Staining of Viable and Nonviable Myotubes and of Myofibrils by the Fluorescent Dye Merocyanine 540

Merocyanine 540 (MC 540) has been reported to interact specifically with excitable plasma membranes in live cells [3]. Here we show that the MC 540 fluorescence staining pattern previously believed to be characteristic of viable myotubes [3] is observed in formaldehyde-fixed cells. In contrast, viable myotubes show an MC 540 fluorescence staining pattern that is characteristic of cell surface staining (no internal structures fluoresce). The specific I-band and H-zone fluorescence of isolated myofibrils is also consistent with the interpretation that the fluorescence patterns previously reported for viable myotubes are in fact characteristic of cells with disrupted plasma membranes. Time-course observations of MC 540 and trypan blue staining of myotubes suggest that when plasma membrane integrity is lost, MC 540 fluorescence can be visualized inside the cell 5–10 min before trypan blue absorbance. Thus the trypan blue viability assay can be misleading when applied to myotubes.     

A COMPARISON OF THE GROWTHS OF WILD AND OF CULTIVATED OATS IN MANGANESE-DEFICIENT SOILS

Two species of wild oats, Avena fatua and A. ludoviciana , and two varieties of cultivated oats for comparison, were grown to maturity in pots of manganese-deficient soil with and without added manganese.
A. ludoviciana showed different leaf symptoms of manganese deficiency from A. fatua and the cultivated varieties. In other respects wild and cultivated oats were similar in their responses to the level of manganese supply, though differences were observed in the severity of the effects. These are summarized in Table 6. The relative susceptibility to manganese deficiency of the wild and cultivated oats differed according to the effect considered. The total dry weight at harvest is an integration of all the preceding effects on growth and is therefore presumably the best single criterion for determining susceptibility to manganese deficiency. On the basis of per cent loss of total dry weight at harvest due to lack of manganese, A. fatua is judged less susceptible and A. ludoviciana more susceptible than the two cultivated varieties and no distinction can be made between the latter.
The most interesting differences between wild and cultivated oats in response to the level of manganese supply occurred in seed production. Manganese-deficient wild oats showed a smaller reduction in the number of seeds formed but a greater reduction in the size and manganese content of individual seeds than the cultivated varieties.
Manganese deficiency lowered the percentage of viable seeds and the percentage of dormant seeds produced by both A. fatua and A. ludoviciana.     

Leaf responses to drought stress in Mediterranean accessions of Solanum lycopersicum: anatomical adaptations in relation to gas exchange parameters

In a previous study, important acclimation to water stress was observed in the Ramellet tomato cultivar (TR) from the Balearic Islands, related to an increase in the water‐use efficiency through modifications in both stomatal (g_s) and mesophyll conductances (g_m). In the present work, the comparison of physiological and morphological traits between TR accessions grown with and without water stress confirmed that variability in the photosynthetic capacity was mostly explained by differences in the diffusion of CO₂ through stomata and leaf mesophyll. Maximization of g_m under both treatments was mainly achieved through adjustments in the mesophyll thickness and porosity and the surface area of chloroplasts exposed to intercellular airspace (S_c). In addition, the lower g_m/S_c ratio for a given porosity in drought‐acclimated plants suggests that the decrease in g_m was due to an increased cell wall thickness. Stomatal conductance was also affected by drought‐associated changes in the morphological properties of stomata, in an accession and treatment‐dependent manner. The results confirm the presence of advantageous physiological traits in the response to drought stress in Mediterranean accessions of tomato, and relate them to particular changes in the leaf anatomical properties, suggesting specific adaptive processes operating at the leaf anatomical level.     

Cell Surface Carbohydrates in Tritrichomonas foetus1

The cell surface of Tritrichomonas foetus was characterized by using 18 highly purified lectins with specificities for N-acetyl glucosamine, N-acetyl galactosamine, galactose, mannose, and sialic acid. The specificity of the lectin-induced cell agglutination was verified by inhibition of the agglutination with the specific sugars. By using cytochemical techniques associated with electron microscopy, carbohydrates were detected on the cell surface of T. foetus. The following techniques were used: periodic acid-thiosemicarbazide-silver proteinate, concanavalin A-horseradish peroxidase, and ruthenium red. Anionic sites were detected on the cell surface of the protozoan at pH's 1.8 and 7.2 with the use of colloidal iron hydroxide and cationized ferritin particles, respectively. The binding of colloidal iron particles, as well as the agglutination induced by the lectin from Limulus polyphemus, indicated the presence of sialic acid on the cell surface of T. foetus.     

Organic matter availability structures microbial biomass and activity in a Mediterranean stream

1. We compared microbial biomass (bacteria, fungi, algae) and the activity of extracellular enzymes used in the decomposition of organic matter (OM) among different benthic substrata (leaves, coarse and fine substrata) over one hydrological year in a Mediterranean stream.
2. Microbial heterotrophic biomass (bacteria plus fungi) was generally higher than autotrophic biomass (algae), except during short periods of high light availability in the spring and winter. During these periods, sources of OM shifted towards autochthonous sources derived mainly from algae, which was demonstrated by high algal biomass and peptidase activity in benthic communities.
3. Heterotrophic activity peaked in the autumn. Bacterial and fungal biomass increased with the decomposition of cellulose and hemicellulose compounds from leaf material. Later, lignin decomposition was stimulated in fine (sand, gravel) and coarse (rocks, boulders and cobbles) substrata by the accumulation of fine detritus.
4. The Mediterranean summer drought provoked an earlier leaf fall. The resumption of the water flow caused the weathering of riparian soils and subsequently a large increase in dissolved organic carbon and nitrate, which led to growth of bacteria and fungi.     

Ecologically relevant cryptic species in the highly polymorphic Amazonian butterfly Mechanitis mazaeus s.l. (Lepidoptera: Nymphalidae; Ithomiini)

The understanding of mimicry has relied on a strong biosystematic framework ever since early naturalists first recognized this textbook example of natural selection. We follow in this tradition, applying new biosystematics information to resolve problems in an especially difficult genus of tropical butterflies. Mechanitis species are important components of Neotropical mimetic communities. However, their colour pattern variability has presented challenges for systematists, and has made it difficult to study the very mimicry they so nicely illustrate. The South American Mechanitis mazaeus and relatives have remained particularly intractable. Recent systematists have recognized one highly polytypic species, whereas earlier work recognized the melanic Andean foothill races as a distinct species: Mechanitis messenoides. Recent molecular evidence suggests M. mazaeus and M. messenoides are genetically well differentiated, but evidence of morphological and ecological differences indicative of separate species was still lacking. Thus, it remains to be conclusively demonstrated whether this is an extreme case of a polymorphic mimetic species, or whether distinct co‐mimetic lineages are involved. Here we provide evidence that M. mazaeus and M. messenoides are ecologically distinct and identify consistent morphological differences in both adult and immature stages. These ecological and morphological differences are correlated with mitochondrial sequence data. In spite of some overlap in almost all traits, wing shape, adult colour pattern, and larval colour pattern differ between the two species, in addition to clutch size and larval host use in local sympatry. Although three well‐differentiated mitochondrial DNA (mtDNA) haplogroups were identified within these two species, one for M. mazaeus and two within M. messenoides, no morphological or ecological differences were found between two mtDNA haplogroups, both of which appear to belong to M. messenoides. We conclude that M. mazaeus and M. messenoides are distinct although highly polymorphic species, each with multiple sympatric co‐mimetic forms, and suggest that further work is needed to clarify the identity of other phenotypes and subspecies of Mechanitis. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 106 , 540–560.     

Early Growth of the Developing Ear of Spring Barley

The change in fresh weight of the developing ear and of groupsof spikelets of Maris Mink spring barley was measured for plantsgrown under three different conditions. Growth was closely describedby a third order polynomial equation. Instantaneous values ofrelative growth rate derived from these equations showed a declineto a minimum value, which was dependent on the growing conditionsand on position along the spike, followed by a rapid rise. Theminimum value was reached about 5 d before spikelet initiationceased and the subsequent rise in relative growth rate coincidedwith the period of rapid elongation of the rachis. Within thecar there was a tendency for younger spikelets to have a slightlyhigher relative growth rate than those initiated earlier. Possiblereasons for these growth changes are discussed in relation tospikelet differentiation, vascularization of the ear and nutrientavailability. Hordeum vulgare L., Barley, shoot apex, spikelets, ear, initiation, growth rate