Rodina auxinových přenašečů PIN: funkční a evoluční pohled

Growth and development of plant body is dependent on correct and effective integration of information about current deployment of its body parts, as well as on perception and transduction of inputs from environment. Multiple developmental processes within plant body are determined by specific and tightly controlled distribution of molecule with unique signaling mission within plant development - auxin. Spatial distribution of auxin is co-determined by plethora of tightly controlled processes, and the polar auxin transport plays unique role among them. PIN proteins are the plant-specific family of secondary transporters driving movement of auxin across membranes. With their frequent asymmetrical localization within cells, specific expression patterns in developing tissues and their reactiveness to external cues they secure unique, dynamic and asymmetric distribution of auxin within the plant body. This dissertation thesis is focused on characterization of the role different PIN proteins play in determining cellular auxin homeostasis and consequent formation of auxin gradients. Controlled overexpression of PIN proteins in tobacco cells showed, that PIN4 and to some extent also PIN6, function as the direct auxin efflux carriers. In the cellular auxin transport they play the role analogous to other...SOUHRN Pro růst rostlin a vývoj rostlinného těla má zásadní význam kvalitní a efektivní zpracování signálů, resp. informací pocházejících jednak přímo z rostlinného těla, ohledně rozložení jednotlivých rostlinných částí a jednak integrace podnětů z prostředí, umožňující rostlině reagovat na vnitřní vývojové podmínky i vnější hrozby vhodnou korekcí svého růstového programu. Velké množství vývojových procesů rostlinného těla je tak determinováno charakteristickým prostorovým rozmístěním auxinu - molekuly s informační hodnotou. Prostorového rozmístění auxinu v prostoru je dosaženo kombinací a bilancí velkého množství striktně regulovaných procesů, mezi nimiž významnou a jedinečnou roli hraje polární transport auxinu. Rodina auxinových přenašečů PIN pak představuje přenašeče auxinu z buňky, které díky své asymetrické lokalizaci, specifickým expresním vzorcům a modulovatelné transportní funkci zajišťují tvorbu specifických auxinových gradientů. Předmětem této disertační práce je charakterizace rolí, kterou jednotlivé PINy hrají při tvorbě a udržování vnitřních buněčných hladin auxinu a následně auxinových gradientů v rostlině. Kontrolovatelná nad-exprese proteinů PIN v tabákových buněčných liniích ukázala, že PIN4 a do určité míry také PIN6, fungují přímo jako přenašeče auxinu ven z buňky. V buněčném auxinovém...Department of Experimental Plant BiologyKatedra experimentální biologie rostlinFaculty of SciencePřírodovědecká fakult

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

Rodina auxinových přenašečů PIN: funkční a evoluční pohled

Growth and development of plant body is dependent on correct and effective integration of information about current deployment of its body parts, as well as on perception and transduction of inputs from environment. Multiple developmental processes within plant body are determined by specific and tightly controlled distribution of molecule with unique signaling mission within plant development - auxin. Spatial distribution of auxin is co-determined by plethora of tightly controlled processes, and the polar auxin transport plays unique role among them. PIN proteins are the plant-specific family of secondary transporters driving movement of auxin across membranes. With their frequent asymmetrical localization within cells, specific expression patterns in developing tissues and their reactiveness to external cues they secure unique, dynamic and asymmetric distribution of auxin within the plant body. This dissertation thesis is focused on characterization of the role different PIN proteins play in determining cellular auxin homeostasis and consequent formation of auxin gradients. Controlled overexpression of PIN proteins in tobacco cells showed, that PIN4 and to some extent also PIN6, function as the direct auxin efflux carriers. In the cellular auxin transport they play the role analogous to other..

New knowledge in the field of mechanism of action and use of auxin herbicides

The methodology summarizes all the latest findings in the field of mechanism of auxin effects in the plant, which give users a new perspective on their use in agricultural practice

Methods in Molecular Biology

Exogenous application of biologically important molecules for plant growth promotion and/or regulation is very common both in plant research and horticulture. Plant hormones such as auxins and cytokinins are classes of compounds which are often applied exogenously. Nevertheless, plants possess a well-established machinery to regulate the active pool of exogenously applied compounds by converting them to metabolites and conjugates. Consequently, it is often very useful to know the in vivo status of applied compounds to connect them with some of the regulatory events in plant developmental processes. The in vivo status of applied compounds can be measured by incubating plants with radiolabeled compounds, followed by extraction, purification, and HPLC metabolic profiling of plant extracts. Recently we have used this method to characterize the intracellularly localized PIN protein, PIN5. Here we explain the method in detail, with a focus on general application

Interaction of PIN and PGP transport mechanisms in auxin distribution-dependent development

The signalling molecule auxin controls plant morphogenesis via its activity gradients, which are produced by intercellular auxin transport. Cellular auxin efflux is the rate-limiting step in this process and depends on PIN and phosphoglycoprotein (PGP) auxin transporters. Mutual roles for these proteins in auxin transport are unclear, as is the significance of their interactions for plant development. Here, we have analysed the importance of the functional interaction between PIN- and PGP-dependent auxin transport in development. We show by analysis of inducible overexpression lines that PINs and PGPs define distinct auxin transport mechanisms: both mediate auxin efflux but they play diverse developmental roles. Components of both systems are expressed during embryogenesis, organogenesis and tropisms, and they interact genetically in both synergistic and antagonistic fashions. A concerted action of PIN- and PGP-dependent efflux systems is required for asymmetric auxin distribution during these processes. We propose a model in which PGP-mediated efflux controls auxin levels in auxin channel-forming cells and, thus, auxin availability for PIN- dependent vectorial auxin movement

Light-induced STOMAGEN-mediated stomatal development in Arabidopsis

The initiation of stomata, microscopic valves in the epidermis of higher plants that control of gas exchange, requires a co-ordinated sequence of asymmetric and symmetric divisions, which is under tight environmental and developmental control. Arabidopsis leaves grown under elevated photosynthetic photon flux density have a higher density of stomata. STOMAGEN encodes an epidermal patterning factor produced in the mesophyll, and our observations indicated that elevated photosynthetic irradiation stimulates STOMAGEN expression. Our analysis of gain and loss of function of STOMAGEN further detailed its function as a positive regulator of stomatal formation on both sides of the leaf, not only in terms of stomatal density across the leaf surface but also in terms of their stomatal index. STOMAGEN function was rate limiting for the light response of the stomatal lineage in the adaxial epidermis. Mutants in pathways that regulate stomatal spacing in the epidermis and have elevated stomatal density, such as stomatal density and distribution (sdd1) and too many mouth alleles, displayed elevated STOMAGEN expression, suggesting that STOMAGEN is either under the direct control of these pathways or is indirectly affected by stomatal patterning, suggestive of a feedback mechanism. These observations support a model in which changes in levels of light irradiation are perceived in the mesophyll and control the production of stomata in the epidermis by mesophyll-produced STOMAGEN, and whereby, conversely, stomatal patterning, either directly or indirectly, influences STOMAGEN levels

PIN6 auxin transporter at endoplasmic reticulum and plasma membrane mediates auxin homeostasis and organogenesis in Arabidopsis

Plant development mediated by the phytohormone auxin depends on tightly controlled cellular auxin levels at its target tissue that are largely established by intercellular and intracellular auxin transport mediated by PIN auxin transporters. Among the eight members of the Arabidopsis PIN family, PIN6 is the least characterized candidate. In this study we generated functional, fluorescent protein-tagged PIN6 proteins and performed comprehensive analysis of their subcellular localization and also performed a detailed functional characterization of PIN6 and its developmental roles. The localization study of PIN6 revealed a dual localization at the plasma membrane (PM) and endoplasmic reticulum (ER). Transport and metabolic profiling assays in cultured cells and Arabidopsis strongly suggest that PIN6 mediates both auxin transport across the PM and intracellular auxin homeostasis, including the regulation of free auxin and auxin conjugates levels. As evidenced by the loss- and gain-of-function analysis, the complex function of PIN6 in auxin transport and homeostasis is required for auxin distribution during lateral and adventitious root organogenesis and for progression of these developmental processes. These results illustrate a unique position of PIN6 within the family of PIN auxin transporters and further add complexity to the developmentally crucial process of auxin transport