The extraradical proteins of Rhizophagus irregularis: A shotgun proteomics approach

Arbuscular Mycorrhizal fungi (AMF, Glomeromycota) form obligate symbiotic associations with the roots of most terrestrial plants. Our understanding of the molecular mechanisms enabling AMF propagation and AMF-host interaction is currently incomplete. Analysis of AMF proteomes could yield important insights and generate hypotheses on the nature and mechanism of AMF-plant symbiosis. Here, we examined the extraradical mycelium proteomic profile of the arbuscular mycorrhizal fungus Rhizophagus irregularis grown on Ri T-DNA transformed Chicory roots in a root organ culture setting. Our analysis detected 529 different peptides that mapped to 474 translated proteins in the R. irregularis genome. R. irregularis proteome was characterized by a high proportion of proteins (9.9 % of total, 21.4 % of proteins with functional prediction) mediating a wide range of signal transduction processes, e.g. Rho1 and Bmh2, Ca-signaling (calmodulin, and Ca channel protein), mTOR signaling (MAP3K7, and MAPKAP1), and phosphatidate signaling (phospholipase D1/2) proteins, as well as members of the Ras signaling pathway. In addition, the proteome contained an unusually large proportion (53.6 %) of hypothetical proteins, the majority of which (85.8 %) were Glomeromycota-specific. Forty-eight proteins were predicted to be surface/membrane associated, including multiple hypothetical proteins of yet-unrecognized functions. However, no evidence for the overproduction of specific proteins, previously implicated in promoting soil health and aggregation was obtained. Finally, the comparison of R. irregularis proteome to previously published AMF proteomes identified a core set of pathways and processes involved in AMF growth. We conclude that R. irregularis growth on chicory roots requires the activation of a wide range of signal transduction pathways, the secretion of multiple novel hitherto unrecognized Glomeromycota-specific proteins, and the expression of a wide array of surface-membrane associated proteins for cross kingdom cell-to-cell communications.     

Acoustic and Morphological Differentiation in the Frog Allobates femoralis: Relationships with the Upper Madeira River and Other Potential Geological Barriers

We studied patterns of call acoustics and external morphological differentiation in populations of the dart-poison frog Allobates femoralis occurring in forested areas along a 250-km stretch of the upper Madeira River, Brazil. Multivariate analyses of variance using principal components representing shared acoustic and morphological parameters distinguished three groups in relation to call structure and external morphology: (1) populations belonging to a two-note call morphotype; (2) populations with four-note calls inhabiting the left riverbank; and (3) populations with four-note calls inhabiting the right riverbank. Our results report a case of Amazonian anuran diversity hidden by current taxonomy and provide evidence for the upper Madeira River being a boundary between distinct populations of A. femoralis , and suggest a new taxonomic interpretation for these groups. Samples that did not fit into the general differentiation pattern and the existence of a well-defined contact zone between two morphotypes on the left riverbank indicate that mechanisms complementary to river-barrier hypotheses are necessary to explain the phenotypic differentiation between populations. Our study shows that at least one anuran species shows congruence between population differentiation and separation by a large Amazonian river, as documented for birds and mammals. Conservation efforts should not consider the taxon now known as A. femoralis as a homogeneous entity. There is much within-taxon variability, which can be probably explained partly by the existence of cryptic species, partly by geological barriers and part of which currently has no obvious explanation.     

DasFestuca vivipara-Problem in den Alpen

The differences betweenFestuca vivipara (tetraploid) andF. ovina subsp.supina (diploid), two often confused taxa, are demonstrated in regard to morphology, leaf anatomy, ecology and distribution. New maps illustrate the distribution ofF. vivipara in the Alps and the Northern Hemisphere. The development of different polyploidy levels withinF. vivipara as a consequence of occasional sexual processes is discussed. (English Summary on p. 39.)

     

Distribution of PERIOD-immunoreactive neurons and temporal change of the immunoreactivity under long-day and short-day conditions in the larval brain of the flesh fly Sarcophaga similis

The flesh fly Sarcophaga similis show a clear photoperiodic response; they develop into adults under long days, whereas they arrest their development at the pupal stage under short days. Although the involvement of a circadian clock in photoperiodic time measurement is suggested in this species, the anatomical location of the clock neurons responsible for the time measurement has been unknown. We detected two PERIOD-immunoreactive cell clusters in the larval brain; one cluster was located at the dorsoanterior region and the other at the medial region. We further investigated their temporal changes in PERIOD-immunoreactivity and compared their patterns under different photoperiods.     

Preparation and Properties of Human Kappa-casein-like Fraction

A κ-casein-like fraction was prepared from human whole casein by gel filtration with Sephadex G-150 and Biogel A-150 m. The fraction was calcium-insensitive and its solution became turbid by rennin. In polyacrylamide gel electrophoretic (PAE) analysis, the fraction gave 11~12 bands after reduction with 2-mercaptoethanol. It appeared to exist as a disulfide- bound complex of many components. The occurrence of human para-IC-caseins from the fraction after rennin treatment was confirmed by PAE. When the reduced, alkylated human κ-casein-like fraction was chromatographed by DEAE-cellulose, several fractions were obtained. After rennin treatment, they formed either a para-SCM-κ-casein band moving toward the cathode on PAE pattern or the one which moved toward the anode. These results suggest that two para-κ-caseins were formed from human whole casein or human κ-casein-like fraction by the action of rennin.     

Free-Radical Scavenging Activity of Submerged Mycelium Extracts from Higher Basidiomycetes Mushrooms

Twenty-four Basidiomycetes strains were evaluated to determine their free-radical scavenging capacity in submerged cultivation. The scavenging capacity of the extracts varied from 1 to 85% depending on the mushroom species, solvent used, and concentration. A calculation of EC₅₀ of extracts from several wood-rotting Basidiomycetes showed high scavenging abilities at low effective concentration.     

Mechanisms of host discrimination and intraspecific competition in the aphid parasitoid Ephedrus cerasicola

The aphid parasitoid Ephedrus cerasicola Starý (Hymenoptera: Aphidiidae) discriminates between parasitized and unparasitized aphid hosts (Myzus persicae (Sulzer), Homoptera: Aphididae) in two different ways. By antennal contact with the parasitized host, it can detect an external marking pheromone which is effective only for a few hours. The behaviour of the parasitoid also indicated that it responded to an internal marker, probably by means of chemoreceptors on the ovipositor. The parasitoid behaviour was studied in encounters with parasitized hosts 0–10 min, 2–3 h, 6–7 h and 23–24 h after the first parasitization. Aphids were superparasitized within 1 h or after 24 h, and aphids in each experiment were later dissected at two different times. It is suggested that the supernumerary offspring were eliminated by chemical means, whether the time between the two ovipositions was 0–1 h or 24 h.

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Résumé Le parasitoïde de puceron, E. cerasicola Starý (Hym.: Aphidiidae) sélectionne les pucerons, Myzus persicae (Sulzer), parasités ou non, de deux façons différentes. Il peut percevoir une phéromone externe de marquage perceptible seulement pendant quelques heures par contact antennaire. Son comportement montre qu'il réagit aussi à un marquage interne, probablement par des chémorécepteurs de sa tarière. Le comportement du parasitoïde a été étudié en le mettant en contact avec des hôtes parasités depuis 0–10 min, 2–3 h, 6–7 h et 23–24 h. Les pucerons ont été surparasités pendant la première heure ou après 24 heures. Les pucerons de toutes les expériences ont été disséqués à deux moments différents. Ceci a conduit à estimer que les descendants en surnombre ont été éliminés chimiquement quand la période entre deux pontes successives était inférieure à 1 h ou supérieure à 24 h.

     

Development of cranial muscles in the actinopterygian fish Senegal bichir,Polypterus senegalus Cuvier, 1829

Polypterus senegalus Cuvier, 1829 is one of the most basal living actinopterygian fish and a member of the Actinopterygii. We analyzed the spatial and temporal pattern of cranial muscle development of P. senegalus using whole‐mount immunostaining and serial sectioning. We described the detailed structure of the external gill muscles which divided into dorsal and ventral parts after yolk exhaustion. The pattern of the division is similar to that of urodeles. We suggest that, the external gill muscles of P. senegalus are involved in spreading and folding of the external gill stem and the branches. The fibers of the external gill muscles appear postero‐lateral to the auditory capsule. In addition, the facial nerve passes through the external gills. Therefore, the external gill muscles are probably derived from the m. constrictor hyoideus dorsalis. In contrast to previous studies, we described the mm. interhyoideus and hyohyoideus fibers as independent components in the yolk‐sac larvae. The m. hyohyoideus fibers appear lateral to the edge of the ventral portion of the external gill muscles, which are probably derived from the m. constrictor hyoideus dorsalis. These findings suggest that the m. hyohyoidues is derived from the m. constrictor hyoideus dorsalis in P. senegalus . In other actinopterygians, the m. hyohyoideus is derived from the m. constrictor hyoideus ventralis; therefore, the homology of the m. hyohyoidues of P. senegalus and other actinopterygians remains unclear. J. Morphol. 278:450–463, 2017. © 2017 Wiley Periodicals, Inc.