Isolation of Capsicum annuum leaf nitrate reductase and characterization of the effect of adenine nucleotides and NADH on its activity

Abstract. In the preliminary purification of Capsicum leaf nitrate reductase (EC 1.6.6.1), treatment of the crude extract on Sephadex G-25 was necessary to prevent a gelling of the extract and sedimentation of the enzyme. Its K_m values for NADH and nitrate were estimated to be 9.3 and 105mmol m⁻³ ADP and ATP gave hyperbolic competitive inhibition, with respect to NADH, while the inhibition by AMP was linear competitive. K_i values calculated were: ADP and ATP approximately lmol m⁻³ and AMP 2.3 mol m⁻³. Inhibition by ADP was not altered by reduced glutathione.
The Capsicum nitrate reduclase was very susceptible to inhibition by NADH (in the absence of nitrate) and an in vivo assay showed that the activity of the enzyme was limited by the supply of nitrate. NADH and adenine nucleotide levels measured in the Capsicum leaf were used to estimate inhibition of nitrate reductase and a prediction was made of the nitrate reductase activity at different times in the photoperiod. This was shown to follow the same trend as the measured in vivo activity of the enzyme. Changes in adenine nucleotide levels had little effect on nitrate reductase activity.     

New species and new records of recently described species of the coral genus Acropora (Scleractinia: Astrocoeniina: Acroporidae) from Indonesia

Six new species of the coral genus Acropora arc described from Indonesia. These include a species which is remarkable for tubercular cocnostcal structures similar to those of the confamilial genus Montipora. The new species include three regional endemics (A. togianensis and A. batunai from central east Sulawesi and A. derawanensis from east Kalimantan), one species with broad distribution across the southern island chains (A. sukarnoi) and two species which occur throughout most of the Indonesian archipelago (A. Indonesia and A. hoeksemai). A further two species described from Western Australia and Papua New Guinea in 1994 (A. turaki and A. jacquelineae respectively) are recorded from Indonesia for the first time, as common members of an unusual assemblage type in the Togian Islands. The range of another species described from Lombok in 1994 (A. suharsonoi) is extended into Bali. With A. desalwii, A. lokani and A. indiana , this brings to 12 the number of Acropora species newly recorded as being endemic to the Indonesian archipelago or to Indonesia and one adjoining region (either the Indian Ocean or the western Pacific).     

Benthic macroinvertebrate community structure, function and production with respect to habitat type, reach and drainage basin in the southern Appalachians (U.S.A.)

1. Benthic macroinvertebrates were sampled for 1 year to assess functional and taxonomic differences in invertebrate biomass and production with respect to habitat types, reaches and catchments in Wine Spring Basin, western North Carolina. Quantitative samples were collected from depositional, cobble-riffle and bedrock outcrop habitats at four stream reaches (two headwater sites, one second order, and one third order). Other measures included physical parameters, periphyton and organic matter standing crops. Invertebrate data from the Wine Spring catchment were also compared with data from another catchment (Ball Creek) within the same region. 2. The three habitat types had different current velocities and mean substratum particle sizes; both measures were greatest in bedrock outcrop habitats and lowest in depositional habitats. Organic matter standing crops, invertebrate functional group productivity and biomass also differed significantly with respect to habitat type. Cobble-riffle areas had the lowest standing crops of organic matter, invertebrate productivity and biomass. 3. Both invertebrate communities and organic matter standing crops differed significantly between the two headwater reaches. First- to third-order reaches differed in taxonomic composition at the genus level, yet had similar relative functional group productivity and biomass. 4. Annual mean invertebrate biomass and secondary production were greater in the Wine Spring Basin than in Ball Creek. Sites in both the Wine Spring and Ball Creek catchments, however, exhibited similar functional group distributions per habitat type. 5. Local geomorphology and related physical parameters influenced the structure of invertebrate functional group composition, and the distribution of organic matter standing crops. Furthermore, comparison of community structure in Wine Spring with that in Ball Creek suggested that taxonomic composition was more related to catchment-specific parameters (e.g. thermal regime, evolutionary history) than stream size.     

MOVEMENT OF EELWORMS

The optimum crumb sizes for movement of potato-root eelworm larvae in a sandy loam, a heavy clay and a peat soil were 150–250 and 250–400 μ. Mobility was very similar in clay and sandy loam, in both of which there was an optimum suction for movement. In the peat, however, mobility increased with suction and no optimum suction was established. Larvae may be able to move in peat at high suctions because friction between the larvae and the peat crumbs is less than between clay or sand crumbs. Larvae moved to the wet end of a moisture gradient in sand, the number increasing with the steepness of the gradient. The rate of spread of larvae in sand 150–250 μ diameter varied between 2 and 3 cm. a day, depending on suction. As pore size increases, any upward movement in a moisture gradient is opposed by falling under gravity. Larvae do not respond to a moisture gradient or fall under gravity in sand where the width of the pore approximates to the diameter of the larva. The presence of host roots also counteracted the response to a moisture gradient; the degree of orientation to the roots increased with the time the roots were in the sand. Direct observation on larvae, newly emerged from cysts, in the presence of host plant roots, suggests that larvae orientate themselves at a distance from the root and do not reach the root by random movement. Many of the movements of eelworms are explicable by considering the relationship between pore size, eelworm diameter and water distribution, and a diagram relates movement and various soil factors.     

Bacterial swimming strategies and turbulence

Most bacteria in the ocean can be motile. Chemotaxis allows bacteria to detect nutrient gradients, and hence motility is believed to serve as a method of approaching sources of food. This picture is well established in a stagnant environment. In the ocean a shear microenvironment is associated with turbulence. This shear flow prevents clustering of bacteria around local nutrient sources if they swim in the commonly assumed "run-and-tumble" strategy. Recent observations, however, indicate a "back-and-forth" swimming behavior for marine bacteria. In a theoretical study we compare the two bacterial swimming strategies in a realistic ocean environment. The "back-and-forth" strategy is found to enable the bacteria to stay close to a nutrient source even under high shear. Furthermore, rotational diffusion driven by thermal noise can significantly enhance the efficiency of this strategy. The superiority of the "back-and-forth" strategy suggests that bacterial motility has a control function rather than an approach function under turbulent conditions.     

Estrogen-Induced Synthesis of Yolk Proteins in Roosters

Vitellogenin is the serum precursor of the yolk proteins -lipovitellin,rß-lipovitellin, and phosvitin. The precursor canbe dissociated to produce the yolk proteins only by proteolyticenzymatic action, to which it is very susceptible. Denaturationin sodium dodecyl sulfate, combined with reduction of disulfidebridges and blocking of thiols, yields a complex with a molecularweight of 200,000 to 250,000. -Lipovitellin contains three polypeptides,with molecular weights of about 135,000, 105,000, and 40,000,and rß-lipovitellin is composed of two polypeptidechains with molecular weights of 135,000 and 30,000. The 40,000subunit of -lipovitellin and both rß-lipovitellinsubunits are phosphopeptides We tested RNA isolated from the liver of estrogen-treated roostersfor mRNA activity in a cell-free reticulocyte system. The vitellogeninmRNA has a sedimentation coefficient greater than 28S and thuscontains enough information to code for a long polypeptide chain.Estrogen administration to roosters induces the appearance ofvitellogenin and a lowdensity lipoprotein, the syntheses ofwhich are not coordinated. The course of vitellogenin synthesiswas calculated from accumulation and turnover data, and it wasfound that from about 25 hr after estradiol-17rß administrationthe rate of vitellogenin synthesis increases linearly for severaldays, paralleling an increase in vitellogenin-synthesizing polysomes.Thus, we estimate a constant translation rate of about 8 aminoacids per ribosome per sec. A "memory" effect is observed when a second hormone dose isgiven some time after the vitellogenin induced by the firstdose has disappeared from the blood. After the second dose vitellogeninsynthesis is detected sooner, and its initial increase is morerapid, than after the first dose. Although the synthesis ofvitellogenin starts 3 to 4 hr after the second as well as afterthe first injection, the rate of synthesis after the first injectionincreases much more slowly during the first 15 hr than duringthe subsequent period of linear accumulation, whereas afterthe second injection the linear increase in the rate of synthesisbegins immediately after the lag period of 3 to 4 hr. The "memory"effect is undiminished even 50 days after the first hormonedose; thus, the causative factor either is very stable or issynthesized in great excess during the first stimulation. Whenthe second injection is given during the descending part ofthe turnover curve, an increase in vitellogenin synthesis isobserved within 3.5 hr. There are thus at least three different effects of estradiol;(i) the "memory" effect, which probably is due to commitmentor differentiation of vitellogenin-synthesizing cells; (ii)the effect that causes the committed cells to give full responseafter the 3- to 4-hr lag period; and (iii) the effect that causesthe immediate response. To explain these results we suggestthat committed cells can synthesize vitellogenin mRNA only duringa certain period of the cell cycle.     

Laboratory experiments on factors affecting the activity of Gammarus pseudolimnaeus Bousfield

Factors such as current, light, food, substrate and time of year were found to influence the activity of specimens of Gammarus pseudolimnaeus maintained in stream tanks in the laboratory. Although there was seasonal variation, levels of diurnal and nocturnal activity were generally much higher when there was no current than when the water was running. Diurnal periodicity occurred but was not as pronounced as had been indicated in the field studies of previous workers. Although there was activity during the day, nocturnal levels were higher especially in specimens collected during the summer. When there was no current, and the lights were left on for 24 h, peak activity occurred at precisely the times when the lights normally would have been off. This strongly suggests that there is an endogenous component in the activity. The nocturnal activity showed a bigeminus pattern, and there was a seasonal change, with the general activity at its highest levels in summer. Activity was significantly greater during daylight both with and without current when there was no substrate present in the tanks. The behaviour of Gammarus appears to be complex and greatly influenced by combinations of the various factors.