Temperature Compensation of Circadian Period Length in Clock Mutants of Neurospora crassa

Temperature compensation of circadian period length in 12 clock mutants of Neurospora crassa has been examined at temperatures between 16 and 34°C. In the wild-type strain, below 30°C (the “breakpoint” temperature), the clock is well-compensated (Q₁₀ = 1), while above 30°C, the clock is less well-compensated (Q₁₀ = 1.3). For mutants at the frq locus, mutations that shorten the circadian period length (frq-1, frq-2, frq-4, and frq-6) do not alter this temperature compensation response. In long period frq mutants (frq-3, frq-7, frq-8), however, the breakpoint temperature is lowered, and the longer the period length of the mutants the lower the breakpoint temperature. Long period mutants at other loci exhibit other types of alterations in temperature compensation—e.g. chr is well-compensated even above 30°C, while prd-3 has a Q₁₀ significantly less than 1 below 30°C. Prd-4, a short period mutant, has several breakpoint temperatures. Among four double mutants examined, the only unusual interaction between the individual mutations occurred with chr prd, which had an unusually low Q₁₀ value of 0.86 below 27°C. There was no correlation between circadian period length and growth rate. These strains should be useful tools to test models for the temperature compensation mechanism.     

Modeling temperature entrainment of circadian clocks using the Arrhenius equation and a reconstructed model from Chlamydomonas reinhardtii

Endogenous circadian rhythms allow living organisms to anticipate daily variations in their natural environment. Temperature regulation and entrainment mechanisms of circadian clocks are still poorly understood. To better understand the molecular basis of these processes, we built a mathematical model based on experimental data examining temperature regulation of the circadian RNA-binding protein CHLAMY1 from the unicellular green alga Chlamydomonas reinhardtii, simulating the effect of temperature on the rates by applying the Arrhenius equation. Using numerical simulations, we demonstrate that our model is temperature-compensated and can be entrained to temperature cycles of various length and amplitude. The range of periods that allow entrainment of the model depends on the shape of the temperature cycles and is larger for sinusoidal compared to rectangular temperature curves. We show that the response to temperature of protein (de)phosphorylation rates play a key role in facilitating temperature entrainment of the oscillator in Chlamydomonas reinhardtii. We systematically investigated the response of our model to single temperature pulses to explain experimentally observed phase response curves.     

A circadian oscillator in Aspergillus spp. regulates daily development and gene expression

We have established the presence of a circadian clock in Aspergillus flavus and Aspergillus nidulans by morphological and molecular assays, respectively. In A. flavus, the clock regulates an easily assayable rhythm in the development of sclerotia, which are large survival structures produced by many fungi. This developmental rhythm exhibits all of the principal clock properties. The rhythm is maintained in constant environmental conditions with a period of 33 h at 30°C, it can be entrained by environmental signals, and it is temperature compensated. This endogenous 33-h period is one of the longest natural circadian rhythms reported for any organism, and this likely contributes to some unique responses of the clock to environmental signals. In A. nidulans, no obvious rhythms in development are apparent. However, a free running and entrainable rhythm in the accumulation of gpdA mRNA (encoding glyceraldehyde-3-phosphate dehydrogenase) is observed, suggesting the presence of a circadian clock in this species. We are unable to identify an Aspergillus ortholog of frequency, a gene required for normal circadian rhythmicity in Neurospora crassa. Together, our data indicate the existence of an Aspergillus circadian clock, which has properties that differ from that of the well-described clock of N. crassa.     

Comparative genomics of Chlamydomonas

Despite its role as a reference organism in the plant sciences, the green alga Chlamydomonas reinhardtii entirely lacks genomic resources from closely related species. We present highly contiguous and well-annotated genome assemblies for three unicellular C. reinhardtii relatives: Chlamydomonas incerta, Chlamydomonas schloesseri, and the more distantly related Edaphochlamys debaryana. The three Chlamydomonas genomes are highly syntenous with similar gene contents, although the 129.2 Mb C. incerta and 130.2 Mb C. schloesseri assemblies are more repeat-rich than the 111.1 Mb C. reinhardtii genome. We identify the major centromeric repeat in C. reinhardtii as a LINE transposable element homologous to Zepp (the centromeric repeat in Coccomyxa subellipsoidea) and infer that centromere locations and structure are likely conserved in C. incerta and C. schloesseri. We report extensive rearrangements, but limited gene turnover, between the minus mating type loci of these Chlamydomonas species. We produce an eight-species core-Reinhardtinia whole-genome alignment, which we use to identify several hundred false positive and missing genes in the C. reinhardtii annotation and >260,000 evolutionarily conserved elements in the C. reinhardtii genome. In summary, these resources will enable comparative genomics analyses for C. reinhardtii, significantly extending the analytical toolkit for this emerging model system.

High-quality genome assemblies and annotations for three of the closest relatives of Chlamydomonas reinhardtii enable comparative genomics analyses.     

Circadian Rhythm of the Prokaryote Synechococcus sp. RF-1

The prokaryotic Synechococcus sp. RF-1 exhibited a nitrogen fixation circadian rhythm with characteristics remarkably similar to the circadian rhythm of eukaryotes. The rhythm had a free-running period of about 24 hours when the length of the preen-trained cycle did not differ too much from 24 hours, and it was insensitive to changes in temperature from 22°C to 33°C. Because the endogenous rhythm of nitrogen fixation was not affected by a phase-shift of its previous cycles, the circadian rhythm in Synechococcus sp. RF-1 was not considered to be controlled simply by a feedback mechanism.     

Physical and Perceptual Cooling with Beverages to Increase Cycle Performance in a Tropical Climate

Purpose

This study compares the effects of neutral temperature, cold and ice-slush beverages, with and without 0.5% menthol on cycling performance, core temperature (T_co) and stress responses in a tropical climate (hot and humid conditions).

Methods

Twelve trained male cyclists/triathletes completed six 20-km exercise trials against the clock in 30.7°C±0.8°C and 78%±0.03% relative humidity. Before and after warm-up, and before exercise and every 5 km during exercise, athletes drank 190 mL of either aromatized (i.e., with 0.5 mL of menthol (5 gr/L)) or a non-aromatized beverage (neutral temperature: 23°C±0.1°C, cold: 3°C±0.1°C, or ice-slush: −1°C±0.7°C). During the trials, heart rate (HR) was continuously monitored, whereas core temperature (T_co), thermal comfort (TC), thermal sensation (TS) and rate of perceived exertion (RPE) were measured before and after warm-up, every 5 km of exercise, and at the end of exercise and after recovery.

Results

Both the beverage aroma (P<0.02) and beverage temperature (P<0.02) had significant and positive effects on performance, which was considerably better with ice-slush than with a neutral temperature beverage, whatever the aroma (P<0.002), and with menthol vs non-menthol (P<0.02). The best performances were obtained with ice-slush/menthol and cold/menthol, as opposed to neutral/menthol. No differences were noted in HR and T_co between trials.

Conclusion

Cold water or ice-slush with menthol aroma seems to be the most effective beverage for endurance exercise in a tropical climate. Further studies are needed to explore its effects in field competition.     

Temperature-Sensitive, Photosynthesis-Deficient Mutants of Chlamydomonas reinhardtii

Mutants of the unicellular, green alga Chlamydomonas reinhardtii were recovered by screening for the absence of photoautotrophic growth at 35°C. Whereas nonconditional mutants required acetate for growth at both 25 and 35°C, the conditional mutants have normal photoautotrophic growth at 25°C. The conditional mutants consisted of two classes: (a) Temperature-sensitive mutants died under all growth conditions at 35°C, but (b) temperature-sensitive, acetate-requiring mutants were capable of heterotrophic growth at 35°C when supplied with acetate in the dark. The majority of mutants within the latter of these two classes had defects in photosynthetic functions. These defects included altered pigmentation, reduced whole-chain electron-transport activity, reduced ribulosebis-phosphate carboxylase activity, or pleiotropic alterations in a number of these photosynthetic components. Both nuclear and chloroplast mutants were identified, and a correlation between light-sensitive and photosynthesis-deficient phenotypes was observed.     

TIME-TEMPERATURE RELATIONS IN THE INCUBATION OF THE WHITEFISH,COREGONUS CLUPEAFORMIS (MITCHILL)

1. Whitefish eggs incubated in aerated lake water at controlled tempera tures of 0°, 0.5°, 2°, 4°, 6°, 8°, 10°, and 12°C., failed to hatch at either 0° or 12°C. 0.6 per cent hatched alive at 10°C., 72.67 per cent hatched alive at 0.5°C., and an intermediate proportion hatched at intermediate temperatures. 2. The percentage of abnormal embryos which developed to the hatching stage varied directly with temperature between 4° and 12°, all embryos being abnormal at 12°C.; but none were abnormal at either 0.5°, or 2°C. Normal development predominated from 0.5 to 6°C. The highest proportion of embryos to hatch alive was 72.67 per cent at 0.5°C., which is, hence, the optimum temperature. 3. Total incubation time ranged from 29.6 days at 10°C. to 141 days at 0.5°C. 4. The time (T) required to attain any given stage of development is expressed in equations See PDF for Equation where temperature, t, is a negative exponent of the constant, A, whose value differs above or below 6°C., a critical temperature. Values of A above 6° fluctuate about 1.13; those of A below 6° fluctuate about 1.19 as a mean. 5. Applying Arrhenius'' equation µ values for the total incubation period are 27,500 below 6° and 27,100 above it. 6. The relative magnitude of A values of the exponential equation and µ values of Arrhenius'' equation show corresponding changes from one developmental period to another. 7. When plotted, thermal increments show cyclic variations, with maxima during periods of cleavage and of organogenesis. These may indicate the interaction of two separate sets of embryonic processes, which give a maximal response to temperature differences during these two separate periods. 8. Above 6°, µ values during the hatching process are distinct from those of developmental stages and are regarded as being due to the action of hatching enzymes.