Pseudomonas chlororaphis strain PA23 is a biocontrol agent capable of suppressing disease caused by the fungal pathogen Sclerotinia sclerotiorum. This bacterium produces the diffusible antibiotics phenazine-1-carboxylic acid, 2-hydroxyphenazine and pyrrolnitrin (PRN). Because the individual contribution of these antibiotics to PA23 biocontrol has not been defined, mutants deficient in the production of phenazine (PHZ), PRN or both antibiotics were created. Analysis of the PHZ mutant revealed enhanced antifungal activity in vitro and wild-type levels of Sclerotinia disease suppression. Conversely, the PRN- and the PRN/PHZ-deficient strains exhibited decreased antifungal activity in vitro and markedly reduced the ability to control Sclerotinia infection of canola in the greenhouse. These findings suggest that PRN is the primary antibiotic mediating biocontrol of this pathogen. Analysis of prnA-lacZ and phzA-lacZ transcriptional fusions revealed that PRN and PHZ are not subject to autoregulation; moreover, they do not cross-regulate each other. However, HPLC showed a twofold increase in PRN levels in the PHZ(-) background. Finally, PHZ, but not PRN production, is involved in biofilm development in P. chlororaphis PA23.

Pseudomonas chlororaphis PA23 is a biocontrol agent that protects against the fungal pathogen Sclerotinia sclerotiorum. Employing transposon mutagenesis, we isolated a gacS mutant that no longer exhibited antifungal activity. Pseudomonas chlororaphis PA23 was previously reported to produce the nonvolatile antibiotics phenazine 1-carboxylic acid and 2-hydroxyphenazine. We report here that PA23 produces additional compounds, including protease, lipase, hydrogen cyanide, and siderophores, that may contribute to its biocontrol ability. In the gacS mutant background, generation of these products was markedly reduced or delayed with the exception of siderophores, which were elevated. Not surprisingly, this mutant was unable to protect canola from disease incited by S. sclerotiorum. The gacS mutant was able to sustain itself in the canola phyllosphere, therefore, the loss of biocontrol activity can be attributed to a reduced production of antifungal compounds and not a declining population size. Competition assays between the mutant and wild type revealed equivalent fitness in aged batch culture; consequently, the gacS mutation did not impart a growth advantage in the stationary phase phenotype. Under minimal nutrient conditions, the gacS-deficient strain produced a tenfold less biofilm than the wild type. However, no difference was observed in the ability of the mutant biofilm to protect cells from lethal antibiotic challenge.

Chronically elevated GAL may regulate body weight, metabolic rate, and lipid and carbohydrate metabolism through a mechanism that is independent of feeding regulation. The obese phenotype in the GAL-Tg mice is related to the reduced energy expenditure and insulin resistance. These findings support the hypothesis that increased circulating GAL levels contribute to the development of metabolic syndrome.

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Objective:Recent genome-wide association studies have identified a strong association between obesity and common variants in the fat mass and obesity associated (FTO) gene. FTO has been detected in the hypothalamus, but little is known about its regulation in that particular brain structure. The present study addressed the hypothesis that hypothalamic FTO expression is regulated by nutrients, specifically by glucose, and that its regulation by nutrients is impaired in obesity.Research design and methods:The effect of intraperitoneal (i.p.) or intracerebroventricular (i.c.v.) administration of glucose on hypothalamic Fto mRNA levels was examined in fasted mice. Additionally, the effect of glucose on Fto mRNA levels was also investigated ex vivo using mouse hypothalamic explants. Lastly, the effect of i.p. glucose injection on hypothalamic Fto immunoreactivity and food intake was compared between lean wild-type and obese ob/ob mice.Results:In wild-type mice, fasting reduced both Fto mRNA levels and the number of Fto-immunoreactive cells in the hypothalamus, whereas i.p. glucose treatment reversed this effect of fasting. Furthermore, i.c.v. glucose treatment also increased hypothalamic Fto mRNA levels in fasted mice. Incubation of hypothalamic explants at high glucose concentration increased Fto mRNA levels. In ob/ob mice, both fasting and i.p. glucose treatment failed to alter the number of Fto-immunoreactive cells in the hypothalamus. Glucose-induced feeding suppression was abolished in ob/ob mice.Conclusion:Reduction in hypothalamic Fto expression after fasting likely arises at least partly from reduced circulating glucose levels and/or reduced central action of glucose. Obesity is associated with impairments in glucose-mediated regulation of hypothalamic Fto expression and anorexia. Hypothalamic Fto-expressing neurons may have a role in the regulation of metabolism by monitoring metabolic states of the body.