Aerial development in Streptomyces coelicolor requires sortase activity

Streptomyces coelicolor is a multicellular bacterium whose life cycle encompasses three differentiated states: vegetative hyphae, aerial hyphae and spores. Among the factors required for aerial development are the 'chaplins', a family of eight secreted proteins that coat the surface of aerial hyphae. Three chaplins (the 'long' chaplins, ChpA, B and C) possess an LAXTG-containing C-terminal sorting signal and are predicted sortase substrates. The five remaining 'short' chaplins are presumed to be associated with the cell surface through interactions with the long chaplins. We show here that two sortase enzymes, SrtE1 and SrtE2, cleave LAXTG-containing peptides at two distinct positions in vitro, and are required for cell wall anchoring of ChpC in vivo. srtE1/E2 double mutants are delayed in aerial hyphae formation, do not sporulate and fail to display all short chaplins on their aerial surfaces. Surprisingly, these mutant characteristics were not shared by a long chaplin mutant, which exhibited only modest delays in aerial development, leading us to revise the current model of chaplin-mediated aerial development. The sortase mutant phenotype, instead, appears to stem from an inability to transcribe aerial hyphae-specific genes, whose products have diverse functions. This suggests that sortase activity triggers an important, and previously unknown, developmental checkpoint.     

The problem of precision and trend detection posed by small elephant populations in West Africa

The precision of elephant estimates from aerial sample surveys and dung counts is inversely proportional to abundance. West African elephant populations are already small, and the power of a monitoring programme to detect changes in abundance diminishes as the population shrinks in size. Thus it will be difficult to evaluate the effects on elephant numbers of new management policies in West Africa. The same will be true of monitoring schemes for antelope and primate populations that are hunted for bushmeat. Elephant estimates from dung counts are more precise than those from aerial sample surveys, and changes in elephant numbers are more likely to be detected in the subregion by dung counts than by aerial sample surveys.     

Aerodynamic advantages of upside down take-off for aerial dispersal in <Emphasis Type="Italic">Tetranychus</Emphasis> spider mites

Aerial dispersal may be important for redistribution of spider mites into new habitats. Evidence for behavioral control of aerial take-off has been well documented for Tetranychus urticae Koch. Before aerial dispersal they exhibit the aerial take-off posture that involves lifting the forelegs upright and raising the forebody. However, whether the aerial take-off posture functions to increase drag has remained unclear. The objectives of this study were to clarify: (i) aerodynamic effects of the aerial take-off posture; and (ii) actual aerial take-off behavior in T. urticae. To evaluate the aerodynamic forces experienced by grounded spider mites in different postures, we constructed three-dimensional models of T. urticae, exhibiting the aerial take-off posture and the normal posture, using computer graphics. We found that the aerial take-off posture was effective in receiving greater rearward forces from wind rather than upward forces. As a result, aerial take-off from a horizontal platform is unlikely. Instead, inverted departure surfaces, e.g., lower leaf surfaces, with inclines are likely to be effective sites for take-off. Laboratory experiments and field observations indicated that the mites preferentially adopted such a position for orientation and take-off. Our findings provided a rationale for the take-off behavior of Tetranychus spider mites.     

Conservation of invertebrate biodiversity on a mountain in a global biodiversity hotspot,Cape Floral Region

Mountains present particular challenges for biodiversity conservation. Table Mountain is a significant mountain in a global biodiversity hotspot, the Cape Floristic Region. It has outstanding angiosperm diversity and endemism. Yet, aerial and foliage invertebrates in the area have been poorly studied, despite their importance as pollinators and predators. These plant and invertebrate assemblages are under great pressure from human disturbance. Aerial and foliage invertebrates were sampled with a range of techniques. Sites were chosen to make comparisons between vegetation structure and type, elevation and aspect. In total, 216 species from 63 families and 14 orders were recorded. Vegetation structure (fynbos or forest) and elevation were the most important environmental variables for both aerial and foliage invertebrates. Peak time for aerial invertebrate abundance was spring and summer in the fynbos and spring in the forests, while the foliage invertebrates showed very little seasonal variation. There was no correlation between the diversity of aerial and foliage invertebrates. When these results were compared with others on epigaeic invertebrates, it became clear that epigaeic and aerial invertebrates are not correlated, while epigaeic and foliage invertebrates were only partially correlated, but not sufficiently so to consider one as a reliable estimator of the other. The management pointer from this study is that sites at all elevations are vital for the conservation of biodiversity on Table Mountain. Both the aerial and epigaeic/foliage invertebrate assemblages will need to be monitored separately to maintain the mountain’s conservation status.     

Protostelids from tropical forests, woodlands and deserts in Australia

Most of what is known about the distribution of protostelids is limited to results from surveys carried out in North and Central America. To increase our knowledge about protostelid diversity and distribution we surveyed protostelids from 12 study sites in northern Queensland and the Northern Territory of Australia during May-Jun 2003. Aerial litter and ground litter samples were randomly collected along a 200 m transect at each site. Study sites ranged from tropical forests to deserts. We recovered 10 species and two apparently undescribed species from samples of aerial (dead but still attached plant parts) and ground litter. Samples from a woodland site characterized by intermediate moisture conditions had the greatest species richness, followed by samples from dry woodland, tropical forest and desert sites. When species richness for a particular microhabitat was considered, samples of aerial litter yielded more species than samples of ground litter. Percentages of samples colonized with protostelids were similar for the aerial and ground litter microhabitats within a given habitat type except for dry woodlands, in which aerial litter samples were characterized by higher numbers of species than ground litter samples. Two species (Protostelium mycophaga and Soliformovum irregularis) that in temperate North America are associated with aerial litter microhabitats also were recovered from aerial litter in dry habitats in Australia. Schizoplasmodiopsis pseudoendospora, a North American temperate ground litter species, was equally abundant in aerial litter and ground litter in Australia. This study is the first of its kind for protostelid ecology in Australia and the most extensive study of protostelids in the southern hemisphere. These data complement ongoing research on protostelid distribution from around the world.     

SapB and the chaplins: connections between morphogenetic proteins in Streptomyces coelicolor

Morphogenesis in the streptomycetes features the differentiation of substrate-associated vegetative hyphae into upwardly growing aerial filaments. This transition requires the activity of bld genes and the secretion of biosurfactants that reduce the surface tension at the colony-air interface enabling the emergence of nascent aerial hyphae. Streptomyces coelicolor produces two classes of surface-active molecules, SapB and the chaplins. While both molecules are important for aerial development, nothing is known about the functional redundancy or interaction of these surfactants apart from the observation that aerial hyphae formation can proceed via one of two pathways: a SapB-dependent pathway when cells are grown on rich medium and a SapB-independent pathway on poorly utilized carbon sources such as mannitol. We used mutant analysis to show that while the chaplins are important, but not required, for development on rich medium, they are essential for differentiation on MS (soy flour mannitol) medium, and the corresponding developmental defects could be suppressed by the presence of SapB. Furthermore, the chaplins are produced by conditional bld mutants during aerial hyphae formation when grown on the permissive medium, MS, suggesting that the previously uncharacterized SapB-independent pathway is chaplin dependent. In contrast, a bld mutant blocked in aerial morphogenesis on all media makes neither SapB nor chaplins. Finally, we show that a constructed null mutant that lacks all chaplin and SapB biosynthetic genes fails to differentiate in any growth condition. We propose that the biosurfactant activities of both SapB and the chaplins are essential for normal aerial hyphae formation on rich medium, while chaplin biosynthesis and secretion alone drives aerial morphogenesis on MS medium.     

Glutamine metabolism during aerial mycelium growth of Neurospora crassa

During vegetative growth, glutamine is accumulated in the mycelium of Neurospora crassa. This high pool of glutamine seems to be required for aerial mycelium growth. Enzymes responsible for the synthesis and catabolism of glutamine were measured before and during the partial transformation of a mycelial mat into aerial mycelium. In the transforming mycelial mat,considerable activities of the biosynthetic NADP-glutamate dehydrogenase and glutamine synthetase (predominantly β polypeptide) and also some activity of glutamate synthase were observed. In the aerial mycelium, glutamine synthetase (predominantly β polypeptide) was detected, but very low activities of NADP-glutamate dehydrogenase and glutamate mycelium could derive from glutamine. No glutaminase activity could be detected. It is suggested that glutamate is formed through the activities of the glutamine transaminase-ω -amidase pathway and another transaminase. High activities of glutamine and alanine transaminases were observed in the aerial mycelium. These results are discussed in terms of the possible role of glutamine as a nitrogen carrier from the mycelium to the growing aerial hyphae.     

Evaluating performance of aerial survey data in elephant habitat modelling

Aerial survey data are widely used to model distribution of wildlife. However, their performance in habitat modelling remains largely untested. We used aerial survey and satellite‐linked Global Positioning System (GPS) collar data for elephants, to test (i) whether there is an optimal spatial resolution of predictor variables at which habitat models based on aerial survey data that are uncorrected for locational error can accurately predict elephant habitat and (ii) whether habitat models based on these data sets can accurately predict the presence of elephants in closed woodland habitats. We applied maximum entropy modelling (Maxent) to these data sets and used the Normalised Difference Vegetation Index (NDVI) as well as distance from water points as the habitat predictors to answer these questions. Our results demonstrate better ability of aerial survey data to predict elephant presence at the coarser spatial resolution of 1000 m of both predictor variables. Habitat models derived from aerial survey data underpredicted elephant presence in more closed woodland habitats than those derived from GPS collar data. This result implies that elephants located under dense tree canopies are likely missed during an aerial survey. Our study is one of the first to empirically test and report results on the poor performance of aerial survey data in habitat modelling especially in dense woodlands.     

Fine structure of the outer sheath of the aerial mycelium in Actinomyces levoris

The outer sheath of the aerial mycelium of Actinomyces levoris 64 bears groove-like and granular submicroscopic structures. They are very susceptible to a short-term treatment of the aerial mycelium with aqueous acetone which causes their disintegration. Under specified conditions, submicroscopic structures, viz. granules and threads, assemble from the acetone extract of the aerial mycelium of Actinomyces levoris. These structures possess the anti-yeast activity. The granular structures reorganized from the acetone extract of the aerial mycelium of Actinomyces levoris somewhat resemble in morphology the structures seen in the outer sheath of the aerial mycelium of this actinomycete. The results obtained are discussed within the framework of the hypothesis on a possible participation of polyene antibiotics in the formation of the outer sheath of the aerial mycelium of actinomycetes which produce these antibiotics.     

Glutamine requirement for aerial mycelium growth in Neurospora crassa

Five amino acids are accumulated during vegetative growth of Neurospora crassa, particularly.during the prestationary growth phase. Alanine, glutamine, glutamate, arginine and ornithine.comprised over 80% of the total amino acid pool in the mycelium. Amino acid pools of different amino acid auxotrophs were followed during the partial transformation of a mycelial mat into an aerial mycelium. The mycelial mat under starvation and in direct contact with air rapidly formed aerial mycelium, which produced thereafter a burst of conidia. During this process,glutamine and alanine in the mycelial mat were consumed more rapidly than other amino acids;in the growing aerial mycelium, glutamate and glutamine were particularly accumulated. Of the amino acids that were initially accumulated in the mycelial mat, only a high glutamine pool was required for aerial mycelium growth induced by starvation. This requirement for glutamine could not be satisfied by a mixture of the amino compounds that are synthesized via glutamine amidotransferase reactions. It is proposed that glutamine serves as a nitrogen carrier from the mycelial mat to the growing aerial mycelium.     

Clonal diversity and conservation genetics of the medicinal plant Carapichea ipecacuanha (Rubiaceae)

The roots of the understorey shrub Carapichea ipecacuanha (ipecac) have medicinal properties, and the uprooting of wild plants has supplied most of the world demand for this species. Although under severe population decline, C. ipecacuanha lacks legal protection. In the wild, the aerial stems of ipecac clump together to form clusters with well-defined borders. Cluster size may range from several to hundreds of aerial stems. To investigate the extent of clonality among aerial stems in ipecac clusters, we sampled 50 wild clusters (a total of 291 aerial stems) and screened them with 89 inter-simple sequence repeat (ISSR) markers. The 291 aerial stems were grouped into 42 putative clones. The clonal groups generally consisted of aerial stems from the same cluster, and there was little or no genetic differentiation among aerial stems at the cluster level. These findings suggest that strategies designed to conserve ipecac in situ should not rely upon census data, which are based on the number of aerial stems per cluster and the number of clusters per population, because such data greatly underestimate the species effective population size and genetic diversity. Our results also indicate that this species needs protection at a federal level.     

Life History Evolution in Amphicarpic Plants

Abstract Plants with dimorphic flowers or seeds provide excellent material for the study of life history evolution because the dimorphism often involves measurable differences in morphology, size, number, or genetic relatedness. For amphicarpic plants, the proportion of aerial: subterranean morphs produced is highly variable (from 0 to > 100) and related to both environmental and genetic factors. Plants from aerial seeds produce lower ratios of aerial: subterranean morphs than those from subterranean seeds. Despite substantial variation, subterranean seeds are generally heavier than aerial seeds (but fewer) and produce vigorous seedlings with high survivorship and high fitness. Adaptive advantages of subterranean seeds include retention of offspring in favorable parental microhabitats, protection of seeds from herbivory, predation, or fire, and avoidance of desiccating conditions on the soil surface; potential disadvantages include lack of gene exchange, high energy costs, limited dispersal, and sibling competition. For the few species studied, aerial reproduction is more plastic than subterranean reproduction and more likely to be affected by environmental conditions. Quantitative genetic analyses of a population of the annual grass Amphicarpum purshii have revealed lower heritabilites for subterranean relative to aerial reproductive traits. Subterranean seed number and mass show genetic correlations with shoot mass while aerial seed number and mass do not; seed set percentages of both seed types as well as percentage allocation to both reproductive morphs show negative genetic correlations with shoot mass. In this Amphicarpum population, directional selection on shoot mass may indirectly select for increased subterranean (but not aerial) seed output.     

Consequences of a mixed reproductive system in the hog peanut,Amphicarpaea bracteata, (Fabaceae)

Summary The forest annual, Amphicarpaea bracteata L. can reproduce via aerial chasmogamous, aerial cleistogamous, and subterranean cleistogamous flowers. Both plant size and light intensity influenced the utilization of the three modes of reproduction. chasmogamous and aerial cleistogamous flower number and the ratio of chasmogamous flowers to the total number of aerial flowers increased with plant size. The latter demonstrated a shift to xenogamy and outbreeding in larger plants. Light intensity indirectly influenced reproductive modes through its infuence on plant size. Seed set by both types of aerial flowers was low and unrelated to plant size. Subterranean seed number and the total dry weight of subterranean seeds per plant increased with size. The subterranean seeds of Amphicarpaea bracteata are thirty-four times larger than the aerial seeds (fresh weight). Under field conditions, subterranean seeds had greater germination after one year than acrial seeds. The plants arising from subterranean seeds were significantly larger and more fecund than those from aerial seeds. Seeds produced by aerial cleistogamous, hand selfpollinated chasmogamous, and naturally pollinated chasmogamous flowers had equivalent germination rates and produced plants of equal size and fecundity. This suggests that the outbred progeny from chasmogamous flowers have no advantage over the inbred progeny from aerial cleistogamous flowers.     

Aerial mycelia of Aspergillus oryzae accelerate α-amylase production in a model solid-state fermentation system

Aspergillus oryzae is commonly used in solid-state fermentation (SSF) and forms abundant aerial mycelia. Previously, we have shown that aerial mycelia are extremely important for the respiration of this fungus during growth on a wheat-flour model substrate. In this paper, we show that aerial mycelia of this fungus give a strong increase in fungal biomass and α-amylase production. Cultures of A. oryzae on wheat-flour model substrate produced twice the amounts of fungal biomass and α-amylase, when aerial mycelia were formed. Utilization of these findings in commercial solid-state fermenters requires further research; results from packed beds of grain indicate that aerial mycelia are of limited importance there. Probably, substrate pre-treatment and an increase in bed voidage are required.     

Visual and acoustic surveys for North Atlantic right whales,Eubalaena glacialis,in Cape Cod Bay,Massachusetts, 2001–2005: Management implications

North Atlantic right whales, Eubalaena glacialis, remain endangered, primarily due to excessive anthropogenic mortality. Current management protocols in US waters are triggered by identifying the presence of at least one right whale in a management area. We assessed whether acoustic detection of right whale contact calls can work as an alternative to visual aerial surveys for establishing their presence. Aerial survey and acoustic monitoring were conducted in Cape Cod Bay, Massachusetts, in 2001–2005 and used to evaluate and compare right whale detections. Over the 58 d with simultaneous aerial and acoustic coverage, aerial surveys saw whales on approximately two-thirds of the days during which acoustic monitoring heard whales. There was no strong relationship between numbers of whales seen during aerial surveys and numbers of contact calls detected on survey days. Results indicate acoustic monitoring is a more reliable mechanism than aerial survey for detecting right whales. Because simple detection is sufficient to trigger current management protocols, continuous, autonomous acoustic monitoring provides information of immediate management utility more reliably than aerial surveillance. Aerial surveys are still required to provide data for estimating population parameters and for visually assessing the frequency and severity of injuries from shipping and fishing and detecting injured and entangled right whales.     

Streptomyces genes involved in aerial mycelium formation

Abstract Cloning of genes as suppressors of the aerial mycelium-defective phenotype of Streptomyces griseus HH1 resulting from A-factor-deficiency has led to the identification of several genes, including amfR, amfAlamfB, amfC , and orf1590 . These genes are involved in aerial mycelium formation independent of secondary metabolic function. Among these, AmfR which belongs to the family of response regulators of two-component regulatory systems and AmfA/AmfB similar to ATP-dependent membrane translocators are analogous to the multicomponent phosphorelay and the Spo0K system, respectively, both of which are required for the initiation of sporulation in Bacillus subtilis . Involvement of a protein serine/threonine kinase in aerial mycelium formation is also suggested, because the Streptomyces coelicolor A3(2) afsK gene encoding a 'eukaryotic'-type protein kinase reverses the aerial mycelium-defective phenotype of strain HH1, independent of secondary metabolic function.     

Aphid aerial density profiles are consistent with turbulent advection amplifying flight behaviours: abandoning the epithet 'passive'

Seminal field studies led by C. G. Johnson in the 1940s and 1950s showed that aphid aerial density diminishes with height above the ground such that the linear regression coefficient, b, of log density on log height provides a single-parameter characterization of the vertical density profile. This coefficient decreases with increasing atmospheric stability, ranging from -0.27 for a fully convective boundary layer to -2.01 for a stable boundary layer. We combined a well-established Lagrangian stochastic model of atmospheric dispersal with simple models of aphid behaviour in order to account for the range of aerial density profiles. We show that these density distributions are consistent with the aphids producing just enough lift to become neutrally buoyant when they are in updraughts and ceasing to produce lift when they are in downdraughts. This active flight behaviour in a weak flier is thus distinctly different from the aerial dispersal of seeds and wingless arthropods, which is passive once these organisms have launched into the air. The novel findings from the model indicate that the epithet 'passive' often applied to the windborne migration of small winged insects is misleading and should be abandoned. The implications for the distances traversed by migrating aphids under various boundary-layer conditions are outlined.     

Gas Exchange between Equisetum limosum and its Environment

Equisetum limosum grows in standing water. Its perennating rhizomesramify in the bottom mud, which is a markedly oxygen-deficientmedium. During the summer months aerial shoots are present andextend above the surface of the water to a height of about 1m. The concentration of oxygen in the internal atmosphere ofplants in the field decreases progressively from the stem tothe perennating rhizomes. There is a reverse gradient of carbondioxide concentrations. The structure of the nodal diaphragms of the aerial stem isdescribed and the progressive occlusion of the pores of thediaphragms between May and November is demonstrated. This occlusionof the diaphragms has been correlated with a reduction in thecase of gaseous diffusion along lengths of aerial stems. Oxygen is present in the rhizomes throughout the winter, despitethe fact that the dead aerial stems have broken off and thestumps have been submerged by the rise in water level, thusisolating the rhizome from atmospheric oxygen. The results are discussed in relation to the ability of theplant to grow in an oxygen-deficient environment.     

Streptomyces inside-out: a new perspective on the bacteria that provide us with antibiotics

Many of the antibiotics used today are made by a group of bacteria called Streptomyces. Streptomycetes evolved about 450 million years ago as branched filamentous organisms adapted to the utilization of plant remains. They reproduce by sending up specialized aerial branches, which form spores. Aerial growth is parasitic on the primary colony, which is digested and reused for aerial growth. The reproductive phase is coordinated with the secretion of antibiotics, which may protect the colony against invading bacteria during aerial growth. A clue to the integration of antibiotic production and aerial growth is provided by bldA mutants, which are defective in both processes. These mutants lack the ability to translate a particularly rare codon, UUA, in the genetic code. The UUA codon (TTA in DNA) is present in several regulatory genes that control sets of antibiotic production genes, and in one, bldH that controls aerial mycelium formation. The regulatory genes for antibiotic production are all involved in self-reinforcing regulatory systems that potentially amplify the regulatory significance of small changes in the efficiency of translation of UUA codons. One of the regulatory targets of bldH is an extracellular protease inhibitor protein that is likely to delay the digestion of the primary biomass until the colony is ready for aerial growth. The use of the UUA codon to orchestrate different aspects of extracellular biology appeared very early in Streptomyces evolution.