A gene loop containing the floral repressor FLC is disrupted in the early phase of vernalization.

Gene activation in eukaryotes frequently involves interactions between chromosomal regions. We have investigated whether higher-order chromatin structures are involved in the regulation of the Arabidopsis floral repressor gene FLC, a target of several chromatin regulatory pathways. Here, we identify a gene loop involving the physical interaction of the 5' and 3' flanking regions of the FLC locus using chromosome conformation capture. The FLC loop is unaffected by mutations disrupting conserved chromatin regulatory pathways leading to very different expression states. However, the loop is disrupted during vernalization, the cold-induced, Polycomb-dependent epigenetic silencing of FLC. Loop disruption parallels timing of the cold-induced FLC transcriptional shut-down and upregulation of FLC antisense transcripts, but does not need a cold-induced PHD protein required for the epigenetic silencing. We suggest that gene loop disruption is an early step in the switch from an expressed to a Polycomb-silenced state.

RNA 3' processing functions of Arabidopsis FCA and FPA limit intergenic transcription.

The RNA-binding proteins FCA and FPA were identified based on their repression of the flowering time regulator FLC but have since been shown to have widespread roles in the Arabidopsis thaliana genome. Here, we use whole-genome tiling arrays to show that a wide spectrum of genes and transposable elements are misexpressed in the fca-9 fpa-7 (fcafpa) double mutant at two stages of seedling development. There was a significant bias for misregulated genomic segments mapping to the 3' region of genes. In addition, the double mutant misexpressed a large number of previously unannotated genomic segments corresponding to intergenic regions. We characterized a subset of these misexpressed unannotated segments and established that they resulted from extensive transcriptional read-through, use of downstream polyadenylation sites, and alternative splicing. In some cases, the transcriptional read-through significantly reduced expression of the associated genes. FCA/FPA-dependent changes in DNA methylation were found at several loci, supporting previous associations of FCA/FPA function with chromatin modifications. Our data suggest that FCA and FPA play important roles in the A. thaliana genome in RNA 3' processing and transcription termination, thus limiting intergenic transcription.

Transcription beyond borders has downstream consequences.

The realization that non-coding RNAs and antisense transcription are pervasive in many genomes has emphasized our relatively poor understanding of what limits transcription and how initiation and termination are linked to processing and turnover of the RNA. In genomes where the density of genes is high it is clearly important to efficiently terminate transcription to prevent read-through into adjacent genes. In a recent paper published in PNAS, we showed that two RNA binding proteins in Arabidopsis thaliana, FCA and FPA, play important roles in limiting intergenic transcription in the A. thaliana genome. Their absence leads to transcriptional read-through over many kilobases (kb), which influences expression, and in some cases chromatin modifications, of associated genes.

Evaluation of novel thermo-resistant Micractinium and Scenedesmus sp. for efficient biomass and lipid production under different temperature and nutrient regimes.

Despite the vast interest in microalgae as feedstock for biodiesel production, relatively few studies examined their response to diurnal temperature fluctuation. Here, we describe biomass and lipid productivities and fatty acid profiles of thermo-resistant Micractinium sp. and Scenedesmus sp. grown in batch cultures in a laboratory set-up that mimics a typically warm summer day in Central Anatolia with a 16-h light temperature of 30°C and 8-h dark temperature of 16°C (30°C (day)/16°C (night)). Both strains can survive a temperature range of 10-50°C. We found the lipid productivities of Micractinium sp. and Scenedesmus sp. as 30/21mgL(-1)d(-1) and 6/7mgL(-1)d(-1), respectively during the 30°C (day)/16°C (night) cycle. Saturated fatty acid content increased with increasing temperature. Additionally, we cultured Micractinium sp. under Nitrogen (N) and Phosphorus (P) limiting conditions. Highest lipid productivity of 85.4±2mgL(-1)d(-1) was obtained under P-depletion during exponential growth phase. Oleic acid amount also increased eight fold during P-deplete.

Thermo-resistant green microalgae for effective biodiesel production: isolation and characterization of unialgal species from geothermal flora of Central Anatolia.

Oil content and composition, biomass productivity and adaptability to different growth conditions are important parameters in selecting a suitable microalgal strain for biodiesel production. Here, we describe isolation and characterization of three green microalgal species from geothermal flora of Central Anatolia. All three isolates, namely, Scenedesmus sp. METUNERGY1402 (Scenedesmus sp. ME02), Hindakia tetrachotoma METUNERGY1403 (H. tetrachotoma ME03) and Micractinium sp. METUNERGY1405 (Micractinium sp. ME05) are adaptable to growth at a wide temperature range (25-50 °C). Micractinium sp. ME05, particularly has superior properties for biodiesel production. Biomass productivity, lipid content and lipid productivity of this isolate are 0.17 g L(-1) d(-1), 22.7% and 0.04 g L(-1) d(-1), respectively. In addition, Micractinium sp. ME05 and Scenedesmus sp. ME03 mainly contain desirable fatty acid methyl esters (i.e. 16:0, 16:1, 18:0 and 18:1) for biodiesel production. All isolates can further be improved via genetic and metabolic engineering strategies.