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2.
PLoS Genet ; 12(12): e1006490, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27973599

RESUMO

Environmental fluctuations affect distribution, growth and abundance of diatoms in nature, with iron (Fe) availability playing a central role. Studies on the response of diatoms to low Fe have either utilized continuous (24 hr) illumination or sampled a single time of day, missing any temporal dynamics. We profiled the physiology, metabolite composition, and global transcripts of the pennate diatom Phaeodactylum tricornutum during steady-state growth at low, intermediate, and high levels of dissolved Fe over light:dark cycles, to better understand fundamental aspects of genetic control of physiological acclimation to growth under Fe-limitation. We greatly expand the catalog of genes involved in the low Fe response, highlighting the importance of intracellular trafficking in Fe-limited diatoms. P. tricornutum exhibited transcriptomic hallmarks of slowed growth leading to prolonged periods of cell division/silica deposition, which could impact biogeochemical carbon sequestration in Fe-limited regions. Light harvesting and ribosome biogenesis transcripts were generally reduced under low Fe while transcript levels for genes putatively involved in the acquisition and recycling of Fe were increased. We also noted shifts in expression towards increased synthesis and catabolism of branched chain amino acids in P. tricornutum grown at low Fe whereas expression of genes involved in central core metabolism were relatively unaffected, indicating that essential cellular function is protected. Beyond the response of P. tricornutum to low Fe, we observed major coordinated shifts in transcript control of primary and intermediate metabolism over light:dark cycles which contribute to a new view of the significance of distinctive diatom pathways, such as mitochondrial glycolysis and the ornithine-urea cycle. This study provides new insight into transcriptional modulation of diatom physiology and metabolism across light:dark cycles in response to Fe availability, providing mechanistic understanding for the ability of diatoms to remain metabolically poised to respond quickly to Fe input and revealing strategies underlying their ecological success.


Assuntos
Diatomáceas/metabolismo , Ferro/metabolismo , Fotoperíodo , Transcriptoma/genética , Ciclo Celular/efeitos dos fármacos , Ciclo Celular/genética , Divisão Celular/efeitos dos fármacos , Divisão Celular/genética , Cloroplastos/genética , Diatomáceas/efeitos dos fármacos , Diatomáceas/crescimento & desenvolvimento , Expressão Gênica , Ferro/farmacologia , Redes e Vias Metabólicas/genética , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Biossíntese de Proteínas/efeitos dos fármacos
3.
Sci Rep ; 6: 19252, 2016 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-26786712

RESUMO

Although sexual reproduction is believed to play a major role in the high diversification rates and species richness of diatoms, a mechanistic understanding of diatom life cycle control is virtually lacking. Diatom sexual signalling is controlled by a complex, yet largely unknown, pheromone system. Here, a sex-inducing pheromone (SIP(+)) of the benthic pennate diatom Seminavis robusta was identified by comparative metabolomics, subsequently purified, and physicochemically characterized. Transcriptome analysis revealed that SIP(+) triggers the switch from mitosis-to-meiosis in the opposing mating type, coupled with the transcriptional induction of proline biosynthesis genes, and the release of the proline-derived attraction pheromone. The induction of cell cycle arrest by a pheromone, chemically distinct from the one used to attract the opposite mating type, highlights the existence of a sophisticated mechanism to increase chances of mate finding, while keeping the metabolic losses associated with the release of an attraction pheromone to a minimum.


Assuntos
Pontos de Checagem do Ciclo Celular , Diatomáceas/fisiologia , Atrativos Sexuais/metabolismo , Comportamento Sexual Animal , Animais , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Ácido Glutâmico/metabolismo , Guanilato Ciclase/genética , Guanilato Ciclase/metabolismo , Meiose , Redes e Vias Metabólicas , Metaboloma , Metabolômica/métodos , Mitose , Diester Fosfórico Hidrolases/genética , Diester Fosfórico Hidrolases/metabolismo , Prolina/metabolismo , Atrativos Sexuais/farmacologia , Transcrição Gênica
4.
Nat Commun ; 6: 6925, 2015 Apr 21.
Artigo em Inglês | MEDLINE | ID: mdl-25897682

RESUMO

Eukaryotic microalgae hold great promise for the bioproduction of fuels and higher value chemicals. However, compared with model genetic organisms such as Escherichia coli and Saccharomyces cerevisiae, characterization of the complex biology and biochemistry of algae and strain improvement has been hampered by the inefficient genetic tools. To date, many algal species are transformable only via particle bombardment, and the introduced DNA is integrated randomly into the nuclear genome. Here we describe the first nuclear episomal vector for diatoms and a plasmid delivery method via conjugation from Escherichia coli to the diatoms Phaeodactylum tricornutum and Thalassiosira pseudonana. We identify a yeast-derived sequence that enables stable episome replication in these diatoms even in the absence of antibiotic selection and show that episomes are maintained as closed circles at copy number equivalent to native chromosomes. This highly efficient genetic system facilitates high-throughput functional characterization of algal genes and accelerates molecular phytoplankton research.


Assuntos
Conjugação Genética , Diatomáceas/genética , Escherichia coli/fisiologia , Plasmídeos , DNA/genética , Eletroporação , Vetores Genéticos , Plasmídeos/genética , Polietilenoglicóis , Recombinação Genética , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo
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