Critical role of LuxS in the virulence of Campylobacter jejuni in a guinea pig model of abortion.

Previous studies on Campylobacter jejuni have demonstrated the role of LuxS in motility, cytolethal distending toxin production, agglutination, and intestinal colonization; however, its direct involvement in virulence has not been reported. In this study, we demonstrate a direct role of luxS in the virulence of C. jejuni in two different animal hosts. The IA3902 strain, a highly virulent sheep abortion strain recently described by our laboratory, along with its isogenic luxS mutant and luxS complement strains, was inoculated by the oral route into both a pregnant guinea pig virulence model and a chicken colonization model. In both cases, the IA3902 luxS mutant demonstrated a complete loss of ability to colonize the intestinal tract. In the pregnant model, the mutant also failed to induce abortion, while the wild-type strain was highly abortifacient. Genetic complementation of the luxS gene fully restored the virulent phenotype in both models. Interestingly, when the organism was inoculated into guinea pigs by the intraperitoneal route, no difference in virulence (abortion induction) was observed between the luxS mutant and the wild-type strain, suggesting that the defect in virulence following oral inoculation is likely associated with a defect in colonization and/or translocation of the organism out of the intestine. These studies provide the first direct evidence that LuxS plays an important role in the virulence of C. jejuni using an in vivo model of natural disease.

Peptidoglycan induces necrosis and regulates cytokine production in murine trophoblast stem cells.

PROBLEM: Intrauterine bacterial infection during pregnancy may lead to adverse outcome. The objective of this study was to assess whether peptidoglycan (PGN) derived from Gram-positive bacteria induces trophoblast stem (TS) cell death or alters TS cell cytokine production. METHOD OF STUDY: Toll-like receptor (TLR) transcript expression was assessed by RT-PCR. Protein expression was determined by confocal microscopy or flow cytometry. 7-Aminoactinomycin D (7-AAD) staining was used to assess TS cell death. Morphological features of cell death were evaluated by transmission electron microscopy. The presence of cleaved caspase-3 and high mobility group box 1 (HMGB1) protein was examined by Western blot. Cytokine levels in cell supernatants were determined using a mouse cytokine 23-plex panel. RESULTS: Toll-like receptor 2 and TLR4 protein was expressed from the 1-cell stage through the blastocyst stage of murine embryo development. Murine TS cells expressed TLR2 and TLR6 but not TLR1 or TLR4 RNA. Only TLR2 protein was detected at the plasma membrane of TS cells. PGN induced TS cell death by a caspase-3-independent mechanism. The cell death pathway induced by PGN was morphologically consistent with necrosis. Finally, PGN induced HMGB1 release and increased MIP-1ß secretion while inhibiting the constitutive release of RANTES. CONCLUSION: Peptidoglycan-induced TS cell necrosis and the subsequent release of HMGB1 and MIP-1ß may regulate an infection-induced inflammatory response at the maternal-fetal interface and thus may play a role in the pathogenesis of infection-associated pregnancy complications.

MECHANISMS OF TOLERANCE TO FLORAL LARCENY IN TWO WILDFLOWER SPECIES.

Tolerance of foliar damage is widely recognized as an effective defense against herbivores and pathogens. However, tolerance of the impacts of antagonists on pollination success is less well understood. Here, we extend the framework of tolerance to foliar damage to understand how plants mitigate the pollination and fitness costs of floral larceny (i.e., the consumption of floral nectar often without pollination). We focused on two mechanisms: high nectar rewards per flower to feed all floral visitors and high flower production to compensate for reproductive losses under reduced pollination and seed set. We compared the efficacy of these mechanisms in two plant species: Polemonium viscosum and Ipomopsis aggregata. In Polemonium, ants acting as larcenists reduce nectar accumulation but do not completely empty flowers. When nectar reserves were augmented, ant consumption increased, negating the efficacy of this putative tolerance mechanism. Similarly, in Ipomopsis, nectar addition had little effect on tolerance to larceny by bumble bees, perhaps because residual intact flowers do not have enough nectar to compensate for lost rewards. Flower production in both species mitigated some of the negative impacts of larceny on seed set. In Polemonium, flower number was not plastic in response to larceny, but large inflorescences enhanced female fitness only when larcenists were present, suggesting that "surplus" flowers in large inflorescences can function to replace reproductive losses due to larceny. In Ipomopsis, high rates of larceny induced flower production, but the fecundity benefits of making more flowers declined inversely to larcenist intensity. Overall, our results suggest (1) that tolerance to floral larceny involves "banking" extra flowers to replace lost reproduction rather than maintaining pollination of ones with larceny, and (2) that the efficacy of flower production as a tolerance mechanism varies inversely to larceny rate.

Light-sensing in roots.

Light gradients in the soil have largely been overlooked in understanding plant responses to the environment. However, roots contain photoreceptors that may receive ambient light through the soil or piped light through the vascular cylinder. In recent experiments we demonstrated linkages between phototropin-1 photoreceptor production, root growth efficiency, and drought tolerance, suggesting that root plasticity in response to light signals contributes to the ecological niche of A. thaliana. However, the availability of light cues in natural soil environments is poorly understood, raising questions about the relevance of light-mediated root growth for fitness in nature. Additionally, photoreceptor expression is characterized by pleiotropy so unique functions cannot be clearly ascribed to root vs. shoot sensory mechanisms. These considerations show that challenges exist for resolving the contribution of light-sensing by roots to plant adaptation. We suggest that blue-light sensing in roots of A. thaliana provides a model system for addressing these challenges. By calibrating blue light gradients in soils of diverse A. thaliana habitats and comparing fitness of phot1 mutant and wild-type controls when grown in presence or absence of soil light cues, it should be possible to elucidate the ecological significance of light-mediated plasticity in roots.

Functional ecology of a blue light photoreceptor: effects of phototropin-1 on root growth enhance drought tolerance in Arabidopsis thaliana.

* The blue light photoreceptor phototropin-1 has been shown to enhance fitness in Arabidosis thaliana under field conditions. Here, we ask whether performance consequences of phototropin-1 reflect its impact on root growth and drought tolerance. * We used a PHOT1-GFP gene construct to test whether phototropin-1 abundance in roots is highest at shallow soil depths where light penetration is greatest. We then compared root growth efficiency and size at maturity between individuals with and without functional phototropin-1. Comparisons were made under wet and dry conditions to assess the impact of phototropin-1 on drought tolerance. * Phototropin-1 was most abundant in upper root regions and its impact on root growth efficiency decreased with soil depth. Roots of plants with functional phototropin-1 made fewer random turns and traveled further for a given length (higher efficiency) than roots of phot1 mutants. In dry (but not wet) soil, enhancement of root growth efficiency by phototropin-1 increased plant size at maturity. * Results indicate that phototropin-1 enhances performance under drought by mediating plastic increases in root growth efficiency near the soil surface.