Pseudomonas syringae pv. tomato DC3000 polymutants deploying coronatine and two type III effectors produce quantifiable chlorotic spots from individual bacterial colonies in Nicotiana benthamiana leaves

Primary virulence factors of Pseudomonas syringae pv. tomato DC3000 include the phytotoxin coronatine (COR) and a repertoire of 29 effector proteins injected into plant cells by the type III secretion system (T3SS). DC3000 derivatives differentially producing COR, the T3SS machinery and subsets of key effectors were constructed and assayed in leaves of Nicotiana benthamiana. Bacteria were inoculated by the dipping of whole plants and assayed for population growth and the production of chlorotic spots on leaves. The strains fell into three classes. Class I strains are T3SS⁺ but functionally effectorless, grow poorly in planta and produce faint chlorotic spots only if COR⁺. Class II strains are T3SS^– or, if T3SS⁺, also produce effectors AvrPtoB and HopM1. Class II strains grow better than class I strains in planta and, if COR⁺, produce robust chlorotic spots. Class III strains are T3SS⁺ and minimally produce AvrPtoB, HopM1 and three other effectors encoded in the P. syringae conserved effector locus. These strains differ from class II strains in growing better in planta, and produce chlorotic spots without COR if the precursor coronafacic acid is produced. Assays for chlorotic spot formation, in conjunction with pressure infiltration of low‐level inoculum and confocal microscopy of fluorescent protein‐labelled bacteria, revealed that single bacteria in the apoplast are capable of producing colonies and associated leaf spots in a 1 : 1 : 1 manner. However, COR makes no significant contribution to the bacterial colonization of the apoplast, but, instead, enables a gratuitous, semi‐quantitative, surface indicator of bacterial growth, which is determined by the strain's effector composition.     

Inactivation ofSalmonella enteritidis on shell eggs by novelN-halamine biocidal compounds

Summary Several newN-halamine compounds have been evaluated as potential replacements for free chlorine as disinfectants for the egg-processing industry. The compounds were tested againstSalmonella enteritidis on the surfaces of egg shells. Test procedure included spraying inoculated egg shells with solutions of several of theN-halamine compounds and free chlorine for comparison, suspending the most stableN-halamine compound in a thin coating of mineral oil on the egg shell and subsequent inoculation, and measuring the rates of diffusion of the compounds and free chlorine through the egg shells. Compounds DBC (1-bromo-3-chloro-2,2,5,5-tetramethylimidazolidin-4-one) and DC (1,3-dichloro-2,2,5,5,-tetramethylimidazolidin-4-one) were significantly more efficacious than free chlorine in inactivatingSalmonella in the spray experiments, while compound MC (1-chloro-2,2,5,5-tetramethylmidazolidin-4-one), in a mineral oil suspension, provided disinfection of the egg shells within 72 h of contact. None of the disinfectant compounds penetrated egg shells at a rate greater than 1 mg/l over a period of 6 h. Compound MC is recommended as a possible replacement for unstable, corrosive-free chlorine as a bactericide for the egg-processing industry.     

Efficacy of Tower Medfly Eclosion Systems

A 16-repetition experiment was conducted to evaluate the performance of the “tower” system for eclosion of sterile medflies, Ceratitis capitata (Wiedemann). This system has now replaced the PARC system previously used in Florida S.I.T. programs. In addition to testing the efficacy of these eclosion systems, as compared to the PARC system, quality control was also monitored and evaluated. No significant differences were found between either system in regards to C. capitata yield, weight or flight ability (p=0.05). Based on these comparative trials, the tower eclosion system appears to be an efficient alternative to the PARC system.     

Oviposition site choice under conflicting risks demonstrates that aquatic predators drive terrestrial egg-laying

Laying eggs out of water was crucial to the transition to land and has evolved repeatedly in multiple animal phyla. However, testing hypotheses about this transition has been difficult because extant species only breed in one environment. The pantless treefrog, Dendropsophus ebraccatus, makes such tests possible because they lay both aquatic and arboreal eggs. Here, we test the oviposition site choices of D. ebraccatus under conflicting risks of arboreal egg desiccation and aquatic egg predation, thereby estimating the relative importance of each selective agent on reproduction. We also measured discrimination between habitats with and without predators and development of naturally laid aquatic and arboreal eggs. Aquatic embryos in nature developed faster than arboreal embryos, implying no cost to aquatic egg laying. In choice tests, D. ebraccatus avoided habitats with fish, showing that they can detect aquatic egg predators. Most importantly, D. ebraccatus laid most eggs in the water when faced with only desiccation risk, but switched to laying eggs arboreally when desiccation risk and aquatic predators were both present. This provides the first experimental evidence to our knowledge that aquatic predation risk influences non-aquatic oviposition and strongly supports the hypothesis that it was a driver of the evolution of terrestrial reproduction.     

Efficacies of Novel N-Halamine Disinfectants against Salmonella and Pseudomonas Species

Six novel N-halamine compounds of potential importance as disinfectants to the food-processing industry were tested against Salmonella enteritidis, Salmonella gallinarum, Salmonella typhimurium, and Pseudomonas fluorescens in aqueous solution. Inactivation times for 10⁶-fold reductions were determined as a function of water quality at pH 6.5 and 25°C. Phenol coefficients for the efficacies of the compounds against S. enteritidis have been reported also. When both stability and efficacy data are considered, as well as cost of production, two compounds, 1,3-dichloro-2,2,5,5-tetramethylimidazolidin-4-one and 1-chloro-2,2,5,5-tetramethylimidazolidin-4-one, offer the greatest potential as biocides for the food-processing industry.     

Increased expression of the immediate-early gene arc/arg3.1 reduces AMPA receptor-mediated synaptic transmission

Arc/Arg3.1 is an immediate-early gene whose expression levels are increased by strong synaptic activation, including synapse-strengthening activity patterns. Arc/Arg3.1 mRNA is transported to activated dendritic regions, conferring the distribution of Arc/Arg3.1 protein both temporal correlation with the inducing stimulus and spatial specificity. Here, we investigate the effect of increased Arc/Arg3.1 levels on synaptic transmission. Surprisingly, Arc/Arg3.1 reduces the amplitude of synaptic currents mediated by AMPA-type glutamate receptors (AMPARs). This effect is prevented by RNAi knockdown of Arc/Arg3.1, by deleting a region of Arc/Arg3.1 known to interact with endophilin 3 or by blocking clathrin-coated endocytosis of AMPARs. In the hippocampal slice, Arc/Arg3.1 results in removal of AMPARs composed of GluR2 and GluR3 subunits (GluR2/3). Finally, Arc/Arg3.1 expression occludes NMDAR-dependent long-term depression. Our results demonstrate that Arc/Arg3.1 reduces the number of GluR2/3 receptors leading to a decrease in AMPAR-mediated synaptic currents, consistent with a role in the homeostatic regulation of synaptic strength.