Mycoplasma pneumoniae Protein P30 Is Required for Cytadherence and Associated with Proper Cell Development

The attachment organelle of Mycoplasma pneumoniae is a polar, tapered cell extension containing an intracytoplasmic, electron-dense core. This terminal structure is the leading end in gliding motility, and its duplication is thought to precede cell division, raising the possibility that mutations affecting cytadherence also confer a defect in motility or cell development. Mycoplasma surface protein P30 is associated with the attachment organelle, and P30 mutants II-3 and II-7 do not cytadhere. In this study, the recombinant wild-type but not the mutant II-3 p30 allele restored cytadherence when transformed into P30 mutants by recombinant transposon delivery. The mutations associated with loss of P30 in mutant II-3 and reacquisition of P30 in cytadhering revertants thereof were identified by nucleotide sequencing of the p30 gene. Morphological abnormalities that included ovoid or multilobed cells having a poorly defined tip structure were associated with loss of P30. Digital image analysis confirmed quantitatively the morphological differences noted visually. Transformation of the P30 mutants with the wild-type p30 allele restored a normal morphology, as determined both visually and by digital image analysis, suggesting that P30 plays a role in mycoplasma cell development. Finally, the P30 mutants localized the adhesin protein P1 to the terminal organelle, indicating that P30 is not involved in P1 trafficking but may be required for its receptor-binding function.     

Convergent adaptation of Saccharomyces uvarum to sulfite,an antimicrobial preservative widely used in human-driven fermentations

Different species can find convergent solutions to adapt their genome to the same evolutionary constraints, although functional convergence promoted by chromosomal rearrangements in different species has not previously been found. In this work, we discovered that two domesticated yeast species, Saccharomyces cerevisiae, and Saccharomyces uvarum, acquired chromosomal rearrangements to convergently adapt to the presence of sulfite in fermentation environments. We found two new heterologous chromosomal translocations in fermentative strains of S. uvarum at the SSU1 locus, involved in sulfite resistance, an antimicrobial additive widely used in food production. These are convergent events that share similarities with other SSU1 locus chromosomal translocations previously described in domesticated S. cerevisiae strains. In S. uvarum, the newly described VII^XVI and XI^XVI chromosomal translocations generate an overexpression of the SSU1 gene and confer increased sulfite resistance. This study highlights the relevance of chromosomal rearrangements to promote the adaptation of yeast to anthropic environments.     

Inverse gene-for-gene interactions contribute additively to tan spot susceptibility in wheat

Key message

Tan spot susceptibility is conferred by multiple interactions of necrotrophic effector and host sensitivity genes.

Abstract

Tan spot of wheat, caused by Pyrenophora tritici-repentis, is an important disease in almost all wheat-growing areas of the world. The disease system is known to involve at least three fungal-produced necrotrophic effectors (NEs) that interact with the corresponding host sensitivity (S) genes in an inverse gene-for-gene manner to induce disease. However, it is unknown if the effects of these NE–S gene interactions contribute additively to the development of tan spot. In this work, we conducted disease evaluations using different races and quantitative trait loci (QTL) analysis in a wheat recombinant inbred line (RIL) population derived from a cross between two susceptible genotypes, LMPG-6 and PI 626573. The two parental lines each harbored a single known NE sensitivity gene with LMPG-6 having the Ptr ToxC sensitivity gene Tsc1 and PI 626573 having the Ptr ToxA sensitivity gene Tsn1. Transgressive segregation was observed in the population for all races. QTL mapping revealed that both loci (Tsn1 and Tsc1) were significantly associated with susceptibility to race 1 isolates, which produce both Ptr ToxA and Ptr ToxC, and the two genes contributed additively to tan spot susceptibility. For isolates of races 2 and 3, which produce only Ptr ToxA and Ptr ToxC, only Tsn1 and Tsc1 were associated with tan spot susceptibility, respectively. This work clearly demonstrates that tan spot susceptibility in this population is due primarily to two NE–S interactions. Breeders should remove both sensitivity genes from wheat lines to obtain high levels of tan spot resistance.

     

Yellow leaf syndrome modifies the composition of sugarcane juices in polysaccharides, phenols and polyamines

Some ultrastructural changes can be observed in diseased Saccharum officinarum L. (cv. Cuba 120-78) plants with visual symptoms of yellow leaf syndrome (YLS), used to discriminate between healthy and diseased plants. Abaxial epidermis of diseased leaves shows a large amount of adhered superficial bodies, which partially occluded some stomata. Bundle sheath cells surrounding the bottom of phloem of diseased leaves are separated from the conducting tissues by a large layer of an amorphous matrix similar to wax. Debris of the end wall can be observed in large xylem vessels. Sometimes, spherical bodies similar to phytoplasma can be observed in the intercellular spaces of bundle sheath cells. These particles have never been observed in healthy plants. YLS was also associated to an increase of the concentration of reducing sugars, glucose index, and glycoproteins recovered in juices whereas the amount of sucrose decreases. Sugarcane juices obtained from both healthy and YLS-affected Cuba 120-78 cultivars of sugarcane contained putrescine (PUT), cadaverine (CAD), spermidine and spermine (SPM) as free and macromolecules-conjugated compounds. Only CAD and SPM appeared as acid-soluble conjugates to small molecules whereas PUT and CAD are the major polyamines (PAs) conjugated to macromolecules, mainly to high molecular mass glycoproteins. The disease was associated to an increase in total PA fraction. Arginase and ornithine decarboxylase activities, responsible for the synthesis of PUT, were higher in YLS juices than in those obtained from healthy plants. CAD and SPM presumably conjugated mostly to chlorogenic, syringic and ferulic acids in juices from YLS plants.     

Are bioleaching rates determined by the available particle surface area concentration?

It is well known that pulp density and particle size determine the available surface area concentration and have an influence in the overall rate of bioleaching of minerals. As metal solubilization takes place through the surface area of the particles, it can be expected that different combinations of pulp densities and particle sizes giving the same surface area concentration would determine the same leaching rate. The objective of this work was to test this hypothesis on the effect of surface area concentration, pulp density and particle size of the biooxidation of a pyritic gold concentrate by the thermophilic Archaeon Sulfolobus metallicus in shake flasks. The gold concentrate was used at 2.5%, 5%, 10%, and 15% w/v pulp density and at four size fractions: 150–106, 106–75, 75–38 and –38 μm. Temperature was 68°C and the initial pH was 2.0. Results showed that the volumetric productivities of iron and sulfate depend not only on the surface area concentration but also on pulp density and particle size considered separately. These two variables not only determine surface area but also exert additional effects on the process, so the hypothesis was not confirmed. Maximum attained iron productivity was 1.042 g/l day with the 75–38 μm fraction at 5% pulp density. Maximum sulfate productivity was 4.279 g/l day with the 75–38 μm fraction at 10% pulp density.     

Identification and characterization of novel TRPV4 modulators

TRPV4, a close relative of the vanilloid receptor TRPV1, is activated by diverse modalities such as endogenous lipid ligands, hypotonicity, protein kinases and, possibly, mechanical inputs. While its multiple roles in vivo are being explored with KO mice and selective agonists, there is a dearth of selective antagonists available to examine TRPV4 function. Herein we detail the use of a focused library of commercial compounds in order to identify RN-1747 and RN-1734, a pair of structurally related small molecules endowed with TRPV4 agonist and antagonist properties, respectively. Their activities against human, rat and mouse TRPV4 were characterized using electrophysiology and intracellular calcium influx. Significantly, antagonist RN-1734 was observed to completely inhibit both ligand- and hypotonicity-activated TRPV4. In addition, RN-1734 was found to be selective for TRPV4 in a TRP selectivity panel including TRPV1, TRPV3 and TRPM8, and could thus be a valuable pharmacological probe for TRPV4 studies.     

Automated classification of bird and amphibian calls using machine learning: A comparison of methods

We compared the ability of three machine learning algorithms (linear discriminant analysis, decision tree, and support vector machines) to automate the classification of calls of nine frogs and three bird species. In addition, we tested two ways of characterizing each call to train/test the system. Calls were characterized with four standard call variables (minimum and maximum frequencies, call duration and maximum power) or eleven variables that included three standard call variables (minimum and maximum frequencies, call duration) and a coarse representation of call structure (frequency of maximum power in eight segments of the call). A total of 10,061 isolated calls were used to train/test the system. The average true positive rates for the three methods were: 94.95% for support vector machine (0.94% average false positive rate), 89.20% for decision tree (1.25% average false positive rate) and 71.45% for linear discriminant analysis (1.98% average false positive rate). There was no statistical difference in classification accuracy based on 4 or 11 call variables, but this efficient data reduction technique in conjunction with the high classification accuracy of the SVM is a promising combination for automated species identification by sound. By combining automated digital recording systems with our automated classification technique, we can greatly increase the temporal and spatial coverage of biodiversity data collection.     

Development of tetranucleotide microsatellite markers for the cushion star, Asterina gibbosa, and cross-species amplification

We describe nine polymorphic tetranucleotide microsatellite loci from the starfish, Asterina gibbosa. Loci were isolated from a partial genomic library that had been enriched for AAAC repeat sequences. Number of alleles per locus ranged from two to 14 in a sample of 85 individuals from three populations (two from Spain and one from the UK). Observed and expected heterozygosities per population ranged from 0.000 to 0.400 and from 0.040 to 0.784, respectively. All loci presented significant heterozygote deficits in one or more populations. Eight of these loci were amplified and variable in A. pancerii and A. phylactica. These loci will be used to study population structure in A. gibbosa.     

In Vivo Generation of Neurotoxic Prion Protein: Role for Hsp70 in Accumulation of Misfolded Isoforms

Prion diseases are incurable neurodegenerative disorders in which the normal cellular prion protein (PrP^C) converts into a misfolded isoform (PrP^Sc) with unique biochemical and structural properties that correlate with disease. In humans, prion disorders, such as Creutzfeldt-Jakob disease, present typically with a sporadic origin, where unknown mechanisms lead to the spontaneous misfolding and deposition of wild type PrP. To shed light on how wild-type PrP undergoes conformational changes and which are the cellular components involved in this process, we analyzed the dynamics of wild-type PrP from hamster in transgenic flies. In young flies, PrP demonstrates properties of the benign PrP^C; in older flies, PrP misfolds, acquires biochemical and structural properties of PrP^Sc, and induces spongiform degeneration of brain neurons. Aged flies accumulate insoluble PrP that resists high concentrations of denaturing agents and contains PrP^Sc-specific conformational epitopes. In contrast to PrP^Sc from mammals, PrP is proteinase-sensitive in flies. Thus, wild-type PrP rapidly converts in vivo into a neurotoxic, protease-sensitive isoform distinct from prototypical PrP^Sc. Next, we investigated the role of molecular chaperones in PrP misfolding in vivo. Remarkably, Hsp70 prevents the accumulation of PrP^Sc-like conformers and protects against PrP-dependent neurodegeneration. This protective activity involves the direct interaction between Hsp70 and PrP, which may occur in active membrane microdomains such as lipid rafts, where we detected Hsp70. These results highlight the ability of wild-type PrP to spontaneously convert in vivo into a protease-sensitive isoform that is neurotoxic, supporting the idea that protease-resistant PrP^Sc is not required for pathology. Moreover, we identify a new role for Hsp70 in the accumulation of misfolded PrP. Overall, we provide new insight into the mechanisms of spontaneous accumulation of neurotoxic PrP and uncover the potential therapeutic role of Hsp70 in treating these devastating disorders.