Recycling of CO2 via Crassulacean acid metabolism in the rock outcrop succulent Sedum pulchellum Michx. (Crassulaceae)

Under well-watered conditions in the laboratory, Sedum pulchellum assimilated CO₂ only during the day, yet exhibited small nocturnal increases in tissue acid content followed by deacidification in the light (CAM-cycling). When drought-stressed, little CO₂ was fixed in the day and none at night, yet even greater acid fluctuations were observed (CAM-idling). Calculations indicate that water savings associated with CAM-cycling when water is available are small. Water saving is more likely to be significant during CAM-idling when water supply is limited and stomata are closed day and night. Thus, in this species, CAM-idling may be of greater benefit to the plant, relative to CAM-cycling, in surviving habitats prone to frequent drought stress.Abbreviations A CO₂ exchange rate - CAM Crassulacean acid metabolism - c_i shoot internal CO₂ concentration - g_c shoot conductance to CO₂ - PPFD photosynthetic photon flux density - WUE water-use efficiency Supported by National Science Foundation Grant No. DMB 8506093.     

IMMUNOLOCALIZATION OF NITRATE REDUCTASE IN THE MARINE DINOFLAGELLATE GONYAULAX POLYEDRA (PYRROPHYTA)1

A polyclonal antibody, raised against nitrate reductase (NR) purified from the photosynthetic dinoflagellate Gonyaulax polyedra Stein, was used as a probe in immunogold-labeling experiments on thin sections prepared from cells harvested both during day and night phases. Previous experiments have shown that both NR activity and the amount of immunoreactive NR in cell extracts is greater when day-phase cells are examined, and this property was exploited as an internal control for the cytochemical labeling. We observed that in day-phase cells, chloroplasts contained approximately three times more gold particles than night-phase cells (highly significant difference; P < 0.0001), whereas cytoplasmic labeling levels remained relatively level between day and night. We conclude from the diurnal difference in labeling that our antibody faithfully reflects the distribution of NR in Gonyaulax cells. Thus, as in to some other higher plants and green algae, Gonyaulax compartmentalizes active NR in its chloroplasts.     

Diel vertical migrations of invertebrate predators (Leptodora kindtii ,Thermocyclops taihokuensis, andMesocyclops sp .) in a shallow, eutrophic lake

We studied vertical distribution patterns of three invertebrate predators – Leptodora kindtii, Mesocyclopssp., and Thermocyclops taihokuensis– in a shallow eutrophic lake, Lake Suwa , Japan. From June to October in 2000 and 2001, we collected samples in the lake center in order to examine the vertical distribution patterns and the densities of the predators in the water column during the day (0900) and at night (2330). We also examined phototactic behavior of Leptodora in the laboratory. The three invertebrate predators showed clear migration patterns. Leptodora and Thermocyclops displayed a typical migration, avoiding the surface and maintaining a high abundance in deeper water during the day, and being distributed uniformly during the night. Mesocyclops, on the other hand, showed no clear vertical distribution pattern in the water column. However, Mesocyclops showed higher densities in the water column during the night than during the day. It suggests that they stayed just above the bottom during the day and migrated upward during the night. Leptodora also showed such a density difference between day and night. In the laboratory, Leptodora showed strong negative phototactic behavior. The observed density changes between day and night in Leptodora and Mesocyclops suggests the possible underestimation of their population density by usual sampling methods, and thus the impact of predation on populations of prey zooplankton species may also be underestimated in shallow water bodies.     

Habitat overlap of enemies: temporal patterns and the role of spatial complexity

Environmental heterogeneity can promote coexistence of conflicting species by providing spatial or temporal refuges from strong interactions (e.g., intraguild predation, competition). However, in many systems, refuge availability and effectiveness may change through time and space because of variability in habitat use by either species. Here I consider how the intensity of intraguild predation risk varies from day to night for aquatic insects that use both vegetated and open water habitats. Large (1,265 l) and small (42 l) mesocosms were used to test the hypothesis that Buenoa would choose an open-water habitat that minimized predation by the ambush predator Notonecta during the day, but that at night Buenoa would safely use both vegetated and open water. Regardless of container size, Notonecta remained in vegetated water during the day and exploited both habitats at night, despite exhibiting greatest instantaneous predation rates in open water during the day. In contrast, Buenoa maintained an even distribution throughout the mesocosms during day and night, even though habitat-specific predation risks were fivefold lower in open waters than in vegetation during the day and habitat-specific predation risk would have been reduced threefold by fully exploiting open waters. Thus, temporal heterogeneity was both beneficial and detrimental to Buenoa; darkness of night reduced predation, but spatial refuges also disappeared. Together, these patterns suggest that while environmental heterogeneity can dampen intense biotic interactions, enemies do not select habitats solely on the basis of conflict avoidance. Instead, it appears that habitat-specific variation in other biotic (e.g., visual predators) or physical factors (e.g., UV radiation) may also mediate species interactions by influencing habitat selection.     

Antioxidative protection in the inducible CAM plant Sedum album L. following the imposition of severe water stress and recovery

The antioxidative protection during the C₃-CAM shift induced by water stress was investigated in the temperate succulent Sedum album L. The C₃-CAM shift was characterized in terms of CO₂ exchange, titratable acidity and phosphoenolpyruvate carboxylase activity. Well-watered plants displayed C₃-like patterns of gas exchange and exhibited a mild day-night acid fluctuation indicating that those plants were performing CAM-cycling metabolism. Imposed drought highly stimulated CAM cycling, decreasing the net CO₂ uptake during the day, eliminating net CO₂ efflux at night and stimulating tissue acid fluctuations. As water deficit developed, chlorophyll fluorescence measurements showed a decrease in the Fv/Fm ratio, indicating that photoinhibition could follow after severe drought. Protection might be performed by the increased activity of enzymes involved in the destruction of free radicals and oxidants, but their response depended on the water status of the plant. Ascorbate peroxidase and superoxide dismutase activities increased in plants subjected to mild stress but declined during severe water stress. Catalase activity, however, was quite stable under mild water stress and was clearly inhibited under severe water stress. At this stage, glutathione reductase and monodehydroascorbate reductase seemed to be very important in the protection against oxidants, both increasing considerably their activities under severe water stress. Even if recycling has been shown to alleviate photoinhibition, our results clearly demonstrate that antioxidative enzymes play an important role in the protection of plants from oxidants during the C₃-CAM shift induced by water stress.     

Comparison of Day and Night Fyke Netting, Electrofishing and Snorkelling for Monitoring a Population of the Threatened Golden Galaxias (Galaxias auratus)

The littoral zone of small off-stream water storage containing a translocated population of Galaxias auratus was sampled fortnightly at day and night with fyke nets, electrofishing and snorkelling over 3 months. Variation in population data provided by each method, including relative abundance indices, size structure, and habitat preferences, were examined. Aspects of behaviour and activity patterns were also investigated. Night sampling using all methods consistently yielded larger catches than day sampling. The size structure of catches varied, with electrofishing at night and fyke netting during the day having higher proportions of juveniles, whilst snorkelling at night and electrofishing during the day had higher proportions of adults. Fyke netting at night yielded by far the largest catches (~3-fold more than other methods) and also captured balanced proportions of juveniles and adults. Galaxias auratus had a strong diel activity pattern and were most active at night. The majority of the population migrated into the littoral zone during the night and back into deeper water during the day. A small number of juveniles remained in the littoral zone and some adults sheltered in the dense cover of species-rich littoral vegetation during the day. Shores with shallow depth profiles appeared to be preferred due to higher catches in these areas using all methods. Based on the results of this study, fyke netting at night in littoral habitats is recommended for monitoring populations of G. auratus. Fyke netting is likely to be an effective method for monitoring other lacustrine galaxiid species; however, further work is required to investigate the effects of habitat variables and fish community structure on activity patterns of galaxiids, and hence their catchability with various methods, in more extensive lentic environments.     

Maltose is the major form of carbon exported from the chloroplast at night

Transitory starch is formed in chloroplasts during the day and broken down at night. We investigated carbon export from chloroplasts resulting from transitory-starch breakdown. Starch-filled chloroplasts from spinach (Spinacia oleracea L. cv. Nordic IV) were isolated 1 h after the beginning of the dark period and incubated for 2.5 h, followed by centrifugation through silicone oil. Exported products were measured in the incubation medium to avoid measuring compounds retained inside the chloroplasts. Maltose and glucose made up 85% of the total exported products and were exported at rates of 626 and 309 nmol C mg^–1 chlorophyll h^–1, respectively. Net export of phosphorylated products was less than 5% and higher maltodextrins were not detected. Maltose levels in leaves of bean (Phaseolus vulgaris L. cv. Linden), spinach, and Arabidopsis thaliana (L.) Heynh. were low in the light and high in the dark. Maltose levels remained low and unchanged during the light/dark cycle in two starch-deficient Arabidopsis mutants, stf1, deficient in plastid phosphoglucomutase, and pgi, deficient in plastid phosphoglucoisomerase. Through the use of nonaqueous fractionation, we determined that maltose was distributed equally between the chloroplast and cytosolic fractions during darkness. In the light there was approximately 24% more maltose in the cytosol than the chloroplast. Taken together these data indicate that maltose is the major form of carbon exported from the chloroplast at night as a result of starch breakdown. We hypothesize that the hydrolytic pathway for transitory-starch degradation is the primary pathway used when starch is being converted to sucrose and that the phosphorolytic pathway provides carbon for other purposes.Abbreviations CAM crassulacean acid metabolism - Chl chlorophyll - DHAP dihydroxyacetone phosphate - FBPase fructose bisphosphatase - GAP glyceraldehyde-3-phosphate - G6P glucose 6-phosphate - PGA 3-phosphoglycerate - TPT triose phosphate translocator - WT wild type     

Starch and apparent rate of photosynthesis of leaves of Lolium perenne grown at different temperatures

Summary Leaves from Lolium perenne plants grown at day/night temperatures of 15°/10° had slower rates of apparent photosynthesis at 15° than leaves from plants grown at 25°/20°. Electron microscopy showed a higher starch concentration in chloroplasts from low- compared with the higher temperature-grown plants. However, all differences in apparent photosynthesis were negatively associated with differences in size of mesophyll cells. It is suggested that the presence of starch per se had no effect on photosynthesis rate and that temperature during growth influences subsequent rates because leaves from the higher temperature have smaller cells and chloroplasts than those from the lower one.     

Diel activity and thermoregulatory behavior of a fully aquatic frog: Xenopus laevis

Ten adult Xenopus laevis were tested individually for 48-hr periods, following an initial 24-hr introductory period, in electronic shuttleboxes which allowed them to control water temperatures without operant conditioning. Locomotor activity was recorded via photocell-monitored light beams. The frogs were nocturnal, being nearly twice as active at night as during the day. The mean preferred temperature was 22.4°C, with no significant difference between night (22.5°C) and day (22.3°C), although the modal preferendum shifted from 24°C by day to 22°C at night, with a corresponding change in skewness. The range of voluntarily occupied temperatures was 14–32°C by day and 14–29°C at night. The median thermal preferendum was 22°C both day and night.     

Diurnal feeding as a potential mechanism of osmoregulation in aphids

Diurnal variation in phloem sap composition has a strong infuence on aphid performance.The sugar-rich phloem sap serves as the sole diet for aphids and a suite of physiological mechanisms and behaviors allowv them to tolerate the high osmotic stress.Here,we tested the hypothesis that night-time feeding by aphids is a behavior that takes advantage of the low sugar diet in the night to compensate for osmotic stress incurred while feeding on high sugar diet during the day.Using the electrical penetration graph(EPG)technique.we examined the eiects of diurmal rhythm on feeding behaviors of bird cherry-oat aphid(Rhopalosiphurm padi L.)on wheat.A strong diurmal rhythm in aphids as indicated by the presence of a cyclical pattern of expression in a core clock gene did not impact aphid feeding and similar feeding behaviors were observed during day and night.The major difference observed between day and night feeding was that aphids spent significantly longer time in phloem salivation during the night compared to the day.In contrast,aphid hydration was reduced at the end of the day-time feeding compared to end of the night-time fepding.Gene expression analysis of R.padi osmoregulatory genes indicated that sugar break down and water transport into the aphid gut was reduced at night.These data suggest that while diumal variation occurs in phloem sap composition,aphids use night time feeding to overcome the high osmotic stress incurred while feeding on sugar-rich phloem sap during the day.     

Diel Habitat Partitioning by Bull Charr and Cutthroat Trout During Fall and Winter in Rocky Mountain Streams

We used underwater observation to determine diel habitat partitioning between bull charr, Salvelinus confluentus, and cutthroat trout, Oncorhynchus clarki, during fall and winter (0.1–8.3°C) in two Rocky Mountain streams that differed in habitat availability. The majority (>70%) of both species emerged from concealment cover at night, though bull charr exhibited a greater tendency for nocturnal behavior than cutthroat trout. Differences in day and night counts were most pronounced at temperatures <3°C, when very few fish of either species were observed in the water column during the day, but both species were common at night. Both species used concealment cover of large woody debris and boulder substrate crevices in deep pools during the day. At night, fish emerged from cover and habitat use shifted to shallow water with low cover. Microhabitat partitioning among species and size classes occurred at night, cutthroat trout moving into shallower, faster water that was farther from cover compared to bull charr. Smaller fish of both species occupied focal positions in slower, shallower water closer to the substrate than larger fish. Large, mixed-species aggregations also were common in beaver ponds both day and night. High variation in diel and site-specific winter habitat use suggests the need for caution in developing habitat suitability criteria for salmonids based solely on daytime observations or on observations from a few sites. Our results support the need to incorporate nocturnal habitat use and partitioning in studies of salmonid ecology.     

Seasonal variation in diel activity and microhabitat use of an endemic New Zealand stream-dwelling galaxiid fish

1. Seasonal variation in microhabitat use and activity of 14 giant kokopu Galaxias argenteus, a drift‐feeding galaxiid fish, was compared using radiotelemetry. 2. During winter giant kokopu predominantly used low velocities and intermediate depths by night and day. Activity recorded during 24 and 72 h periods indicated that fish were consistently active at night and inactive during the day. Activity data corresponded with point‐in‐time habitat use data, both of which indicated that fish were concealed amongst cover during the day and used open water habitats at night. 3. During summer, giant kokopu used higher water velocities, shallower depths and coarser substrata, particularly at night but also occasionally during the day relative to winter. Giant kokopu were active by both day and night in summer, although periods of activity were less defined and less predictable than during winter. 4. Adults used predictable home reaches at base‐flow, with most individuals repeatedly using of one or two cover locations within their ‘home’ reach. Reaches used by fish were relatively short (rarely exceeding 26 m) irrespective of season and always included a single pool‐riffle sequence. 5. Diel and seasonal behaviour of giant kokopu was generally comparable with that exhibited by other drift feeding fish species in small temperate streams. However, the nocturnal activity of giant kokopu contrasts with activity patterns in various salmonids, indicating that the impact of predation by different drift feeding fish may vary considerably.     

The role of amylomaltase in maltose metabolism in the cytosol of photosynthetic cells

Transitory starch is stored during the day inside chloroplasts and then broken down at night for export. Recent data indicate that maltose is the major form of carbon exported from the chloroplast at night but its fate in the cytosol is unknown. An amylomaltase gene (malQ) cloned from Escherichia coli is necessary for maltose metabolism in E. coli. We investigated whether there is an amylomaltase in the cytosol of plant leaves and the role of this enzyme in plants. Two mutants of Arabidopsis thaliana (L) Heynh. were identified in which the gene encoding a putative amylomaltase enzyme [disproportionating enzyme 2, DPE2 (DPE1 refers to the plastid version of this enzyme)] was disrupted by a T-DNA insertion. Both dpe2-1 and dpe2-2 plants exhibited a dwarf phenotype and accumulated a large amount of maltose. In addition, dpe2 mutants accumulated starch and a water-soluble, ethanol/KCl-insoluble maltodextrin in their chloroplasts. At night, the amount of sucrose in dpe2 plants was lower than that in wild-type plants. These results show that Arabidopsis has an amylomaltase that is involved in the conversion of maltose to sucrose in the cytosol. We hypothesize that knocking out amylomaltase blocks the conversion from maltose to sucrose, and that the higher amount of maltose feeds back to limit starch degradation reactions in chloroplasts. As a result, dpe2 plants have higher maltose, higher starch, and higher maltodextrin but lower nighttime sucrose than wild-type plants. Finally, we propose that maltose metabolism in the cytosol of Arabidopsis leaves is similar to that in the cytoplasm of E. coli.Abbreviations F6P fructose 6-phosphate - G1P glucose 1-phosphate - G6P glucose 6-phosphate - GTase glucanotransferase     

Diurnal shifts in use of summer habitat by age-0 brown trout in a regulated mountain stream

Diurnal shifts in habitat use by age-0 (< 50 mm total length) brown trout (Salmo trutta) were observed from June to July 1989 and 1990 in low-gradient (< 1% channel slope) reaches of Douglas Creek, Medicine Bow National Forest, Wyoming. Fish were visually located during day and night, and water depth, water velocity, distance from the stream edge, substrate, habitat type, and cover were determined at each location. The fish were found predominantly in locations with slow water velocities in stream-margin and backwater-pool habitats. They were observed in slower water, closer to the stream edge at night than during the day.     

Diel differences in fish assemblages in nearshore eelgrass and kelp habitats in Prince William Sound,Alaska

The importance of a particular habitat to nearshore fishes can be best assessed by both diurnal and nocturnal sampling. To determine diel differences in fish assemblages in nearshore eelgrass and understory kelp habitats, fishes were sampled diurnally and nocturnally at six locations in western Prince William Sound, Alaska, in summer 2007. Abundance of fish between day and night were similar, but species composition and mean size of some fish changed. Species richness and species diversity were similar in eelgrass during the day and night, whereas in kelp, species richness and species diversity were greater at night than during the day. In eelgrass, saffron cod (Eleginus gracilis) was the most abundant species during the day and night. In kelp, the most abundant species were Pacific herring (Clupea pallasii) during the day and saffron cod at night. Diel differences in fish size varied by species and habitat. Mean length of saffron cod was similar between day and night in eelgrass but was greatest during the day in kelp. Pacific herring were larger at night than during the day in kelp. Diel sampling is important to identity nearshore habitats essential to fish and help manage fish stocks at risk.     

The daily activity pattern of Octopus vulgar is in a natural habitat

The homes of Octopus vulgaris in shallow Mediterranean water were found and marked by divers using SCUBA; hourly inspection by day and night was maintained for eleven days. It was shown’ that O. vulgaris in this habitat generally hunts throughout the night, with only short excursions from the hole by day.     

Effects of trout on the diel periodicity of drifting in baetid mayflies

Some benthic invertebrates in streams make frequent, short journeys downstream in the water column (=drifting). In most streams there are larger numbers of invertebrates in the drift at night than during the day. We tested the hypothesis that nocturnal drifting is a response to avoid predation from fish that feed in the water column during the day. We surveyed diel patterns of drifting by nymphs of the mayfly Baetis coelestis in several streams containing (n=5) and lacking (n=7) populations of rainbow trout, Oncorhynchus mykiss. Drifting was more nocturnal in the presence of trout (85% of daily drift occurred at night) than in their absence (50% of daily drift occurred at night). This shift in periodicity is due to reduced daytime drifting in streams with trout, because at a given nighttime drift density, the daytime drift density of B. coelestis was lower in streams occupied by trout than in troutless streams. Large size classes of B. coelestis were underrepresented in the daytime drift in trout streams compared to nighttime drift in trout streams, and to both day and night drift in troutless streams. Differences in daytime drift density between streams with and without trout were the result of differences in mayfly drift behaviour among streams because predation rates by trout were too low to significantly reduce densities of drifting B. coelestis. We tested for rapid (over 3 days) phenotypic responses to trout presence by adding trout in cages to three of the troutless streams. Nighttime drifting was unaffected by the addition of trout, but daytime drift densities were reduced by 28% below cages containing trout relative to control cages (lacking trout) placed upstream. Drift responses were measured 15 m downstream of the cages suggesting that mayflies detected trout using chemical cues. Overall, these data support the hypothesis that infrequent daytime drifting is an avoidance response to fish that feed in the water column during the day. Avoidance is more pronounced in large individuals and is, at least partially, a phenotypic response mediated by chemical cues.     

Water flow and water storage in Agave deserti: osmotic implications of crassulacean acid metabolism

Abstract Water flow and water storage were investigated for Agave deserti, a desert succulent showing crassulacean acid metabolism (CAM). The anatomy and water relations of the peripheral chlorenchyma, where CAM occurs, and the central water-storage parenchyma were investigated for its massive leaves so that these tissues could be incorporated as discrete elements into an electrical-circuit analogue of the whole plant. The daily cycling of osmotic pressure was represented by voltage sources in series with the storage capacitors. With soil water potential and leaf transpiration rate as input variables, axial water flow through the vascular bundles and radial flows into and out of storage during the day/night cycle were determined. The predominantly nocturnal transpiration was coincident with increases in cell osmotic pressure and in titratable acid of the leaf chlorenchyma. In the outer layers of the chlorenchyma, water potential was most negative at the beginning of the night when transpiration was maximum, while the water-storage parenchyma reached its minimal water potential 9 h later. The roots plus stem contributed 7% and the leaves contributed 50% to the total water flow during maximal transpiration; peak water flow from the soil to the roots occurred at dawn and was only 58% of the maximal transpiration rate. Over each 24-h period, 39% of the water lost from the plant was derived from storage, with flow into storage occurring mainly during the daytime. Simulations showed that the acid accumulation rhythm of CAM had little impact on water uptake from the soil under the conditions employed. In the outer chlorenchyma, water potential and water flows were more sensitive to the day/night changes in transpiration than in osmotic pressure. Nevertheless, cell osmotic pressure had a large influence on turgor pressure in this tissue and determined the extent to which storage was recharged during the latter part of the night.     

Heat Shock Effects on the Circadian Rhythm of Protein Synthesis and Phosphorylation of Ribosomal Proteins in Gonyaulax polyedra

Circadian changes in protein synthesis and phosphorylation of ribosomal and cytoplasmic proteins in the marine dinoflagellate Gonyaulax polyedra were analyzed by radioactive labeling and polyacrylamide gel electrophoresis. Maximal rates of protein synthesis were found during the subjective night and minimal rates during the subjective day. Protein synthesis was inhibited by heat shock to a different extent at different circadian phases—maximally during the subjective night. Heat shock proteins (HSPs) having molecular weights of approximately 105, 89, 83, 66, 35, and 18 kDa were induced by these treatments. Induction of HSP89 and HSP35 showed circadian differences with maximal synthesis rates at CT 15, whereas most HSPs maintained a constant constitutive and induced synthesis. Recovery of normal protein synthesis after heat shock occurred faster during the subjective night than during the subjective day. Ribosomal proteins with molecular weights of 16 and 18 kDa were highly phosphorylated by [³⁵S] thio gamma adenosine triphosphate during day phase in a light-dark cycle or at CT 6 in constant dim light and labeled only to a minor degree during night phase or at CT 18. A ribosome-associated protein (35 kDa) was labeled during the day and not during the night, but after heat shock during both day and night. In the 200,000 g cytosolic fraction, a 35-kDa protein was found to be more intensely labeled at night than during the day phase after heat shock. The results of this study show a correlation between circadian changes in the overall protein synthesis and ribosomal protein phosphorylation. The rhythm of protein synthesis and phosphorylation of a ribosome-associated protein are drastically altered by heat shock and dependent on the circadian phase.     

Measurement of,and Some Reasons for,Differences in Eating Habits Between Night and Day Workers

A questionnaire was designed to assess the following: why working people chose to eat or not to eat at a particular time of day; the factors that influenced the type of food eaten; and subjective responses to the meal (hunger before, enjoyment during, satiety afterward). Self-assessments were done every 3h during a typical week containing work and rest days, by one group of 50 day workers and another group of 43 night workers. During the night work hours compared to rest days, night workers evidenced a significantly altered food intake, with a greater frequency of cold rather than hot food (p < 0.001). The type and frequency of meals were influenced significantly more (p < 0.05) by habit and time availability and less by appetite. This pattern continued into the hours immediately after the night shift had ended. In day workers food intake during work hours, compared to rest days, was also influenced significantly more often (p < 0.05) by time availability than hunger, but less so than with night workers. Moreover, day workers were less dependent than night workers upon snacks (p = 0.01), and any significant differences from rest days did not continue beyond work hours. Not only did night workers change their eating habits during work days more than did day workers but also they looked forward to their meals significantly less (p < 0.001) and felt more bloated after consuming them (p < 0.05), such effects being present to some extent during their rest days also. These findings have clear implications for measures designed to ease eating problems that are commonly problematic in night workers.