Rhizosphere microbial community composition shifts diurnally and in response to natural variation in host clock phenotype.

Plant-associated microbial assemblages are known to shift at time scales aligned with plant phenology, as influenced by the changes in plant-derived nutrient concentrations and abiotic conditions observed over a growing season. But these same factors can change dramatically in a sub-24-hour period, and it is poorly understood how such diel cycling may influence plant-associated microbiomes. Plants respond to the change from day to night via mechanisms collectively referred to as the internal "clock," and clock phenotypes are associated with shifts in rhizosphere exudates and other changes that we hypothesize could affect rhizosphere microbes. The mustard Boechera stricta has wild populations that contain multiple clock phenotypes of either a 21- or a 24-hour cycle. We grew plants of both phenotypes (two genotypes per phenotype) in incubators that simulated natural diel cycling or that maintained constant light and temperature. Under both cycling and constant conditions, the extracted DNA concentration and the composition of rhizosphere microbial assemblages differed between time points, with daytime DNA concentrations often triple what were observed at night and microbial community composition differing by, for instance, up to 17%. While we found that plants of different genotypes were associated with variation in rhizosphere assemblages, we did not see an effect on soil conditioned by a particular host plant circadian phenotype on subsequent generations of plants. Our results suggest that rhizosphere microbiomes are dynamic at sub-24-hour periods, and those dynamics are shaped by diel cycling in host plant phenotype. IMPORTANCE We find that the rhizosphere microbiome shifts in composition and extractable DNA concentration in sub-24-hour periods as influenced by the plant host's internal clock. These results suggest that host plant clock phenotypes could be an important determinant of variation in rhizosphere microbiomes.

Whole-Genome Duplication and Host Genotype Affect Rhizosphere Microbial Communities.

The composition of microbial communities found in association with plants is influenced by host phenotype and genotype. However, the ways in which specific genetic architectures of host plants shape microbiomes are unknown. Genome duplication events are common in the evolutionary history of plants and influence many important plant traits, and thus, they may affect associated microbial communities. Using experimentally induced whole-genome duplication (WGD), we tested the effect of WGD on rhizosphere bacterial communities in Arabidopsis thaliana. We performed 16S rRNA amplicon sequencing to characterize differences between microbiomes associated with specific host genetic backgrounds (Columbia versus Landsberg) and ploidy levels (diploid versus tetraploid). We modeled relative abundances of bacterial taxa using a hierarchical Bayesian approach. We found that host genetic background and ploidy level affected rhizosphere community composition. We then tested to what extent microbiomes derived from a specific genetic background or ploidy level affected plant performance by inoculating sterile seedlings with microbial communities harvested from a prior generation. We found a negative effect of the tetraploid Columbia microbiome on growth of all four plant genetic backgrounds. These findings suggest an interplay between host genetic background and ploidy level and bacterial community assembly with potential ramifications for host fitness. Given the prevalence of ploidy-level variation in both wild and managed plant populations, the effects on microbiomes of this aspect of host genetic architecture could be a widespread driver of differences in plant microbiomes. IMPORTANCE Plants influence the composition of their associated microbial communities, yet the underlying host-associated genetic determinants are typically unknown. Genome duplication events are common in the evolutionary history of plants and affect many plant traits. Using Arabidopsis thaliana, we characterized how whole-genome duplication affected the composition of rhizosphere bacterial communities and how bacterial communities associated with two host plant genetic backgrounds and ploidy levels affected subsequent plant growth. We observed an interaction between ploidy level and genetic background that affected both bacterial community composition and function. This research reveals how genome duplication, a widespread genetic feature of both wild and crop plant species, influences bacterial assemblages and affects plant growth.

Greenhouse gases and performance of growing pigs fed wheat-based diets containing wheat millrun and a multi-carbohydrase enzyme.

The objective of this project was to determine the impact of feeding growing pigs with high wheat millrun diets supplemented with a multi-carbohydrase enzyme (amylase, cellulase, glucanase, xylanase, and invertase activities) on nutrient digestibility, growth performance, and greenhouse gas (GHG) output (carbon dioxide, CO2; nitrous oxide, N2O; methane, CH4). Three experiments were conducted utilizing six treatments arranged as a 3 × 2 factorial (0%, 15%, or 30% wheat millrun; with or without enzyme) for the digestibility experiment or as a 2 × 2 factorial (0% or 30% wheat millrun; with or without enzyme) for the performance and GHG experiments. The digestibility, performance, and GHG experiments utilized 48 individually housed pigs, 180 pigs housed 5 per pen, or 96 pigs housed 6 per chamber, respectively. Increasing wheat millrun up to 30% in the diet of growing pigs resulted in decreased energy, nitrogen (N) and phosphorus (P) apparent total tract digestibility and net energy content (P < 0.01). Overall, average daily gain (ADG) and gain to feed ratio were reduced in pigs fed wheat millrun (P < 0.05). Enzyme supplementation had minimal effects on the digestibility or performance parameters measured. Feeding diets with 30% millrun did not affect GHG output (CH4: 4.7 and 4.9; N2O: 0.45 and 0.42; CO2: 1,610 and 1,711 mg/s without or with millrun inclusion, respectively; P > 0.78). Enzyme supplementation had no effect on GHG emissions (CH4: 4.5 and 5.1; N2O: 0.46 and 0.42; CO2: 1,808 and 1,513 mg/s without or with enzymes, respectively; P > 0.51). Overall, the carbohydrase enzyme had minimal effects on parameters measured, regardless of wheat millrun inclusion (P > 0.10). Although energy, N and P digestibility, and ADG were reduced, the inclusion of up to 30% wheat millrun in the diet has no effect on GHG emissions from growing pigs (P > 0.10).

The effect of rhizosphere microbes outweighs host plant genetics in reducing insect herbivory.

Rhizosphere microbes affect plant performance, including plant resistance against insect herbivores; yet, a direct comparison of the relative influence of rhizosphere microbes versus plant genetics on herbivory levels and on metabolites related to defence is lacking. In the crucifer Boechera stricta, we tested the effects of rhizosphere microbes and plant population on herbivore resistance, the primary metabolome, and select secondary metabolites. Plant populations differed significantly in the concentrations of six glucosinolates (GLS), secondary metabolites known to provide herbivore resistance in the Brassicaceae. The population with lower GLS levels experienced ~60% higher levels of aphid (Myzus persicae) attack; no association was observed between GLS and damage by a second herbivore, flea beetles (Phyllotreta cruciferae). Rhizosphere microbiome (disrupted vs. intact native microbiome) had no effect on plant GLS concentrations. However, aphid number and flea beetle damage were respectively about three- and seven-fold higher among plants grown in the disrupted versus intact native microbiome treatment. These differences may be attributable to shifts in primary metabolic pathways previously implicated in host defence against herbivores, including increases in pentose and glucoronate interconversion among plants grown with an intact microbiome. Furthermore, native microbiomes with distinct community composition (as estimated from 16s rRNA amplicon sequencing) differed two-fold in their effect on host plant susceptibility to aphids. The findings suggest that rhizosphere microbes, including distinct native microbiomes, can play a greater role than population in defence against insect herbivores, and act through metabolic mechanisms independent of population.

The plant circadian clock influences rhizosphere community structure and function.

Plants alter chemical and physical properties of soil, and thereby influence rhizosphere microbial community structure. The structure of microbial communities may in turn affect plant performance. Yet, outside of simple systems with pairwise interacting partners, the plant genetic pathways that influence microbial community structure remain largely unknown, as are the performance feedbacks of microbial communities selected by the host plant genotype. We investigated the role of the plant circadian clock in shaping rhizosphere community structure and function. We performed 16S ribosomal RNA gene sequencing to characterize rhizosphere bacterial communities of Arabidopsis thaliana between day and night time points, and tested for differences in community structure between wild-type (Ws) vs clock mutant (toc1-21, ztl-30) genotypes. We then characterized microbial community function, by growing wild-type plants in soils with an overstory history of Ws, toc1-21 or ztl-30 and measuring plant performance. We observed that rhizosphere community structure varied between day and night time points, and clock misfunction significantly altered rhizosphere communities. Finally, wild-type plants germinated earlier and were larger when inoculated with soils having an overstory history of wild-type in comparison with clock mutant genotypes. Our findings suggest the circadian clock of the plant host influences rhizosphere community structure and function.

Activation of the NLRP3 inflammasome by IAV virulence protein PB1-F2 contributes to severe pathophysiology and disease.

The ability for a host to recognize infection is critical for virus clearance and often begins with induction of inflammation. The PB1-F2 of pathogenic influenza A viruses (IAV) contributes to the pathophysiology of infection, although the mechanism for this is unclear. The NLRP3-inflammasome has been implicated in IAV pathogenesis, but whether IAV virulence proteins can be activators of the complex is unknown. We investigated whether PB1-F2-mediated activation of the NLRP3-inflammasome is a mechanism contributing to overt inflammatory responses to IAV infection. We show PB1-F2 induces secretion of pyrogenic cytokine IL-1ß by activating the NLRP3-inflammasome, contributing to inflammation triggered by pathogenic IAV. Compared to infection with wild-type virus, mice infected with reverse engineered PB1-F2-deficient IAV resulted in decreased IL-1ß secretion and cellular recruitment to the airways. Moreover, mice exposed to PB1-F2 peptide derived from pathogenic IAV had enhanced IL-1ß secretion compared to mice exposed to peptide derived from seasonal IAV. Implicating the NLRP3-inflammasome complex specifically, we show PB1-F2 derived from pathogenic IAV induced IL-1ß secretion was Caspase-1-dependent in human PBMCs and NLRP3-dependent in mice. Importantly, we demonstrate PB1-F2 is incorporated into the phagolysosomal compartment, and upon acidification, induces ASC speck formation. We also show that high molecular weight aggregated PB1-F2, rather than soluble PB1-F2, induces IL-1ß secretion. Furthermore, NLRP3-deficient mice exposed to PB1-F2 peptide or infected with PB1-F2 expressing IAV were unable to efficiently induce the robust inflammatory response as observed in wild-type mice. In addition to viral pore forming toxins, ion channel proteins and RNA, we demonstrate inducers of NLRP3-inflammasome activation may include disordered viral proteins, as exemplified by PB1-F2, acting as host pathogen 'danger' signals. Elucidating immunostimulatory PB1-F2 mediation of NLRP3-inflammasome activation is a major step forward in our understanding of the aetiology of disease attributable to exuberant inflammatory responses to IAV infection.

Analysis of the attenuation of railway squeal noise by preloaded rings inserted in wheels.

Squeal from railway wheels occurring in short radius curves produces a very intense and highly annoying noise in the range 400-8000 Hz. When the excitation, due to lateral forces acting on the wheel, cannot be avoided, additional systems can be added on the wheel to limit acoustic emission. A very economical approach is the use of metal rings inserted into grooves machined in the wheels. Unfortunately the effectiveness of these so called damping rings varies from one wheel to another and for different rings. Because the mechanisms of attenuation are not well understood, these variations have not to date been explained. The aim of this paper is to clarify the attenuation mechanisms for damping rings especially for the first three axial wheel modes, which are the predominant sound radiated ones in curve passage and for which the effectiveness of the treatment is lower. It has been generally assumed that friction between the ring and the groove has been the mechanism for squeal noise attenuation. Here it is shown that the vibration attenuation is due to modal coupling between the wheel and the ring. The validity of this proposed mechanism is investigated using experimental measurements and theoretical and numerical models. The results presented here will provide an avenue for optimization of the damping ring noise control treatment to obtain significant levels of squeal noise attenuation notably for the first three axial modes.

The chloroplast genomes of the green algae Pyramimonas, Monomastix, and Pycnococcus shed new light on the evolutionary history of prasinophytes and the origin of the secondary chloroplasts of euglenids.

Because they represent the earliest divergences of the Chlorophyta and include the smallest known eukaryotes (e.g., the coccoid Ostreococcus), the morphologically diverse unicellular green algae making up the Prasinophyceae are central to our understanding of the evolutionary patterns that accompanied the radiation of chlorophytes and the reduction of cell size in some lineages. Seven prasinophyte lineages, four of which exhibit a coccoid cell organization (no flagella nor scales), were uncovered from analysis of nuclear-encoded 18S rDNA data; however, their order of divergence remains unknown. In this study, the chloroplast genome sequences of the scaly quadriflagellate Pyramimonas parkeae (clade I), the coccoid Pycnococcus provasolii (clade V), and the scaly uniflagellate Monomastix (unknown affiliation) were determined, annotated, and compared with those previously reported for green algae/land plants, including two prasinophytes (Nephroselmis olivacea, clade III and Ostreococcus tauri, clade II). The chlorarachniophyte Bigelowiella natans and the euglenid Euglena gracilis, whose chloroplasts originate presumably from distinct green algal endosymbionts, were also included in our comparisons. The three newly sequenced prasinophyte genomes differ considerably from one another and from their homologs in overall structure, gene content, and gene order, with the 80,211-bp Pycnococcus and 114,528-bp Monomastix genomes (98 and 94 conserved genes, respectively) resembling the 71,666-bp Ostreococcus genome (88 genes) in featuring a significantly reduced gene content. The 101,605-bp Pyramimonas genome (110 genes) features two conserved genes (rpl22 and ycf65) and ancestral gene linkages previously unrecognized in chlorophytes as well as a DNA primase gene putatively acquired from a virus. The Pyramimonas and Euglena cpDNAs revealed uniquely shared derived gene clusters. Besides providing unequivocal evidence that the green algal ancestor of the euglenid chloroplasts belonged to the Pyramimonadales, phylogenetic analyses of concatenated chloroplast genes and proteins elucidated the position of Monomastix and showed that the Mamiellales, a clade comprising Ostreococcus and Monomastix, are sister to the Pyramimonadales + Euglena clade. Our results also revealed that major reduction in gene content and restructuring of the chloroplast genome occurred in conjunction with important changes in cell organization in at least two independent prasinophyte lineages, the Mamiellales and the Pycnococcaceae.

Aberration correction in an adaptive free-space optical interconnect with an error diffusion algorithm.

Aberration correction within a free-space optical interconnect based on a spatial light modulator for beam steering and holographic wavefront correction is presented. The wavefront sensing technique is based on an extension of a modal wavefront sensor described by Neil et al. [J. Opt. Soc. Am. A 17, 1098 (2000)], which uses a diffractive element. In this analysis such a wavefront sensor is adapted with an error diffusion algorithm that yields a low reconstruction error and fast reconfigurability. Improvement of the beam propagation quality (Strehl ratio) for different channels across the input plane is achieved. However, due to the space invariancy of the system, a trade-off among the beam propagation quality for channels is obtained. Experimental results are presented and discussed.

Cross-talk analysis in a telecentric adaptive free-space optical relay based on a spatial light modulator.

We present an analysis of the performance limit of an adaptive multichannel free-space optical interconnect based on a spatial light modulator (SLM). The SLM function is to provide an active alignment of the signal beam in the detector plane. A thorough cross-talk analysis based on the diffractive properties of an ideal SLM in an isoplanatic optical system is shown. We analyze the performance in terms of the bit-error rate (BER) due to cross talk between different channels in the optical interconnect for different alignment states and for different phase-modulation schemes.

Involvement of the pineal gland in rats with experimental autoimmune uveitis.

Pineal glands of rats with experimentally induced autoimmune uveitis (EAU) were studied histologically. Inflammatory changes, characterized by mononuclear infiltration, were found in the pineal glands of one-third of the Lewis rats that developed EAU by active immunization with S-antigen. No changes in the pineal gland were observed in AVN rats which are "low responders" for EAU and did not develop ocular disease. Frequency and severity of both pineal gland and ocular involvement clearly were elevated by intravenous injection of Bordetella pertussis along with the S-antigen immunization; all B. pertussis-treated rats of both Lewis and AVN strains developed pineal and ocular changes. Inflammatory changes of the pineal gland also were found in rats in which EAU was induced passively by transfer of lymphocytes from S-antigen-immunized donors. The frequency of involvement of the pineal gland was found to be lower than that of the retinas in rats where EAU was induced by active immunization or by adoptive transfer of lymphocytes.

Immune response of sheep to Haemonchus contortus: serum antibodies against cross reacting antigens between parasites.

Normal and H. contortus infected sera were studied by ELISA technique against different stages of the parasites. In all cases antibody activity was detected. This activity in serum is partially eliminated after absorption with an adult worm extract of N brasiliensis. N. brasiliensis and H. contortus antigens were analysed by TCIEP with a rabbit anti-N. brasiliensis serum to examine shared antigens of H. contortus. A minimum of seven cross reacting antigens were detected. H. contortus adult worm extract was absorbed by the rabbit anti-N. brasiliensis serum. After absorption all cross reacting antigens were removed but at least one antigen reacting with a rabbit serum anti-H. Contortus is maintained. When this antigen is tested in elisa technique only a weak antibody activity is found in normal serum.

[Immune response in sheep experimentally infected with Haemonchus contortus, Comparative study in male and female (author's transl)]. / Réponse immunitaire chez les ovins infestés expérimentalement par Haemonchus contortus. Etude comparative chez le mále et chez la femelle.

Merino breed sheep eight months old at the beginning of the experiment and carriers of hemaglobin A were used. They were infected one or more times with H. contortus larvae and then challenged with 500 or 5000 larvae L3. Self-cure was reproduced and animals were found immune after infectation. Self-cure was more evident in females than in males and immunity was also better and appeared earlier in females than in males.

Effects of autogamy in Paramecium tetraurelia on catalase activity and on radiosensitivity to natural ionizing radiations.

Catalase activity of Paramecium tetraurelia decreased during autogamy and recovered to normal 5 days later. Autogamy also caused changes in the ciliate's sensitivity to natural ionizing radiations--the decrease in cell growth rate previously described in shielded cultures did not occur when autogamous cells were used. Maximum effect of shielding was observed in 11-day-old postautogamous cells. The role of the catalase in the mechanism of natural iradiation effect is discussed.

Phosphorylcholine antigens from Nippostrongylus brasiliensis. II.--Isolation and partial characterization of phosphorylcholine antigens from adult worm.

The phosphorylcholine antigens (C substance) were specifically isolated from Nippostrongglus brasiliensis adult worms. They formed a gorup of fairly closely related molecules, but it was not possible to evidence that the carrier molecule was unique. An indirect immunoenzymatic test using immobilized lectins (concanavalin A, phytohaemagglutinin Els, wheat germ agglutinin, recin types I and II, peanut agglutinin) gave some light on the carbohydrate composition of the carrier molecules, whereas the amino acid part of these molecules seemed to indicate an unique oligopeptide, the composition of which would be: Asx (4), Thr (2), Ser (4), Glux (6), Gly (6) Ala (3), Val (2), Ile (1), Leu (2), Phe (8), Lys (2), Arg (1). An epitope of the carrier molecules was demonstrated with anti-N. brasiliensis egg antisera. It was shared with various pathogens including Haemonchus contortus, Schistosoma mansoni and Dipetalonema vitae. It was also found on the purified pneumococcal C. polysaccharide. The C. substance from a variety of parasites can now be isolated by a combination of specific reactions using both anti-phosphorylcholine and anti-carrier molecule antibodies.

Phosphorylcholine antigens from Nippostrongylus brasiliensis. I.--Anti-phosphorylcholine antibodies in infected rats and location of phosphorylcholine antigens.

The anti-phosphorylcholine (PC) antibody synthesis was investigated in the rat after infection with the nematode Nippostrongylus brasiliensis. Serum IgM and IgG antibodies were demonstrated from day 2 or 3 post-infection. Intestinal IgA-antibody synthesis began shortly after the worms reached the intestine. Antigens containing PC were located with the fluorescent antibody technique in L3 infective larvae, adult worms and eggs of the parasites. They were always found to internal structures such as intestinal tract or gonads. It seems, therefore, that the anti-PC antibody synthesis resulted from the release of PC antigens by the parasite (moulting fluids, excretion, secretion or breakage products).

Cytidine-5'-diphospho-choline conjugates. I.--Synthesis and fixation to phosphorylcholine-binding proteins.

The oxidation of the ribose ring of cytidine-5'-diphospho-choline (CDPC) by periodate to produce reactive dialdehydes was used to couple this activated reagent onto various proteins, onto Ficoll 400 and onto Sepharose 4B. Careful control of parameters of the different steps of the reaction enabled us to synthesize conjugates with a graded number of nucleotide residues. Human serum albumin conjugates with a relatively high degree of substitution were used to demonstrate and to measure anti-phosphorylcholine antibodies and the acute phase reactant, C-reactive protein by precipitation or passive haemagglutination techniques. These methods for measuring C-reactive protein in serum of patient suffering from acute inflammation may be useful for clinicians. CDPC-AH Sepharose was used to purify the phosphorylcholine-binding myeloma protein HOPC8 and to separate C-reactive protein from the bulk of serum proteins. Improvements of this technique will certainly lead to the complete purification of C-reactive protein.

Cytidine-5'-diphospho-choline conjugates. II.--Immunogenicity in rats.

Cytidine diphospho-choline (CDPC) derivatives of human serum albumin (HSA) are immunogenic in the rat when injected in adjuvants. They induce a very rapid synthesis of IgM anti-phosphorylcholine antibodies which was followed by the synthesis of anti-CDPC antibodies and of anti-HSA antibodies. The activated nucleotide was able to induce specific skin sensitivity in rats after one skin painting. Gastric intubations of rats with low doses of HSA conjugates 8 days before an infection with the nematode Nippostrongylus brasiliensis were followed by a reduction of the resulting intestinal worm burden.