Influence of the experimental setup on the determination of enzyme kinetic parameters

For the design of bioconversion processes parallel experimentation in microtiter plates is commonly applied to reduce the experimental load, although data accuracy and reproducibility are often reduced. In an effort to quantify the impact of different microscale experimental systems on the estimation of enzyme kinetic parameters from progress curves, we comprehensively evaluated the enzymatic reduction of acetophenone in both open and closed polystyrene and quartz microtiter plates as well as quartz cuvettes. Differences in conversion of up to 50% over time were observed increasing from polystyrene MTPs to quartz MTPs to quartz cuvettes. Initial reaction velocities increased systematically from polystyrene to quartz MTPs and cuvettes. The experimental errors decreased in the same order showing highest experimental error of about 20% in polystyrene. We further evaluated reasons causing the deviations within one system as well as between the systems. The choice of reaction vessel material, temperature effects and substrate cross contaminations in MTPs were shown to be of importance in the experimental results. Although the experimental data differed between the reaction vessels, no distinct trends in estimated kinetic parameters were found. While the microkinetic parameters vary up to an order of magnitude between different systems, the corresponding macrokinetic parameters lie in the same range for all systems varying by 29–118%. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:87–95, 2017     

Alternative Confidence Regions for Bonferroni‐Based Closed‐Testing Procedures that are not Alpha‐Exhaustive

This article complements the results in Guilbaud (Biometrical Journal 2008; 50 :678–692). Simultaneous confidence regions were derived in that article that correspond to any given multiple testing procedure (MTP) in a fairly large class of consonant closed‐testing procedures based on marginal p‐values and weighted Bonferroni tests for intersection hypotheses. This class includes Holm's MTP, the fixed‐sequence MTP, gatekeeping MTPs, fallback MTPs, multi‐stage fallback MTPs, and recently proposed MTPs specified through a graphical representation and associated rejection algorithm. More general confidence regions are proposed in this article. These regions are such that for certain underlying MTPs which are not alpha‐exhaustive, they lead to confidence assertions that may be sharper than rejection assertions for some rejected null hypotheses H when not all Hs are rejected, which is not the case with the previously proposed regions. In fact, various alternative confidence regions may be available for such an underlying MTP. These results are shown through an extension of the previous direct arguments (without invoking the partitioning principle), and under the same general setup; so for instance, estimated quantities and marginal confidence regions are not restricted to be of any particular kinds/dimensions. The relation with corresponding confidence regions of Strassburger and Bretz (Statistics in Medicine 2008; 27 :4914–4927) is described. The results are illustrated with fallback and parallel‐gatekeeping MTPs.     

Simultaneous confidence regions for closed tests,including Holm‐, Hochberg‐, and Hommel‐related procedures

The derivation of simultaneous confidence regions for some multiple‐testing procedures (MTPs) of practical interest has remained an unsolved problem. This is the case, for example, for Hochberg's step‐up MTP and Hommel's more powerful MTP that is neither a step‐up nor a step‐down procedure. It is shown in this article how the direct approach used previously by the author to construct confidence regions for certain closed‐testing procedures (CTPs) can be extended to a rather general setup. The general results are then applied to a situation with one‐sided inferences and CTPs belonging to a class studied by Wei Liu. This class consists of CTPs based on ordered marginal p‐values. It includes Holm's, Hochberg's, and Hommel's MTPs. A property of the confidence regions derived for these three MTPs is that no confidence assertions sharper than rejection assertions can be made unless all null hypotheses are rejected. Briefly, this is related to the fact that these MTPs are quite powerful. The class of CTPs considered includes, however, also MTPs related to Holm's, Hochberg's, and Hommel's MTPs that are less powerful but are such that confidence assertions sharper than rejection assertions are possible even if not all null hypotheses are rejected. One may thus choose and prespecify such an MTP, though this is at the cost of less rejection power.     

The yeast mitochondrial transport proteins: new sequences and consensus residues, lack of direct relation between consensus residues and transmembrane helices, expression patterns of the transport protein genes, and protein-protein interactions with other proteins

Mitochondrial transport proteins (MTP) typically are homodimeric with a 30-kDa subunit with six transmembrane helices. The subunit possesses a sequence motif highly similar to Pro X Asp/Glu X X Lys/Arg X Arg within each of its three similar 10-kDa segments. Four (YNL083W, YFR045W, YPR021C, YDR470C) of the 35 yeast (S. cerevisiae) MTP genes were resequenced since the masses of their proteins deviate significantly from the typical 30 kDa. We now find these four proteins to have 545, 285, 902, and 502 residues, respectively. Together with only four other MTPs, the sequences of YPR021C and YDR470C show substitutions of some of the five residues that are absolutely conserved among the 12 MTPs with identified transport function and 17 other MTPs. We do now find these five consensus residues also in the new sequences of YNL083W and YFR045W. Additional analyses of the 35 yeast MTPs show that the location of transmembrane helix sequences do not correlate with the general consensus residues of the MTP family; protein segments connecting the six transmembrane helices and facing the intermembrane space are not uniformly short (about 20 residues) or long (about 40 residues) when facing the matrix; most MTPs have at least one transmembrane helix for which the sum of the negative hydropathy values of all residues yields a very small negative value, suggesting a membrane location bordering polar faces of other transmembrane helices or a non-transmembrane location. The extra residues of the three large MTPs are hydrophilic and at the N-terminal. The 200-residue N-terminal segment of YNL083W has four putative Ca2+-binding sites. The 500-residue N-terminal segment of YPR021C shows sequence similarity to enzymes of nucleic acid metabolism. cDNA microarray data show that YNL083W is expressed solely during sporulation, while the expressions of YFR045W, YPR021C, and YDR470C are induced by various stress situations. These results also show that the 35 MTP genes are expressed under a rather diverse set of metabolic conditions that may help identify the function of the proteins. Interestingly, yeast two-hybrid screens, that will also be useful in identifying the function of MTPs, indicate that MIR1, AAC3, YOR100C, and YPR011C do interact with non-MTPs.     

Thermally induced plasticity of body shape in adult zebrafish Danio rerio (Hamilton, 1822)

We examined the effect of temperature during the early development on the phenotypic plasticity of Danio rerio. The effect of temperature was examined during two different early developmental periods of 280°d (the product of days × temperature) each, 28‐308°d or 280‐560°d, by subjecting the experimental populations to three different water temperatures (22°C, 28°C, and 32°C). Before and after the end of the 280°d period of the different thermal exposure, all populations were cultured in standard temperature (28°C). Five to 10 months after exposure to the different thermal regimes, the body shape of the adults was analyzed by geometric morphometrics. In both ontogenetic windows and experimental repetitions, the results showed that developmental temperature and sex significantly affected the body shape of adult zebrafish. Thermally induced shape variation discriminated the fish that developed at 22°C from those developed at 28°C–32°C. In the early developmental period (DP1, 28–308°d postfertilization), dorsal, anal, and caudal fin structures differed between the animals that developed at 22°C and 28°C–32°C. In the later developmental period (DP2, 280–560°d postfertilization), caudal, anal, pectoral, and pelvic fins, as well as the gill cover and lower jaw, were affected when animals developed at different temperatures. These results show that thermal history during a short period of embryonic and larval life affects the body form of adult zebrafish with potentially functional consequences. Based on previous data on the effects of temperature on fish development, we suggest thermally induced muscle and bone remodelling as possible mechanism underlying the observed plasticity. J. Morphol., 2010. © 2010 Wiley‐Liss, Inc.     

Effects of temperature and soil moisture on survival of eggs of the mosquito Psorophora columbiae (Diptera: Culicidae)

The results of laboratory tests indicated the average survival rates for Psorophora columbiae eggs remained quite high for all of the egg populations exposed to a temperature of 27°C (range 83.0–100.0% survival) after 96 days of exposure, except for the non‐diapausing eggs on dry soil (66.3%). In regard to the exposure of egg populations to moderately cold temperatures (i.e. 8°C, 4°C and ?2°C) for periods of up to 16 days, survival rates for egg populations exposed to 8°C continued to remain relatively high (average >85%) for the remainder of the experimental exposure period (i.e. 96 days). Diapausing Ps. columbiae eggs were more tolerant (82.0% survival) to low temperatures (?2°C) than non‐diapausing eggs (2.4% survival) for 64 days, particularly at temperatures of and below 4°C. Diapausing and non‐diapausing eggs were similar in their ability to survive under high temperatures (34°C and 38°C). High soil moisture (30–40%) or substrate moisture (95% relative humidity) content appeared to enhance the ability of the mosquito eggs to survive both low and high temperature extremes.     

Not so robust: Robusta coffee production is highly sensitive to temperature

Coffea canephora (robusta coffee) is the most heat‐tolerant and ‘robust’ coffee species and therefore considered more resistant to climate change than other types of coffee production. However, the optimum production range of robusta has never been quantified, with current estimates of its optimal mean annual temperature range (22–30°C) based solely on the climatic conditions of its native range in the Congo basin, Central Africa. Using 10 years of yield observations from 798 farms across South East Asia coupled with high‐resolution precipitation and temperature data, we used hierarchical Bayesian modeling to quantify robusta's optimal temperature range for production. Our climate‐based models explained yield variation well across the study area with a cross‐validated mean R² = .51. We demonstrate that robusta has an optimal temperature below 20.5°C (or a mean minimum/maximum of ≤16.2/24.1°C), which is markedly lower, by 1.5–9°C than current estimates. In the middle of robusta's currently assumed optimal range (mean annual temperatures over 25.1°C), coffee yields are 50% lower compared to the optimal mean of ≤20.5°C found here. During the growing season, every 1°C increase in mean minimum/maximum temperatures above 16.2/24.1°C corresponded to yield declines of ~14% or 350–460 kg/ha (95% credible interval). Our results suggest that robusta coffee is far more sensitive to temperature than previously thought. Current assessments, based on robusta having an optimal temperature range over 22°C, are likely overestimating its suitable production range and its ability to contribute to coffee production as temperatures increase under climate change. Robusta supplies 40% of the world's coffee, but its production potential could decline considerably as temperatures increase under climate change, jeopardizing a multi‐billion dollar coffee industry and the livelihoods of millions of farmers.     

Effects of temperature and pollination site on pollen performance in Betula pendula Roth – evidence for genotype-environment interactions

We studied whether the differences between genetically different pollen donors (Betula pendula Roth clones) with respect to pollen-tube growth rate were consistent under different thermal conditions during pollen germination in vivo and in vitro. We conducted a single-donor hand-pollination experiment with same pollen donors and recipients in a plastic house seed orchard and at an outdoor clone collection. The prevailing daily mean temperature during pollen germination was 13°C higher in the plastic house than outdoors. The pollen-tube growth rate of each pollen donor was additionally determined in vitro on agar medium at five temperatures (10°, 15°, 22°, 30° and 35°C). A significant interaction between paternal clone and pollination site as well as between paternal clone and temperature was found, which provides evidence for genotype-environment interactions. Genotype-environment interactions can have evolutionary significance in maintaining the variation in pollen-tube growth rates. At seed orchards, genotype-environment interactions can cause deviations from the expected genetic composition of the seed crop depending on the prevailing environmental conditions during pollen-tube growth. Received: 24 September 1999 / Accepted: 29 September 1999     

Effects of photoperiod and temperature on the cholinesterase activity in the ganglia of Schistocerca gregaria

The presence of acetyl cholinesterases, cholinesterases, and non-specific esterases in four different ganglia of Schistocerca gregaria was demonstrated using various substrates, as well as by means of inhibition with physostigmine salicylate and BW 284 C51. The contribution of these enzymes to the total esterase activity in each ganglion was also determined quantitatively.In animals kept under a L:D = 12:12 regime with concurrent changes of temperature (about 30°C during the light period and 20°C during the dark period), the activity of the AChE displays a maximum at about the time of light change or 1 to 2 hr later. The activity peak lasts for about 2 hr. If average enzyme activities are calculated, no statistically significant differences between ‘day’ and ‘night’ animals are apparent.An elevation of the environmental temperature from 20°C to 30°C during the light period led to a significant rise in enzyme activity in all ganglia after 1 to 3 hr. Lowering the environmental temperature from 30°C to 20°C during the light period led to a significant decrease in enzyme activity only in the suboesophageal ganglion after 2 hr. Artificial stimulation of the animals in a wind tunnel decreases the acetyl cholinesterase activity in the suboesophageal ganglion, whereas in the other ganglia no significant change in enzyme activity was observed. Further, no correlation was found between the enzyme activity and O₂ consumption of the experimental animals.     

Biogeographic variation in temperature sensitivity of decomposition in forest soils

Determining soil carbon (C) responses to rising temperature is critical for projections of the feedbacks between terrestrial ecosystems, C cycle, and climate change. However, the direction and magnitude of this feedback remain highly uncertain due largely to our limited understanding of the spatial heterogeneity of soil C decomposition and its temperature sensitivity. Here we quantified C decomposition and its response to temperature change with an incubation study of soils from 203 sites across tropical to boreal forests in China spanning a wide range of latitudes (18°16′ to 51°37′N) and longitudes (81°01′ to 129°28′E). Mean annual temperature (MAT) and mean annual precipitation primarily explained the biogeographic variation in the decomposition rate and temperature sensitivity of soils: soil C decomposition rate decreased from warm and wet forests to cold and dry forests, while Q_10‐MAT (standardized to the MAT of each site) values displayed the opposite pattern. In contrast, biological factors (i.e. plant productivity and soil bacterial diversity) and soil factors (e.g. clay, pH, and C availability of microbial biomass C and dissolved organic C) played relatively small roles in the biogeographic patterns. Moreover, no significant relationship was found between Q_10‐MAT and soil C quality, challenging the current C quality–temperature hypothesis. Using a single, fixed Q_10‐MAT value (the mean across all forests), as is usually done in model predictions, would bias the estimated soil CO₂ emissions at a temperature increase of 3.0°C. This would lead to overestimation of emissions in warm biomes, underestimation in cold biomes, and likely significant overestimation of overall C release from soil to the atmosphere. Our results highlight that climate‐related biogeographic variation in soil C responses to temperature needs to be included in next‐generation C cycle models to improve predictions of C‐climate feedbacks.     

Temperature effects on egg development of the rosy apple aphid and forecasting of egg hatch

The development of Dysaphis plantaginea (Pass.) (Homoptera: Aphididae) winter eggs was studied at six different constant temperatures ranging from 7.5 to 16.5 °C in order to improve the basis for phenological forecasts in early spring. The mortality was generally low at temperatures below 13.5 °C but increased considerably at 16.5 °C. The effect of temperature on development rates could be described with linear regression within the temperature range under study. The lower temperature threshold for development was estimated to be 4.0 °C and the thermal constant 140 day‐degrees. A time‐varying distributed delay approach was used to establish a temperature driven phenology model for winter egg hatch of D. plantaginea considering the intrinsic variability in development time. The model parameters such as temperature‐dependent development times and corresponding variances were quantified based on the experimental data. When compared with independent observations on egg hatch under semifield conditions, the model gave satisfactory validation results. It can be used as forecasting tool for the optimal timing of monitoring and control measures for D. plantaginea in early spring.     

Effects of rearing and induction temperature on the temporal dynamics of heat shock protein 70 expression in a butterfly

The temporal dynamics of heat shock protein 70 (HSP70) expression in response to longer‐term acclimation and rapid hardening in the butterfly Lycaena tityrus is investigated. After a 1‐h exposure to 1 °C or 37 °C, HSP70 is quickly up‐regulated within 1 h and down‐regulated within 2 h. The fast dynamic of HSP70 expression is in contrast to the patterns found in organisms inhabiting more stable thermal environments, and is interpreted as an adaptation to the large and rapid temperature variation experienced by flying ectotherms. HSP70 expression is higher in males than in females, as well as in animals reared at 27 °C than at 20 °C, although it is very similar across the high and low induction temperatures. Animals reared at the higher temperature, however, respond less strongly to high‐temperature stress.     

Enzymatic properties of phenoloxidase from Pieris rapae (Lepidoptera) larvae

The kinetic parameters of partially purified phenoloxidase (PO, EC. 1.14.18.1) from the 5th instar larvae of Pieris rapae (Lepidoptera) were determined, using L‐3, 4‐dihydroxyphenylalanine (L‐DOPA) as substrate. The optimal pH and temperature of the enzyme for the oxidation of L‐DOPA were determined to be at pH 7.0 and at 42°C, respectively. The enzyme was stable between pH 6.5 and 7.4 and at temperatures lower than 37°C. At pH 6.8 and 37°C, the Michaelis constant (K_m) and maximal velocity (V_m) of the enzyme for the oxidation of L‐DOPA were determined to be 0.80 μmol/L and 1.84 μmol/ L/min, respectively. Tetra‐hexylresorcinol and 4‐dodecylresorcinol effectively inhibited activity of phenoloxidase and this inhibition was reversible and competitive, with the IC₅₀ of 1.50 and 1.12 μmol/L, respectively. The inhibition constants were estimated to be 0.50 and 0.47 μmol/L, respectively.     

Climate change and the effect on the biomass growth in steppe communities in Kherlon River Basin, Northern China

The bacterial community composition and diversity in rock varnish of Turpan Basin were investigated by restriction fragment length polymorphism (RFLP) and clone library of the 16S rRNA gene. 114 positive clones were screened, which could be grouped into 28 phylotypes and then further divided into 23 different operational taxonomic units (OTUs). These were affiliated into 5 phyla (Acidobacteria, Proteobacteria, Chloroflexi, Firmicutes and Cyanobacteria). Clones from actinobacteria were the dominant, accounting for 67.5% of total clones in the library, followed by Proteobacteria (15.8%), Chloroflexi (13.2%), Firmicutes (2.6%) and Cyanobacteria (0.9%). Rubrobacter (accounts for 35%) in the phylum Actinobacteria was the dominant genus and contained many species which might be resistant to gamma radiation. A 70% of the library clone sequences showed less 97% similarity to 16S rRNA gene sequences of standard strains obtained by pure culture. Shannon–Wiener index value of this study is 2.52 and is lower than deep-sea sediments, soils, lakes and other environments. Results of this study showed that bacterial diversity in rock varnishes of Turpan Basin was low, but maybe exist a large number of new unknown taxons, especially species that could well adapted to drought and resist radiation.     

The effect of storage on whole blood chemiluminescence measurement of equine neutrophils

The aim of this study was to determine the effect of duration and temperature of sample storage on whole blood chemiluminescence measurement results. Venous blood from 18 clinically healthy Polish half‐bred horses aged 4 to 11 years were used in the study. Luminol dependent chemiluminescence (CL) was used to measure neutrophil oxygen metabolism in whole blood. Blood samples were examined for spontaneous CL and stimulated by a surface receptor stimulus as well as extra‐receptor stimulus. The assay was performed in two parallel experimental sets with samples stored at 4 and 22 °C, respectively. Whole blood CL was estimated at 2, 6, 24, 48, 72, 96 and 120 h after collection. The study demonstrated that temperature and duration of sample storage are factors that determine the quality of CL measurements of whole blood in horses. The study concluded that samples should be stored at 4 °C and the assay should be performed as early as possible. It was also shown that the viability period of horse blood for CL assays is relatively long. Material stored at room temperature for 24 h and even up to 48 h at 4 °C did not show any significant decrease in spontaneous or stimulated chemiluminescence. Copyright © 2012 John Wiley & Sons, Ltd.     

Protocols for dry DNA storage and shipment at room temperature

The globalization of DNA barcoding will require core analytical facilities to develop cost‐effective, efficient protocols for the shipment and archival storage of DNA extracts and PCR products. We evaluated three dry‐state DNA stabilization systems: commercial Biomatrica^® DNAstable^® plates, home‐made trehalose and polyvinyl alcohol (PVA) plates on 96‐well panels of insect DNA stored at 56 °C and at room temperature. Controls included unprotected samples that were stored dry at room temperature and at 56 °C, and diluted samples held at 4 °C and at ?20 °C. PCR and selective sequencing were performed over a 4‐year interval to test the condition of DNA extracts. Biomatrica^® provided better protection of DNA at 56 °C and at room temperature than trehalose and PVA, especially for diluted samples. PVA was the second best protectant after Biomatrica^® at room temperature, whereas trehalose was the second best protectant at 56 °C. In spite of lower PCR success, the DNA stored at ?20 °C yielded longer sequence reads and stronger signal, indicating that temperature is a crucial factor for DNA quality which has to be considered especially for long‐term storage. Although it is premature to advocate a transition to DNA storage at room temperature, dry storage provides an additional layer of security for frozen samples, protecting them from degradation in the event of freezer failure. All three forms of DNA preservation enable shipment of dry DNA and PCR products between barcoding facilities.     

Plasticity of photosynthetic heat tolerance in plants adapted to thermally contrasting biomes

In many biomes, plants are subject to heatwaves, potentially causing irreversible damage to the photosynthetic apparatus. Field surveys have documented global, temperature‐dependent patterns in photosynthetic heat tolerance (P _HT); however, it remains unclear if these patterns reflect acclimation in P _HT or inherent differences among species adapted to contrasting habitats. To address these unknowns, we quantified seasonal variations in T _crit (high temperature where minimal chlorophyll‐a fluorescence rises rapidly, reflecting disruption to photosystem II) in 62 species native to 6 sites from 5 thermally contrasting biomes across Australia. T _crit and leaf fatty acid (FA) composition (important for membrane stability) were quantified in three temperature‐controlled glasshouses in 20 of those species. T _crit was greatest at hot field sites and acclimated seasonally (summer > winter, increasing on average 0.34 °C per °C increase in growth temperature). The glasshouse study showed that T _crit was inherently higher in species from warmer habitats (increasing 0.16 °C per °C increase in origin annual mean maximum temperature) and acclimated to increasing growth temperature (0.24 °C °C^?1). Variations in T _crit were positively correlated with the relative abundance of saturated FAs, with FAs accounting for 40% of T _crit variation. These results highlight the importance of both plastic adjustments and inherent differences determining contemporary continent‐wide patterns in P _HT.     

Microtransponders, the miniature RFID electronic chips, as platforms for cell growth in cytotoxicity assays.

BACKGROUND: An electronic radio frequency (RF) microchip, the microtransponder (MTP), has been developed as a platform for assays in the fields of genomics and proteomics. Upon activation by light, each MTP provides a unique RF identification (ID) signal that matches a chip to the specific biological material attached to it. The MTP is powered by a photocell and has an antenna that transmits the signal. The aim of the present study was to explore utility of MTPs as a platform for cell growth in cytotoxicity assays. METHODS: The MCF-7, MCF-116, A549, or T-24 cells growing on MTPs placed in petri dishes or slide chambers were cultured untreated or exposed to antitumor drugs topotecan, mitoxantrone, or onconase for up to 4 days. Their attachment to- and growth on- MTPs was assessed by fluorescence microscopy and laser scanning cytometry (LSC) and compared with growth on the dish surface in the MTP neighborhood. The MTPs were fixed in ethanol, stained with propidium iodide (PI), and interrogated in flow in the instrument capable to rapidly (up to 103 MTPs/s) identify their ID signal and measure fluorescence. RESULTS: The cells plated on MTPs exhibited similar attachment properties to those plated in culture dishes. When measured by LSC, they had similar mitotic activity, growth rate, and cell cycle distributions as the cells adhering to the culture dish in the neighborhood of MTPs. The fluorescence intensity of MTPs provided information about the cell number per MTP, which made it possible to assess cell growth rate and monitor the cytostatic/cytotoxic effects of the tested drugs. CONCLUSIONS: The MTP-based system holds promise for the multiplexed cell assays in which numerous different cell lines can be screened for their growth rate or sensitivity while exposed to particular agents in the same vessel. Other advantages of the system are the rapidity of the screening and a very large number of ID codes. Because many cell lines/types can be assayed in a single dish, the system also offers cost savings on tissue culture reagents.     

The influence of thermal history on upper thermal limits of two species of riverine insects: the stonefly, <Emphasis Type="Italic">Aphanicerca capensis</Emphasis>, and the mayfly, <Emphasis Type="Italic">Lestagella penicillata</Emphasis>

The upper thermal limits of two cold-water stenotherms: the mayfly, Lestagella penicillata (Teloganodidae), and the stonefly, Aphanicerca capensis (Notonemouridae), were determined from six rivers in the Western Cape, South Africa. Limits were estimated using the Critical Thermal Method (expressed as Critical Thermal maximum) and the Incipient Lethal Temperature method (expressed as Incipient Lethal Upper Limit). Hourly water temperatures recorded in these rivers were used to characterise thermal signatures. Median CT_max and 96 h ILUT varied significantly amongst rivers for both species (≤5.7°C for CT_max and ≤4.0°C for 96 h ILUT) and variation was similar for both species. Differences in water temperature amongst rivers during the experimental period (spring) were insufficient (<2.0°C) to confirm the relationship between upper thermal limits and thermal history, expressed as an averaging statistic derived from in situ water temperatures. Greatest thermal range was over the warm summer period (>8.0°C) and it is likely that this is when thermal history may influence thermal limits. Maximum Weekly Allowable Temperature thresholds averaged for all rivers were lower for A. capensis (17.0°C) compared to L. penicillata (19.0°C). Both species have life cycles that allow them to avoid the thermally stressful summer period.     

Cold tolerance in obligate and cyclical parthenogens of the peach-potato aphid, Myzus persicae

Abstract. 1. Many aphids form mixed populations of cyclical and obligate parthenogens. This is puzzling, because all else being equal, obligate parthenogens should outcompete cyclical parthenogens due to the two‐fold cost of sex. Yet cyclical parthenogens produce frost‐resistant, diapausing eggs in autumn, while obligate parthenogens spend the winter as active stages. Frost resistance thus represents a short‐term advantage to sexual reproduction mediated by winter temperatures, which may promote this coexistence. 2. Because obligate parthenogens overwinter as active stages, there may be selection for increased cold tolerance compared to cyclical parthenogens. This has the potential to gradually erode the advantage of sexually producing eggs. 3. Four obligately and four cyclically parthenogenetic lines of Myzus persicae (Sulzer) (Hemiptera: Aphididae) were collected from each of two areas differing in winter severity, and their survival after exposure to a severe experimental frost (14 h at ?9 °C), as well as their reproductive performance at a low (10 °C) and a high (20 °C) temperature were compared. 4. There was significant variation among lines in survival after the experimental frost, but this variation was neither related to their reproductive mode, nor to their area of origin. Similarly, neither reproductive mode nor origin had a significant effect on reproductive performance, independent of temperature. The average slope of the response to variation in temperature was also similar for both reproductive modes, despite the fact that slopes differed significantly among lines. 5. Within the limits of extrapolating from laboratory experiments, it is concluded that in M. persicae, the active stages of obligate parthenogens are not better adapted to cold temperatures than those of cyclical parthenogens.