Phylogeny of the Rhizobium–Allorhizobium–Agrobacterium clade supports the delineation of Neorhizobium gen. nov.

The genera Agrobacterium, Allorhizobium, and Rhizobium belong to the family Rhizobiaceae. However, the placement of a phytopathogenic group of bacteria, the genus Agrobacterium, among the nitrogen-fixing bacteria and the unclear position of Rhizobium galegae have caused controversy in previous taxonomic studies. To resolve uncertainties in the taxonomy and nomenclature within this family, the phylogenetic relationships of generic members of Rhizobiaceae were studied, but with particular emphasis on the taxa included in Agrobacterium and the “R. galegae complex” (R. galegae and related taxa), using multilocus sequence analysis (MLSA) of six protein-coding housekeeping genes among 114 rhizobial and agrobacterial taxa. The results showed that R. galegae, R. vignae, R. huautlense, and R. alkalisoli formed a separate clade that clearly represented a new genus, for which the name Neorhizobium is proposed. Agrobacterium was shown to represent a separate cluster of mainly pathogenic taxa of the family Rhizobiaceae. A. vitis grouped with Allorhizobium, distinct from Agrobacterium, and should be reclassified as Allorhizobium vitis, whereas Rhizobium rhizogenes was considered to be the proper name for former Agrobacterium rhizogenes. This phylogenetic study further indicated that the taxonomic status of several taxa could be resolved by the creation of more novel genera.     

The detoxification of α-tomatine by Fusarium oxysporum f. sp. lycopersici

Fusarium oxysporum f. sp. lycopersici detoxifies α-tomatine by producing an inducible extra-cellular enzyme which cleaves the glycoalkaloid into the tetrasaccharide lycotetraose and tomatidine.     

Evolutionary relationships between Fusarium oxysporum f. sp. lycopersici and F. oxysporum f. sp. radicis-lycopersici isolates inferred from mating type, elongation factor-1α and exopolygalacturonase sequences

Fusarium oxysporum is a ubiquitous species complex of soilborne plant pathogens that comprises many different formae speciales, each characterized by a high degree of host specificity. In this study, the evolutionary relationships between different isolates of the F. oxysporum species complex have been examined, with a special emphasis on the formae speciales lycopersici and radicis-lycopersici, sharing tomato as host while causing different symptoms. Phylogenetic analyses of partial sequences of a housekeeping gene, the elongation factor-1α (EF-1α) gene, and a gene encoding a pathogenicity trait, the exopolygalacturonase (pgx4) gene, were conducted on a worldwide collection of F. oxysporum strains representing the most frequently observed vegetative compatibility groups of these formae speciales. Based on the reconstructed phylogenies, multiple evolutionary lineages were found for both formae speciales. However, different tree topologies and statistical parameters were obtained for the cladograms as several strains switched from one cluster to another depending on the locus that was used to infer the phylogeny. In addition, mating type analysis showed a mixed distribution of the MAT1-1 and MAT1-2 alleles in the F. oxysporum species complex, irrespective of the geographic origin of the tested isolates. This observation, as well as the topological conflicts that were detected between EF-1α and pgx4, are discussed in relation to the evolutionary history of the F. oxysporum species complex.     

Hidden diversity uncovered in Hygrophorus sect. Aurei (Hygrophoraceae), including the Mediterranean H. meridionalis and the North American H. boyeri, spp. nov.

For many years, the binomial Hygrophorus hypothejus was widely applied to collections from various geographical regions in different continents, assuming a circum-boreal and circum-mediterranean distribution for this species. This hypothesis, however, had never been put to the test. To assess the diversity and species-limits within this complex of yellow-coloured waxcaps, a phylogenetic, morphological and taxonomical investigation into Hygrophorus sect. Aurei and similar species in sect. Olivaceoumbrini was carried out, including material of pan-European origin, as well as the east and west coasts of North America. Following sequencing of the ITS rDNA locus, nine lineages are confirmed in sect. Aurei, most of them highly continentalised. Of these, two are new to science, introduced here as Hygrophorus boyeri sp. nov., from Pinus banksiana and P. rigida forests in eastern North America and from P. muricata and P. contorta forests in western North America, and Hygrophorus meridionalis sp. nov., from Pinus brutia and Pinus halepensis forests in the island of Cyprus and mainland Greece. H. hypothejus is lectotypified and epitypified, and here resolved as a strictly European species, with the old forgotten taxon Hygrophorus siccipes revived as its North American vicariant. The placement of Hygrophorus fuligineus in sect. Aurei is phylogenetically confirmed and detailed comparisons between morphologically similar and phylogenetically affiliated taxa in sect. Aurei and sect. Olivaceoumbrini are provided. The chronic confusion associated with Hygrophorus fuscoalbus, a highly controversial taxon described from Germany nearly two centuries ago and variously interpreted since, is discussed, concluding that this name is too ambiguous to be applied to any currently recognized species.     

In vitro, ex vivo and in vivo models: A comparative analysis of Paracoccidioides spp. proteomic studies

Members of the Paracoccidioides complex are human pathogens that infect different anatomic sites in the host. The ability of Paracoccidioides spp. to infect host niches is putatively supported by a wide range of virulence factors, as well as fitness attributes that may comprise the transition from mycelia/conidia to yeast cells, response to deprivation of micronutrients in the host, expression of adhesins on the cell surface, response to oxidative and nitrosative stresses, as well as the secretion of hydrolytic enzymes in the host tissue. Our understanding of how those molecules can contribute to the infection establishment has been increasing significantly, through the utilization of several models, including in vitro, ex vivo and in vivo infection in animal models. In this review we present an update of our understanding on the strategies used by the pathogen to establish infection. Our results were obtained through a comparative proteomic analysis of Paracoccidioides spp. in models of infection.     

Competitiveness of endophytic Phialocephala fortinii s.l. – Acephala applanata strains in Norway spruce roots

Dark septate endophytes of the Phialocephala fortinii s.l. – Acephala applanata species complex (PAC) are presumed to be the most abundant root colonizing endophytes of conifers across the Northern hemisphere. To test the competitiveness of different PAC strains, PAC-free Picea abies saplings were inoculated with five different PAC strains by planting them in pre-colonized substrates. Saplings were left to grow for six weeks and then transplanted crosswise into a substrate colonized by one of the other four strains for a further two weeks. PAC were isolated and genotyped using microsatellite markers. The power of colonization, i.e. the ability of colonizing roots already colonized by another PAC strain, and the power of retention, i.e. the ability of a resident strain of not being suppressed by an invading PAC strain, were calculated for each strain in every combination. The experiment was run twice under two different climatic conditions. Our results show that PAC strains differ (1) in their ability to colonize PAC-free, non-sterile roots, (2) in resistance against being suppressed by another PAC strain and (3) in their ability to invade roots already colonized by another PAC strain. In addition, both the PAC–PAC and the PAC-host interactions depend on the climatic conditions.     

Alcaloïdes d'Alstonia vitiensis var. Novo ebudica monachino

Five alkaloids have been isolated from Alstonia vitiensis: pleïocarpamine, vincorine, cabucraline, alstovine (11-methoxycompactinervine) and quaternoxine; the latter two are new alkaloids.     

E. coli sabotages the in vivo production of O-linked β-N-acetylglucosamine-modified proteins

The O-linked β-N-acetylglucosamine (O-GlcNAc) post-translational modification is an important, regulatory modification of cytosolic and nuclear enzymes. To date, no 3-dimensional structures of O-GlcNAc-modified proteins exist due to difficulties in producing sufficient quantities with either in vitro or in vivo techniques. Recombinant co-expression of substrate protein and O-GlcNAc transferase in Escherichia coli was used to produce O-GlcNAc-modified domains of human cAMP responsive element-binding protein (CREB1) and Abelson tyrosine-kinase 2 (ABL2). Recombinant expression in E. coli is an advantageous approach, but only small quantities of insoluble O-GlcNAc-modified protein were produced. Adding β-N-acetylglucosaminidase inhibitor, O-(2-acetamido-2-dexoy-d-glucopyranosylidene)amino-N-phenylcarbamate (PUGNAc), to the culture media provided the first evidence that an E. coli enzyme cleaves O-GlcNAc from proteins in vivo. With the inhibitor present, the yields of O-GlcNAc-modified protein increased. The E. coli β-N-acetylglucosaminidase was isolated and shown to cleave O-GlcNAc from a synthetic O-GlcNAc-peptide in vitro. The identity of the interfering β-N-acetylglucosaminidase was confirmed by testing a nagZ knockout strain. In E. coli, NagZ natively cleaves the GlcNAc-β1,4-N-acetylmuramic acid linkage to recycle peptidoglycan in the cytoplasm and cleaves the GlcNAc-β-O-linkage of foreign O-GlcNAc-modified proteins in vivo, sabotaging the recombinant co-expression system.     

On barriers to hybridization between Strongylocentrotus droebachiensis (O.F. Müller) and S. Pallidus (G.O. Sars)

Strongylocentrotus droebachiensis (O. F. Müller) and Strongylocentrotus pallidus (G. O. Sars) occur together over much of their ranges in both the Atlantic and Pacific Oceans and remain distinct species, but no strong barrier to genetic exchange has been found in a study in the San Juan Islands. Co-occurrence of embryos of both species in the plankton indicates similar times of spawning. Ova of S. droebachiensis are fertilized by sperm of S. pallidus but the reciprocal cross gives a percent fertilization near zero. Hybrids of both crosses are viable with pigmentation resembling S. pallidus in the larval stage and S. droebachiensis in the adult stage. Ova of first generation female hybrids are more readily fertilized by sperm of S. pallidus, so that gene flow through female hybrids is more likely to be from S. droebachiensis to S. pallidus than the reverse. Male hybrids were not tested. S. droebachiensis and S. pallidus from deeper waters have lower fecundity than shallow water S. droebachiensis, which suggests that in the deeper waters individuals are food limited. The hypothesis that S. pallidus uses the sterile male technique to reduce reproductive output of S. droebachiensis is considered and rejected.     

How does a Psittacanthus robustus Mart. population structure relate to a Vochysia thyrsoidea Pohl. host population?

Distribution of parasitic plants is directly linked with the distribution of host species and behavioral patterns of seed dispersers. Psittacanthus robustus (Loranthaceae) is a neotropical hemiparasite that mainly colonizes species of the family Vochysiaceae. Vochysia thyrsoidea is the main host of P. robustus and is commonly found in areas of cerrado rupestre (rocky savanna), an abundant vegetation in our study site. We conducted the study in the ecological park Parque Ecológico Quedas do Rio Bonito (PEQRB), over an area of 2.82 ha of cerrado rupestre. The objective of this work was to investigate population structure, parasitic behavior (mistletoe grip height and circumference of host branch), and spatial distribution of Psittacanthus robustus on a population of Vochysia thyrsoidea. We sampled 267 V. thyrsoidea individuals and found that the population had a random distribution pattern. Seventy-nine individuals (29.6% of the sample population) hosted the hemiparasite, to a total of 193 P. robustus individuals. The number of mistletoe individuals per host plant ranged between 1 and 12. The V. thyrsoidea individuals most infested with mistletoes were those reaching greater heights. The correlation between height of host plant and preferred grip height was highly significant, with the preferred grip height being the uppermost portions of host plants. The crown size of P. robustus individuals ranged between 10 and 230 cm. The main disperser of P. robustus fruit was found to be swallow-tanager Tersina viridis viridis. Its activities led to a clumped pattern of spatial distribution of the hemiparasite along with higher infestation in larger trees.     

In vitro susceptibilities of Neoscytalidium spp. sequence types to antifungal agents and antimicrobial photodynamic treatment with phenothiazinium photosensitizers

Neoscytalidium spp. are ascomycetous fungi consisting of pigmented and hyaline varieties both able to cause skin and nail infection. Their color-based identification is inaccurate and may compromise the outcome of the studies with these fungi. The aim of this study was to genotype 32 isolates morphologically identified as Neoscytalidiumdimidiatum or N. dimidiatum var. hyalinum by multilocus sequence typing (MLST), differentiate the two varieties by their sequence types, evaluate their susceptibility to seven commercial antifungal drugs [amphotericin B (AMB), voriconazole (VOR), terbinafine (TER), 5-flucytosine (5FC), ketoconazole (KET), fluconazole (FLU), and caspofungin (CAS)], and also to the antimicrobial photodynamic treatment (APDT) with the phenothiazinium photosensitizers (PS) methylene blue (MB), new methylene blue (NMBN), toluidine blue O (TBO) and the pentacyclic derivative S137. The efficacy of each PS was determined, initially, based on its minimal inhibitory concentration (MIC). Additionally, the APDT effects with each PS on the survival of ungerminated and germinated arthroconidia of both varieties were evaluated. Seven loci of Neoscytalidium spp. were sequenced on MLST revealing eight polymorphic sites and six sequence types (ST). All N. dimidiatum var. hyalinum isolates were clustered in a single ST. AMB, VOR and TER were the most effective antifungal agents against both varieties. The hyaline variety isolates were much less tolerant to the azoles than the isolates of the pigmented variety. APDT with S137 showed the lowest MIC for all the isolates of both varieties. APDT with all the PS killed both ungerminated and germinated arthroconidia of both varieties reducing the survival up to 5 logs. Isolates of the hyaline variety were also less tolerant to APDT. APDT with the four PS also increased the plasma membrane permeability of arthroconidia of both varieties but only NMBN and S137 caused peroxidation of the membrane lipids.     

Kayserops? cantarmoricus nov. sp., Trilobite Asteropyginae du Devonien moyen armoricain et cantabrique

The discovery in rocks of Middle Devonian age of a new Asteropyginae, Kayserops? cantarmoricus n. sp., common to both Central Armorican Syncline (Rade de Brest, France) and the south flank of the Cantabrian Mountains (Spain), suggests a geographical connection, perhaps temporary, between these two regions. The particular morphological characteristics of this species shed new light on present knowledge of the sub-family Asteropyginae.     

Are the diatoms Navicula sp. and Thalassiosira fluviatilis suitable to be fed to the benthic harpacticoid copepod Tisbe biminiensis?

The objective of this study was to compare the influence of the diets of two diatoms, Navicula sp. (benthic) and Thalassiosira fluviatilis (planctonic), on the development, fecundity and survival of the harpacticoid copepod Tisbe biminiensis. In order to determine the optimal concentration of food, 35 egg-bearing females were submitted to six algal concentrations and controls (without food). After 24 h, the content of the recipients was fixed with 4% formalin and then fecal pellets produced by each female were counted and measured. The larval development was studied by surveying 50 nauplii on each diet individually until the adult stage, at intervals of 6 h. The cast exoskeletons were removed to count the number of segments and for measurement. The fecundity was obtained counting the naupliar production every 48 h of groups containing 10 females in different algal concentrations in both diets. The diet influence on fecundity was tested by submitting four groups of 10 females fed on optimal algal concentrations based on the fecal pellet experiments. In the Navicula sp. concentration of 0.4 μg Chl-a/ml, considered to be optimal for fecal pellets production, the diatoms were shown toxic, resulting in a low survival rate and inhibiting the egg production of copepods. The optimal concentration considering fecundity was estimated to be 0.1 μg Chl-a/ml for both diets. Copepods fed on Navicula sp. presented a faster development rate and higher naupliar production compared to copepods fed on T. fluviatilis. Size and survival did not vary significantly among diets. The algal concentration interfered significantly in the reproductive success of females. Both very low and very high algal concentrations reduced reproductive success. Concluding, the benthic diatom Navicula sp. was more favorable to the copepod T. biminiensis than the planktonic diatom T. fluviatilis.     

Experimental analysis of the ‘Paarkultureffekt’ in the protandric polychaete, Ophryotrocha puerilis Clap. Mecz.

Female Ophyrotrocha puerilis Clap. Mecz. were coupled. Certain parts of the prostomium and of the pygidium in one or both partners of a couple were amputated in order to prove that they are responsible for the mutual influence which partners normally exert on their sexual differentiation. The results demonstrate that the palps and the ventrolateral prostomial cirri are the transmitters and the ciliated lateral pits on the prostomium the receivers of the mutual influence. The antennae and the pygidial cirri are not necessary as far as the ‘Paarkultureffekt’ is concerned. The nature of the stimulus is still unknown. At the present stage of these investigations, the favoured conception is of a pheromone transmitted during the contact of the partners.