Host‐ and stage‐dependent secretome of the arbuscular mycorrhizal fungus Rhizophagus irregularis

Arbuscular mycorrhizal fungi form the most wide‐spread endosymbiosis with plants. There is very little host specificity in this interaction, however host preferences as well as varying symbiotic efficiencies have been observed. We hypothesize that secreted proteins (SPs) may act as fungal effectors to control symbiotic efficiency in a host‐dependent manner. Therefore, we studied whether arbuscular mycorrhizal (AM) fungi adjust their secretome in a host‐ and stage‐dependent manner to contribute to their extremely wide host range. We investigated the expression of SP‐encoding genes of Rhizophagus irregularis in three evolutionary distantly related plant species, Medicago truncatula, Nicotiana benthamiana and Allium schoenoprasum. In addition we used laser microdissection in combination with RNA‐seq to study SP expression at different stages of the interaction in Medicago. Our data indicate that most expressed SPs show roughly equal expression levels in the interaction with all three host plants. In addition, a subset shows significant differential expression depending on the host plant. Furthermore, SP expression is controlled locally in the hyphal network in response to host‐dependent cues. Overall, this study presents a comprehensive analysis of the R. irregularis secretome, which now offers a solid basis to direct functional studies on the role of fungal SPs in AM symbiosis.     

The Plasmodium knowlesi MAHRP2 ortholog localizes to structures connecting Sinton Mulligan's clefts in the infected erythrocyte

During development within the host erythrocyte malaria parasites generate nascent membranous structures which serve as a pathway for parasite protein transport to modify the host cell. The molecular basis of such membranous structures is not well understood, particularly for malaria parasites other than Plasmodium falciparum. To characterize the structural basis of protein trafficking in the Plasmodium knowlesi-infected erythrocyte, we identified a P. knowlesi ortholog of MAHRP2, a marker of the tether structure that connects membranous structures in the P. falciparum-infected erythrocyte. We show that PkMAHRP2 localizes on amorphous structures that connect Sinton Mulligan's clefts (SMC) to each other and to the erythrocyte membrane. Three dimensional reconstruction of the P. knowlesi-infected erythrocyte revealed that the SMC is a plate-like structure with swollen ends, reminiscent of the morphology of the Golgi apparatus. The PkMAHRP2-localized amorphous structures are possibly functionally equivalent to P. falciparum tether structure. These findings suggest a conservation in the ultrastructure of protein trafficking between P. falciparum and P. knowlesi.     

Temporal and Spatial Diversity and Distribution of Arboreal Carabidae (Coleoptera) in a Western Amazonian Rain Forest1

Diversity of arboreal carabid beetles was sampled by fumigation in 100 3 × 3 m stations within a 100 × 1000 m terra firme forest plot in Ecuadorian Amazonia. Nine sampling dates from January 1994 to October 1996 yielded 2329 individuals belonging to 318 species of which more than 50 percent were undescribed species. A high percentage of the species sampled were rare; the proportion that occurred once per sampling date (singletons) ranged from 50.0 to 62.5 percent. Estimates of species richness were from 82 to 282 species of arboreal carabids in the study plot on a given sampling date. Most richness values were greater than 173 species. Species accumulation curves attained asymptotes for all but one sampling date, indicating that an adequate level of sampling effort was used to characterize the diversity of carabid fauna. Total accumulation curves based on pooled data failed to reach asymptotes. There was a high turnover in species composition between sampling dates; less than 50 percent of the species between the majority of sampling dates were shared, suggesting that the total species pool may be extremely large. Although species composition changed seasonally, species richness varied little. Spatial autocorrelation analysis revealed that the structure of this species assemblage was significantly patterned at distances below 280 m. Taken together, the large percentage of undescribed species, die failure of the overall species accumulation curves to level off, and the high turnover in species composition indicate that the species diversity of carabid beetles is far higher than previously thought.     

Growth and survival of Bacillus cereus in mageu,a sour maize beverage

The growth and survival of Bacillus cereus, a known pathogen commonly found in cereals, during lactic acid fermentation of mageu, a sour maize beverage, was studied. In the mageu base inoculated with both the starter culture and B. cereus, the acidity developed to pH 4.00 and 0.10% titratable acidity after 24 h; the growth of B. cereus was reduced from 10⁶ c.f.u./ml to 10² c.f.u./ml within 24 h; after the first 6 h of fermentation, the rate of inhibition of B. cereus was correlated to the rate of decrease in pH (r = 0.85, p < 0.05); the redox potential (E_h) decreased from 463 to 149 mV within the first 12 h. The control mageu base to which neither starter nor lactic acid was added, had a pH of 6.50, titratable acidity of 0.015% and lowest E_h of 244 mV. In the mageu base to which lactic acid and B. cereus were added, the pathogen was inhibited to < 10¹ c.f.u./ml. The B. cereus in the mageu base to which no starter culture nor lactic acid was added, grew to over 10⁷ c.f.u./ml after 12 h. The decrease in E_h seemed to have no inhibitory effect on the growth and survival of B. cereus. No strains of lactic acid bacteria were found to produce bacteriocins antagonistic to B. cereus. Low pH and acidity were found to be the major factors inhibiting growth of B. cereus in mageu.     

Genetic characterization of cassava (Manihot esculenta Crantz) genotypes using agro-morphological and single nucleotide polymorphism markers

Physiology and Molecular Biology of Plants - Dearth of information on extent of genetic variability in cassava limits the genetic improvement of cassava genotypes in Sierra Leone. The aim of this...     

Rickettsia africae infection rates and transovarial transmission in Amblyomma hebraeum ticks in Mnisi,Bushbuckridge, South Africa

Rickettsia africae is a gram-negative bacterium, which causes African tick bite fever (ATBF) in humans. ATBF is a febrile disease mainly affecting travellers to southern Africa. This bacterium is known to be transmitted by Amblyomma hebraeum and Amblyomma variegatum ticks. In southern Africa, the principal vector is A. hebraeum. Febrile disease is a serious issue in the study area. There is a high prevalence of non-malaria illness caused by Rickettsia, so there is a need to have more knowledge on these species. Infection rates and transovarial transmission efficiency of R. africae in A. hebraeum ticks were investigated in a rural area of Mpumalanga province, South Africa. Adult and engorged A. hebraeum female ticks were collected from cattle. Larvae were collected by dragging a cloth at ground level using 100 steps, equivalent to an area of 100 m². Tick identification was performed according to standard taxonomic keys using a microscope. Engorged ticks were incubated to oviposit and egg masses were collected. DNA was extracted from the ticks, larvae and egg masses, and screened for gltA and ompA genes, using quantitative real-time PCR and conventional PCR, respectively. Positive ompA amplicons were sequenced and phylogenetic analysis showed 99.8-100% identity with R. africae. Infection rates were 13.7 and 12.7% for adults and larvae, respectively. Transovarial transmission of R. africae in A. hebraeum from this study was 85.7%. The results provide a clear indication that people living in the study area and travellers that visit the area are at risk of contracting ATBF.

     

Electron microscopy of human skin fibroblasts in situ during growth in culture

We have studied the electron microscopic (EM) appearance of cultured diploid human skin fibroblasts during logarithmic and confluent stages of growth using a simple technique which permits in situ visualization of individual cells in monolayer. By comparison, cells disrupted from a monolayer and pelleted showed drastic distortion of the cell surface and appearance of organelles: mechanical scraping produced massive dilatation of the rough endoplasmic reticulum (RER); and trypsin-produced multiple blebs of the plasma membrane and cytoplasmic vacuoles. In situ, these changes in trypsinized cells disappeared within 1 h after plating. Six hours later, microtubules and microfilaments had surrounded the nucleus and were oriented in longitudinal bundles beneath the plasma membrane during rapid growth. At confluence these cytoskeletal elements seemed to extend beyond the plasma membrane at intercellular junctions. Pinocytotic vesicles were abundant at those surfaces devoid of filaments. Mitochondria, aligned with the long axis of the cell, were extremely long and narrow. During logarithmic growth there were many free ribosomes, polysomes, and some flat cisternae of RER. At confluence most ribosomes were found in spirals on dilated saccules of RER which contained electron-dense material. Lysosomes of several types were present during all phases of growth and varied in number from cell to cell. Late in culture the lysosomes tended to be larger, often occupying whole areas of cytoplasm or even extruding from the cell, and resembled those seen in lysosomal storage diseases. Understanding the in situ ultrastructure of normal human fibroblasts during growth in culture will permit systematic examination of such cells in a variety of pathological states.