Understanding the dynamical bases of the interaction between the plant mineral nutrition and the circadian clock could contribute to improve crop yield and resistance to adverse conditions, such as mineral element deficiencies. Magnesium is an essential element that catalyzes more than six hundred enzymatic reactions and occupies the center of the chlorophyll structure in plants. Physiological targets of magnesium deficiency are generally better described in aerial than in belowground organs. In this thesis, we first characterized the root morphology of the model species Arabidopsis thaliana (Columbia-0) in response to magnesium supply during in vitro culture. The length of primary root and the number and length of lateral roots decreased during magnesium depletion. A local magnesium-rich source does not enhance the root foraging capacity, unlike some other major nutrients. Auxin and abscisic acid emerged as two hormones shaping root morphology in response to magnesium deficiency. Second, we investigated the natural variation of the root morphology response to magnesium supply in Arabidopsis. Thirty-six accessions were screened in vitro. Compared to the reference Columbia-0, some accessions had higher number and length of lateral roots at low magnesium supply. Root or shoot magnesium concentrations did not implicitly correlate with the root morphological traits. However, shoot calcium and root phosphorus concentrations correlated positively with the lateral root number and length, while root iron negatively with the length of primary root. Third, we focused on the interaction between the plant magnesium nutrition and the circadian clock circuit. We tested for a possible involvement of PHYTOCHROME INTERACTING FACTOR 3-LIKE 1 (PIL1) - a clock-associated gene that is down-regulated during magnesium depletion - in morphological and physiological responses, and for a circadian connection of PIL1 in the context of magnesium deficiency. The phenotypes of loss-of-function and overexpression lines did not reveal a major role of PIL1 in the magnesium deficiency symptom manifestation but rather in the plant mineral profile. The expression of PIL1 was apparently not under any circadian control. However, PIL1 seemed to regulate the expressions of some core clock genes (CCA1, LHY and PRR9), which were also targets during magnesium deficiency. In conclusion, PIL1 has a link with the circadian rhythm machinery but it does not emerge as a pivotal regulator of magnesium stress responses. / Doctorat en Sciences / info:eu-repo/semantics/nonPublished

京都大学 / 0048 / 新制・課程博士 / 博士(薬科学) / 甲第21717号 / 薬科博第108号 / 新制||薬科||12(附属図書館) / 京都大学大学院薬学研究科医薬創成情報科学専攻 / (主査)教授 土居 雅夫, 教授 竹島 浩, 教授 中山 和久 / 学位規則第4条第1項該当 / Doctor of Pharmaceutical Sciences / Kyoto University / DGAM

Internal clocks, or circadian rhythms, are nearly ubiquitous across taxa (e.g., animals, plants, fungi, and cyanobacteria), and it is widely believed that a biological clock benefits organisms by enabling them to schedule behavioral and physiological changes in anticipation of predictable changes in environmental conditions. Theory and evidence suggest it is important that the internal clock resonate closely with the 24-h daily cycle. Recently, however, Cyclosa turbinata (Walckenaer, 1841) (Araneidae) was revealed to have a circadian clock with a period of about 19 h, which was presumed to be anomalous. Here, we report on the behavioral rhythms of a nocturnal orbweaver, Metazygia wittfeldae (McCook, 1894), from the same family. Under laboratory conditions of a 12:12 h light:dark cycle, we found that locomotor activity initiates shortly after dark, reaching a peak early in the dark phase, continuing at a lower level throughout the remaining dark phase, and then diminishing shortly after lights-on. Locomotor activity continued to cycle under constant dark conditions with a mean free-running period of 22.7 h. We also found a second component in the free-running activity (mean 11.5 h) which correlated very tightly with the free-running period. Thus, M. wittfeldae has what can be considered a typical circadian clock resonating with the 24-h day. Notably, however, there were two outliers close to the 19-h period observed in C.Turbinata, suggesting that there may be sufficient variation in clock period among araneid spiders upon which selection could act leading to the short-period clocks in C.Turbinata.

京都大学 / 0048 / 新制・課程博士 / 博士(薬科学) / 甲第18923号 / 薬科博第37号 / 新制||薬||5(附属図書館) / 31874 / 京都大学大学院薬学研究科医薬創成情報科学専攻 / (主査)教授 岡村 均, 教授 中山 和久, 教授 竹島 浩 / 学位規則第4条第1項該当 / Doctor of Pharmaceutical Sciences / Kyoto University / DFAM

京都大学 / 新制・課程博士 / 博士(薬科学) / 甲第23842号 / 薬科博第157号 / 新制||薬科||17(附属図書館) / 京都大学大学院薬学研究科医薬創成情報科学専攻 / (主査)教授 土居 雅夫, 教授 竹島 浩, 教授 中山 和久 / 学位規則第4条第1項該当 / Doctor of Pharmaceutical Sciences / Kyoto University / DFAM

In this study, the magnitude of stomatal water loss was compared between different Arabidopsis thaliana mutants. The stomatal water loss among different mutants of Populus sp. was also examined in this study. Fourteen different Arabidopsis thaliana mutants were sown and later genotyped using DNA-extraction, PCR, and gel electrophoresis. Leaf water loss analysis was made on both Arabidopsis thaliana and Populus sp. The data was then used in statistical tests, to find if there was a significant difference in rate of stomatal water loss between different genotypes. The result showed a significant difference in water loss when comparing the toc1-5 and T89 genotypes and the toc1-5 and 1119-4 genotypes in Populus sp. There was no significant difference between the 1119-4 and T89 genotypes in Populus sp. The result also showed that there was a significant difference between 25 combinations of the Arabidopsis thaliana genotypes. A conclusion was that TOC1 reduces water loss in Arabidopsis thaliana, indicating a role in stomatal clousure. Future research is needed to examine differences between genotypes among both species included in the study.

No description available.

Suprachiasmatic nuclei (SCN) is the master circadian pacemaker that generates coordinated rhythms and drives oscillations in other peripheral tissues. Extracellular vesicles (exosomes) have been implicated in cell-to-cell communication and the regulation of circadian clock. However, mammalian clock-derived exosomes have not been characterized. This thesis examine the contents of exosome released from SCN2.2 cells in vitro using a combination of proteomics, next-generation sequencing, and bioinformatic analyses. SCN2.2 cells-derived exosomes, that carry unique microRNAs and proteins, could be taken up by fibroblast cells in vitro. Interestingly, several unique microRNAs and proteins found in SCN2.2 cells-derived exosomes have shown circadian rhythmicity in other cells. In addition, differential expressed microRNAs secreted by SCN cells were also observed outside of exosomes. Taken together, these studies demonstrate that exosomes, containing small RNAs, RNAs and proteins, are released from SCN2.2 cells and likely have a biological role in circadian regulation of metabolism in downstream cells.

Trees, like many other organisms, decrease their rate of metabolic activities to cope up with harsh environments. This stage of "dormancy" is marked by shedding of leaves and bud-set in deciduous trees. Recent studies have revealed the role of the circadian clock in synchronizing the timing of dormancy and physiology for conferring fitness in trees. To better understand the possible role of natural selection on circadian clock-related genes in climatic adaptation, I took a candidate gene approach, selecting circadian clock genes, some of which had been functionally validated, and others hypothesized, to identify signatures of natural selection in Populus trichocarpa and P. deltoides. Using both frequency spectrum based tests and tests of heterogeneity, I identified genetic variants deviating from selective neutrality. Results reveal that photoreceptors and dormancy regulator genes may have been the targets of natural selection. Nearly the same levels of selective constraints were found in different functional groups of genes irrespective of pleiotropy. Further, upstream regions of all genes showed high selective constraint, with some of them (FT-2, PIF-4, FRIGIDA) showing significantly higher variation than the other genes, hinting at the role of non-coding regulatory regions in local adaption. In some cases, the same genes in both species appeared as outliers, including PIF-6, FRI, FT-2, SRR1, TIC, and CO, which might reflect their common role in adaptation across species boundaries. All of these results indicate a complex nature of phenology regulation and local adaptation in Populus species with photoreceptors and dormancy regulator genes playing key roles. / Master of Science

De part ses propriétés physico-chimiques, le fer est un cofacteur de choix pour de nombreuses enzymes, impliquées dans de multiples processus biologiques, comme la photosynthèse ou la respiration. Cependant, sa capacité à gagner/perdre des électrons le rend très réactif, et potentiellement toxique. Son homéostasie doit donc être finement régulée. Chez A. thaliana, le gène de ferritine AtFER1 est régulé transcriptionnellement par le fer, et son expression est régulée par le rythme circadien. Au début de ce travail, aucun facteur de transcription impliqué dans cette régulation n'était identifié. Des cribles simple hybride en levure ont été réalisés, permettant l'identification de deux facteurs de transcription régulant le gène AtFER1: AtPHR1 et AtPIF7.PHR1 (Phosphate starvation Response1) et son homologue PHL1 (PHr1 Like 1) sont des facteurs de transcription impliqués dans la réponse à la carence en phosphate. Ils régulent directement le gène AtFER1, en se fixant sur l'élément 2 du promoteur d'AtFER1. Cette régulation ne fait pas intervenir l'IDRS et est indépendante du statut en fer des plantes. Par ailleurs, l'homéostasie du fer est affectée dans le double mutant phr1 phl1. Ces résultats montrent l'existence d'un lien moléculaire direct entre les homéostasies du fer et du phosphate.PIF7 (Phytochrome Interacting Factor 7) est un facteur de transcription de type bHLH impliqué dans la régulation circadienne des gènes DREB. Il se fixe probablement sur la G-box présente dans l'élément 5 du promoteur d'AtFer1. Dans un mutant perte de fonction pour le gène PIF7, l'amplitude des oscillations de l'expression du gène AtFer1 en cycles jour/nuit est augmentée. Un résultat similaire est obtenu lorsque l'élément 5 est muté dans des lignées trasngéniques exprimant le gène rapporteur LUC sous le contrôle du promoteur d'AtFer1. Ces résultats montrent que PIF7 est un répresseur de l'expression d'AtFer1. / Due to its redox properties, iron is a major cofactor for numerous proteins involved in many biological processes such as photosynthesis or respiration. Nevertheless, its ability to easily gain or lose electrons makes it highly reactive with oxygen and potentially toxic. Iron homeostasis has to be tightly regulated. In A. thaliana, AtFER1 ferritin gene is regulated at the transcriptional level by iron, and its expression is regulated by the circadian clock. Before this work, no transcription factor involved AtFer1 regulation has been identified. A yeast one hybrid was performed and allowed us to identify PHR1 and PIF7 as transcription factors involved in AtFER1 regulation. PHR1 (Phosphate starvation Response1) and its homolog PHL1 (PHr1 Like 1) are transcription factors involved in phosphate starvation response. They directly regulate AtFER1 expression and bind to the element 2 present in AtFER1 promoter. This regulation does not involve the IDRS sequence and is independent of iron status of plants. Moreover, iron homeostasis is affected in phr1phl1 double mutant. These results highlight a direct molecular link between iron and phosphate homeostasis.PIF7 (Phytochrome Interacting Factor 7) is a bHLH transcription factor involved in the circadian regulation of DREB genes. PIF7 probably interacts with the G-box found in element 5 in AtFER1 promoter region. In a pif7 knock-out mutant, amplitude of AtFer1 oscillations during light dark cycles are increased. Such a result was also obtained with transgenic lines expressing LUC reporter gene under the control of AtFER1 promoter region harboring a mutation in element 5. These results show that PIF7 is a repressor of AtFer1 expression.