Metabolism of auxin in pine tissues: Indole-3-acetic acid conjugation

Incubation of sections of various tissues of Pinus pinea L. with a relatively low concentration (3.6 μM) of indole-3-acetic acid-2-¹⁴C (IAA) resulted in the formation of two major metabolites. The first, which has not been identified, seemed to be a polar acidic compound and the second was identified as indole-3-acetylaspartic acid (IAAsp). The polar acidic metabolite has been found to be the major metabolite in needles, shoot wood and roots, while IAAsp has been found to be the major metabolite in shoot bark. Increasing the concentration of IAA in the incubation medium resulted in an increase in the formation of a third metabolite which proved to be l-O-(indole-3-acetyl)-β-d -glucose (IAGlu) and a concomitant decrease in the amount of the polar acidic metabolite. This phenomenon was prominent particularly in needles. IAGlu was isolated from needles and IAAsp was isolated from shoot bark by means of polyvinylpolypyrrolidone column chromatography and preparative thin-layer chromatography. IAGlu was identified by comparison with authentic material by co-chromatography in three different solvent systems and by ¹H-nuclear magnetic resonance analysis. IAAsp was identified by comparison with authentic material by gas-liquid chromatography and ¹H-nuclear magnetic resonance analysis. Several aspects of formation, separation and isolation of IAA metabolites are discussed.     

Effect of predatory fish presence on habitat use and diel movement of the stream amphipod, Gammarus minus

SUMMARY.

• 1 We examined how the presence of predatory fish affected macro—and microhabitat use and movement patterns of the amphipod, Gammarus minus Say, in a second order stream.
• 2 Among macrohabitats, amphipods were consistently more abundant in runs than in pools. Densities in pools were low regardless of fish presence, whereas low densities in runs were correlated with fish presence. Siltation and the lack of coarse substrate particles probably accounted for the scarcity of G. minus in pools. Among microhabitats, G. minus was more abundant in leaf litter and gravel than in silt/sand substrates. Fish presence did not affect microhabitat use in the field. Laboratory manipulations showed, however, that predation rates by green sunfish (Lepomis cyanellus Rafinesque) were significantly lower on amphipods in leaf litter than in other microhabitats.
• 3 Most movement by G. minus in runs occurred at night and was directed downstream. In the laboratory, amphipods significantly reduced swimming activity at night in response to water conditioned with green sunfish. In the field, however, amphipods were only slightly less active in runs at night when exposed to chemical cues of confined green sunfish. The distribution of amphipods in streams is influenced by the presence of fish, but chemical cues alone may not be important in triggering predator avoidance behaviours in nature.

     

Site of Nitrogenase Activity in the Blue-Green Alga <Emphasis Type="Italic">Anabaena</Emphasis> sp. L-31

MANY blue-green algae fix nitrogen, assimilate carbon dioxide and evolve oxygen and as algal nitrogenase is inhibited^1–3 by high oxygen pressure, enhanced nitrogen fixation accompanying photosynthesis is surprising. Heterocysts do not contain⁴ or have comparatively less amounts^4–7 of photosystem II (PS II) pigments, which are responsible for the evolution of oxygen. This tends to favour the suggestion of Fay et al.⁸ that these cells are the sites of nitrogenase activity. Until now, however, attempts at obtaining unequivocal evidence for heterocysts as principal loci for nitrogenase activity have yielded conflicting results. Stewart et al.⁷ first demonstrated nitrogenase activity in heterocysts incubated aerobically, a finding confirmed by Wolk and Wojciuch⁹ and Van Gorkom and Donze¹⁰. By contrast, Smith and Evans^3,11 and Kurz and La Rue¹² reported results favouring vegetative cells as the major site of nitrogenase activity. Other evidence^2,13 showed high nitrogenase activity in cell-free preparations of Anabaena cylindrica and the non-heterocystous alga Plectonema boryanum strain 594.     

Low Pressure and Ethylene in Lettuce Seed Germination

The extent and manner of ethylene involvement in germination of lettuce (Lactuca Sativa L. cv. Mesa 659) seed at a moderate temperature (20°C) were investigated. Inhibition of germination at low pressure of 150 mmHg in an oxygen flow-through system was alleviated to a marked extent by ethylene. Carbon dioxide was ineffective by itself but caused further alleviation of inhibition in presence of ethylene and oxygen. Other seed treatments which partially alleviated the inhibition caused by low pressure included soaking in 10μM of fusicoccin and a prior treatment with acetone. Of the two ethylene adsorbents used, Purafil was more effective in inhibiting germination in a closed container. Although the ethylene biosynthesis inhibitor, 8-hydroxyquinoline (1.0 mM). showed no effect on ethylene production, it markedly inhibited germination and the effect was partially reversed by ethylene and GA₃. An ethoxy analog of rhizobitoxine, on the other hand, had little or no effect on germination but strongly inhibited the ethylene production. Although no causal relation of ethylene to germination was established, the evidence presented here implicates ethylene, together with other gases, in the regulation of germination.     

Materials and methods for carbon dioxide and water exchange analysis§

Abstract Some of the materials which are commonly used in equipment for carbon dioxide and water exchange analysis can adsorb substantial amounts of carbon dioxide and water and, thus, may cause large experimental errors. Also presented are procedures for determining the influence of light intensity, carbon dioxide concentration, and temperature upon photosynthesis and transpiration.     

The Nutrition of Tetrahymena setifera HZ-1; Sterol and Alcohol Requirements

SYNOPSIS. In a chemically-defined medium Tetrahymena setifera HZ-1 required a sterol, an alcohol, eleven amino acids, a purine, a pyrimidine, and six B-complex vitamins. The sterol requirement was met by a variety of 3β-OH, C₂₇-C₂₉ sterols including cholesterol and stigmasterol, but not by precursors of cholesterol which precede desmosterol or Δ⁷-cholestenol. Some combinations of long-chain fatty acids with a synthetic dipalmitoyl phosphorylethanolamine partly substituted for sterol. Ethyl and methyl alcohols (but not a variety of other alcohols and organic acids) satisfied the alcohol requirement.     

Has the importance of the amniote egg been overstated?

The evolution of the amniote egg is commonly regarded as an important milestone in the history of the vertebrates, an innovation that completed the transition from aquatic to fully terrestrial existence by permitting eggs to be laid away from standing water. This view derives ultimately from the recapitulationist theories of Haeckel, and rests on the assumption that extant frogs and salamanders are good models for the reproductive habits of early tetrapods and the ancestors of the amniotes. It also assumes that it is more difficult to lay eggs on land than in water, and that the amniote egg is an adaptation to the physical rigours that eggs encounter in terrestrial environments. Taken together, these assumptions comprise what may be termed the 'Haeckelian framework' for the origin of vertebrate terrestriality. Several independent lines of evidence suggest that the assumptions of the Haeckelian framework are false. There appear to be no theoretical reasons to assume that the evolution of terrestrial egg-laying was difficult, or required a structure as elaborate as the amniote egg. The physical conditions eggs encounter in the terrestrial environments where they are actually laid are quite mild. Land may in fact be an easier place to lay eggs than water. In addition, analysis of the distribution of key reproductive character states among vertebrates provides no evidence that the 'typical amphibian' reproductive mode is primitive for tetrapods. Amniotes are as likely as frogs or salamanders to retain primitive reproductive character states.     

THE USE OF STREPTAVIDIN COATED MAGNETIC BEADS FOR DETECTING PATHOGENIC BACTERIA BY LIGHT ADDRESSABLE POTENTIOMETRIC SENSOR (LAPS)

A modified procedure for magnetic capture of antibody-conjugated bacteria for light addressable potentiometric sensor (LAPS) detection using the Threshold System was developed. Streptavidin coated magnetic beads, partially labeled with biotinylated anti Escherichia coli O157 antibodies, were used to capture Escherichia coli O157:H7. Captured bacteria were further labeled with fluorescein-conjugated anti -E. coli O157:H7 antibodies and urease-labeled. anti-fluorescein antibody. Magnetically concentrated bacteria-containing complexes were then immobilized through streptavidin-biotin interactions on 0.45 μ biotinylated nitro-cellulose membranes assembled as sample sticks for the Threshold instrument. The rate of pH change associated with the production of NH₃ by the urease in urea-containing solution was measured by a LAPS incorporated in the Threshold instrument. This approach allowed us to detect 10³ to 10⁴ CPU of cultured E. coli O157:H7 in PBS solutions. Furthermore, detectable LAPS signals of the sample sticks remained relatively constant for at least 24 h at 4C. The developed approach was applied to detect the E. coli in beef hamburger spiked with the bacteria. After a 5 to 6-h enrichment at 37C, as low as 1 CFU/g of E. coli O157:H7 in beef hamburger could be detected.     

A STATISTICAL ANALYSIS OF DIFFERENCE TESTS WITH REPLICATIONS

This paper proposes a statistical method for difference tests with repetitions. Classical methods for difference tests are based upon the binomial distribution, and are not concerned with the number of repetitions per judge. But when more than one replication of a difference test is required, judgements from different judges are more independent than replicates from the same judge; these two cannot be combined in the classical methods. In this paper, we propose another approach that takes into account two points: the number of repetitions per judge, and the differences within subjects. Two examples are presented to illustrate this approach.     

Trophic interactions and benthic animal community structure in the Colorado River, Arizona, U.S.A.

1. Cladophora glomerata is the dominant filamentous green alga in the tailwaters of the Colorado River, U.S.A., below Glen Canyon Dam, but becomes co-dominant with filamentous cyanobacteria, Oscillatoria spp., below the confluence of the Paria River (26km below the dam) where suspended sediments are elevated. 2. Benthic algal assemblages played an important role in the distribution of the amphipod, Gammarus lacustris, in the dam-controlled Colorado River through Grand Canyon National Park, Arizona. Cladophara and G. lacustris showed a weak positive relationship at ten cobble-riffle habitats in the Colorado River from Lees Ferry (25km below the dam) to Diamond Creek (362km downstream), while no relationship was found between Oscillatoria and G. lacustris. 3. The relationship between algal substrata and G. lacustris was tested by a series of in situ habitat choice experiments. G. lacustris showed a significant preference for Cladophora (with epiphytes) over Oscillatoria spp., detritus and gravel in treatment pans at Lees Ferry. 4. Epiphytic diatoms (i.e. food) were the overriding determinant of subtratum choice by G. lacustris in laboratory experiments. Gammarus chose the Cladophora/epiphytic diatom community over sonicated Cladophora with few diatoms. The amphipods also chose string soaked in diatom extract over string without diatom extract. 5. Importance of mutualistic interactions in aquatic benthic community structure is discussed.