Effect of cycloheximide on the process of "glucose-bleaching" in Chlorella protothecoides

The process of bleaching of Chlorella protothecoides inducedby the addition of glucose was strongly inhibited by cycloheximide,an inhibitor of protein synthesis, whereas it was suppressedonly weakly by chloramphenicol, puromycin and ethionine. Whencycloheximide was added simultaneously with glucose at the beginningof die bleaching experiment, no bleaching of algal cells occurredduring the subsequent incubation. When it was added after glucose,the bleaching of algal cells proceeded for a period of timeas actively as in the control, then gradually ceased. Cycloheximidewas found to suppress the uptake of glucose by algal cells,and to severely inhibit the assimilation of glucose into lipidswhen added at the beginning of the bleaching experiment. Theseinhibitory effects of cycloheximide are discussed in relationto the induction of "glucose-bleaching" in algal cells. (Received December 16, 1968; )     

Difference in nitrate reduction in "light" and "dark" stages of synchronously grown Chlorella pyrenoidosa and resultant metabolic changes

Using synchronized cells of Chlorella pyrenoidosa, the incorporationpatterns of ¹⁴C into various metabolites with and without nitrogensources were studied under steady-state and non steady-stateconditions. From the patterns it was found that the smallestcells which are divided in the dark utilize nitrate and nitritevery little, if at all. The importance of ammonia for regulation of secondary flow forChlorella is discussed and the suggested regulatory points aredescribed. ¹This work was sponsored, in part, by the U.S. Atomic EnergyCommission (Received January 26, 1970; )     

Behavioral Thermoregulation and the "Final Preferendum" Paradigm

Wider attention to Fry's (1947) "final preferendum" paradigmwould facilitate comparative studies of temperature preference(behavioral thermoregulation) among different animal groups.According to Fry's bipartite definition, the final preferendumis that temperature at which preference and acclimation areequal, and to which an animal in a thermal gradient will finallygravitate regardless of its prior thermal experience (acclimation).This paradigm is helpful in distinguishing between acute thermalpreferenda (measured within 2 hr or less after placing an animalin a thermal gradient), which are influenced by acclimationtemperature, and the species-specific final preferendum (measured24–96 hr after placement in the gradient), which is essentiallyindependent of prior acclimation because reacclimation occursduring the gravitation process. The paradigm does not take intoaccount non-thermal acclimatization influences (e.g., season,photoperiod, age, light intensity, salinity, disease, nutrition,pollutants, biotic interactions) which can also affect temperaturepreference. However, a graph of acutely preferred temperaturesversus acclimation temperatures can be employed to determinean equivalent acclimation temperature for any given acclimatizationstate, as a simple means of quantifying acclimatization statesresulting from interactions of many influences. This paradigm,developed for use with fishes, can also be applied to otherectothermic taxa, although it is most easily employed with aquaticorganisms because of the simplicity of specifying aquatic thermalenvironments in terms of water temperature alone. Methodologiesused in studies of behavioral thermoregulation should take theparadigm into account (especially with respect to length oftests) to enhance the comparative value of data across taxa.     

Effect of Auxin and Gibberellin on Conidial Germination in Neurospora crassa II. "Conidial Density Effect" and Auxin

Conidial germination in liquid shake culture of Neurospora crassawas affected by the number of conidia in the medium (conidialdensity effect) with the optimum density being around 2?10⁶conidia/ml. The conidial density effect at 2?10⁴ and 2?10⁵ conidia/mlwas eliminated by the addition of auxin, IAA or 2,4-D with theoptimum concentrations of IAA and 2,4-D being around 10^–6M. IAA and 2,4-D had no effect on the effect at 2?10⁷ conidia/ml.An active substance(s) for conidial germination which was relatedto the conidial density effect was detected in the cell-freefiltrate of the germination medium, and was found to be non-dialyzableand thermolabile. At the early stage of germination, the concentrationof active substance(s) in the medium increased in proportionto the conidial density and reached a supraoptimum amount forgermination at 2?10⁷ conidia/ml. IAA (10^–6M) enhancedthe concentration. Endogenous auxin concentrations in filtratesof the germination media containing 2?10⁵, 2?10⁶ and 2?10⁷ conidia/mlwere 5.8?10^–12, 4.6?10^–11 and 1.7?10^–10M IAAequivalent, respectively. The conclusion reached was that auxinmay control conidial germination with mediation by the activesubstance(s). (Received June 8, 1982; Accepted September 27, 1982)     

Use of the "SMURF" in taste analysis

An improved moving chart recording of intensity/time of tasteresponse has been achieved using a potentiometer ‘dialbox’ linked by a cable to a Telsec recorder. The deviceallows rates of taste response to be determined and is describedas a Sensory Measuring Unit for Recording Flux (SMURF) on theassumption that the flux of stimuli at the taste receptor isresponsible for the time course of response. Fourteen trained and sixteen untrained panellists evaluatedone standard and four unknown sucrose solutions using the SMURFand determined their intensity and persistence time of responsefor each of these same solutions by conventional interval scalingand use of a stop-clock. The SMURF gave results which were higher(but not significantly so) than the conventional method. Trainedpanellists tended to prefer the SMURF and found it quicker andeasier to use than the conventional method. Untrained panelliststended to prefer the conventional method but these results weregenerally not significant. The SMURF is therefore an extremelyuseful device in reducing time and effort whilst still maintainingaccuracy in the measurement of intensity and time of taste response. The SMURF was also used to obtain intensity/time data for threeother sugars so that a comparison between the sugars could bemade.     

Juvenile Hormone: The Status of "Status Quo"

SYNOPSIS. Cuticular proteins show specificity for stage, age,and anatomical region. Analysis of the cuticular proteins ofsecond pupae created by application of juvenile hormone demonstratesthat the hormone prevents the onset of new sequences of synthesesand favors repetition of the region-specific, temporal patternof syntheses used in the previous stage. The argument made isthat juvenile hormone might exert this "status quo" action bypreventing alterations in chromatin configuration. Evidencefrom a wide variety of systems shows that polyamines might beinvolved in reprogramming chromatin. Ecdysterone induces ornithinedecarboxylase, the rate limiting enzyme in animal polyaminesynthesis. I suggest that juvenile hormone might be exertingits status quo action by inhibiting this induction. Preliminarystudies of ornithine decarboxylase induction support this specifichypothesis; experiments with an inhibitor of this enzyme, -difluoromethylornithine, however, do not show the expected juvenile hormonemimicry. Further studies are needed to define the control ofpolyamine biosynthesis in insects and to discover whether juvenilehormone plays a role in this control.     

The "Lost Year" Question in Young Sea Turtles

An adequate understanding of young sea turtle dispersal patternsis necessary for effective management of threatened or endangeredspecies. Such patterns are poorly understood, and the term "lostyear" has been adopted to emphasize this gap in sea turtle lifehistory information. Tag returns from pen-reared yearling seaturtles indicate ocean current dispersal. Evidence indicateshatchlings would be similarly dispersed by ocean currents. Feedingstudies with tank-held animals suggest that food resources areavailable in ocean currents for long-term sea turtle survival.Green turtle (Chelonia mydas) growth appears slow in nature.     

Feeding of freshwater filter-feeders at very low food concentrations: Poor evidence for "threshold feeding" and "optimal foraging" in Daphnia longispina and Eudiaptomus gracilis

The feeding behaviour of two freshwater zooplankton species,the calanoid copepod Eudiaptomus gracilis and the cladoceranDaphnia longispina, at extremely low food concentrations hasbeen studied in order to test whether the animals react as predictedby the models of optimal foraging. Three different sized algae were offered in concentrations downto 1µg C/litre at 7°C during winter and 19°C duringsummer. Ingestion rates were measured by use of a flow-through¹⁴C-technique In contrast to the findings of several authors working withmarine copepods a threshold concentration where the animalsceased filtering could be detected only in one experiment byregression analysis (Daphnia, 19°C, food: Stichococcus).A depression of the filtering rate at low concentrations wasobserved only in Daphnia at 19°C with Stichococcus and Scenedesmusas food. It is supposed that in this case the reduction of thefiltering rate is not a direct response of the filtrator, butis caused by exhaustion due to the experimental conditions. As freshwater zooplankton is very rarely exposed to food abundancesas low as in the marine environment, there seems to be no selectionpressure favouring the evolution of an "optimal forager" behaviour. ⁽¹⁾Supported by Deutsche Forschungsgemeinschaft     

Development of Sink Capacity of the "Storage Root" in a Radish Cultivar with a High Ratio of "Storage Root" to Shoot

The mechanisms that control sink capacity are poorly understood.in radish, a major sink is the "storage root", which beginsto thicken early in development, mainly as a result of thickeningof the hypocotyl. We investigated changes in the accumulationof dry matter, sink activity (increase in dry weight of thehypocotyl per unit of dry weight present per unit of time),carbohydrate content, levels of metabolites, activities of enzymesrelated to the breakdown of sucrose, and the profile of solubleproteins, as well as changes in anatomy, using hypocotyls ofa cultivar with a high ratio of "storage root" to shoot. Wefound that sink activity was strongly related to the level andactivity of sucrose synthase but not to the activity of invertase.We also found a significant correlation between sucrose contentand the level and activity of sucrose synthase. Our resultssuggest that sucrose synthase, but not invertase, might be criticalfor the development of the sink activity of the radish hypocotyland that the level of sucrose might regulate the expressionof sucrose synthase. A discussion of sink capacity is presentedthat includes consideration of structural changes in the hypocotyl. (Received December 14, 1998; Accepted January 27, 1999)     

Development of Sink Capacity of the "Storage Root" in a Radish Variety with a Low Ratio of "Storage Root" to Shoot

Controling mechanisms of sink capacity are poorly understood.Previously we suggested that sucrose synthase (SuSy), but notinvertase, plays an important role for sink capacity of theradish "storage root" in a variety, Raphanus sativus L. (cv.White Cherish) [plant Cell Physiol. (1999) 40: 369]. With thisvariety about 50% of the total dry weight (DW) was in the "storageroot" at 21 d after sowing (DAS). We investigated the sink capacityof another radish variety, R. sativus L. (cv. Kosena) with alow ratio of "storage root" to shoot. With the latter varietyonly 3% of the total DW was in the "storage root" at 21 DAS.Sink activity (increase in DW of the "storage root" per unitof DW present per unit of time) of the "storage root" in Kosenaas well as White Cherish was strongly related to the level andactivity of SuSy but not to the activity of invertase. Theseresults confirmed that SuSy rather than invertase may be criticalfor the development of the sink activity of the radish "storageroot" and that the reaction products of UDP-glucose and fructoseare utilized for sink growth including biosynthesis of the cellwall. In Kosena photosynthates seemed to be partitioned mainlyinto developing leaves and fibrous roots. Differences in partitioningof photosynthates among various sinks with these two varietiesare discussed including anatomical considerations. (Received July 19, 1999; Accepted September 30, 1999)     

Growth of grana from "primary" thylakoids in Phaseolus vulgaris

The plastids of young dark-grown bean leaves, exposed to periodiclight are agranal, devoid of chlorophyll b and contain primarythylakoids and chlorophyll a. Transfer of these plants to continuousillumination results in synthesis of new chlorophyll a, chlorophyllb and grana. This study was done in order to study whether andhow the grana are formed from preexisting primary thylakoids.¹⁴C--aminolevulinic acid was used to label the chlorophyll aof the primary thylakoids, and its fate was studied after transferof the plants to continuous light. It was found that chlorophyll b and grana become ¹⁴C-labelled.The total radioactivity of chlorophyll b per bean increasedwith the parallel decrease of that of chlorophyll a. All subchloroplastfractions, obtained after digitonin disruption of chloroplasts,contained chlorophyll a of equal specific radioactivity. Thespecific radioactivity of chlorophyll b was lower than thatof chlorophyll a, and, in addition, it was lower in the granathan in the stroma lamellae fraction. The data suggest that chlorophyll b is formed from chlorophylla; the grana are formed by stacking of preexisting primary thylakoids;chlorophyll b is synthesized faster in the grana than the stromalamellae; the newly formed chlorophyll a molecules are distributedat random throughout the developing photosynthetic membraneand not on specific growing sites. (Received April 24, 1976; )     

Effects of nitrogen starvation on "stable" RNA synthesis in Rhodotorula gracilis cells

In exponentially growing cells of Rhodotorula gracilis, transferto a nitrogen-free medium causes the acceleration of RNA breakdownprocesses and the arrest of the synthesis of 5 s and 4 s RNA,followed by a progressive fall in the formation of mature rRNAspecies. At the same time, the accumulation of protein graduallydecreases, and the final block corresponds to the block of theaccumulation of rRNA. Labelling experiments suggest that theprogressive arrest of the formation of mature rRNA species depends,at least to a good extent, on the breakdown of the newly-synthesizedrRNA precursor due to hypomethylation of the molecule. The possiblerelationship between the level of amino acids and the synthesisof 5 s and 4 s RNA is discussed. (Received April 22, 1976; )     

The effect of growth regulators on the growth and pigmentation of "Baccara" rose flowers

"Baccara" rose buds were treated with various growth regulatorsduring late stages of bud development. The effect of these substanceson growth and pigmentation were determined. Growth regulatorswere applied by spray or injection or as a lanolin paste, alsoin the nutrient media on which petals were cultured in vitro.Injection of GA into the base of the receptacle caused elongationof the bud whereas IAA, K, ABA, AMO-1618, CCC, and SADH hadlittle or no effect. CCC and MeCl-F did not reduce the elongationcaused by GA. GA treatments also enhanced flower weight andpetal pigmentation and MeCl-F decreased the gibberellin effecton pigmentation. GA treatments of intact flowers and excisedpetals cultured in vitro, were only effective at low temperatures. Gibberellin treatments increased the size of petals, the receptacleand the pedicel only if applied directly to the receptacle.Treatments at lower positions on the flowering shoot eitherhad no effect at all, or caused elongation of only the receptacle. Endogenous gibberellin levels are higher in the receptacle thanin petals or in the pedicel. Injection of GA into the receptaclesignificantly increased gibberellin activity in all flower partswhereas injection into the flowering-shoot base increased gibberellinactivity only in the receptacle. The possibility is discussed that GA, which is exogenously supplieddirectly to the receptacle, enhances flower dimensions and pigmentationby drawing photosynthates to the flower as a consequence ofintensification of the sink. (Received August 17, 1973; )     

Acetate metabolism in the process of "acetate-bleaching" of Chlorella protothecoides

Green cells of Chlorella protothecoides when incubated in amedium containing acetate but no nitrogen source, have beenshown to be bleached as strongly as in glucose-induced bleaching.Using U-¹⁴C-acetate as tracer, the acetate metabolism of algalcells during the process of acetate-induced bleaching was investigated.Changes in algal cell activities for respiration and assimilationof added ¹⁴C-acetate were followed during bleaching processesin "acetate-adapted" and "non-adapted" green cells. As in glucose-inducedbleaching of algal cells, algal cell activity for incorporating¹⁴C into lipids showed the most characteristic change, suggestingthat lipogenesis is causally related to the occurrence of bleachingin algal cells. (Received March 5, 1969; )     

The waiting game: a "battle of waits" between predator and prey

Many prey respond to the presence of a predator by retreatinginto a shell or burrow, or by taking refuge in some other waythat guarantees their safety but restricts further informationfrom being obtained about the predator's continued presence.When this occurs, the individual predator and prey involvedbecome opponents in a "waiting game." The prey must decide howlong to wait for the predator to depart before re-emerging andpotentially exposing itself to attack. The predator must decidehow long to wait for the prey to re-emerge before departingin search of other foraging opportunities. I use a numericalapproach to determine the evolutionarily stable waiting strategyof both players and examine the effects of various parameterson the ESS. The model predicts that each player's waiting distribution—thedistribution of waiting times one would expect to observe forindividuals in that role—will have a characteristic shape:the predator's distribution should resemble a negative exponentialfunction, whereas the waiting time of the prey is predictedto be more variable and follow a positively skewed distribution.The model also predicts that very little overlap will occurbetween the players' waiting distributions, and that the predatorwill rarely outwait the prey. Empirical studies relating tothe model and comparisons between the waiting game and the asymmetricwar of attrition are discussed.     

Male and female cooperatively breeding fish provide support for the "Challenge Hypothesis"

The idea that territorial aggression is regulated by androgensand that aggression itself can modulate androgen levels is wellestablished in males. In many species, females also displayaggressive behavior, yet little work has been conducted on theeffects of female aggression on hormone levels. In this study,we compared the effects of a simulated territory intrusion (amethod for testing the Challenge Hypothesis) on males and femalesof the fish, Neolamprologus pulcher. This cichlid fish fromLake Tanganyika is a particularly useful species to examinesex differences in the behavioral mediation of hormones as breedingpairs remain in a territory year round and both sexes defendthis territory against conspecific and heterospecific intruders.In our study, both sexes indeed aggressively defended theirterritory against a simulated territory intruder. In responseto intruders, both males and females displayed elevated levelsof circulating 11-ketotestosterone, but only females exhibitedincreases in testosterone. Neither aggressing male nor femalefish showed changes in estradiol levels compared to control(nonaggressing) fish. Residents were more aggressive than theintruders and won most of the interactions. However, residents(or winners) did not show higher hormone levels than intruders(or losers). We suggest that aggression commonly modulates androgenlevels in both male and female teleost fish.     

Intra- and Extra-Cellular Localization of "Cytosolic" CuZn-Superoxide Dismutase in Spinach Leaf and Hypocotyl

Immunogold-electron microscopic analysis of spinach leaves donewith the antibody specific for "cytosolic" CuZn-superoxide dismutase(SOD) indicates that SOD is localized in the apoplast, in thenucleus and in, or near, the tonoplast. The association of CuZn-SODwith the nucleus indicates it has a role in preventing fatalmutation caused by reactive species of oxygen. The localizingsite of CuZn-SOD in the apoplastic region of spinach leaf tissuescorresponds to that of the accumulation of lignin. In spinachhypocotyl "cytosolic" CuZn-SOD is localized in vascular tissueswhere lignification and the generation of superoxide respectivelywere shown by the phloroglucin-HCl reaction and formation offormazane from nitroblue tetrazolium. Because hydrogen peroxideis required for lignification via the peroxidase-catalyzed reaction,the CuZn-SOD in the apoplast appears to function in the biosynthesisof lignin by causing rapid disproportionation of the superoxideanion radical prior to its interaction with cellular componentsand peroxidase. (Received March 25, 1996; Accepted June 17, 1996)     

METABOLISM OF GLUCOSE IN THE PROCESS OF "GLUCOSE-BLEACHING" OF CHLORELLA PROTOTHECOIDES

1. As previously demonstrated, normal cells of Chlorella protothecoidesare bleached with degeneration of chloroplasts when they areincubated, under aerobic conditions—either in the lightor in darkness—, in a glucose-containing medium withoutadded nitrogen source ("glucose-bleaching"). It was found inthe present study that under the atmosphere of N₂, neither bleachingnor growth of algal cells occurs in the dark, while in the lighta significant growth of cells takes place with formation ofa certain amount of chlorophyll.
2. Studies on the effects ofvarious inhibitors (ammonium ion,DNP, CMU, -hydroxysulphonates,arsenate, cyanide, azide, andantimycin A) under different conditionsshowed that oxidativephosphorylation is a necessary processfor the occurrence ofthe glucosebleaching as well as the assimilationof glucose(cellular growth). Under light-anaerobic conditionsin the presenceof glucose, assimilation of glucose (cellulargrowth) takesplace being supported by photophosphorylation,but no bleachingoccurs.
3. When the algal cells in the courseof bleaching were transferredto the glucose-free mineral medium,the cell growth ceased immediatelybut the cell bleaching proceededfor several hours before itscessation. The respiratory activity,which was high in the glucose-containingmedium, became loweron transferring the algal cells into theglucose-free medium.The lowered level of respiration was maintained,for more than8 hr after the transfer of cells to the glucose-freemedium.
4. When the cells in the course of bleaching were placed underthe atmosphere of N₂, the cell bleaching ceased almost instantaneously.
5. Based on these observations and other inhibition experiments,it was inferred that a certain intermediate(s) produced by theaerobic respiration of glucose is closely associated with theoccurrence of cell bleaching, and that an O₂-requiring stepmay be involved in the process of chlorophyll degradation.

(Received September 9, 1965; )     

Ecological Bases of Hormone--Behavior Interactions: The "Emergency Life History Stage"

SYNOPSIS. Superimposed upon seasonal changes in morphology,physiology and behavior, are facultative responses to unpredictableevents known as labile (i.e., short-lived) perturbation factors(LPFs). These responses include behavioral and physiologicalchanges that enhance survival and collectively make up the "emergency"life history stage. There is considerable evidence that glucocorticosteroids,and other hormones in the hypothalamo—pituitary—adrenal(HPA) cascade, initiate and orchestrate the emergency life historystage within minutes to hours. This stage has a number of sub—stagesthat promote survival and avoid potential deleterious effectsof stress that may result from chronically elevated levels ofcirculating glucocorticosteroids over days and weeks. Thesesub—stages may include: redirection of behavior from anormal life history stage to increased foraging, irruptivetypemigration during the day, enhanced restfulness at night, andelevated gluconeogenesis. Once the perturbation passes, glucocorticosteroidsmay also promote recovery. Additional evidence from birds indicatesthat glucocorticosteroid responses to a standardized capture,handling and restraint protocol are modulated both on seasonaland individual levels. Field work reveals that these changesin responsiveness to LPFs have ecological bases, such as reproductivestate, body condition etc., that in turn indicate differenthormonal control mechanisms in the HPA cascade.