Median Lethal Temperature Research Articles

Guijing2501 (Citrus unshiu) Has Stronger Cold Tolerance Due to Higher Photoprotective Capacity as Revealed by Comparative Transcriptomic and Physiological Analysis and Overexpression of Early Light-Induced Protein

Cold is one of the major limiting factors for citrus production, particularly extreme cold waves. Therefore, it is of great importance to develop cold-tolerant varieties and clarify their cold tolerance mechanisms in citrus breeding. In this study, comparative transcriptomic and physiological analyses were performed to dissect the cold tolerance mechanism of Guijing2501 (GJ2501), a new satsuma mandarin (Citrus unshiu) variety with about 1 °C lower LT50 (the median lethal temperature) relative to Guijing (GJ). The physiological analysis results revealed that GJ2501 is more cold-tolerant with less photoinhibition, PSII photodamage, and MDA accumulation, but higher POD activity than GJ under cold stress. Comparative transcriptomic analysis identified 4200 DEGs between GJ and GJ2501, as well as 4884 and 5580 up-regulated DEGs, and 5288 and 5862 down-regulated DEGs in response to cold stress in GJ and GJ2501, respectively. “Photosynthesis, light harvesting” and “photosystem” were the specific and most significantly enriched GO terms in GJ2501 in response to cold stress. Two CuELIP1 genes (encoding early light-induced proteins) related to the elimination of PSII photodamage and photoinhibition were remarkably up-regulated (by about 1000-fold) by cold stress in GJ2501 as indicated by RT-qPCR verification. Overexpression of CuELIP1 from GJ2501 in transgenic Arabidopsis protected PSII against photoinhibition under cold stress. Taken together, the cold tolerance of GJ2501 may be ascribed to its higher photoprotective capacity under cold stress.

Efficient Cold Tolerance Evaluation of Four Species of Liliaceae Plants through Cell Death Measurement and Lethal Temperature Prediction

Although Liliaceae are valuable resource plants with medicinal and edible uses, techniques for evaluating their tolerance to various abiotic stresses are very limited. This study evaluated the cold tolerance using visual inspection, electrolyte leakage, and Evan’s blue assay. Visual inspection of the responses to different temperatures, using a temperature range of 4 to −12 °C, showed that Scilla scilloides was receive the least damage. However, electrolyte leakage tests showed slightly different results from visual inspection. The median lethal temperature (LT50) was expected to be between −4 and −8 °C. The LT50 was considered a measure of damage due to electrolyte leakage in plant cold tolerance evaluation. As a result of predicting the lethal temperature using the logistic regression equation, the average LT50 of the four plants was −9.0 °C. The species with the lowest LT50 was measured for Hosta plantagines (−11.14°C), whereas the highest LT50 was measured for Hemerocallis fulva (−7.14°C). As a result of the Evan’s blue assay, it was found that cell necrosis occurred when the plants were exposed to low temperatures. Visual observation showed that more than 50% of the three plants’ cells, except for H. plantaginea, were stained blue even at 8 °C. From this result, H. plantaginea was judged to have strong low-temperature tolerance. At −12 °C, more than 50% of the four Liliaceae plants were colored blue, and the LT50 value was expected to be below −12 °C. The reducing sugar content, an indicator of plant cold tolerance, was the highest in H. plantaginea, followed by S. scilloide and H. longipes. Combining the three methods, H. plantaginea had the highest cold tolerance, followed by H. longipes, S. scilloides, and H. fulva. The results of this study will be widely used in selecting cold-tolerant useful resource plants.

A review of lethal thermal tolerance among freshwater mussels (Bivalvia: Unionida) within the North American faunal region

Freshwater mussels of the order Unionida are currently one of the most imperiled groups of organisms in the North American faunal region. Accurate risk assessments and development of effective management strategies for remaining populations require knowledge of thermal limits in the face of increasing surface water temperature due to climate change and various anthropogenic factors. We conducted a systematic literature review of unionid mussels (order Unionida, families Margaritiferidae and Unionidae) in the North American faunal region to (1) summarize lethal thermal tolerance data by life stage and taxonomy, (2) discuss ecological and climate change implications of existing lethal tolerance data, and (3) identify needs for future research. We identified lethal tolerance estimates for only 28 of 302 species in the families Unionidae and Margaritiferidae. The mean acute median lethal temperatures were 32.8 °C for glochidia (19 species), 35.0 °C for juveniles (13 species), and 36.3 °C for adults (4 species). Generally, glochidia were less tolerant than juveniles or adults of the same species—but there were several exceptions. Generally, Amblemini had the highest acute and chronic thermal tolerance of all tribes followed by Anodontini, Pleurobemini, Lampsilini, and Quadrilini. Acclimation temperature affected lethal tolerance endpoints in less than half (52 of 145) of comparisons within species. Lethal tolerance data for additional species, combined with a comprehensive database of in situ surface water temperatures, would be useful for modeling the frequency and duration of lethal limit exceedance in North America and identifying populations currently living at or near their upper lethal limits.

Plasticity of upper thermal limits of Australian Paratya spp. (Decapoda, Atyidae) and considerations of climate-change adaptation

Context The ability of ectothermic stream invertebrates to adapt to the predicted increases in mean and extreme stream temperatures is crucial to ensuring they continue to exist. Aims To examine the plasticity of thermal limits of Australian Paratya spp. (Decapoda, Atyidae) from streams in eastern New South Wales (NSW). We hypothesised that the upper lethal temperature (ULT, as indicated by the median lethal temperature, LT50) would be higher for warm water-acclimated shrimp individuals than for winter-acclimatised shrimp individuals because of the importance of acclimatisation temperature. Methods Controlled experiments were undertaken to determine the ULT by using ramping assays for winter field-acclimatised and warm water laboratory-acclimated Paratya spp. Key results Warm water-acclimated shrimp individuals demonstrated a significantly higher LT50 of 36.1°C than did winter-acclimatised shrimp individuals at 34.6°C. Paratya spp. exhibited a limited plasticity for acclimation to warmer temperatures. Conclusions Results demonstrated the potential vulnerability of ectothermic stream invertebrates to climate change if stream temperatures increase as predicted and thermal thresholds are exceeded. Implications Understanding the ULT of stream invertebrates helps predict their ability to respond to temperature variability and response to climate change. Increasing resilience through target management of resorting riparian vegetation for shade and securing environmental flows may reduce the impacts of stream warming.

Elevated Salinity Rapidly Confers Cross-Tolerance to High Temperature in a Splash-Pool Copepod.

SynopsisAccurate forecasting of organismal responses to climate change requires a deep mechanistic understanding of how physiology responds to present-day variation in the physical environment. However, the road to physiological enlightenment is fraught with complications: predictable environmental fluctuations of any single factor are often accompanied by substantial stochastic variation and rare extreme events, and several factors may interact to affect physiology. Lacking sufficient knowledge of temporal patterns of co-variation in multiple environmental stressors, biologists struggle to design and implement realistic and relevant laboratory experiments. In this study, we directly address these issues, using measurements of the thermal tolerance of freshly collected animals and long-term field records of environmental conditions to explore how the splash-pool copepod Tigriopus californicus adjusts its physiology as its environment changes. Salinity and daily maximum temperature—two dominant environmental stressors experienced by T. californicus—are extraordinarily variable and unpredictable more than 2–3 days in advance. However, they substantially co-vary such that when temperature is high salinity is also likely to be high. Copepods appear to take advantage of this correlation: median lethal temperature of field-collected copepods increases by 7.5°C over a roughly 120 parts-per-thousand range of ambient salinity. Complementary laboratory experiments show that exposure to a single sublethal thermal event or to an abrupt shift in salinity also elicits rapid augmentation of heat tolerance via physiological plasticity, although the effect of salinity dwarfs that of temperature. These results suggest that T. californicus’s physiology keeps pace with the rapid, unpredictable fluctuations of its hypervariable physical environment by responding to the cues provided by recent sublethal stress and, more importantly, by leveraging the mechanistic cross-talk between responses to salinity and heat stress.

Temperature-Mortality Response of Eggs, Larvae, Protonymphs, Deutonymphs, and Adult Females of Tetranychus urticae (Acari: Tetranychidae)

Abstract The two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), is one of the most important pests of cultivated plants. Its control is mainly based on the application of acaricides with concomitant adverse environmental and human health effects. Consequently, we examined the potential of using temperature as an alternative. To that end, we conducted bioassays of the temperature-mortality response of eggs, larvae, protonymphs, deutonymphs, and adult females of T. urticae. Groups of 20 individuals from each stage were placed on 4-cm diameter bean leaf (Phaseolus vulgaris L.) discs. A stream of air at the designated temperature was passed over the disc at a rate of 3.7 ± 1 m s–1) for 2 ± 1 s. Mortality was evaluated at 24 h (larva, protonymph, deutonymph, and female) or 72 h (egg) after exposure. Based on preliminary mortality responses, we selected 10 temperatures with a response range between 0 and 100% mortality. Those were 30, 35, 40, 45, 47, 50, 53, 55, 60, and 65°C [SD, ± 1.5 for each]). In total, 30 replications were conducted, and each repetition included these temperatures and untreated controls. The median lethal temperature ranged from 41.8°C for adult females to 46.9°C for eggs. The lethal temperature to kill 95% of the exposed individuals ranged from 55°C for females to 62.5°C for eggs. There were no survivors at 65°C (eggs), 60°C (larva, protonymph, and deutonymph), or 55°C (adult females).

Effect of water temperature on walleye pollock (Gadus chalcogrammus) embryos, larvae and juveniles: Survival, HSP70 expression, and physiological responses

This study investigated the survival rates, physiological responses, and the induction of heat shock protein 70 (HSP70) expression in size-specific walleye pollock (Gadus chalcogrammus) at different water temperatures. Four groups consisting of walleye pollocks in various lifecycle stages—embryos, larvae, small juveniles, and large juveniles—were used. Embryos had the highest hatching rate at 5 °C, which decreased as the water temperature increased. Larval survival rate was the highest at 5 °C. The median lethal temperatures (LT50) for small and large juveniles were 19.1 and 18.9 °C for 7 d, respectively. HSP70 was cloned using the pET21b + vector and expressed in Escherichia coli to analyze HSP70 expression in walleye pollock. Polyclonal antibodies were obtained from BALB/c mice after purification on a His-Trap HP column and used for protein analysis using enzyme-linked immunosorbent assay. HSP70 mRNA and proteins were expressed in similar patterns at both 5 °C and 8 °C, with a drastic increase in HSP70 expression induction at ≥11 °C. The gill tissues of small and large juveniles had the highest HSP70 mRNA and protein levels. Physiological responses of large juveniles' plasma showed that glutamic pyruvic transaminase, glutamic oxaloacetic transaminase, and glucose levels were highest at 14 °C, whereas total cholesterol and triglyceride levels were lowest. Plasma superoxide dismutase and cortisol levels were highest at 14 °C, whereas growth hormone and insulin-like growth factor-1 levels were highest at 8 °C. These results suggest that elevated water temperatures increased stress in the early lifecycle stages (embryos and larvae) of walleye pollock. HSP70 and stress-related indicator levels also increased with an increase in water temperature. Subsequently, the optimal water temperature for walleye pollock growth was determined to be below 8 °C.

Physiological and Transcriptional Responses to Acute and Chronic Thermal Stress in the Ark Shell Scapharca subcrenata

Ark shells (Scapharca subcrenata) grown on the tidal flats are often exposed to high temperature stresses in summer. In order to better understand their adaption to extreme or natural high temperature, we first determined the 96-h upper lethal temperature of ark shell and then investigated their physiological and transcriptional responses to acute or chronic thermal stress at the 96-h upper median lethal temperature (32°C). A significantly higher cumulative mortality (52% in 96 h) was observed in the acute heating treatment (AHT) group than that (22% in 7 days) in the chronic heating treatment (CHT) group. The apoptosis and necrosis rates of hemocytes were increased significantly in a time-dependent manner under both thermal stress strategies. Activities of antioxidant enzymes [superoxide dismutase (SOD) and catalase (CAT)] increased dramatically in a short time followed by a quick decline and reached to a lower level within 12 h in the AHT group, but maintain relatively high levels over a long period in the CHT group. The contents of malondialdehyde (MDA) were increased significantly firstly and restored to the original later in both acute and chronic thermal stress. Moreover, expression of the genes related to heat shock proteins (HSPs; HSP90, HSP70, HSP20, and sHSP), apoptosis [TNF receptor-associated factor 6 (TRAF6), glucose regulated protein 78 kD (GRP78), and caspase-3 (Casp-3)] and antioxidant responses [glutathione S-transferase (GST) and multidrug resistance protein (MRP)] could be induced and up-regulated significantly by thermal stress, however, expression of regucalcin (RGN), metallothionein (MT), and peroxiredoxin (PRX) was down-regulated dramatically under the two heating treatments. These results suggested that anti-apoptotic system, antioxidant defense system and HSPs could play important roles in thermal tolerance of ark shells, and the heat-resistant ark shell strains could be selected continuously by properly chronic thermal stress.

Molecular adaptations to heat stress in the thermophilic ant genus Cataglyphis.

Over the last decade, increasing attention has been paid to the molecular adaptations used by organisms to cope with thermal stress. However, to date, few studies have focused on thermophilic species living in hot, arid climates. In this study, we explored molecular adaptations to heat stress in the thermophilic ant genus Cataglyphis, one of the world's most thermotolerant animal taxa. We compared heat tolerance and gene expression patterns across six Cataglyphis species from distinct phylogenetic groups that live in different habitats and experience different thermal regimes. We found that all six species had high heat tolerance levels with critical thermal maxima (CTmax ) ranging from 43℃ to 45℃ and a median lethal temperature (LT50) ranging from 44.5℃ to 46.8℃. Transcriptome analyses revealed that, although the number of differentially expressed genes varied widely for the six species (from 54 to 1118), many were also shared. Functional annotation of the differentially expressed and co-expressed genes showed that the biological pathways involved in heat-shock responses were similar among species and were associated with four major processes: the regulation of transcriptional machinery and DNA metabolism; the preservation of proteome stability; the elimination of toxic residues; and the maintenance of cellular integrity. Overall, our results suggest that molecular responses to heat stress have been evolutionarily conserved in the ant genus Cataglyphis and that their diversity may help workers withstand temperatures close to their physiological limits.

Effect of acclimation on thermal limits and hsp70 gene expression of the New Zealand sea urchin Evechinus chloroticus

Seawater temperature is projected to increase globally due to climate change, affecting physiological responses, fitness and survival of marine organisms. Thermal tolerance studies are critical to determine the ability of animals to adapt to future environmental conditions. In this study, we aimed to determine if the thermal limits of the New Zealand Evechinus chloroticus would shift with animal's thermal history. We tested the effect of six thermal regimes on the righting ability, temperature of loss of righting (TLOR), median lethal temperature (LT50), lethal temperature (LT) and the gene expression of the heat shock protein 70 (hsp70) of the New Zealand sea urchin E. chloroticus when exposed to a thermal shock of 1 °C day−1 (duration of 7–16 days depending on the treatment). Treatments consisted of laboratory acclimation for one and four weeks to 18 °C and 24 °C (mean winter (15 °C) and summer temperature (21 °C) + 3 °C of warming, respectively), compared to non-acclimated sea urchins collected during winter (14.6 °C) and summer seasons (20.4 °C). Thermal history did not have a significant effect on the righting ability of E. chloroticus (TLOR ranged between 28 and 29 °C for all treatments) and LT50 (ranged between 29 and 30 °C for all treatments). However, LT of E. chloroticus collected during winter season was significantly lower than animals acclimated for one week at 18 °C. Maximum expression of hsp70 mRNA (Tmax) was observed at around 27–28 °C regardless of treatment; however, relative hsp70 mRNA levels were significantly higher in animals acclimated for four weeks at 24 °C. Despite proving to be a thermotolerant species with LTs around 30 °C, E. chloroticus was unable to increase thermal tolerance and Tmax when acclimated to high temperatures, suggesting that E. chloroticus may have a limited adaptive capacity to modify its phenotype; however, evolutionary adaptations may allow E. chloroticus to adapt to future ocean temperatures.

Linking flow and upper thermal limits of freshwater mussels to inform environmental flow benchmarks

Abstract Freshwater ecosystems are experiencing shifts in the natural range and variation of water temperatures due to anthropogenic activity, and these shifts can negatively affect survival, growth, and reproduction of aquatic species. Among the groups most affected are freshwater mussels of the family Unionidae. Knowledge of sublethal and lethal effects on mussels from changes in water temperature are largely unknown, especially for species from arid and semi‐arid regions such as the south‐western U.S.A. This limits the ability to assess, forecast, and adaptively manage this threat for those species and to understand how temperature influences population performance and community structure. To determine the effects of elevated water temperature on mussels from the south‐western U.S.A., we evaluated the upper thermal tolerances of adults of three species (Amblema plicata, Cyclonaias necki, and Fusconaia mitchelli) from the Guadalupe River. Mussels were acclimated to 27°C and then tested across a range of experimental temperatures (30–39°C) in standard acute (96‐hr) and chronic (10‐day) laboratory tests. The acute and chronic thresholds identified in thermal tolerance testing were then related to in situ water temperature and flows using a uniform continuous above‐threshold analysis, which evaluates the duration and frequency of continuous events above a specified temperature threshold. Median lethal temperature in 96‐hr tests averaged 36.4°C and ranged from 33.7 to 37.5°C, while the chronic 10‐day tests averaged 35.9°C and ranged from 32.4 to 37.5°C. Thermal tolerances of F. mitchelli were significantly lower than both A. plicata and C. necki, and the uniform continuous above‐threshold analysis showed that temperature affecting 5% of the population thresholds were exceeded for F. mitchelli in the Guadalupe River at both acute (96‐hr) and chronic (10‐day) values (30.5 and 28.4°C, respectively). Findings from this study indicate that freshwater mussels from the arid and semi‐arid regions of the south‐west U.S.A. are already at risk from rising environmental temperatures and altered hydrology. However, by incorporating laboratory thermal tolerance estimates with in situ temperature and discharge data, we provide a range of hydrologic thresholds to inform environmental flow recommendations and potentially mitigate thermal stress occurring during periods of low flow. In addition, this method can be readily adapted to other arid regions to guide flow recommendations or assess whether flow standards are sufficient to protect freshwater mussel populations during severe droughts and low flow periods.

Transcriptome Analysis of Genes Involved in Cold Hardiness of Peach Tree (Prunus persica) Shoots during Cold Acclimation and Deacclimation.

We analyzed the transcriptomes in the shoots of five-year-old ‘Soomee’ peach trees (Prunus persica) during cold acclimation (CA), from early CA (end of October) to late CA (middle of January), and deacclimation (DA), from late CA to late DA (middle of March), to identify the genes involved in cold hardiness. Cold hardiness of the shoots increased from early to late CA, but decreased from late CA to late DA, as indicated by decreased and increased the median lethal temperature (LT50), respectively. Transcriptome analysis identified 17,208 assembled transcripts during all three stages. In total, 1891 and 3008 transcripts were differentially expressed with a |fold change| > 2 (p < 0.05) between early and late CA, and between late CA and late DA, respectively. Among them, 1522 and 2830, respectively, were functionally annotated with gene ontology (GO) terms having a greater proportion of differentially expressed genes (DEGs) associated with molecular function than biological process or cellular component categories. The biochemical pathways best represented both periods from early to late CA and from late CA to late DA were ‘metabolic pathway’ and ‘biosynthesis of secondary metabolites’. We validated these transcriptomic results by performing reverse transcription quantitative polymerase chain reaction on the selected DEGs showing significant fold changes. The relative expressions of the selected DEGs were closely related to the LT50 values of the peach tree shoots: ‘Soomee’ shoots exhibited higher relative expressions of the selected DEGs than shoots of the less cold-hardy ‘Odoroki’ peach trees. Irrespective of the cultivar, the relative expressions of the DEGs that were up- and then down-regulated during CA, from early to late CA, and DA, from late CA to late DA, were more closely correlated with cold hardiness than those of the DEGs that were down- and then up-regulated. Therefore, our results suggest that the significantly up- and then down-regulated DEGs are associated with cold hardiness in peach tree shoots. These DEGs, including early light-induced protein 1, chloroplastic, 14-kDa proline-rich protein DC2.15, glutamate dehydrogenase 2, and triacylglycerol lipase 2, could be candidate genes to determine cold hardiness.

Comparative Transcriptome Analysis of the Heat Stress Response in Monochamus alternatus Hope (Coleoptera: Cerambycidae).

Temperature is a critical factor of insect population abundance and distribution. Monochamus alternatus Hope (Coleoptera: Cerambycidae) is a significant concern since it is transmitted vector of the pinewood nematode posing enormous economic and environmental losses. This pest shows tolerance to heat stress, especially extremely high temperatures. Exposing for 6, 12, 24, 48, or 96 h, the 50% median lethal temperatures (Ltem50) for fourth-instar larvae were 47.5, 45.5, 43.9, 43.4, and 42.3°C, respectively. A total of 63,360 unigenes were obtained from complementary DNA libraries of M. alternatus fourth-instar larvae (kept at 25°C and exposed to 40°C for 3 h) and annotated with six databases. Five hundred sixty-one genes were significantly upregulated, and 245 genes were downregulated after heat stress. The Gene Ontology enrichment analysis showed that most different expression genes are categorized into “protein folding” and “unfold protein binding” terms. In addition, “Longevity regulating pathway-multiple species,” “Antigen processing and presentation” as well as “MAPK signaling pathway” were significantly enriched Kyoto Encyclopedia of Genes and Genomes pathways. Further analysis of different expression genes showed that metabolism processes were suppressed, while ubiquitin proteolytic system, heat shock proteins, immune response, superoxide dismutase, cytochrome P450s, and aldehyde dehydrogenase were induced after heat shock. The stress signaling transduction pathways such as MAPK, Hippo, and JAK-STAT might be central convergence points in M. alternatus heat tolerance mechanism. The expression levels from quantitative real-time PCR of 13 randomly selected genes were consistent with the transcriptome results. These results showed that M. alternatus possessed strong heat tolerance and genes related to protein activity, immune response, and signal transduction composed of a complicated heat tolerance mechanism of M. alternatus. This research provided new insights into the mechanisms of thermal tolerance in other insects and aided in exploring the function of heat resistance-related genes.

Upper thermal tolerance of early juveniles of six terrestrial hermit crab species

Abstract Because temperature strongly influences biological processes of ectotherms, they must adapt to the thermal conditions of their habitats. We evaluated the upper thermal tolerance of early juveniles of six terrestrial hermit crab species in the family Coenobitidae (genera Birgus and Coenobita), B. latro, C. brevimanus, C. cavipes, C. purpureus, C. rugosus, and C. violascens, that occur in the northwestern Pacific region of Japan. A total of 30 laboratory-raised juveniles (approximately 1 mm in shield length) carrying gastropod shells were individually stocked in small plastic cups with sandy bottoms in temperature-controlled incubation chambers at ∼27°C. The temperature was increased by 1°C every 48 h, and the juveniles were observed until all the crabs had died. The median upper lethal temperature was estimated as the temperature at which 50% of the test juveniles had died. The median upper lethal temperature estimates significantly varied among the species. Coenobita violascens, which mainly occurs in mangrove estuaries with lower thermal conditions, had the lowest median upper lethal temperature values. The median upper lethal temperature values estimated for B. latro were slightly lower than those for the other Coenobita species, probably reflecting its cryptic nature in natural habitats during the juvenile stage. The most northerly distributed species, C. purpureus, had the highest median upper lethal temperature values, suggesting the existence of common physiological mechanisms that regulate both upper and lower thermal tolerance abilities.

Heat tolerance and acclimation capacity in subterranean arthropods living under common and stable thermal conditions.

Cave‐dwelling ectotherms, which have evolved for millions of years under stable thermal conditions, could be expected to have adjusted their physiological limits to the narrow range of temperatures they experience and to be highly vulnerable to global warming. However, most of the few existing studies on thermal tolerance in subterranean invertebrates highlight that despite the fact that they show lower heat tolerance than most surface‐dwelling species, their upper thermal limits are generally not adjusted to ambient temperature. The question remains to what extent this pattern is common across subterranean invertebrates. We studied basal heat tolerance and its plasticity in four species of distant arthropod groups (Coleoptera, Diplopoda, and Collembola) with different evolutionary histories but under similar selection pressures, as they have been exposed to the same constant environmental conditions for a long time. Adults were exposed at different temperatures for 1 week to determine upper lethal temperatures. Then, individuals from previous sublethal treatments were transferred to a higher temperature to determine acclimation capacity. Upper lethal temperatures of three of the studied species were similar to those reported for other subterranean species (between 20 and 25°C) and widely exceeded the cave temperature (13–14°C). The diplopod species showed the highest long‐term heat tolerance detected so far for a troglobiont (i.e., obligate subterranean) species (median lethal temperature after 7 days exposure: 28°C) and a positive acclimation response. Our results agree with previous studies showing that heat tolerance in subterranean species is not determined by environmental conditions. Thus, subterranean species, even those living under similar climatic conditions, might be differently affected by global warming.

Evaluating the upper thermal limits of glochidia for selected freshwater mussel species (Bivalvia: Unionidae) in central and east Texas, and the implications for their conservation

Abstract Understanding the temperature tolerances of organisms is critical because the thermal regimes of freshwater ecosystems are changing globally. Native freshwater mussels are sensitive to increasing water temperatures because of their physiology and unique life history. Detailed knowledge on lethal temperatures for mussels has been limited to less than 5% of the species known to occur in North America, and little is known about the thermal tolerances of mussel species from rivers within the south‐western USA. To determine the effects of elevated water temperature on mussels, the upper thermal tolerances of larvae (glochidia) for the following species across four basins in Texas (Neches, Guadalupe, San Antonio, and Colorado) were tested: Amblema plicata, Cyclonaias necki, Fusconaia mitchelli, Lampsilis bracteata, Lampsilis hydiana, Lampsilis satura, Lampsilis teres, and Obovaria arkansasensis. Glochidia were acclimated to 27°C across a range of experimental temperatures (30–39°C) in 24‐h standard acute laboratory tests. The median lethal temperature (LT50) among glochidia averaged 32.4°C and ranged from 26.9 to 36.4°C. Thermal tolerances differed significantly among and within species, and by season. Comparing these results with current water temperatures in central and east Texas indicated that populations of the focal species studied are at risk from rising environmental temperatures and, as a consequence, their long‐term viability will be challenging in future years.

Physiological response and miRNA-mRNA interaction analysis in the head kidney of rainbow trout exposed to acute heat stress.

The rainbow trout is a cold-water fish cultured in China. Heat stress has a serious impact on the summer survival and the yield of rainbow trout. A better understanding of the regulatory response of rainbow trout to heat stress will help in determining the relationship between heat stress signaling pathways and adaption mechanisms and help contribute to breeding new high-temperature tolerant strains of rainbow trout. In this study, the 48-h median lethal temperature (48h-LT50) of rainbow trout was determined as 22.5°C. We developed control (16°C) and heat-treated (22.5°C) groups and extracted RNA from the head kidney tissues for high-throughput sequencing to study the microRNA (miRNA) expression profiles. Twelve up-regulated and five down-regulated miRNAs were identified between the control and heat-treated groups. A total of 22 target genes were predicted for 6 of the differentially expressed miRNAs, including 31 negative miRNA-mRNA interactions. Important regulatory pathways under heat stress are related to the metabolism and immune responses of the rainbow trout. Our findings provide preliminary data for investigating the high-temperature molecular mechanism of the rainbow trout and can help producers to reduce the economic losses caused by high temperature weather.

Low-temperature tolerance of early juveniles of six terrestrial hermit crab species

Abstract Because temperature strongly influences biological processes of ectotherms, it also plays a fundamental role in determining their geographical distribution. We evaluated the low-temperature tolerance of early juveniles of six terrestrial hermit crab species in the family Coenobitidae (genera Birgus and Coenobita), B. latro, C. brevimanus, C. cavipes, C. purpureus, C. rugosus, and C. violascens that occur in the northwestern Pacific region, Japan. A total of 30 laboratory-raised juveniles (about 1 mm in shield length) carrying gastropod shells were individually stocked in small plastic cups with sandy bottoms in temperature-controlled incubatory chambers at ∼27°C. The temperature was reduced by 1°C every 48 h, and the juveniles were observed until all the crabs had died; the median lethal temperature (MLT) was estimated as the temperature at which 50% of the test juveniles had died. The MLT estimates varied significantly among the species, and the most northward distributed species, C. purpureus, had the lowest MLT values. The phylogeny, paleoceanography, paleogeography, and paleoclimatology suggest that cooler thermal regimes might have acted as an evolutionary force for the divergence of C. purpureus in the Pliocene. A negative correlation was found between the northern latitudinal limit of distribution and the MLT values, even after controlling for the phylogenetic relationships in the six coenobitids. A temperature-dependent biogeography was thus recognized in terrestrial hermit crab species in the northwestern Pacific region, and global warming is expected to affect their geographical distributions.

Nonlinear Regression on Cold Tolerance Data for Brassica Napus

This study purposes to derive the predictive model for the cold tolerance of Brassica napus, using the data collected in the Tree Breeding Lab of Gyeongsang National University during July and August of 2016. Three Brassica napus samples were treated at each of low temperatures from 4℃ to -12℃ by decrement of 4℃, step by step, and electrolyte leakage levels were measured at each stage. Electrolyte leakages were observed tangibly from -4℃. We tried to fit the six nonlinear regression models to the electrolyte leakage data of Brassica napus: 3-parameter logistic model, baseline logistic model, 4-parameter logistic model, (4-1)-parameter logistic model, 3-parameter Gompertz model, and (3-1)-parameter Gompertz model. The baseline levels of the electrolyte leakage estimated by these models were 4.81%, 4.07%, 4.19%, 4.07%, 4.55%, and 0%, respectively. The estimated median lethal temperature, LT50, were -5.87℃, -6.31℃, -6.05℃, -6.35℃, -4.98℃, and -5.15℃, respectively. We compared and discussed the measures of goodness of fit to select the appropriate nonlinear regression model.

Influence of respiratory mode on the thermal tolerance of intertidal limpets.

Predicting ecological responses to climate change requires an understanding of the mechanisms that influence species’ tolerances to temperature. Based on the idea that air and water breathing animals are differentially suited to life in either medium due to differences in their respiratory morphology, we examined the possibility that the thermal tolerances of co-existing intertidal pulmonate and patellogastropod limpets may differ in different breathing media. We tested this by determining each species’ median lethal temperature (LT50) and cardiac Arrhenius breakpoint temperature (ABT) as measures of upper thermal tolerance limits, in air and water. Although all these species can survive in air and water, we hypothesised that the pulmonate limpets, Siphonaria capensis and S. serrata, would have higher thermal limits than the patellogastropod limpets, Cellana capensis and Scutellastra granularis, in air and vice versa in water. The results did not support our hypotheses, since C. capensis had similar thermal tolerance limits to the pulmonate limpets in air and the pulmonate limpets had thermal tolerance limits similar to or higher than S. granularis in water. Thus, considering pulmonate and patellid limpets as groups, we found no differences in their collective upper thermal tolerance limits in either medium. We conclude that differences between these two limpet groups in their respiratory morphology do not influence thermal tolerance, but that tolerances are species-specific.