Responses to acute and chronic desiccation stress in <Emphasis Type="Italic">Enchytraeus</Emphasis> (Oligochaeta: Enchytraeidae)

Enchytraeids are small soil living oligochaete worms with high sensitivity to low soil moisture. The effects of acute and chronic desiccation on survival and reproduction were determined in Enchytraeus albidus and Enchytraeus crypticus. Further, effects of acute drought stress on the water balance physiology and accumulation of osmolytes were investigated in E. albidus. Survival of E. crypticus and E. albidus was significantly influenced by exposure time. Reproduction was much more sensitive to desiccation than survival and was significantly reduced from −0.06 bar, which was surprising because no dehydration or change in the body fluid osmolality of E. albidus occurred until much harsher drought regimes occurred. The body fluid osmolality of E. albidus was relatively high, about 500 mOsm. Congruent with this no water loss or changes in osmotic pressure occurred until equivalent or higher water potential values of the environment were reached. Two osmolytes, glucose and alanine, were up-regulated in drought exposed E. albidus. Even though enchytraeids display moderate physiological protection to rapid changes in soil moisture (by having a high osmotic pressure) in the short term, populations subjected to long-term drought stress can be severely reduced even under moderate drought levels.     

Correlation of Local Changes in the Temperature-Dependent Conformational Flexibility of Thioredoxins with Their Thermostability

For the development of a method capable of predicting single point mutations substantially affecting protein thermostability, we studied the effect of the E85R and R82E mutations on the thermostability of thioredoxins from Escherichia coli (Trx) andBacillus acidocaldarius (BacTrx), respectively. The basic method of investigation was the molecular dynamics simulation of 3D protein models in an explicit solvent at different temperatures (300 and 373 K). Some thermolabile regions in Trx, BacTrx, and their mutants were revealed by analyzing the temperature effect on the molecular dynamics of the protein molecule. The effect of single point mutations on the temperature changes of the protein conformation flexibility in several thermolabile regions was found. The results of the simulations are in accord with experimental data indicating that the mutation E85R increases Trx thermostability, whereas the mutation R82E decreases BacTrx thermostability. The thermostability of these proteins was revealed to depend on ionic interactions between the thermolabile regions. The single point mutations change the parameters of these interactions and make them more favorable in the E85R-Trx mutant and less favorable in the R82E-BacTrx mutant.     

Thermal proteome profiling in bacteria: probing protein state in vivo

Increasing antibiotic resistance urges for new technologies for studying microbes and antimicrobial mechanism of action. We adapted thermal proteome profiling (TPP) to probe the thermostability of Escherichia coli proteins in vivo. E. coli had a more thermostable proteome than human cells, with protein thermostability depending on subcellular location—forming a high‐to‐low gradient from the cell surface to the cytoplasm. While subunits of protein complexes residing in one compartment melted similarly, protein complexes spanning compartments often had their subunits melting in a location‐wise manner. Monitoring the E. coli meltome and proteome at different growth phases captured changes in metabolism. Cells lacking TolC, a component of multiple efflux pumps, exhibited major physiological changes, including differential thermostability and levels of its interaction partners, signaling cascades, and periplasmic quality control. Finally, we combined in vitro and in vivo TPP to identify targets of known antimicrobial drugs and to map their downstream effects. In conclusion, we demonstrate that TPP can be used in bacteria to probe protein complex architecture, metabolic pathways, and intracellular drug target engagement.     

Aquatic toxicity test for enchytraeids

It is the purpose of this report to present a simple method for testing the toxicity of chemical substances by using enchytraeids in an aquatic environment. Up to eight different environmental chemicals were applied to various species, mainly of the genus Enchytraeus. The results were compared with those achieved for D. magna. Significant differences, however, between the LC₅₀ values of the various enchytraeid species and the LC₅₀ values for enchytraeids and daphnids could not be observed. For E. cf. buchholzi the toxicological sensitivity of discrete ontogenetic stages was tested. The Aquatic Enchytraeid Test results were compared with those obtained from the Terrestrial Enchytraeid Test. It was found that in soil a chemical could be 600 times less toxic than in water, although the same species (E. albidus) was used in both environments. Even more pronounced were the discrepancies between the terrestrial and aquatic toxicities when the LC₅₀ values for earthworms and daphnids were compared.     

Effectiveness of phytoplankton control by large-bodied and small-bodied zooplankton

Employingin situ enclosures containing inocula of the lake zooplankton (mainlyDaphnia galeata, Daphnia cucullata andBosmina spp.) from a moderately eutrophic Lake Ros (Northern Poland) or large-bodiedDaphina magna, the following observations on succession of phytoplankton were made: 1) whereasD. magna could control the density of all the photoplankton size classes, the lake zooplankton could not suppress the large-sized phytoplankters or net phytoplankton; 2) the lake zooplankton was able to control the density of small algae (< 50μm), but its effect on large algae may be opposite: a promotion of net phytoplankton growth by removing small-sized algae which can out-compete net phytoplankton for limited PO₄-P resources (<5μg P l⁻¹). Since efficiency of phytoplankton density control byD. magna decreased with an increase in net phytoplankton abundance, biomanipulation could not be successful without introducing or maintaining a high population of large-bodied cladoceran species before high densities of large algae would make the control of phytoplankton inefficient.     

Xylan structure,microbial xylanases,and their mode of action

Xylans, the major portion of the hemicellulose of plant cell walls and grasses, are heteropolymers consisting principally of xylose and arabinose. Microbial xylanases with different multiplicities and properties are reported. Most studies on the mode of action of these xylanases have been carried out with fungi and there is very little information available on bacterial xylanases. Fungal xylanases have three or more substrate binding sites: for exampleAspergillus niger, Ceratocytis paradoxa, Cryptococcus albidus andChainia sp. endoxylanases have four to seven subsites with the catalytic site located at the centre of these sub-sites. The analysis of these sub-sites is either by kinetic or end-product analysis studies. Kinetic studies are used for exo-type enzymes while the end-product analysis studies are more convenient for endo-type enzymes. This review covers microbial xylanases with special emphasis on studies of sub-site mapping. The industrial applications of the microbial xylanases are also discussed.     

Laboratory studies on the life-histories of four enchytraeid worms (Oligochaeta) which inhabit sewage percolating filters

The life-histories of four enchytraeid worms, Lumbricillus rivalis, Enchy-traeus coronatus, E. buchholzi, and E. albidus which occur in sewage percolating filters, were studied under laboratory conditions at 8 , 15 and 20°C. The number of ova per cocoon varied from 0 to 50 (L. rivalis), 0 to 33 (E. coronatus), 1 to 9 (E. buchholzi) and 0 to 22 (E. albidus). The mean number of ova per cocoon was highest at 15°C for all species except E. coronatus which had a highest mean value at 8°C. The number of ova in cocoons was correlated with cocoon length (P < 0.001) for all species. Cocoon production usually increased with temperature ranging from 0.8 cocoons per adult per week at 8°C to 2.0 at 20°C for L. rivalis, and from 1–4 to about 2.6 for E. coronatus and E. buchholzi. The total number of ova produced by each E. coronatus (350 at 8°C to 550 at 20°C) was similar to that produced by each L. rivalis (600 at 8°C to 350 at 20°C) and was about five times greater than the total numbers produced by the other two species. Cocoon and ova production and the number of ova per cocoon varied with the age of the adult, usually reaching a peak soon after maturity. Hatching success was low and generally 40–50 % of ova failed to develop; subsequent mortality among immature worms was about 10–20%. Growth was more rapid at the higher temperatures; L. rivalis matured in about 26 days at 20°C, the clitellum forming when the worm was 13–14 mm long; data for the other species are 13 days and 5–6 mm (E. coronatus); 16 days and 3–4 mm (E. buchholzi); 28 days and 13–14 mm (E. albidus). The maturation period at 8°C was at least twice that at 20°C. The generation period (cocoon to cocoon) was about a month at 20°C for all species except E. albidus (2 months), but as some species had longer reproductive periods than others the actual number of generations per year was highest in E. buchholzi, 7.0 per year, and lowest in E. albidus, about 3.3 per year, At 8°C all four species had between 1.4 and 2.8 generations a year. A comparison of expected and observed population densities of L. rivalis and E. coronatus in a sewage percolating filter showed that neither achieved values approaching their potential summer densities although ample food was apparently available. Of the four species studied only E. buchholzi produced viable ova without pairing.     

Evolutionary history of the scorpionfly Dicerapanorpa magna (Mecoptera,Panorpidae)

Climate changes can have fundamental impacts on the distributional patterns of montane species, and range shifts frequently lead to allopatric divergence followed by the establishment of secondary contact zones. Many European and North American organisms have retreated to southern refugia during glacial periods and colonized northward during postglacial periods, but little is known about the evolutionary response of cold‐adapted insects to Pleistocene climate changes in eastern Asia. The scorpionfly Dicerapanorpa magna (Chou), with cold temperate habitat preference and weak dispersal ability, provides a good model system to explore how climate changes have influenced the distribution and divergence of cold‐adapted insects in eastern Asia. This study reconstructed the demographic dynamics and evolutionary history of D. magna with phylogeographic approaches, and predicted the species’ suitable areas under the Last Glacial Maximum (LGM) and current scenarios with the ecological niche modelling analysis. The mitochondrial cytochrome c oxidase subunit I resolved three phylogenetic lineages in D. magna dating back to Pleistocene, corresponding well with the geographically isolated Qinling, Bashan and Minshan Mountains. The ecological niche modelling recovered the suitable habitats for D. magna were the Qinling and Bashan Mountains under LGM and current conditions. The three lineages of D. magna might be in a process of incipient speciation, and likely derived their current distribution from separate glacial origins, followed by vicariance and divergence.     

Rapid local adaptation mediates zooplankton community assembly in experimental mesocosms

Adaptive evolution can occur over similar timescales as ecological processes such as community assembly, but its particular effects on community assembly and structure and their magnitude are poorly understood. In experimental evolution trials, Daphnia magna were exposed to varying environments (presence and absence of fish and artificial macrophytes) for 2 months. Then, in a common gardening experiment, we compared zooplankton community composition when either experimentally adapted or D. magna from the original population were present. Local adaptation of D. magna significantly altered zooplankton community composition, leading to a suppression of abundances for some zooplankton taxa and facilitation for others. The effect size of D. magna adaptation was similar to that of adding fish or macrophytes to mesocosms, two important drivers of zooplankton community structure. Our results suggest that substantial amounts of variation in community composition in natural systems may be unexplained if evolutionary dynamics are ignored.     

Improvement in the biocontrol of postharvest diseases of apples with the use of yeast mixtures

Mixtures of yeasts were tested for theirability to control Penicillium expansum andBotrytis cinerea on Red Delicious apple fruits. The occurrence of synergistic or antagonisticinteractions between yeast strains in differentmixtures was also evaluated. Two strains ofRhodotorula (R. glutinis SL 1 and R. glutinisSL 30) and two strains of Cryptococcus (C. albidus SL 43 and C. laurentii SL 62) were selected fordeveloping yeasts mixtures.The R. glutinis SL 1–R. glutinis SL 30 mixtureexhibited a lower effectiveness than eachstrain alone, against both moulds. Othermixtures (R. glutinis SL 1–C. albidus SL 43 and R. glutinis SL 30–C. albidus SL 43) showedsynergism against P. expansum but not against B. cinerea. The R. glutinis SL 1–C. laurentii SL62 mixture was the only mixture which showedsynergism against gray mould. There was not anymixture, which showed high effectivenessagainst both moulds at the same time. Differentresults could be explained by the dynamics ofthe population of the yeasts.By using yeast mixtures, it was possible toimprove biocontrol without increasing theamount of antagonists applied. The synergismobserved could be useful in enhancingbiological control.     

Osmoregulation in two aquatic oligochaetes from habitats with different salinity and comparison to other annelids

The osmoregulatory capacity of two oligochaete species, Enchytraeus albidus Henle, 1837, and Heterochaeta costata (Claparède, 1863), was investigated by direct measurements of the osmolality of the coelomic fluid. Terrestrial and marine (28 S) populations of Enchytraeus albidus and a brackish water population (14 S) of H. costata were used in the study. The range of salinity acclimation investigated was 0–40. The response to osmotic stress was measured (a) after a long-term maintenance (>14 days) in various salinities (E. albidus only), and (b) after a hyperosmotic shock as a short-term time-course sequence. The rate of water loss following a hyperosmotic shock was measured for E. albidus. Long-term acclimation. E. albidus maintained a hyperosmotic coelomic fluid over all salinities tested. In low salinities the osmolality of the coelomic fluid of the marine population was significantly higher than that of the terrestrial population. Possible genetic discrepancies or long-term acclimation may account for this difference. The coelomic fluid of H. costata was hyperosmotic at 15%. S and isoosmotic at 30 S. Short-term acclimation (hyperosmotic shock). Both species investigated, kept at 15 S and then exposed to a salinity of 30, showed fast responses: within the first two hours the internal concentrations were adjusted to the new external condition with only small subsequent changes. Regulation of the body-water content after an exposure to a hyperosmotic shock was much slower: individuals of terrestrial E. albidus, acclimated for two weeks to either 0 or 15 S, had the same water content; hence, they showed a 100% regulation. However, after exposure to 30 S, a 100% regulation was still not attained 4 days after the hyperosmotic shock. Enchytraeus albidus is capable of actively reducing water loss following the hyperosmotic shock: the observed loss of water was only 40% of that expected for a passive osmotic flow. The observed reactions are compared with those in other annelids. It seems that an active transport of ions combined with a changeable permeability of the body wall play a major role in the regulation of body fluids.     

Freeze tolerance and accumulation of cryoprotectants in the enchytraeid Enchytraeus albidus (Oligochaeta) from Greenland and Europe

The freeze tolerance and accumulation of cryoprotectants was investigated in three geographically different populations of the enchytraeid Enchytraeus albidus (Oligochaeta). E. albidus is widely distributed from the high Arctic to temperate Western Europe. Our results show that E. albidus is freeze tolerant, with freeze tolerance varying extensively between Greenlandic and European populations. Two populations from sub Arctic (Nuuk) and high Arctic Greenland (Zackenberg) survived freezing at −15 °C, whereas only 30% of a German population survived this temperature. When frozen, E. albidus responded by catabolising glycogen to glucose, which likely acted as a cryoprotectant. The average glucose concentrations were similar in the three populations when worms were frozen at −2 °C, approximately 50 μg glucose mg⁻¹ tissue dry weight (DW). At −14 °C the glucose concentrations increased to between 110 and 170 μg mg⁻¹ DW in worms from Greenland. The average glycogen content of worms from Zackenberg and Nuuk were about 300 μg mg⁻¹ DW, but only 230 μg mg⁻¹ DW in worms from Germany showing that not all glycogen was catabolised during the experiment. Nuclear magnetic resonance spectrometry (NMR) was used to screen for other putative cryoprotectants. Proline, glutamine and alanine were up regulated in frozen worms at −2 °C but only in relatively small concentrations suggesting that they were of little significance for freeze survival. The present study confirms earlier reports that freeze tolerant enchytraeids, like other freeze tolerant oligochaete earthworms, accumulate high concentrations of glucose as a primary cryoprotectant.     

Resource-niche complementarity and autotrophic compensation determines ecosystem-level responses to increased cladoceran species richness

This study examines the relationship between cladoceran species richness and ecosystem functioning. I conducted an experiment in which four cladocerans, Daphnia. magna, D. longispina, D. pulex and Chydorus sphaericus, were cultured in microcosms using different species combinations and levels of species richness. The results demonstrate that even within this closely related group of organisms the effects on ecosystem-level variables, such as total algae and zooplankton biomass, per capita productivity, and nutrient concentrations, as well as phytoplankton community structure, were highly variable between different combinations of these species. Since only four species where involved in this study, species-specific effects dominated the general relationship between species richness and ecosystem functioning. Particular combinations of species resulted in effects that indicated more efficient grazing. These effects, which were most pronounced in combinations including both D. magna and C. sphaericus, were manifested as an indirect effect as the prey community shifted towards grazing-resistant species. As a result, the productivity of the prey community decreased, because phytoplankton species with lower per capita productivity became more dominant. I suggest that the primary mechanism that caused this significant effect was complementarity in prey-size use of D. magna and C. sphaericus. In terms of prey-size range, D. pulex and D. longispina were redundant when D. magna was present and were quickly out-competed by the latter despite higher per capita filtering efficiency. The results show that different mechanisms are important for different combinations of species. Furthermore, the ability of the prey community to respond to changes of consumer species composition is an important factor in experiments in which consumer species richness is experimentally manipulated. Received: 2 November 1998 / Accepted: 8 September 1999     

Absorption of neutral and basic amino acids across the body surface of two annelid species

Summary 1. Absorption of L-arginine across the body surface of the oligachaeteEnchytraeus albidus and the polychaeteNereis diversicolor from 10 µM ambient concentrations proceeds at only few percentages of glycine and L-valine uptake rates. Distribution ratios do not exceed equilibrium. InE. albidus L-lysine is transported at the same low rates as observed for L-arginine.2. InE. albidus the concentration dependent uptake of L-arginine consits of a saturable component and a diffusion component. The saturable component is susceptible to inhibition by structurally analogous substances. This inhibition is strongest in the presence of other basic amino acids and is still significant in the presence of neutral amino acids. Slight inhibition of L-arginine absorption is caused by L-glutamine and L-asparagine, while inhibition of arginine is only inconsiderable in the presence of acidic and- and-amino acids. Arginine uptake rates are unchanged by organic acids and carbohydrates.3. Because of nonsusceptibility of arginine transport to metabolic inhibitors (NaCN and ouabain) and Na⁺-depletion inE. albidus, the saturable component of arginine uptake is regarded to fulfill the requirements for applying the term facilitated diffusion, which is discussed in relation to the active transport of neutral amino acids.4. The mode of basic amino acid absorption is discussed in comparison to that in other animal species and absorbing tissues.

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Absorption von neutralen und basischen Aminosäuren durch die Körperoberfläche zweier Annelidenarten

Kurzfassung In Versuchen zur Aufnahme verschiedener Aminosäuren durch die Körperoberfläche bei dem OligochaetenEnchytraeus albidus und dem PolychaetenNereis diversicolor betrug die Absorptionsrate von¹⁴C-L-Arginin aus einer Umgebungskonzentration von 10 µM nur wenige Prozent der für Glycin und L-Valin bestimmten Werte. Nach einer halbstündigen Versuchsdauer waren neutrale¹⁴C-Aminosäuren im Organismus 9- bis 15fach gegenüber dem Medium angereichert, die L-Argininverteilung blieb dagegen unterhalb des Äquilibriums. Die konzentrationsabhängige Aufnahmekurve für L-Arginin bestand beiE. albidus aus einer Diffusions- und einer Sättigungskomponente, die durch andere Aminosäuren hemmbar war. Die stärkste Inhibition erfolgte durch basische Aminosäuren. Eine erhelbiche Hemmung wurde auch durch neutrale Aminosäuren bewirkt. L-Glutamin und L-Asparagin verminderten die L-Argininaufnahme nur geringfügig. Die Gegenwart von sauren,- und-Aminosäuren sowie organischen Säuren und Kohlenhydraten hatte nur einen unbedeutenden oder gar keinen Einfluß zur Folge. Da die L-Argininaufnahme nicht dem Einfluß von Stoffwechselinhibitoren (NaCN und Ouabain) unterlag und Na⁺-unabhängig war, erfüllt die Sättigungskomponente des L-Arginintransports die Merkmale der erleichterten Diffusion. Der Unterschied zu neutralen Aminosäuren, welche aktiv transportiert werden, wird diskutiert.

     

ATP and related purines stimulate motility,spatial congregation,and coalescence in red algal spores

Adenosine 5′‐triphosphate (ATP) is a versatile extracellular signal along the tree of life, whereas cAMP plays a major role in vertebrates as an intracellular messenger for hormones, transmitters, tastants, and odorants. Since red algal spore coalescence may be considered analogous to the congregation process of social amoeba, which is stimulated by cAMP, we ascertained whether exogenous applications of ATP, cAMP, adenine, or adenosine modified spore survival and motility, spore settlement and coalescence. Concentration‐response studies were performed with carpospores of Mazzaella laminarioides (Gigartinales), incubated with and without added purines. Stirring of algal blades released ADP/ATP to the cell media in a time‐dependent manner. 10–300 μM ATP significantly increased spore survival; however, 1,500 μM ATP, cAMP or adenine induced 100% mortality within less than 24 h; the exception was adenosine, which up to 3,000 μM, did not alter spore survival. ATP exposure elicited spore movement with speeds of 2.2–2.5 μm · s^?1. 14 d after 1,000 μM ATP addition, spore abundance in the central zone of the plaques was increased 2.7‐fold as compared with parallel controls. Likewise, 1–10 μM cAMP or 30–100 μM adenine also increased central zone spore abundance, albeit these purines were less efficacious than ATP; adenosine up to 3,000 μM did not influence settlement. Moreover, 1,000 μM ATP markedly accelerated coalescence, the other purines caused a variable effect. We conclude that exogenous cAMP, adenine, but particularly ATP, markedly influence red algal spore physiology; effects are compatible with the expression of one or more membrane purinoceptor(s), discarding adenosine receptor participation.     

Promotion of Monacolin K Production by <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis>-Mediated Transformation in <Emphasis Type="Italic">Monascus albidus</Emphasis> 9901

The binary vector pCAMBIA3300-gpdA-hph-trpC with hygromycin B phosphotransferase (hph) was constructed and transformed into Monascus albidus 9901 by Agrobacterium tumefaciens-mediated transformation, with gene hph as the selective marker. In order to improve the efficiency of A. tumefaciens-mediated transformation in M. albidus 9901, we optimized various factors including concentration of M. albidus 9901 spores, cell density of A. tumefaciens, co-cultivation time, temperature, and acetosyringone concentration. Most transformants of M. albidus 9901 could grow stably on media containing 50 μg ml⁻¹ hygromycin B up to five generations. The presence of hph was identified by PCR. Two transformants H1 and H2 which produced more Monacolin K than M. albidus 9901 were screened, and the concentration of Monacolin K in the fermented millet by H1 and H2 increased by 42.15% and 40.34% respectively compared with that produced by M. albidus 9901.     

Enhancing thermostability of <Emphasis Type="Italic">Escherichia coli</Emphasis> phytase AppA2 by error-prone PCR

Phytases are used to improve phosphorus nutrition of food animals and reduce their phosphorus excretion to the environment. Due to favorable properties, Escherichia coli AppA2 phytase is of particular interest for biotechnological applications. Directed evolution was applied in the present study to improve AppA2 phytase thermostability for lowering its heat inactivation during feed pelleting (60–80°C). After a mutant library of AppA2 was generated by error-prone polymerase chain reaction, variants were expressed initially in Saccharomyces cerevisiae for screening and then in Pichia pastoris for characterizing thermostability. Compared with the wild-type enzyme, two variants (K46E and K65E/K97M/S209G) showed over 20% improvement in thermostability (80°C for 10 min), and 6–7°C increases in melting temperatures (T _m). Structural predictions suggest that substitutions of K46E and K65E might introduce additional hydrogen bonds with adjacent residues, improving the enzyme thermostability by stabilizing local interactions. Overall catalytic efficiency (k _cat / K _m) of K46E and K65E/K97M/S209G was improved by 56% and 152% than that of wild type at pH 3.5, respectively. Thus, the catalytic efficiency of these enzymes was not inversely related to their thermostability.     

Redescription of Ilyocryptus brevidentatus Ekman, 1905 (Anomopoda,Cladocera, Branchiopoda)

During the wet season of 1999–2000, we studied the effects of the hydroperiod and other physical and chemical variables on planktonic copepod communities from six stations in Everglades National Park. Two stations were located in a slough (Taylor Slough 1, Taylor Slough 2) and four stations in the marl prairies of the Rocky Glades (Long Pine Key 7, Long Pine Key 8, Pa-hay-okee, Chekika). During the period of investigation, Taylor Slough sites had the longest hydroperiods, together with Pa-hay-okee, which is located near the eastern edge of Shark River Slough. Long Pine Key 7 and Long Pine Key 8 had the shortest hydroperiods, and Chekika had an intermediate hydroperiod. The pineland edge sites in the southern Rocky Glades (Long Pine Key) had higher numbers of individuals, and high percentages of larval stages, especially at the end of the wet season. The pineland ecotone is morphologically very heterogeneous, with solution holes in the limestone bedrock that provide below-ground refugia when there is no water on the marsh surface. The slough stations had the lowest numbers of individuals, as well as Chekika in the Rocky Glades, probably as a consequence of the altered water quality and hydropatterns caused by water management structures and operations We collected two species of calanoids, 18 cyclopoids, and three harpacticoids. The most abundant species were Acanthocyclops robustus, Tropocyclops prasinus mexicanus, Arctodiaptomus floridanus, Mesocyclops americanus, Macrocyclops albidus, Osphranticum labronectum, Microcyclops varicans, Microcyclops rubellus, Eucyclops conrowae, and Mesocyclops edax. Of these species, T. prasinus mexicanus and A. floridanus seemed to be adapted to short-hydroperiod habitats, M. rubellus and M. varicans to longer hydroperiod habitats, and E. conrowae to high conductivity habitats. Acanthocyclops robustus, M. albidus, and O. labronectum were dominant regardless of hydroperiod. As regards the temporal distribution, A. robustus was abundant throughout the entire wet season, M. edax, M. rubellus, M. americanus and M. varicans were most abundant in mid-wet season, in September–October, and T. prasinus mexicanus, M. albidus, and E. conrowae were abundant late in the wet season, in winter. The two calanoids only slightly overlapped in time: A. floridanus was abundant at the beginning of the wet season, in July–August, and O. labronectum was abundant at the end of the wet season, in December.     

Studies on Isoleucyl-tRNA Synthetase of an Extreme Thermophile,Thermus thermophilus HB8

Isoleucyl-tRNA synthetase (IRS) was partially purified from an extreme thermophile, T. thermophilus HB 8. The molecular weight (11.5 ×10⁴) and some kinetic constants were obtained and compared with IRS from other sources.

The present IRS catalyzed both isoleucine dependent and valine dependent ATP-PPi exchange reactions (optimum at around 80°C) but not valyl-tRNA formation. The optimum temperature for isoleucyl-tRNA formation was 62°C with E. coli tRNA and 75°C with T. thermophilus tRNA.

The enzyme showed a remarkable thermostability. The addition of E. coli or T. thermophilus tRNA enhanced the thermostability of the enzyme, which was shown to be fully active up to 77°C. When E. coli tRNA was used, the loading activity decreased in parallel to the unfolding of the substrate tRNA molecule. From these results the relation is discussed between tRNA conformation and function.