Ecological genomics of marine Roseobacters.

Bacterioplankton of the marine Roseobacter clade have genomes that reflect a dynamic environment and diverse interactions with marine plankton. Comparative genome sequence analysis of three cultured representatives suggests that cellular requirements for nitrogen are largely provided by regenerated ammonium and organic compounds (polyamines, allophanate, and urea), while typical sources of carbon include amino acids, glyoxylate, and aromatic metabolites. An unexpectedly large number of genes are predicted to encode proteins involved in the production, degradation, and efflux of toxins and metabolites. A mechanism likely involved in cell-to-cell DNA or protein transfer was also discovered: vir-related genes encoding a type IV secretion system typical of bacterial pathogens. These suggest a potential for interacting with neighboring cells and impacting the routing of organic matter into the microbial loop. Genes shared among the three roseobacters and also common in nine draft Roseobacter genomes include those for carbon monoxide oxidation, dimethylsulfoniopropionate demethylation, and aromatic compound degradation. Genes shared with other cultured marine bacteria include those for utilizing sodium gradients, transport and metabolism of sulfate, and osmoregulation.

Requirement for the cell division protein DivIB in polar cell division and engulfment during sporulation in Bacillus subtilis.

During spore formation in Bacillus subtilis, cell division occurs at the cell pole and is believed to require essentially the same division machinery as vegetative division. Intriguingly, although the cell division protein DivIB is not required for vegetative division at low temperatures, it is essential for efficient sporulation under these conditions. We show here that at low temperatures in the absence of DivIB, formation of the polar septum during sporulation is delayed and less efficient. Furthermore, the polar septa that are complete are abnormally thick, containing more peptidoglycan than a normal polar septum. These results show that DivIB is specifically required for the efficient and correct formation of a polar septum. This suggests that DivIB is required for the modification of sporulation septal peptidoglycan, raising the possibility that DivIB either regulates hydrolysis of polar septal peptidoglycan or is a hydrolase itself. We also show that, despite the significant number of completed polar septa that form in this mutant, it is unable to undergo engulfment. Instead, hydrolysis of the peptidoglycan within the polar septum, which occurs during the early stages of engulfment, is incomplete, producing a similar phenotype to that of mutants defective in the production of sporulation-specific septal peptidoglycan hydrolases. We propose a role for DivIB in sporulation-specific peptidoglycan remodelling or its regulation during polar septation and engulfment.

Cell division in Bacillus subtilis: FtsZ and FtsA association is Z-ring independent, and FtsA is required for efficient midcell Z-Ring assembly.

The earliest stage in cell division in bacteria is the assembly of a Z ring at the division site at midcell. Other division proteins are also recruited to this site to orchestrate the septation process. FtsA is a cytosolic division protein that interacts directly with FtsZ. Its function remains unknown. It is generally believed that FtsA localization to the division site occurs immediately after Z-ring formation or concomitantly with it and that FtsA is responsible for recruiting the later-assembling membrane-bound division proteins to the division site. Here, we report the development of an in vivo chemical cross-linking assay to examine the association between FtsZ and FtsA in Bacillus subtilis cells. We subsequently use this assay in a synchronous cell cycle to show that these two proteins can interact prior to Z-ring formation. We further show that in a B. subtilis strain containing an ftsA deletion, FtsZ localized at regular intervals along the filament but the majority of Z rings were abnormal. FtsA in this organism is therefore critical for the efficient formation of functional Z rings. This is the first report of abnormal Z-ring formation resulting from the loss of a single septation protein. These results suggest that in this organism, and perhaps others, FtsA ensures recruitment of the membrane-bound division proteins by ensuring correct formation of the Z ring.

The relationship between low-level benzene exposure and leukemia in Canadian petroleum distribution workers.

This study was conducted to evaluate the relationship between leukemia occurrence and long-term, low-level benzene exposures in petroleum distribution workers. Fourteen cases were identified among a previously studied cohort [Schnatter et al., Environ Health Perspect 101 (Suppl 6):85-89 (1993)]. Four controls per case were selected from the same cohort, controlling for birth year and time at risk. Industrial hygienists estimated workplace exposures for benzene, without knowledge of case-control status. Average benzene concentrations ranged from 0.01 to 6.2 ppm. Company medical records were used to abstract information on other potential confounders such as cigarette smoking. Odds ratios were calculated for several exposure metrics. Conditional logistic regression modeling was used to control for potential confounders. The risk of leukemia was not associated with increasing cumulative exposure to benzene for these exposure levels. Duration of benzene exposure was more closely associated with leukemia risk than other exposure metrics, although results were not statistically significant. A family history of cancer and cigarette smoking were the two strongest risk factors for leukemia, with cumulative benzene exposure showing no additional risk when considered in the same models. This study is consistent with other data in that it was unable to demonstrate a relationship between leukemia and long-term, low-level benzene exposures. The power of the study was limited. Thus, further study on benzene exposures in this concentration range are warranted.

Effects of Phenamiphos, Methyl Bromide, and Fallowing on Pratylenchus Penetrans, Yield of Medicago sativa, and Fusarium Infections.

A field study was made of the effects of a residual nematicide (phenamiphos), a fumigant (methyl bromide), and fallowing on the number of root lesion nematodes (Pratylenchus penetrans), forage yields of alfalfa, and the occurrence of Fusarium spp. in plant roots and soil. Fallowing controlled nematodes initially, but by the end of the second growing season, nematode numbers were as high as in plots which had grown a nematode-susceptible crop. Forage yield was greater in fallowed plots only for the first cut in the year after seeding. Fusarium in alfalfa roots and soil was not reduced by fallowing. Phenamiphos reduced nematode numbers, increased forage yields in 2 of 4 years, and reduced Fusarium infections of taproots. Soil fumigation with methyl bromide gave the best control of nematodes and Fusarium and gave significantly higher forage yields for the 4 years of study following fumigation. The 34% increase in alfalfa yield from fumigated plots over the 4 years indicates that the yield of alfalfa is being reduced significantly by microorganisms. The study does not establish the relative contributions of the root lesion nematodes and Fusarium spp. to the reduction.

Efficacy of multiple applications of oxamyl and phenamiphos for control of Pratylenchus penetrans in birdsfoot trefoil.

Preplant soil applications of granular phenamiphos effectively reduced Pratytenchus penetrans in soil during the seeding year and 1 year after, and in the roots of birdsfoot trefoil 2 years after seeding. Forage yields were increased in the season following application of phenamiphos, but stands of plants/m(2) were not greater (P = 0.05) than those in the checks 1 and 2 years after treatment. Additional spring applications of phenamiphos 1 and 2 years after seeding further reduced numbers of nematodes in the soil but did not improve forage yields or plant stand over that of a single application. Broadcast preplant soil sprays of oxamyl followed by several foliar sprays at different rates and frequencies of application over a 3-year period restricted populations of P. penetrans in the soil and roots of birdsfoot trefoil but did not consistently result in increased forage yields. Stands of birdsfoot trefoil continued to decline each year even with oxamyl treatments.