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Farre, E. M.

Publications and source records attributed to Farre, E. M..

3 recordsLinked to original sources

High resolution diel transcriptomes of autotetraploid potato reveal expression and sequence conservation among rhythmic genes

Photoperiodic changes in diel cycles of gene expression are pervasive in plants. Timing of circadian regulators together with light signals regulate multiple photoperiod dependent responses such as growth, flowering or tuber formation. However, for most genes the importance of cyclic mRNA levels is less clear. We analyzed the diel transcriptome of modern cultivated potato, a highly heterozygous autotetraploid. Clonal propagation and limited meiosis have led to the accumulation of deleterious alleles and therefore tetraploid potato is an ideal model system to investigate the conservation of cyclic expression and cyclic genes during the artificial selection process. We observed that cyclic alleles were more highly expressed than non-cyclic ones and were highly co-expressed not only under diel cycles but also across tissues, developmental stages and stress conditions. Moreover, the smaller ratio of non-synonymous to synonymous differences within cyclic as compared to non-cyclic allelic groups indicates that cyclic genes, in general, have more conserved core functions than those of non-cyclic ones. In accordance with this observation, fully rhythmic allelic groups were highly enriched in photosynthesis and ribosome biogenesis genes, which play core functions in plants. Furthermore, we investigated differences in cyclic expression patterns between photoperiods. We identified transcription factors potentially regulating the strong differences in phase between photoperiods observed in ribosome biogenesis and pathogen response genes. Finally, analyses of genes involved in tuber formation suggests that the regulation of CO gene transcription is not the only factor enabling tuberization under long days in modern cultivated potato. This study not only provides high quality diel transcriptomic datasets of cultivated potato but also important insight on the role of allelic diversity in rhythmic expression in plants.

plant biology↗

Aureochromes are necessary for maintaining polyunsaturated fatty acid content in Nannochloropsis oceanica

Nannochloropsis oceanica, as other stramenopile microalgae, is rich in long-chain polyunsaturated fatty acids (LC-PUFA) such as eiconsapentaenoic acid (EPA). We observed that fatty acid desaturases (FAD) involved in LC-PUFA biosynthesis were among the strongest blue light induced genes in N. oceanica CCMP1779. Blue light was also necessary for maintaining LC-PUFA levels in CCMP1779 cells, and growth under red light led to a reduction in EPA content. Aureochromes are stramenopile specific proteins that contain a light-oxygen-voltage-sensing (LOV) domain that associates with a flavin mononucleotide and is able to sense blue light. These proteins also contain a bZIP DNA binding motif and can act as blue light regulated transcription factors by associating with a E-box like motif, which we found enriched in the promoters of blue light induced genes. We demonstrated that, in vitro, two CCMP1779 aureochromes were able to absorb blue light. Moreover, the loss or reduction of any of the three aureochromes led to a decrease in the blue light specific induction of several FADs in CCMP1779. EPA content was also significantly reduced in NoAureo 2 and NoAureo 4 mutants. Taken together, our results indicate that aureochromes mediate blue light dependent regulation of LC-PUFA content in N. oceanica CCMP1779 cells.

plant biology↗

Identification of circadian rhythms in Nannochloropsis species using bioluminescence reporter lines

Circadian clocks allow organisms to predict environmental changes caused by the rotation of the Earth. Although circadian rhythms are widespread among different taxa, the core components of circadian oscillators are not conserved and differ between bacteria, plants, animals and fungi. Stramenopiles are a large group of organisms in which circadian rhythms have been only poorly characterized and no clock components have been identified. We have investigated cell division and molecular rhythms in Nannochloropsis species. In the four strains tested, cell division occurred principally during the night period under diel conditions, however, rhythms dampened within 2-3 days after transfer to constant light. We developed firefly luciferase reporters for long-term monitoring of in vivo transcriptional rhythms in two Nannochlropsis species, N. oceanica CCMP1779 and N. salina CCMP537. The reporter lines express free-running bioluminescence rhythms with periods of ~21-31 h that dampen within ~3-4 days under constant light. Using different entrainment regimes, we demonstrate that these rhythms are regulated by a circadian-type oscillator. In addition, the phase of free-running luminescence rhythms can be modulated pharmacologically using a CK1 {varepsilon}/{delta} inhibitor, suggesting a role of this kinase in the Nannochloropsis clock. Together with the molecular and genomic tools available for Nannochloropsis species, these reporter lines represent an excellent system for future studies on the molecular mechanisms of stramenopile circadian oscillators. Significance statementStramenopiles are a large and diverse line of eukaryotes in which circadian rhythms have been only poorly characterized and no clock components have been identified. We have developed bioluminescence reporter lines in Nannochloropsis species and provide evidence for the presence of a circadian oscillator in stramenopiles; these lines will serve as tools for future studies to uncover the molecular mechanisms of circadian oscillations in these species.

molecular biology↗