glowworms) use an unusual predatory strategy: larvae bioluminesce to lure positively phototropic insects into their adhesive webs. We investigated the metabolic costs associated with bioluminescence and web maintenance in larval Arachnocampa flava. The mean rate of CO(2) production (VCO(2)) during continuous bioluminescence was 4.38 μl h(-1) ± 0.78 (SEM). The mean VCO(2) of inactive, non-bioluminescing larvae was 3.49 ± 0.35 μl h(-1). The mean VCO(2) during web maintenance when not bioluminescencing was 8.95 ± 1.78 μl h(-1), a value significantly lower than that measured during trap construction by other predatory arthropods. These results indicate that bioluminescence itself is not energetically expensive, in accordance with our prediction that a high cost of bioluminescence would render the Arachnocampa sit-and-lure predatory strategy inefficient. In laboratory experiments, both elevated feeding rates and daily web removal caused an increase in bioluminescent output. Thus, larvae increase their investment in light output when food is plentiful or when stressed through having to rebuild their webs. As light production is efficient and the cost of web maintenance is relatively low, the energetic returns associated with continuing to glow may outweigh the costs of continuing to attract prey.

Larvae of the genus Arachnocampa, known as glowworms, are bioluminescent predatory insects that use light to attract prey. One species, Arachnocampa flava, is known to possess true circadian regulation of bioluminescence: light:dark cycles entrain the rhythm of nocturnal glowing. Given the absence of natural light as a cue in caves, we addressed the question of whether cave populations of Arachnocampa tasmaniensis, a species known to inhabit caves as well as epigean environments, are rhythmic. We found that the major dark-zone cave populations of A. tasmaniensis maintain a high-amplitude 24-hour rhythm of bioluminescence, with the acrophase during external daylight hours. Populations of A. tasmaniensis in caves many kilometers apart show similar, but not exactly the same, timing of the acrophase. Systematic investigation of colonies in the dark zone of a single cave showed that some smaller colonies distant to the main ceiling colony, also in the dark zone, glow in antiphase. Periodic monitoring of a single colony over several years showed that the acrophase shifted from nocturnal to diurnal some time between October 2008 and January 2009. Prey availability was investigated as a possible zeitgeber. The acrophase of prey availability, as measured by light trapping, and the acrophase of bioluminescence do not precisely match, occurring 3 hours apart. Using in-cave artificial light exposure, we show that after LD cycles, cave larvae become entrained to bioluminesce during the foregoing photophase. In contrast, epigean larvae exposed to artificial LD cycles after a period of DD become entrained to bioluminesce during the foregoing scotophase. One explanation is that individuals within colonies in the dark zone synchronize their bioluminescence rhythms through detection and matching of each other 's bioluminescence.