Conditional gene deletion with DiCre demonstrates an essential role for CRK3 in Leishmania mexicana cell cycle regulation

Leishmania mexicana has a large family of cyclin‐dependent kinases (CDKs) that reflect the complex interplay between cell cycle and life cycle progression. Evidence from previous studies indicated that Cdc2‐related kinase 3 (CRK3) in complex with the cyclin CYC6 is a functional homologue of the major cell cycle regulator CDK1, yet definitive genetic evidence for an essential role in parasite proliferation is lacking. To address this, we have implemented an inducible gene deletion system based on a dimerised Cre recombinase (diCre) to target CRK3 and elucidate its role in the cell cycle of L. mexicana. Induction of diCre activity in promastigotes with rapamycin resulted in efficient deletion of floxed CRK3, resulting in G2/M growth arrest. Co‐expression of a CRK3 transgene during rapamycin‐induced deletion of CRK3 resulted in complementation of growth, whereas expression of an active site CRK3^T178E mutant did not, showing that protein kinase activity is crucial for CRK3 function. Inducible deletion of CRK3 in stationary phase promastigotes resulted in attenuated growth in mice, thereby confirming CRK3 as a useful therapeutic target and diCre as a valuable new tool for analyzing essential genes in Leishmania.     

Rapid thermal responses and thermal tolerance in adult codling moth Cydia pomonella (Lepidoptera: Tortricidae)

In order to preserve key activities or improve survival, insects facing variable and unfavourable thermal environments may employ physiological adjustments on a daily basis. Here, we investigate the survival of laboratory-reared adult Cydia pomonella at high or low temperatures and their responses to pre-treatments at sub-lethal temperatures over short time-scales. We also determined critical thermal limits (CTLs) of activity of C. pomonella and the effect of different rates of cooling or heating on CTLs to complement the survival assays. Temperature and duration of exposure significantly affected adult C. pomonella survival with more extreme temperatures and/or longer durations proving to be more lethal. Lethal temperatures, explored between −20 °C to −5 °C and 32 °C to 47 °C over 0.5, 1, 2, 3 and 4 h exposures, for 50% of the population of adult C. pomonella were −12 °C for 2 h and 44 °C for 2 h. Investigation of rapid thermal responses (i.e. hardening) found limited low temperature responses but more pronounced high temperature responses. For example, C. pomonella pre-treated for 2 h at 5 °C improved survival at −9 °C for 2 h from 50% to 90% (p < 0.001). At high temperatures, pre-treatment at 37 °C for 1 h markedly improved survival at 43 °C for 2 h from 20% to 90% (p < 0.0001). We also examined cross-tolerance of thermal stressors. Here, low temperature pre-treatments did not improve high temperature survival, while high temperature pre-treatment (37 °C for 1 h) significantly improved low temperature survival (−9 °C for 2 h). Inducible cross-tolerance implicates a heat shock protein response. Critical thermal minima (CTmin) were not significantly affected by cooling at rates of 0.06, 0.12 and 0.25 °C min⁻¹ (CTmin range: 0.3-1.3 °C). By contrast, critical thermal maxima (CTmax) were significantly affected by heating at these rates and ranged from 42.5 to 44.9 °C. In sum, these results suggest pronounced plasticity of acute high temperature tolerance in adult C. pomonella, but limited acute low temperature responses. We discuss these results in the context of local agroecosystem microclimate recordings. These responses are significant to pest control programmes presently underway and have implications for understanding the evolution of thermal tolerance in these and other insects.     

An interaction switch predicts the nested architecture of mutualistic networks

Nested architecture is distinctive in plant-animal mutualistic networks. However, to date an integrative and quantitative explanation has been lacking. It is evident that species often switch their interactive partners in real-world mutualistic networks such as pollination and seed-dispersal networks. By incorporating an interaction switch into a novel multi-population model, we show that the nested architecture rapidly emerges from an initially random network. The model allowing interaction switches between partner species produced predictions which fit remarkably well with observations from 81 empirical networks. Thus, the nested architecture in mutualistic networks could be an intrinsic physical structure of dynamic networks and the interaction switch is likely a key ecological process that results in nestedness of real-world networks. Identifying the biological processes responsible for network structures is thus crucial for understanding the architecture of ecological networks.     

Basal cold but not heat tolerance constrains plasticity among Drosophila species (Diptera: Drosophilidae)

Thermal tolerance and its plasticity are important for understanding ectotherm responses to climate change. However, it is unclear whether plasticity is traded‐off at the expense of basal thermal tolerance and whether plasticity is subject to phylogenetic constraints. Here, we investigated associations between basal thermal tolerance and acute plasticity thereof in laboratory‐reared adult males of eighteen Drosophila species at low and high temperatures. We determined the high and low temperatures where 90% of flies are killed (ULT₉₀ and LLT₉₀, respectively) and also the magnitude of plasticity of acute thermal pretreatments (i.e. rapid cold‐ and heat‐hardening) using a standardized, species‐specific approach for the induction of hardening responses. Regression analyses of survival variation were conducted in ordinary and phylogenetically informed approaches. Low‐temperature pretreatments significantly improved LLT₉₀ in all species tested except for D. pseudoobscura, D. mojavensis and D. borealis. High‐temperature pretreatment only significantly increased ULT₉₀ in D. melanogaster, D. simulans, D. pseudoobscura and D. persimilis. LLT₉₀ was negatively correlated with low‐temperature plasticity even after phylogeny was accounted for. No correlations were found between ULT₉₀ and LLT₉₀ or between ULT₉₀ and rapid heat‐hardening (RHH) in ordinary regression approaches. However, after phylogenetic adjustment, there was a positive correlation between ULT₉₀ and RHH. These results suggest a trade‐off between basal low‐temperature tolerance and acute low‐temperature plasticity, but at high temperatures, increased basal tolerance was accompanied by increased plasticity. Furthermore, high‐ and low‐temperature tolerances and their plasticity are clearly decoupled. These results are of broad significance to understanding how organisms respond to changes in habitat temperature and the degree to which they can adjust thermal sensitivity.     

False codling moth Thaumatotibia leucotreta (Lepidoptera,Tortricidae) larvae are chill‐susceptible

Abstract This study reports on the low temperature tolerance and cold hardiness of larvae of false codling moth, Thaumatotibia leucotreta. We found that larvae have mean critical thermal minima (lower limits of activity) of 6.7°C which was influenced by feeding status. The effects of low temperature exposure and duration of exposure on larval survival were assessed and showed that the temperature at which 50% of the population survives is ?11.5 ± 0.3°C after 2 h exposure. The supercooling point (SCP, i.e., freezing temperature) was investigated using a range of cooling rates and under different conditions (feeding and hydration status) and using inoculative freezing treatments (in contact with water or orange juice). The SCP decreased significantly from ?15.6°C to ?17.4°C after larvae were fasted for 24 h. Twenty‐four hour treatments at either high or low relative humidity (95.9% or 2.4%) also significantly decreased SCP to ?17.2°C and ?18.2°C respectively. Inoculative freezing (by water contact) raised SCP from ?15.6°C to ?6.8°C which could have important implications for post‐harvest sterilization. Cooling rates did not affect SCP which suggests that there is limited phenotypic plasticity of SCP during the larval life‐stage, at least over the short time‐scales investigated here. In conclusion, larvae of T. leucotreta are chill‐susceptible and die upon freezing. These results are important in understanding this pest's response to temperature variation, understanding pest risk status and improving post‐harvest sterilization efficacy.     

Methods and approaches for the management of arthropod border incursions

Biological invasions are increasing and are strongly associated with negative agricultural, economic and ecological impacts. It is increasingly recognized that the primary focus in minimizing biological invasions should be to prevent initial entry of alien species. However, exclusion of terrestrial arthropods such as insects and mites is difficult, in part because of their relatively small size, cryptic habits, broad physiological tolerances and close association with various internationally traded goods. Here we discuss methods, approaches, management and intervention systems used by border biosecurity agencies to prevent entry of inadvertently transported arthropods. We examine the at-border systems that exist for the detection and identification of and response to alien arthropods, and discuss the constraints and challenges present in these systems. We critically review current border biosecurity systems and discuss their relative efficacy. We then discuss additional measures and key areas that could be addressed that may further improve these systems. These include: (1) the application of appropriate sampling strategies; (2) employment of suitable inspection methods adequate to detect small and hidden arthropods; and (3) thorough recording of methods, organisms detected and both negative and positive results of inspections. We emphasize that more research is needed on taxonomy, biology, genetics, distribution, host and disease associations, impacts and pathways of introductions for invasive arthropods. Of critical importance is the compilation of complete and accurate invasive species lists and high-risk species watch-lists. The adoption of these recommendations will contribute to improved biosecurity systems for the exclusion of alien, invasive and pest arthropods.     

THE EFFECTS OF EXHAUSTIVE EXERCISE ON THE ACTIVITY LEVELS OF CATALASE IN VARIOUS TISSUES OF MALE AND FEMALE RATS

The effects of exhaustive exercise on the activity levels of catalase (EC 1.11.1.6) in various tissues of male and female Sprague—Dawley rats (Rattus norvegicus) were investigated. Both the male and female rats were subdivided into an experimental group and a control group consisting of eight rats each. One group of each sex was subjected to a swimming session of 1h (experimental group) while the other group of each sex served as sedentary control groups. The tissues investigated were liver, heart, kidney and lung. The activity levels of catalase in all the tissues investigated were significantly (P< 0.05) elevated in both male and female rats as a result of exercise. The average increase in the activity levels of catalase in the various tissues investigated for both male and female rats was 417% (males 404%; females 430%). The male and female rats exhibited comparable activity levels of catalase in all the tissues investigated. The higher activity levels of catalase as a result of exercise might be indicative of a compensatory measure to counteract the possible detrimental effects associated with oxidative stress.     

Critical thermal limits and their responses to acclimation in two sub-Antarctic spiders: <Emphasis Type="Italic">Myro kerguelenensis</Emphasis> and <Emphasis Type="Italic">Prinerigone vagans</Emphasis>

Despite the relative richness of spider species across the Southern Ocean islands remarkably little information is available on their biology. Here, the critical thermal limits of an indigenous (Myro kerguelenensis, Desidae) and an introduced (Prinerigone vagans, Linyphiidae) spider species from Marion Island were studied after 7–8 days acclimation to 0, 5, 10 and 15°C. Critical thermal minima (CT_Min) were low in these species by comparison with other spiders and insects measured to date, and ranged from −6 to −7°C in M. kerguelenensis and from −7 to −8°C in P. vagans. In contrast, critical thermal maxima (CT_Max) were similar to other insects on Marion Island (M. kerguelenensis: 35.0–35.6°C; P. vagans: 35.1–36.0°C), although significantly lower than those reported for other spider species in the literature. The magnitude of acclimation responses in CT_Max was lower than those in CT_Min for both species and this suggests decoupled responses to acclimation. Whilst not conclusive, the results raise several important considerations: that oxygen limitation of thermal tolerance needs to be more widely investigated in terrestrial species, that indigenous and alien species might differ in the nature and extent of their plasticity, and that upper and lower thermal tolerance limits might be decoupled in spiders as is the case in insects.     

Diurnal variation in supercooling points of three species of Collembola from Cape Hallett, Antarctica

Daily changes in microclimate temperature and supercooling point (SCP) of Collembola were measured during summer at Cape Hallett, North Victoria Land, Antarctica. Isotoma klovstadi and Cryptopygus cisantarcticus (Isotomidae) showed bimodal SCP distributions, predominantly in the high group during the day and in the low group during the night. There were no concurrent diurnal changes in water content or haemolymph osmolality. By contrast, Friesea grisea (Neanuridae) had a unimodal distribution of SCPs that was invariant between daytime and nighttime. Isotoma klovstadi collected foraging on moss had uniformly high SCPs, which shifted towards the low group when the animals were starved for 2-8 h. When I. klovstadi was acclimated for five days with lichen or algae, SCPs were higher than if they were supplied with moss, while those that were starved (with free water or 100% relative humidity) displayed a trimodal SCP distribution. A variety of pre-treatments, including cold, heat, desiccation and slow cooling were ineffective at inducing SCP shifts in C. cisantarcticus or I. klovstadi. It is postulated that behavioural avoidance of low temperatures by vertical migration may be key in I. klovstadi's short-term survival of nighttime temperatures. These data suggest that the full range of thermal responses of Antarctic Collembola is yet to be elucidated.     

Plant fructose-1,6-bisphosphatases: characteristics and properties.

In this minireview the properties and characteristics of plant fructose-1,6-bisphosphatases (D-fructose-1,6-bisphosphatase 1-phosphohydrolase, EC 3.1.3.11) are discussed. The properties and characteristics of the chloroplastic and cytoplasmic forms of the enzyme are reviewed. For purposes of comparison some reference is made to fructose-1,6-bisphosphatases from other species.