Cyanobacteria and Biosequestration: The Effects of High CO2 Levels on Calcifying Strains of Synechococcus

Increased carbon dioxide levels in the atmosphere and its influence on climate change is a growing concern in the scientific, political, and social communities. Methods of mitigation are being tested to explore carbon capture and storage through the biosynthesis of stable carbon-containing compounds using different strains of calcifying cyanobacteria. By utilizing marine genera, the cyanobacteria could potentially be grown in brine waste waters, conserving valuable fresh water resources. In this experiment, two strains of Synechococcus were cultured in flasks with varying levels of CO2: air, 5% CO2, and 15% CO2. Growth of each culture was monitored by measuring optical density and pH level, as calcification requires an alkaline environment. Morphological characteristics of each culture were analyzed through light microscopy and scanning electron microscopy to compare differences in cell surface and association. Preliminary results have shown inconsistent morphology and growth. Cultures started from previous experiments lacked duplication of observed filamentous morphology, but exhibited better growth in high CO2 levels. The incubation of cultures in media with varying levels of calcium chloride will be used to analyze and compare the sequestration of carbon through calcium carbonate production. Analysis of the chemical composition of precipitates in the media and the S-layer of cells will verify the presence of calcium carbonate. Methods include the use of SEM-EDX (energy-dispersive X-ray spectroscopy) and polarized light microscopy. If experimental outcomes verify efficient production of calcium carbonate from sources of high CO2, these cyanobacteria may be viable systems for capturing carbon

Phylogenetic and comparative gene expression analysis of barley (Hordeum vulgare) WRKY transcription factor family reveals putatively retained functions between monocots and dicots

WRKY proteins belong to the WRKY-GCM1 superfamily of zinc finger transcription factors that have been subject to a large plant-specific diversification. For the cereal crop barley (Hordeum vulgare), three different WRKY proteins have been characterized so far, as regulators in sucrose signaling, in pathogen defense, and in response to cold and drought, respectively. However, their phylogenetic relationship remained unresolved. In this study, we used the available sequence information to identify a minimum number of 45 barley WRKY transcription factor (HvWRKY) genes. According to their structural features the HvWRKY factors were classified into the previously defined polyphyletic WRKY subgroups 1 to 3. Furthermore, we could assign putative orthologs of the HvWRKY proteins in Arabidopsis and rice. While in most cases clades of orthologous proteins were formed within each group or subgroup, other clades were composed of paralogous proteins for the grasses and Arabidopsis only, which is indicative of specific gene radiation events. To gain insight into their putative functions, we examined expression profiles of WRKY genes from publicly available microarray data resources and found group specific expression patterns. While putative orthologs of the HvWRKY transcription factors have been inferred from phylogenetic sequence analysis, we performed a comparative expression analysis of WRKY genes in Arabidopsis and barley. Indeed, highly correlative expression profiles were found between some of the putative orthologs. HvWRKY genes have not only undergone radiation in monocot or dicot species, but exhibit evolutionary traits specific to grasses. HvWRKY proteins exhibited not only sequence similarities between orthologs with Arabidopsis, but also relatedness in their expression patterns. This correlative expression is indicative for a putative conserved function of related WRKY proteins in mono- and dicot species

Antisense oligodeoxynucleotide inhibition as a potent diagnostic tool for gene function in plant biology

Antisense oligodeoxynucleotide (ODN) inhibition emerges as an effective means for probing gene function in plant cells. Employing this method we have established the importance of the SUSIBA2 transcription factor for regulation of starch synthesis in barley endosperm, and arrived at a model for the role of the SUSIBAs in sugar signaling and source-sink commutation during cereal endosperm development. In this addendum we provide additional data demonstrating the suitability of the antisense ODN technology in studies on starch branching enzyme activities in barley leaves. We also comment on the mechanism for ODN uptake in plant cells. Antisense ODNs are short (12-25 nt-long) stretches of single-stranded ODNs that hybridize to the cognate mRNA in a sequence-specific manner, thereby inhibiting gene expression. They are naturally occurring in both prokaryotes and eukaryotes where they partake in gene regulation and defense against viral infection. The mechanisms for antisense ODN inhibition are not fully understood but it is generally considered that the ODN either sterically interferes with translation or promotes transcript degradation by RNase H activation. The earliest indication of the usefulness of antisense ODN technology for the purposes of molecular biology and medical therapy was the demonstration in 1978 that synthetic ODNs complementary to Raos sarcoma virus could inhibit virus replication in tissue cultures of chick embryo fibroblasts. Since then the antisense ODN technology has been widely used in animal sciences and as an important emerging therapeutic approach in clinical medicine. However, antisense ODN inhibition has been an under-exploited strategy for plant tissues, although the prospects for plant cells in suspension cultures to take up single-stranded ODNs was reported over a decade ago. In 2001, two reports from Malho and coworker demonstrated the use of cationic-complexed antisense ODNs to suppress expression of genes encoding pollen-signaling proteins in pollen tubes from the lilly Agapanthus umbellatus. For the uptake of DNA pollen tubes represent a unique system since the growing tip is surrounded by a loose matrix of hemicellulose and pectins, exposing the plasma membrane7 and the first uptake of ODNs by pollen tubes was reported as early as 1994. A breakthrough in the employment of antisense ODN inhibition as a powerful approach in plant biology was recently presented through our work on intact barley leaves. As was illustrated by confocal microscopy and fluorescently labeled ODNs, naked ODNs were taken up through the leaf petiole and efficiently imported into the plant cell and the nucleus. The work portrayed in that study demonstrate the applicability of antisense ODN inhibition in plant biology, e.g. as a rapid antecedent to time-consuming transgenic studies, and that it operates through RNase H degradation. We employed the antisense ODN strategy to demonstrate the importance of the SUSIBA2 transcription factor in regulation of starch synthesis, and to depict a possible mechanism for sugar signaling in plants and how it might confer endosperm-specific gene expression during seed development. We also described the employment of the antisense ODN strategy for studies on in vitro spike cultures of barley. Here we present further evidence as to the value of the antisense ODN approach in plant biology by following the effects on starch branching enzyme (SBE) accumulation in barley leaves after suppression of individual SBE genes. In agreement with transcript analyses of SBE expression in barley leaves, a zymogram assay (Fig. 1) revealed that sucrose treatment of barley leaves increased the number of SBE activity bands as compared to sorbitol treatment. In the presence of antisense SBEI or SBEIIA ODNs, zymograms of sucrose-treated leaves displayed only a subset of these activities with bands in the top portion of the zymogram gel missing or diminished. With antisense SBEIIB ODN, all activity bands in the top portion of the gel as well as the lowest band were absent. Based on these data we provide a tentative annotation for the various SBE activity bands. In animal experiments, naked ODNs are usually not taken up by the cells since both the ODNs and the outside of the plasma membrane carry a net negative charge. Thus the uptake of naked ODNs into barley leaf cells was surprising and called for an explanation. As demonstrated in our subsequent paper, the answer seems to be that the ODNs slip into the cells through sugar translocators as they are activated in the presence of the appropriate sugar (Fig. 2). Whether it is the structural resemblance between the sugar (deoxyribose) backbone of the ODNs and the transported sugars that allows for the ODNs to be transferred, or if other mechanisms are involved, remains to be elucidated

Foodborne infections in Iceland. Relationship to allergy and lung function

Neðst á síðunni er hægt að nálgast greinina í heild sinni með því að smella á hlekkinn View/OpenBACKGROUND: Foodborne or orofecal transmitted infections can have influence on health by direct consequences of the infection and indirectly by modulating the immune system. OBJECTIVES: To investigate the prevalence and risk factors for T. gondii, H. pylori and HAV infection in the Icelandic population and their influence on atopy, allergy related lung symptoms and lung function. MATERIAL AND METHODS: Blood samples were collected in 1999-2001 from 505 subjects in age group 28-52, randomly selected from the Icelandic population. The presence of T. gondii, H. pylori and HAV IgG antibodies was determined by an ELISA method. Allergy related lung symptoms were assessed with questionnaire and IgE sensitization and lung function measured. X(2) test was used to test for trend but unadjusted logistic regression for comparison of IgG prevalence. Multiple logistic regression was used to calculate adjusted odds ratios and 95% confidence intervals for different infections factors. RESULTS: The prevalence of antibodies was 9.8%, for T. gondii, 36.3% for H. pylori and 4.9% for HAV. Attending day care before the age of 3 years was a risk factor for having T. gondii antibodies. The prevalence of H. pylori increased with age and smoking. The infections were not associated with the prevalence of asthma or atopy. Having IgG antibodies against T. gondii was, however, associated with an increased risk of having FEV/FVC ratio below 70%. CONCLUSION: T. gondii, H. pylori and H AV infection does not influence the prevalence of atopy or asthma. The data indicated that infection with T. gondii might be associated with a diminished lung function

Patterns of nucleotide diversity in Meisa1 and G3pdh in wild and cultivated cassava

The distribution and frequency of single nucleotide polymorphisms (snps) is an excellent tool for discerning evolutionary relatedness between cultivated and wild plant genomes. This type of information is scanty for the genus Manihot, and thus limiting systematic approaches in the genetic improvement of cassava. Here, we present a detailed description of the comparative patterns of snps in Isoamylase1 (Meisa1) and Glyceraldehyde-3-phosphate dehydrogenase (G3pdh) in 10 accessions of wild (Manihot esculenta subsp. flabellifolia) and 12 accessions of cultivated cassava (M. esculenta). The results show that Meisa1 is more variable in cultivated cassava than that in subspecies flabellifolia, where the 954 bp sequence region differs at 1 in 111 and 250 nucleotides of cultivated and wild species, respectively. Frequency analysis shows that snp occurs once every 42 bp in cultivated and every 70 bp in wild. Tajima’s D test statistics showed that Meisa1 has been evolving under different selection pressures, diversifying in cultivated and purifying in wild. G3pdh is under diversifying selection in both populations. This may indicate the importance for isoamylase1 in starch quality traits in cassava, a trait that is likely to have been the target for artificial selection by farmers and breeders, in addition to natural selection. This study also suggests that G3pdh may be a good marker for phylogeny study while Meisa1 may be useful for intra and inter-cultivar diversity studies. The non-synonymous snps that changed the amino acid property were identified and the potential implication of the change in protein function was analyzed and discussed

Transcriptomic analysis of field-droughted sorghum from seedling to maturity reveals biotic and metabolic responses.

Drought is the most important environmental stress limiting crop yields. The C4 cereal sorghum [Sorghum bicolor (L.) Moench] is a critical food, forage, and emerging bioenergy crop that is notably drought-tolerant. We conducted a large-scale field experiment, imposing preflowering and postflowering drought stress on 2 genotypes of sorghum across a tightly resolved time series, from plant emergence to postanthesis, resulting in a dataset of nearly 400 transcriptomes. We observed a fast and global transcriptomic response in leaf and root tissues with clear temporal patterns, including modulation of well-known drought pathways. We also identified genotypic differences in core photosynthesis and reactive oxygen species scavenging pathways, highlighting possible mechanisms of drought tolerance and of the delayed senescence, characteristic of the stay-green phenotype. Finally, we discovered a large-scale depletion in the expression of genes critical to arbuscular mycorrhizal (AM) symbiosis, with a corresponding drop in AM fungal mass in the plants' roots

High-throughput mutational screening adds clinically important information in myelodysplastic syndromes and secondary or therapy-related acute myeloid leukemia

Non peer reviewe