A gain-of-function mutation in a cytokinin receptor triggers spontaneous root nodule organogenesis

Legume root nodules originate from differentiated cortical cells that reenter the cell cycle and form organ primordia. We show that perception of the phytohormone cytokinin is a key element in this switch. Mutation of a Lotus japonicus cytokinin receptor gene leads to spontaneous development of root nodules in the absence of rhizobia or rhizobial signal molecules. The mutant histidine kinase receptor has cytokinin-independent activity and activates an Escherichia coli two-component phosphorelay system in vivo. Mutant analysis shows that cytokinin signaling is required for cell divisions that initiate nodule development and defines an autoregulated process where cytokinin induction of nodule stem cells is controlled by shoots.     

A role for interaction of the RNA polymerase flap domain with the sigma subunit in promoter recognition

In bacteria, promoter recognition depends on the RNA polymerase sigma subunit, which combines with the catalytically proficient RNA polymerase core to form the holoenzyme. The major class of bacterial promoters is defined by two conserved elements (the -10 and -35 elements, which are 10 and 35 nucleotides upstream of the initiation point, respectively) that are contacted by sigma in the holoenzyme. We show that recognition of promoters of this class depends on the "flexible flap" domain of the RNA polymerase beta subunit. The flap interacts with conserved region 4 of sigma and triggers a conformational change that moves region 4 into the correct position for interaction with the -35 element. Because the flexible flap is evolutionarily conserved, this domain may facilitate promoter recognition by specificity factors in eukaryotes as well.     

Comparison of digital and optical hand-held refractometers for the measurement of feline urine specific gravity

Measuring urine specific gravity (USG) is an important component of urine analysis as it evaluates renal concentrating capability. The objective of this study was to quantify the difference in USG values between a hand-held optical analogue refractometer and a cat-specific digital instrument. Urine samples from 55 cats were assessed. There was a statistically significant difference between these two refractometers (P<0.001), with the optical refractometer (mean USG=1.031) consistently reading higher than the digital refractometer (mean USG=1.027). Results for a random subset of the samples (n=10) were compared with urine osmolality and both the optical and digital instruments demonstrated excellent correlation. While an accurate USG reading is important, it is unlikely that the statistical significance between the two instruments is clinically significant and, therefore, unlikely to result in a change in patient evaluation or treatment plans. While both the digital and optimal refractometers are highly correlated to the urine osmolality, making both devices valid for assessment of USG in clinical practice, this digital device is easier to read and eliminates the variability of subjective interpretation.     

One- and two-dimensional electrophoretic identification of IgE-binding polypeptides of Lupinus albus and other legume seeds

The prevalence of food allergies in the world population requires integrated approaches to identify new potential allergens, especially those of plant origin. The aim of this work was the allergen in vitro analysis of Lupinus albus seed proteome, a promising food protein source, and the assessment of IgE cross-reactivities with other more diffused legume species. A combination of one- and two-dimensional gel electrophoresis and immunoblotting analyses with specific IgGs for band identification and lupin-sensitized patients' circulating IgEs for allergenicity studies has been used. Two lupin proteins, namely, conglutin gamma and 11S globulin basic subunits, strongly reacted with all patients' sera. Also, cross-reactivities with the homologous polypeptides of other legume species were observed. Otherwise, no reaction at all was detected with a 2S-type lupin protein. This global electrophoretic approach has allowed the identification of a new potential lupin allergen and confirmed the cross-reactivity among the legume 11S globulin basic subunits.     

A Novel Particle Contact Assay with the Yeast Saccharomyces cerevisiae (8 pp)

Goal, Scope and Background   Numerous xenobiotics released into surface waters are transferred to suspended particulate matter and finally attached to sediments. Aquatic organisms may be exposed to them by direct particle feeding, by physical contact with contaminated surfaces as an exposure route, and by the uptake of dissolved contaminants after equilibration via the free water phase. In order to assess potential sediment toxicity, each of these exposure routes has to be addressed. This paper presents a newly developed particle contact assay that uses the fermentation performance of a specific Saccharomyces cerevisiae strain for the assessment of toxic effects in sediments. The test procedure is based on the characteristic feature of growing yeast cells to attach to sediment particles, which are also relevant for the accumulation of contaminants. The physical contact with lipophilic contaminants mirrors an exposition pathway for the direct uptake into the cells. In order to quantitatively characterize the toxic effects of particle attached pollutants on the fermentation performance, unpolluted native reference sediment was spiked with representatives for widely distributed anthropogenic contaminants. Methods   Saccharomyces cerevisiae was established as sensitive eukaryotic microorganism for the ecotoxicological assessment of particle attached anthropogenic contaminants in freshwater sediments. For this purpose, yeast cells were cultivated in sediment samples and the resulting fermentation performance was continuously measured. Sediments artifically spiked with HCB, PCB, g-HCH, DDT, and benzo(a)pyrene and solutions of each contaminant were comparatively investigated by means of their adverse effects on yeast fermentation performance. Additionally, four native river sediments characterized by increasing levels of pollution were assessed by the yeast particle contact assay, and simultaneously by standard aquatic tests with algae, daphniae, and luminescent bacteria using pore water and elutriates. Results of the bioassays were related to specific sediment contamination with respect to metals and organic priority pollutants. Results and Discussion   In sediments spiked with PCB and benzo(a)pyrene fermentation, performance was affected extensively below concentrations inhibiting fermentation in contaminant solutions. This suggests a high efficiency of the exposure route by physical contact. The fermentation performance was only slightly affected by single lipophilic pollutants, whereas mixtures of individually spiked sediments caused critically reduced fermentation performance suggesting additive synergistic effects. Native river sediments modestly to critically polluted by hazardous organic compounds lead to a slightly to dangerously reduced fermentation performance in the yeast contact assay. These inhibitory effects were much less pronounced in the standard bioassays conducted with algae, daphniae and luminescent bacteria, applying pore waters and elutriates as sample matrices. Using pore water, inhibition was measured only in the most polluted sediment, elutriates lead to a slight inhibition of the algal growth in the undiluted sample only. These results indicate an improved sensitivity of the yeast particle contact assay compared to the standard assays, due to uptake and physical cell contact as additional routes of exposure. Conclusion   The yeast particle contact assay is a valuable tool for the assessment of ecotoxicological potential in freshwater sediments. Since the assay addresses physical contact as an exposure route, it indicates bioavailability of lipophilic compounds in sediments. Outlook   The sensitive indication of bioavailable contaminants associated to sediment particles by the newly developed yeast particle contact assay recommends it as a complementary microbial bioassay in a test battery for assessing major pathways of contaminants in whole sediments.     

Interactions of fungicide-insecticide combinations against Rhizoctonia solani in vitro and in soil

The inhibition of mycelial growth of Rhizoctonia solani in vitro was strongest with pencycuron, followed by tolclofos-methyl, carboxin and thiabendazole. Against cowpea seedling rot in soil infested with R. solani, tolclofos-methyl was most effective, followed by pencycuron, thiabendazole and carboxin. Of nine insecticides tested, only parathion-methyl and quinalphos suppressed mycelial growth of R. solani in vitro, although their activity was much lower than that of the four tested fungicides. The inhibition of mycelial growth by fungicide-insecticide mixtures was antagonistic in only two out of 36 combinations; it was additive in most cases or synergistic, e.g. for most mixtures of pencycuron and insecticides. Quinalphos, applied to the soil, was the only insecticide which reduced cowpea seedling rot in soil infested with R. solani. Soil application of parathion-methyl, phorate, aldicarb or carbofuran and seed treatment with phosphamidon, monocrotophos, endosulfan or dimethoate had little or no effect on seedling rot. Carboxin gave better disease control when applied to the seed already coated with phosphamidon, monocrotophos, endosulfan or dimethoate and when carboxin-treated seeds were sown in soil treated with quinalphos, parathion-methyl, aldicarb or carbofuran. Efficacy of thiabendazole seed treatment was slightly higher in the presence of insecticides, particularly dimethoate. Pencycuron and tolclofos-methyl as seed treatment gave nearly 100% disease control both in the presence and absence of insecticides. The synergistic interactions detected between fungicides and insecticides represent interesting opportunities for the control of R. solani.