Transport auxinu v řasách

Fytohormon auxin hraje důležitou roli v regulaci rostlinného vývoje. Směrovaný (polární) transport auxinu mezi buňkami vytváří jeho gradienty v rostlinných pletivech, které spouští specifickou vývojovou odpověď. Valná většina dostupných dat se týká krytosemenných rostlin. Nižší rostliny jsou v tomto ohledu mnohem méně prozkoumány, ale důležité auxinové mechanismy (včetně polárního transportu) jsou přítomny již v meších. Abychom odhalili počátky role auxinu v rostlinách, musíme se zaměřit na zelené řasy - obzvláště řasy ze skupiny Streptophyta, které jsou přímými předchůdci všech rostlin. V této studii byl zkoumán možný vliv auxinů, nativních i syntetických, na dvě řasy: původní, jednobuněčnou Chlorella lobophora a pokročilou, vláknitou Spirogyra sp. (šroubatka). Šroubatka dostála více pozornosti, neboť se řadí do skupiny dnes uznávané jako sesterská k rostlinám. Růst kultur Chlorella lobophora nebyl ovlivněn syntetickým auxinem NAA. Naproti tomu, průměrná délka buněk šroubatky byla ovlivněna auxiny ve vysokých koncentracích. Prostřednictvím akumulačních esejí radioaktivně značených auxinů a HPLC analýzy byl pozorován metabolismus a transport auxinů ve šroubatce. Šroubatka byla schopna metabolizovat nativní auxin IAA, nikoli však syntetické NAA a 2,4-D. Výdej auxinu buňkami, pokud je přítomen, nebyl...Phytohormone auxin plays an important role in regulating plant development. Directional (polar) cell-to-cell auxin transport creates auxin gradients within plant tissues, which trigger a specific developmental response. The vast majority of available data concerns angiosperms. Lower land plants have been much less explored in this regard, but the important auxin-related mechanisms (including polar auxin transport) are already present in mosses. To uncover the origins of auxin action, one must focus on green algae, especially of clade Streptophyta, which are the direct ancestors of all land plants. In this study, the possible effects of auxins, both native and synthetic, were investigated on two algae: basal, unicellular Chlorella lobophora and advanced, filamentous Spirogyra sp. The latter received comparably more attention, since it belongs to a clade now acknowledged as a sister group to land plants. Chlorella lobophora culture growth was irresponsive to synthetic auxin NAA. The average Spirogyra sp. cell length was, however, changed by auxins at high concentrations. By conducting accumulation assays of radioactively labelled auxins and HPLC analysis, auxin metabolism and transport was investigated in Spirogyra sp. This alga was able to metabolize the plant-native IAA, but not synthetic auxins...Department of Experimental Plant BiologyKatedra experimentální biologie rostlinPřírodovědecká fakultaFaculty of Scienc

Evoluce mechanismů transportu auxinu.

Auxin, nejdéle studovaný fytohormon, je v porovnání s ostatními fytohormony unikátní svým směrovaným tzv. polárním transportem. Tato vlastnost napomáhá zprostředkovat širokou škálu působnosti auxinu ve všech stadiích života rostliny. Polární transport auxinu se v evoluci vyvíjel společně s rostlinami. Zkoumáním mechanismů transportu auxinu a jeho biosyntézy, metabolismu a především signalizace můžeme snad lépe objasnit mnoho důležitých milníků evoluce rostlin, jako vývoj složitější mnohobuněčnosti či přesun rostlin na souš. Tato bakalářská práce shrnuje dostupná data a podává základní přehled rysů s auxinem spojených. Jak dosud známo, pokročilé mechanismy transportu auxinu a jeho signalizace, známé z vyšších rostlin, nejsou zřejmě fylogeneticky staré a v řadě řas zcela absentují. Biosyntéza auxinu je ovšem vcelku běžná a mnoho zelených řas obsahuje ortology důležitých enzymů spojených s metabolismem auxinu. Dle dostupných informací se zdá, že kompletní auxinová signalizace, spojená s degradací v proteasomu a ovlivňující genovou expresi, v řasách chybí. Polární transport auxinu, dosud znám nejdříve ze sporofytů mechů, byl nedávno zjištěn v gametofytické stélce parožnatky (Chara) z oddělení zelených řas Streptophyta, které jsou považovány za přímé předchůdce vyšších rostlin. Proteiny sekvenčně podobné...Auxin, the longest studied phytohormone, is distinguished from other phytohormones by its unique directional, so-called polar transport. This feature helps to facilitate the broad range of auxin action at all stages of plant development. The polar auxin transport has been evolving together with plant lineages. By studying the mechanisms of auxin transport, biosynthesis, metabolism and particularly signaling we can perhaps better elucidate many milestones of plant evolution, such as complex multicellularity or transition to land. This bachelor thesis summarizes the available data and gives a basic overview of auxin-related characteristics. As far as we know, the advanced mechanisms of auxin transport and signaling known from land plants are probably not very ancient and are absent in various algae. Auxin biosynthesis, however, is rather common and a lot of green algae contain orthologs of important biosynthetic enzymes from land plants. Based on the available data it seems that a complete auxin signalling pathway coupled with proteasomal degradation and affecting gene expression is not present in algae. The polar auxin transport, so far with the earliest evidence from moss sporophytes, was recently found in the gametophytic thallus of stonewort (Chara) from a green algal clade Streptophyta, which is...Katedra experimentální biologie rostlinDepartment of Experimental Plant BiologyPřírodovědecká fakultaFaculty of Scienc

Evoluce mechanismů homeostáze auxinu

The evolution of auxin homeostasis mechanisms Ph.D. thesis Roman Skokan, 2021 Abstract The streptophyte lineage consists of land plants (embryophytes) and several groups of primarily freshwater green algae called charophytes. While the phytohormone auxin is a conserved regulator of land plant development, little has been known of the possible origins of auxin response mechanisms in charophytes. We found that one of these mechanisms, the cellular auxin efflux via the PIN family of transport proteins, is most likely a deeply conserved feature in streptophytes. Additionally, we investigated the state of conservation in the green lineage (Viridiplantae) of the gene families known to be involved in auxin transport in land plants. We revealed that some families are deeply conserved outside land plants, but not others. We also helped uncover a unique radiation within the PIN family in Charophyceae. Striving to uncover the native significance of auxin transport in charophytes, we discovered a growth response to exogenously-applied auxin in Closterium, though the effort to produce stable mutant lines in the native PIN homolog is still underway. Altogether, we brought important insights into the evolution of auxin transport and response in the streptophyte lineage, though many questions still remain.Evoluce mechanismů homeostáze auxinu Dizertační práce Roman Skokan, 2021 Abstrakt Linie streptofyt obsahuje vyšší rostliny (Embryophyta) a několik skupin primárně sladkovodních řas zvaných charofyti. Přestože fytohormon auxin je konzervovaným vývojovým regulátorem ve vyšších rostlinách, málo je známo o možných evolučních počátcích mechanismů odpovědi na auxin v charofytech. Zjistili jsme, že jeden z těchto mechanismů, tedy transport auxinu z buněk skrze proteiny z rodiny PIN, je velmi pravděpodobně hluboce konzervovanou vlastností streptofyt. Dále jsme studovali stupeň konzervace genových rodin známých rostlinných auxinových přenašečů v zelené linii (Viridiplantae). Objevili jsme hlubokou konzervaci některých z těchto rodin mimo vyšší rostliny. Dále jsme pomohli popsat jedinečné rozrůznění rodiny PIN ve skupině Charophyceae. Ve snaze objevit nativní funkci transportu auxinu v charofytech jsme objevili růstovou odpověď na externě aplikovaný auxin v řase Closterium, ačkoli snaha připravit stabilní mutantní linie v nativním PIN homologu stále probíhá. Celkově naše práce přinesla důležité poznatky o evoluci transportu a odpovědi na auxin ve streptofytech, přestože mnoho otázek zůstává stále nezodpovězených.Department of Experimental Plant BiologyKatedra experimentální biologie rostlinPřírodovědecká fakultaFaculty of Scienc

Evolutionary conserved cysteines function as cis-acting regulators of Arabidopsis PIN-FORMED 2 distribution

Coordination of plant development requires modulation of growth responses that are under control of the phytohormone auxin. PIN-FORMED plasma membrane proteins, involved in intercellular transport of the growth regulator, are key to the transmission of such auxin signals and subject to multilevel surveillance mechanisms, including reversible post-translational modifications. Apart from well-studied PIN protein modifications, namely phosphorylation and ubiquitylation, no further post-translational modifications have been described so far. Here, we focused on root-specific Arabidopsis PIN2 and explored functional implications of two evolutionary conserved cysteines, by a combination of in silico and molecular approaches. PIN2 sequence alignments and modeling predictions indicated that both cysteines are facing the cytoplasm and therefore would be accessible to redox status-controlled modifications. Notably, mutant pin2C−A alleles retained functionality, demonstrated by their ability to almost completely rescue defects of a pin2 null allele, whereas high resolution analysis of pin2C−A localization revealed increased intracellular accumulation, and altered protein distribution within plasma membrane micro-domains. The observed effects of cysteine replacements on root growth and PIN2 localization are consistent with a model in which redox status-dependent cysteine modifications participate in the regulation of PIN2 mobility, thereby fine-tuning polar auxin transport

PIN-driven auxin transport emerged early in streptophyte evolution

PIN-FORMED (PIN) transporters mediate directional, intercellular movement of the phytohormone auxin in land plants. To elucidate the evolutionary origins of this developmentally crucial mechanism, we analysed the single PIN homologue of a simple green alga Klebsormidium flaccidum. KfPIN functions as a plasma membrane-localized auxin exporter in land plants and heterologous models. While its role in algae remains unclear, PIN-driven auxin export is probably an ancient and conserved trait within streptophytes

Evoluce mechanismů transportu auxinu.

Auxin, the longest studied phytohormone, is distinguished from other phytohormones by its unique directional, so-called polar transport. This feature helps to facilitate the broad range of auxin action at all stages of plant development. The polar auxin transport has been evolving together with plant lineages. By studying the mechanisms of auxin transport, biosynthesis, metabolism and particularly signaling we can perhaps better elucidate many milestones of plant evolution, such as complex multicellularity or transition to land. This bachelor thesis summarizes the available data and gives a basic overview of auxin-related characteristics. As far as we know, the advanced mechanisms of auxin transport and signaling known from land plants are probably not very ancient and are absent in various algae. Auxin biosynthesis, however, is rather common and a lot of green algae contain orthologs of important biosynthetic enzymes from land plants. Based on the available data it seems that a complete auxin signalling pathway coupled with proteasomal degradation and affecting gene expression is not present in algae. The polar auxin transport, so far with the earliest evidence from moss sporophytes, was recently found in the gametophytic thallus of stonewort (Chara) from a green algal clade Streptophyta, which is..

Auxin transport in algae

Phytohormone auxin plays an important role in regulating plant development. Directional (polar) cell-to-cell auxin transport creates auxin gradients within plant tissues, which trigger a specific developmental response. The vast majority of available data concerns angiosperms. Lower land plants have been much less explored in this regard, but the important auxin-related mechanisms (including polar auxin transport) are already present in mosses. To uncover the origins of auxin action, one must focus on green algae, especially of clade Streptophyta, which are the direct ancestors of all land plants. In this study, the possible effects of auxins, both native and synthetic, were investigated on two algae: basal, unicellular Chlorella lobophora and advanced, filamentous Spirogyra sp. The latter received comparably more attention, since it belongs to a clade now acknowledged as a sister group to land plants. Chlorella lobophora culture growth was irresponsive to synthetic auxin NAA. The average Spirogyra sp. cell length was, however, changed by auxins at high concentrations. By conducting accumulation assays of radioactively labelled auxins and HPLC analysis, auxin metabolism and transport was investigated in Spirogyra sp. This alga was able to metabolize the plant-native IAA, but not synthetic auxins..

The evolution of auxin homeostasis mechanisms

The evolution of auxin homeostasis mechanisms Ph.D. thesis Roman Skokan, 2021 Abstract The streptophyte lineage consists of land plants (embryophytes) and several groups of primarily freshwater green algae called charophytes. While the phytohormone auxin is a conserved regulator of land plant development, little has been known of the possible origins of auxin response mechanisms in charophytes. We found that one of these mechanisms, the cellular auxin efflux via the PIN family of transport proteins, is most likely a deeply conserved feature in streptophytes. Additionally, we investigated the state of conservation in the green lineage (Viridiplantae) of the gene families known to be involved in auxin transport in land plants. We revealed that some families are deeply conserved outside land plants, but not others. We also helped uncover a unique radiation within the PIN family in Charophyceae. Striving to uncover the native significance of auxin transport in charophytes, we discovered a growth response to exogenously-applied auxin in Closterium, though the effort to produce stable mutant lines in the native PIN homolog is still underway. Altogether, we brought important insights into the evolution of auxin transport and response in the streptophyte lineage, though many questions still remain