Pollination Biology of the Exotic Rattleweed Crotalaria retusa L. (Fabaceae) in NE Brazil1

The rattleweed Crotalaria retusa was introduced in Brazil from Africa, and combines a series of characters that have ensured its establishment in NE Brazil. We focused on its reproductive biology and pollinator behavior to explain its reproductive success. We performed manual pollination and germination experiments, and monitored the behavior of C. retusa's main pollinators in monospecific plots, and in mixed plots where C. retusa occurred together with two congeners, Crotalaria pallida and Crotalaria lanceolata. Crotalaria retusa is self‐compatible and capable of automatic selfing. Inbreeding depression was expressed at the level of percent seed germination, but not seed set. Few insects visited the inflorescences. Legitimate pollinators were two large carpenter bees, Xylocopa frontalis and Xylocopa grisescens which, together, accounted for more than 90 percent of the visits. The former foraged on C. retusa exclusively and has low pollen spread potential. The latter flew longer distances between plants and visited fewer flowers per inflorescence, potentially increasing the extent of pollen carryover, but at the risk of increasing heterospecific pollen transfer, because it visits other Crotalaria species during the same foraging bout. The different foraging strategies, allied to morphological disadvantages represented by pollen overlap on X. grisescens' body, may partially explain the much lower seed germination observed in C. pallida and C. lanceolata. These results are consistent with the hypothesis that a reduction in flower constancy may significantly depress viable seed set by increasing the chances of self‐pollination.     

Aluminium modulation of the photosynthetic carbon reduction cycle in Zea mays

Two-weeks-old maize (Zea mays L. cv. XL-72.3) plants were submitted to Al concentrations of 0-81 g m-3 for 20 d, after which the A1 concentration-dependent effects on CO2 uptake by the mesophyll tissue and subsequent CO2 assimilation in the photosynthetic carbon reduction cycle of bundle sheath cells were investigated. The net photosynthetic rate (PN) and stomatal conductance (gs) increased continuously up to 27 g m-3 Al, whereas the intercellular CO2 concentration showed minimum values with the 27 g m-3 Al treatment. Moreover, the starch and saccharide concentrations, and fructose-1,6-bisphosphatase did not change significantly with increasing Al concentrations. The photosynthetic electron transport rates along with photosystems 2 and 1 started falling from 9 g m-3 Al onwards, while thylakoid acyl lipid composition did not show a clear pattern. With the Al concentration at 81 g m-3, NADP-malate dehydrogenase activity decreased to minimum values, whereas the opposite occurred with those of pyruvate dikinase, NADP-malic enzyme, and phosphoenolpyruvate carboxylase. Thus in vivo Al concentrations modulate the photosynthetic reduction cycle, possibly by interacting with the carbon flow rate exported to the cytosol. Although the inhibition of NADP-malate dehydrogenase activity might limit pyruvate dikinase, NADP-malic enzyme, and phosphoenolpyruvate carboxylase activities, in vivo the balance between phosphoenolpyruvate production and its carboxylation remains unaffected.     

Aluminium Toxicity Modulates Nitrate to Ammonia Reduction

Two weeks-old maize (Zea mays cv. XL-72.3) plants were exposed to Al concentrations 0 (Al0), 9 (Al9), 27 (Al27) or 81 (Al81) g m-3 for 20 d in a growth medium with low ionic strength. Thereafter, the Al concentration-dependent interactions on root nitrate uptake, and its subsequent reduction to ammonia in the leaves were investigated. Al concentrations in the roots sharply increased with increasing Al concentrations while root elongation correspondingly decreased. Root fresh and dry masses, acidification capacity, and nitrate and nitrogen contents decreased from Al27 onwards, whereas leaf nitrogen, nitrate, nitrite, and ammonia concentrations decreased starting with Al9. Electrolytic conductance increased by 60 % in root tissues from Al0 to Al81 but it did not increase significantly in the leaves. In Al9, Al27, and Al81 plants a decrease in shoot fresh and dry masses was observed. Al concentrations between 0 and 27 g m-3 increased net photosynthetic rate, stomatal conductance, and the quantum yield of photosynthetic electron transport, whereas the intercellular CO2 concentration was minimum in Al27 plants. In the leaves, nitrate reductase (E.C. 1.6.6.1) activity increased until Al27, and nitrite reductase (E.C. 1.6.6.4) activity until Al81. Hence there may be an Al mediated extracellular and intracellular regulation of root net nitrate uptake. Nitrate accumulation in the roots affects the translocation rates and, therefore, the nitrate concentration in the leaves. The in vivo reducing power generated by the photosynthetic electron flow does not limit nitrate to ammonia reduction, and the increase of maximum nitrate and nitrite reductase activities parallels the decreasing nitrate, nitrite, and ammonia concentrations.     

Do flood pulses structure amphibian communities in floodplain environments?

Beta diversity can provide insights into the processes that regulate communities subjected to frequent disturbances, such as flood pulses, which control biodiversity in floodplains. However, little is known about which processes structure beta diversity of amphibians in floodplains. Here, we tested the influence of flood pulses on the richness, composition, and beta diversity of amphibians in Amazonian floodplain environments. We also evaluated indicator species for each environment. We established linear transects in three environments: low várzea, high várzea, and macrophyte rafts. Species richness decreased and beta diversity increased according to the susceptibility of habitats to flood pulses. Indicator species differed among environments according to forest succession promoted by the flood pulse. The decrease in species richness between high and low várzea is due to non‐random extinctions. The higher rates of species turnover between várzeas and macrophyte rafts are driven by the colonization of species adapted to open areas. Our results highlight that the maintenance of complex environments is needed to protect biodiversity in floodplains.     

Mitogen activated protein kinases SakAHOG1 and MpkC collaborate for Aspergillus fumigatus virulence

Here, we investigated which stress responses were influenced by the MpkC and SakA mitogen‐activated protein kinases of the high‐osmolarity glycerol (HOG) pathway in the fungal pathogen Aspergillus fumigatus. The ΔsakA and the double ΔmpkC ΔsakA mutants were more sensitive to osmotic and oxidative stresses, and to cell wall damaging agents. Both MpkC::GFP and SakA::GFP translocated to the nucleus upon osmotic stress and cell wall damage, with SakA::GFP showing a quicker response. The phosphorylation state of MpkA was determined post exposure to high concentrations of congo red and Sorbitol. In the wild‐type strain, MpkA phosphorylation levels progressively increased in both treatments. In contrast, the ΔsakA mutant had reduced MpkA phosphorylation, and surprisingly, the double ΔmpkC ΔsakA had no detectable MpkA phosphorylation. A. fumigatus ΔsakA and ΔmpkC were virulent in mouse survival experiments, but they had a 40% reduction in fungal burden. In contrast, the ΔmpkC ΔsakA double mutant showed highly attenuated virulence, with approximately 50% mice surviving and a 75% reduction in fungal burden. We propose that both cell wall integrity (CWI) and HOG pathways collaborate, and that MpkC could act by modulating SakA activity upon exposure to several types of stresses and during CW biosynthesis.     

Myrip uses distinct domains in the cellular activation of myosin VA and myosin VIIA in melanosome transport

Myrip is a Rab27a and MyosinVIIa (MyoVIIa) linking protein that may regulate melanosome transport in the retinal pigment epithelium (RPE). Myrip also binds MyosinVa (MyoVa) in vitro however it is unclear whether this interaction is of sufficient affinity to be physiologically relevant. Here, we addressed the questions of whether Myrip interacts with MyoVa in cells and the molecular basis of cellular activation of MyoVa and MyoVIIa by Myrip. To answer these questions we used melanosome transport in skin melanocytes and RPE cells as read‐outs of MyoVa and MyoVIIa activity. We found that Myrip recruits and activates MyoVa on skin melanosomes with similar efficiency to the established MyoVa activator Melanophilin (Mlph). Mutagenesis showed that a Myrip–Mlph conserved amphipathic helix (MMAH) is essential for MyoVa interaction while other Myrip regions, including the MyoVa exon F binding domain equivalent, play non‐essential roles in this interaction. This suggests that, in contrast to Mlph, Myrip interacts with MyoVa lacking melanocyte‐specific exon F. Parallel studies of RPE melanosome transport reveal that Myrip‐specific inserts, but not the MMAH, are essential for MyoVIIa activation. We conclude that Myrip is a versatile Rab27a‐associated myosin‐activating protein that mediates cellular activation of MyoVa and MyoVIIa via non‐overlapping domains.     

Ubiquitin-mediated internalization of connexin43 is independent of the canonical endocytic tyrosine-sorting signal

Gap junctions are specialized cell-cell contacts that provide direct intercellular communication between eukaryotic cells. The tyrosine-sorting signal (YXX?), present at amino acids 286-289 of Cx43 (connexin43), has been implicated in the internalization of the protein. In recent years, ubiquitination of Cx43 has also been proposed to regulate gap junction intercellular communication; however, the underlying mechanism and molecular players involved remain elusive. In the present study, we demonstrate that ubiquitinated Cx43 is internalized through a mechanism that is independent of the YXX? signal. Indeed, expression of a Cx43-Ub (ubiquitin) chimaera was shown to drive the internalization of a mutant Cx43 in which the YXX? motif was eliminated. Immunofluorescence, cycloheximide-chase and cell-surface-protein biotinylation experiments demonstrate that oligomerization of Cx43-Ub into hemichannels containing wild-type Cx43 or mutant Cx43Y286A is sufficient to drive the internalization of the protein. Furthermore, the internalization of Cx43 induced by Cx43-Ub was shown to depend on its interaction with epidermal growth factor receptor substrate 15.     

Molecular modeling of Mycobacterium tuberculosis dUTpase: docking and catalytic mechanism studies

Mycobacterium tuberculosis is a leading cause of infectious disease in the world today. This outlook is aggravated by a growing number of M. tuberculosis infections in individuals who are immunocompromised as a result of HIV infections. Thus, new and more potent anti-TB agents are necessary. Therefore, dUTpase was selected as a target enzyme to combat M. tuberculosis. In this work, molecular modeling methods involving docking and QM/MM calculations were carried out to investigate the binding orientation and predict binding affinities of some potential dUTpase inhibitors. Our results suggest that the best potential inhibitor investigated, among the compounds studied in this work, is the compound dUPNPP. Regarding the reaction mechanism, we concluded that the decisive stage for the reaction is the stage 1. Furthermore, it was also observed that the compounds with a -1 electrostatic charge presented lower activation energy in relation to the compounds with a -2 charge.     

Methodology to Improve the Efficiency of Evaluation of the Severity of Angular Leaf Spot in Common Bean

Disease severity assessment by means of a scoring scale, especially for angular leaf spot (Pseudocercospora griseola) in common bean, is hindered in experiments for assessment of progenies and/or breeding lines due to lack of uniformity of occurrence of the pathogens and segregation within progenies. The purpose of this study was to estimate the efficiency of the use of one plant per plot in assessing the severity of angular leaf spot in experiments for assessment of progenies and/or breeding lines in the common bean crop. To that end, two experimental strategies were used – one of them using one plant per plot and another using a standard size plot (SPP) (2–4‐m length rows). The experiments were conducted in the period from November 2011 to May 2012 in the municipalities of Lavras and Lambari, state of Minas Gerais, Brazil. Forty‐one lines from the breeding programme of the Universidade Federal de Lavras (UFLA) and from other research institutions were assessed, which differed in regard to their degree of susceptibility to P. griseola. The lines were assessed in regard to the severity of said disease using a five‐degree diagrammatic scale. In all the one plant per plot experiments, severity scores of angular leaf spot from the beginning of its occurrence, and later in intervals ranging from 7 to 12 days, were obtained. In the experiment with the SPP, assessment was made a few days prior to grain harvest. Estimates of the correlations between severity scores and grain yield (GY) were mostly of small magnitude. There was good coincidence between the lines classified as more resistant or more susceptible to the pathogen under the two conditions.     

Carbonate Production by Benthic Communities on Shallow Coralgal Reefs of Abrolhos Bank,Brazil

The abundance of reef builders, non-builders and the calcium carbonate produced by communities established in Calcification Accretion Units (CAUs) were determined in three Abrolhos Bank shallow reefs during the period from 2012 to 2014. In addition, the seawater temperature, the irradiance, and the amount and composition of the sediments were determined. The inner and outer reef arcs were compared. CAUs located on the inner reef shelf were under the influence of terrigenous sediments. On the outer reefs, the sediments were composed primarily of marine biogenic carbonates. The mean carbonate production in shallow reefs of Abrolhos was 579 ± 98 g m^-2 y^-1. The builder community was dominated by crustose coralline algae, while the non-builder community was dominated by turf. A marine heat wave was detected during the summer of 2013–2014, and the number of consecutive days with a temperature above or below the summer mean was positively correlated with the turf cover increase. The mean carbonate production of the shallow reefs of Abrolhos Bank was greater than the estimated carbonate production measured for artificial structures on several other shallow reefs of the world. The calcimass was higher than the non-calcareous mass, suggesting that the Abrolhos reefs are still in a positive carbonate production balance. Given that marine heat waves produce an increase of turf cover on the shallow reefs of the Abrolhos, a decrease in the cover represented by reef builders and shifting carbonate production are expected in the near future.