Release of Membrane-Bound Trails by Trypanosoma cruzi Amastigotes onto Modified Surfaces and Mammalian Cells

ABSTRACT. Upon incubation at 37° C onto glass coverslips coated with Concanavalin A, poly-L-lysine, or a monoclonal antibody (1D9) directed to the parasite major surface glycoprotein Ssp-4, extracellular Trypanosoma cruzi amastigotes release trails of material barely visible by light microscopy. This release is not associated with parasite movements. Immunolabeling studies confirmed that the material is derived from the parasite's membrane since thin section through samples labeled with 1D9 revealed that the trails are membrane-bound structures. Scanning electron microscopy showed that the ∼0.1-μm thick trails of material emerging from the amastigotes can be uniform or beaded, indicating a tendency to vesiculation. The trails are preferentially released from the flagellar pocket region and/or at the opposite posterior end of the parasite body, and seem to be devoid of microtubules. The release is time and temperature-dependent and fixed parasites do not form trails. All attempts to inhibit trail release using drugs (antimycin A, sodium azide, cytochalasin D, nocodazole, genistein, staurosporine, EGTA) failed. The observation of trails associated with intracellular parasites and amastigotes invading Vero cells suggests that this is probably a physiological process.     

Virgin and recently mated males are equally able to achieve new matings in Neoleucinodes elegantalis

Lepidopteran male mating success is recognized to be directly related to physical and behavioural traits such as ability, vigour, activity and persistence in courtship. In the tomato fruit borer Neoleucinodes elegantalis Guenée, the mating system is known to be monogamic and therefore males' sexual investment is apparently low. The hypothesis that recently mated males have a remating probability equal to that of virgin males is tested. The impact of body size in remating success and the cost of remating are also analyzed. Mated males show as much propensity to remate as naïve ones. Copula duration and the time taken to copulate are similar in mated and virgin males. However, spermatophore size is not related to male size. The results suggest the ability of N. elegantalis males to remate within 24 h between mating events and their propensity to remate are not affected by copula investment. © 2013 The Royal Entomological Society     

Differential requirement for NO during ABA-induced stomatal closure in turgid and wilted leaves

Abscisic acid (ABA)-induced stomatal closure is mediated by a complex, guard cell signalling network involving nitric oxide (NO) as a key intermediate. However, there is a lack of information concerning the role of NO in the ABA-enhanced stomatal closure seen in dehydrated plants. The data herein demonstrate that, while nitrate reductase (NR)1-mediated NO generation is required for the ABA-induced closure of stomata in turgid leaves, it is not required for ABA-enhanced stomatal closure under conditions leading to rapid dehydration. The results also show that NO signalling in the guard cells of turgid leaves requires the ABA-signalling pathway to be both capable of function and active. The alignment of this NO signalling with guard cell Ca²⁺-dependent/independent ABA signalling is discussed. The data also highlight a physiological role for NO signalling in turgid leaves and show that stomatal closure during the light-to-dark transition requires NR1-mediated NO generation and signalling.     

Influence of Radiotransmitters on Fecal Glucocorticoid Levels of Free-Ranging Male American Kestrels

ABSTRACT Although radiotelemetry is considered a valuable technique for ornithological field studies, several assumptions have been made about the impact that transmitters may have on the estimation of behavioral, ecological, and reproductive parameters. To assess the potential effects of backpack radiotransmitters, we captured and assigned 8 male American kestrels (Falco sparverius) into 2 groups: radiotagged (n = 6) and control individuals (leg-banded, n = 2). Thereafter, we collected feces approximately 2 hours after capture (day −1), and subsequently during days 0 (releasing day), 4, 7, 15, 30, 40, and 55. Prior to fecal analysis, we validated the corticosterone enzyme immunoassay using standard procedures (e.g., parallelism, dose-response curve), and we confirmed physiological significance of fecal glucocorticoid metabolites through adrenocorticotropin challenge, which induced an increase of 4-fold (446.10 ± 60.73 ng/g) above baseline (114.27 ± 15.23 ng/g) within 4 hours (P < 0.001). Both groups exhibited a significant increase in fecal glucocorticoids during day 0 (P < 0.001), but concentrations returned to preattachment values within 4 days. Fecal glucocorticoid concentrations did not differ between samples of radiotagged and leg-banded kestrels (P > 0.05). In spite of the small number of monitored subjects, these findings suggested that radiotransmitters did not affect adrenocortical activity in these male American kestrels.     

Leaf anatomical variation in Alchornea triplinervia (Spreng) Mull. Arg. (Euphorbiaceae) under distinct light and soil water regimes

Alchornea triplinervia (Spreng.) Müll. Arg. (Euphorbiaceae) is a tree which occurs in a broad range of habitats in Brazil. In the State of Rio de Janeiro, it occurs from montane forests to swamplands at sea level. A quantitative approach was used to examine the role of light and soil water regime on the variations found in anatomical traits of the palisade and spongy parenchyma, outer epidermal cell wall of the abaxial and adaxial surfaces, the percentage of sclerenchymatous area in relation to the total midrib area and the ratio of palisade to spongy parenchyma for five distinct ecological populations: M—late secondary montane forest (shaded, unflooded); M2—early secondary montane forest (semi-exposed, unflooded); SI—primary swamp forest (semi-exposed, flooded); S2—secondary swamp forest (exposed, flooded); and D—deforestation area (exposed, unflooded). Tukey tests were carried out for multiple comparisons, while one-way factor variance analysis was used to test for differences among ecological populations. A principal component analysis was used to order the populations as well as to find the higher variance component. These populations developed different response levels to the environmental factors studied, namely light and soil water regime. Light accounted for the variations found in palisade and spongy parenchyma while the combination of light and soil water determined the variations found regarding the outer epidermal cell wall of the abaxial surface, the percentage of sclerenchymatous area in relation to the total midrib area and the compaction of the spongy parenchyma. The separation of S1/M2 and S2/D populations into two groups was due to similar light regimes, which suggested that light was affecting the leaf anatomical variation of A. triplinervia more than the interaction of light and soil water regime.     

Hybridization and introgression across different ploidy levels in the Neotropical orchids Epidendrum fulgens and E. puniceoluteum (Orchidaceae)

The hypothesis of gene flow between species with large differences in chromosome numbers has rarely been tested in the wild, mainly because species of different ploidy are commonly assumed to be reproductively isolated from each other because of instantaneous and strong postzygotic barriers. In this study, a broad‐scale survey of molecular variation was carried out between two orchid species with different ploidy levels: Epidendrum fulgens (2n = 2x = 24 chromosomes) and Epidendrum puniceoluteum (2n = 4x = 52 chromosomes). To test the strength of their reproductive barriers, we investigated the distribution of genetic variation in sympatric and allopatric populations of these two species and conducted crossing experiments. Nuclear and plastid microsatellite loci were used to genotype 463 individuals from eight populations across the geographical range of both species along the Brazilian coastal plain. All six sympatric populations analysed presented hybrid zones, indicating that hybridization between E. fulgens and E. puniceoluteum is a common phenomenon. Bayesian assignment analysis detected the presence of F₁ and F₂ individuals and also signs of introgression, demonstrating a high potential for interspecific gene flow. Introgression occurs preferentially from E. fulgens to E. puniceoluteum. Pure parental individuals of both species display strong genotype–habitat associations, indicating that environment‐dependent selection could be acting in all hybrid zones. This study suggests that hybridization and introgression are evolutionary processes playing a role in the diversification of Epidendrum and indicates the importance of investigations of hybrid zones in understanding reproductive barriers and speciation processes in Neotropical orchid species.