The TrpA protein of Trichodesmium erythraeum IMS101 is a non-fibril-forming collagen and a component of the outer sheath.

Collagen molecules are structural in nature and primarily found in eukaryotic, multicellular organisms. Recently, a collagen-like protein, TrpA, was identified and characterized in the marine cyanobacterium Trichodesmium erythraeum IMS 101, and it was shown to be involved in maintaining the structural integrity of the trichomes. The TrpA protein contains one glycine interruption in the otherwise perfectly uninterrupted collagenous domain. In this study, we used phylogenetic analysis to determine that the TrpA protein sequence is most closely associated with non-fibril-forming collagen proteins. Structural modelling and circular dichroism data suggest that the glycine insertion decreases the stability of TrpA compared to uninterrupted collagen sequences. Additionally, scanning electron microscopy revealed that TrpA is expressed entirely on the surface of the trichomes, with no specific pattern of localization. These data indicate that the TrpA protein is part of the outer sheath of this organism. As such, this protein may function to promote adhesion between individual T. erythraeum trichomes, and between this organism and heterotrophic bacteria found in the same environment.

Indoor-biofilter growth and exposure to airborne chemicals drive similar changes in plant root bacterial communities.

Due to the long durations spent inside by many humans, indoor air quality has become a growing concern. Biofiltration has emerged as a potential mechanism to clean indoor air of harmful volatile organic compounds (VOCs), which are typically found at concentrations higher indoors than outdoors. Root-associated microbes are thought to drive the functioning of plant-based biofilters, or biowalls, converting VOCs into biomass, energy, and carbon dioxide, but little is known about the root microbial communities of such artificially grown plants, how or whether they differ from those of plants grown in soil, and whether any changes in composition are driven by VOCs. In this study, we investigated how bacterial communities on biofilter plant roots change over time and in response to VOC exposure. Through 16S rRNA amplicon sequencing, we compared root bacterial communities from soil-grown plants with those from two biowalls, while also comparing communities from roots exposed to clean versus VOC-laden air in a laboratory biofiltration system. The results showed differences in bacterial communities between soil-grown and biowall-grown plants and between bacterial communities from plant roots exposed to clean air and those from VOC-exposed plant roots. Both biowall-grown and VOC-exposed roots harbored enriched levels of bacteria from the genus Hyphomicrobium. Given their known capacities to break down aromatic and halogenated compounds, we hypothesize that these bacteria are important VOC degraders. While different strains of Hyphomicrobium proliferated in the two studied biowalls and our lab experiment, strains were shared across plant species, suggesting that a wide range of ornamental houseplants harbor similar microbes of potential use in living biofilters.

Characterization of a novel collagen-like protein TrpA in the cyanobacterium Trichodesmium erythraeum IMS101.

The collagen protein family is diverse and its membership is continually expanding as new collagen-like molecules are identified. Identification of collagen in unicellular eukaryotes and prokaryotes has opened discussion on the function of these collagens and their role in the emergence of multicellularity. The previous identification of a collagen gene in Trichodesmium erythraeum raises the question of function of this structural protein in a prokaryote. In this study, we show that this gene is expressed during all phases of growth, indicating that it may be required for all phases of growth. Using immunofluorescence techniques, we demonstrate that the collagen-like protein is localized in a specific manner between adjacent cells along the trichome of T. erythraeum. Trichomes treated with the enzyme collagenase exhibited fragmentation, supporting our immunofluorescence localization data that this collagen-like protein is found between adjacent cells. Our data strongly suggest that the collagen-like protein found in T. erythraeum functions to maintain the structural integrity of the trichome through the adhesion of adjacent cells.

Microbial quality of food available to populations of differing socioeconomic status.

BACKGROUND: Low SES has been shown to be linked to poorer-quality diets, decreased consumption of fresh produce, and an increased reliance on small retail stores. PURPOSE: The objective of this research was to determine if there is a difference in the microbial quality and potential safety of food available to low-SES versus high-SES populations at the retail level. METHODS: Aerobic plate count (APC); yeast and mold counts (Y & M); and total coliforms were determined in ready-to-eat (RTE) greens, pre-cut watermelon, broccoli, strawberries, cucumbers, milk, and orange juice and compared among products purchased in stores in low- versus those purchased in high-SES neighborhoods between June 2005 and September 2006. APC, fecal coliforms, and E. coli in ground beef and the presence of Salmonella and Campylobacter in chicken were also compared. RESULTS: Results showed higher microbial loads on produce from markets in low-SES areas. Significant differences observed included (1) APC and Y&M in RTE greens, (2) APC and Y&M in strawberries, and (3) YMCs in cucumbers. No difference was detected in the level of pathogens in raw meat and poultry; however, the APC in ground beef available in high-SES markets was significantly higher compared with that found in low-SES markets. CONCLUSIONS: The results presented here indicate that populations of low SES may be more likely to experience produce of poorer microbial quality, which may have an impact on both the appeal and potential safety of the produce.

DNA microarrays detect effects of soil contamination on Arabidopsis thaliana gene expression.

Soil contamination, such as heavy metals and benzene compounds, is a widespread problem on military installations. It is important to be able to determine the effects of soil contamination before any adverse effects appear in organisms in surrounding areas. We examined gene expression in Arabidopsis thaliana grown in soil from three sites at the Radford Army Ammunition Plant in Radford, Virginia, USA, using DNA microarrays. We analyzed soil, germination, and growth rate to compare with the microarray data. Soil contamination affected both external phenotype and gene expression. Plants grown in soil with high levels of contaminants were chloritic and were smaller than control plants grown in potting soil. Plants grown in soil with the highest copper concentration had the lowest growth rates and had genes up-regulated across several functional groups. Plants grown in soils with elevated lead had many genes down-regulated that were related to photosystem II, metabolism, cellular transport, and protein synthesis. Genes consistently up-regulated across most microarrays were genes related to photosystem I, genes related to water deprivation and oxidative stress response, heat shock proteins, and toxin catabolism genes such as glutathiones. DNA microarrays, in concert with a model genetic organism such as A. thaliana, were an effective assessment tool to determine the presence of toxic substances in soil at a site used for the production of military explosives.

Genetic selection scheme for isolation of signal transduction pathway mutants.

Genetic characterization of a signal transduction pathway requires the isolation of mutations in the pathway. Characterization of these mutated genes and their loci enumerates the components of the pathway and leads to an understanding of the role of each gene locus in the pathway under study. We have designed and developed a strategy based on resistance to the chemical flucytosine for the identification of mutations in a given pathway. In this study, the Escherichia coli codA gene, which encodes the enzyme cytosine deaminase, was fused to the light-intensity-regulated gene promoter psbDII. Cytosine deaminase converts 5'-fluorocytosine to the toxic product 5-fluorouracil. Wild-type cells containing an intact signal transduction pathway that regulates the psbDII promoter will die in the presence of this chemical. Cells that carry mutations in the pathway that inactivate the psbDII promoter will not express the codA gene and, consequently, will live on 5'-fluorocytosine, allowing the isolation and subsequent characterization of mutations in this signaling pathway. Utilizing this selection method, we have successfully isolated and characterized mutations in the psbDII pathway. This selection scheme can be used with a tissue-specific or phase-specific promoter fused to the codA gene to direct the timing of expression of codA to obtain mutants defective in temporal or cell-specific expression of a particular pathway. This scheme also allows the isolation of mutants even when a clearly identifiable phenotype is not available. The selection scheme presented here extends the molecular tools available for the genetic dissection of signal transduction pathways.