Biology subjects

Galiana, E.

Publications and source records attributed to Galiana, E..

8 recordsLinked to original sources

bioRxiv2026

Potassium modulates Phytophthora zoospore motility and flagellar beating through dose-dependent effects

Biflagellate zoospores are critical agents in the spread of Phytophthora spp., plant pathogens causing major agronomic and economic losses worldwide. These motile cells navigate soil water films toward plant roots using taxis triggered by environmental cues. Here, we investigate how potassium modulates zoospore motility by analyzing trajectory and flagellar beating parameters across KCl concentrations. Using high-speed imaging, we apply kymograph analysis and Fourier transform of the cell body direction over time to quantify flagellar dynamics, while low-speed imaging provides trajectory characteristics. Our results demonstrate that increasing potassium concentration significantly reduces swimming speed while enhancing directional changes. We establish a direct correlation between altered swimming speed and changes in flagellar beating parameters. Our findings indicate that potassium differentially affects anterior and posterior flagella beating. Furthermore, potassium exerts multiple effects, influencing beat amplitude, wave propagation speed and direction- change frequency. This reveals potassium as a key modulator of zoospore motility, triggering a shift from exploratory swimming to confined, reoriented movement. By uncovering the biomechanical responses of zoospores to potassium, this study advances our understanding of how surrounding signals regulate pathogen dispersal in the environment.

biophysics↗

bioRxiv2026

Zoospore-derived extracellular vesicles in the flagellated stramenopile Phytophthora parasitica

Zoospores are unicellular, wall-less and flagellated cells produced by a number of eukaryotic microorganisms. They allow microbial dispersion, enable the search and location for new sources of nutrients where they aggregate, initiate pathogen-host interactions and communication within microbiota. Little is known about their ability to release bioactive extracellular vesicles (EVs) supporting these adaptations. Here we used electron microscopy to establish that in the biflagellate zoospores of the heterokont and phytopathogenic species Phytophthora parasitica, EV biogenesis occurs from vesicles budding either at cell body plasma membrane or at the front flagellum, in particular from the tubular mastigonemes. Zoospore-conditioned water supernatant was fractionated and characterized by means of morphological, immunochemical, proteomic and lipidomic analyses. Three fractions enriched in cell body, flagella and EVs were obtained by differential ultracentrifugation at 3,000g, 31,000g and 100,000g, respectively, with the EV fraction consisting of small vesicles (100-150 nm). Using mass spectrometry, label-free proteomic analysis of the EV fraction (1,470 proteins) revealed a collection of proteins involved in lipid transport, vesicle membrane and cell wall organization, and tubular mastigoneme architecture, but not in virulence. Label-free quantitative lipidomic analysis (191 lipids) revealed enrichment of EVs in sphingolipids, particularly ceramides compared with the cell body. These findings provide a first hallmark of features characteristic of zoosporic EVs, with ceramides, mastigoneme proteins and the EV-related protein PPTG_13069 containing two tetraspanin MARVELous domains among molecular markers. They define the molecular and cellular principles in understanding zoosporic EV biogenesis within Stramenopiles and intercellular communication between self-aggregating zoospores, with host plant cells or microbiota.

molecular biology↗

bioRxiv2026

Transcriptional responses of Solanum lycopersicum to three distinct parasites reveal host hubs and networks underlying parasitic successes

Crops face attacks from a wide range of pathogens and pests that deploy various parasitic strategies. All aggressors must manipulate major plant functions, such as immune signaling, hormone pathways, metabolism, and development, to successfully establish themselves and complete their life cycle. Here, we compared the tomato plant's response to three evolutionary distant pathogens (nematodes, aphids and oomycetes) during compatible interactions using a transcriptomic approach. We identified differentially expressed genes and biological processes, and highlighted potential key host hubs associated with successful parasitism. By integrating recent published datasets, we refined our understanding of the global and tissue-specific mechanisms targeted during compatible interactions and, through co-expression analysis, identified clusters showing shared dysregulation patterns enriched in specific GO terms. Finally, model-to-crop translational analysis using the Arabidopsis interactome network, repositioned tomato candidates within larger interaction networks and emphasized the key positions occupied by some of them. Identifying these pivotal tomato targets is crucial to decipher processes underlying parasitism and, consequently, offers new opportunities to develop sustainable multi-pathogen control strategies.

plant biology↗

bioRxiv2025

Membrane Proteome of Phytophthora parasitica Zoospores: How Does Sensing Occur?

Phytophthora plant pathogens rely on motile biflagellated zoospores to efficiently locate and colonise host tissues. While rhizospheric signals guiding zoospore movement toward roots are known, the protein composition of membranes mediating these responses remains unclear. Here, we used liquid chromatography with tandem mass spectrometry (LC-MS/MS) and proteomic data mining to analyse membrane fractions from the flagella and cell bodies of Phytophthora parasitica zoospores. Major classes of membrane proteins (receptors, transporters and enzymes) were identified and their subcellular distribution between flagella and cell bodies quantified. Immunolocalization revealed that while most membrane proteins are evenly distributed, a subset localizes to the flagella, suggestive of specialized roles in sensing and movement regulation, particularly for sterol recruitment and ion flux variations. These findings advance our understanding of protein-mediated dispersal and host targeting by zoospores and support the hypothesis that zoospores use polarized signal perception mechanisms for environmental sensing and movement.

microbiology↗

bioRxiv2025

Contractile Vacuole and Papilla drive Cyst/Telotroch transition in Vorticella microstoma

The cyst of the protist ciliate Vorticella efficiently differentiates into teletroch, the free-swimming stage. Here, using video microscopy and quantitative imaging we establish that the cyst differentiation follows two strict temporal and spatial patterns. The temporal pattern is initially marked by the functional formation of the contractile vacuole, which then discharges its fluid into the neoformed cytopharynx and, in a third stage, into a membrane invagination which, to the rhythm of the vacuole, differentiates into an oral cavity exhibiting a polarized cilia array revealed by -tubulin immunostaining. Two poles delineate the spatial pattern. The apical pole is defined by the position of the pre-existing papilla, which determines the site of oral cavity formation directly below it. The second basal pole is placed at the rupture point of the cyst wall. It is characterized by {beta}-actin accumulation and allowed release of the telotroph according to its basal-apical polarity. Both the temporal and spatial patterns are impaired by concanamycin A treatment, a specific inhibitor of vacuolar type H+-ATPases altering functions of contractile vacuoles. The findings indicate that cyst to telotroch transition is predetermined by the location of the papilla on the cyst wall and under the control of a functional contractile vacuole. We propose that Vorticella cyst is a simple single-cell model for investigating basic principles of integrative neogenesis of organelles in Protozoa, and of apical-basal cell polarity in Eukaryotes.

microbiology↗

bioRxiv2023

Kinetics of zoospores approaching a root using a microfluidic device

Phytophthora species are plant pathogens that cause considerable damage to agrosystems and ecosystems, and have a major impact on the economy. Infection occurs when their biflagellate zoospores move and reach a root on which they aggregate. However, the communication between the plant and the zoospores and how this communication modifies the behavior of the swimming zoospores is not yet well characterized. Here we show that using a microfluidic device comprising a growing Arabidopsis thaliana root, we are able to study the kinetics of Phytophthora parasitica zoospores approaching the root and aggregating on a specific area, in real time. We show that the kinetics of zoospores is modified only below a distance of about 300 m from the center of aggregation, with a decrease in the speed coupled with an increase in the number of turns made. In addition, we show that the rate of aggregation is constant throughout the experiment, approximately one hour, and depends on the density of zoospores. The rate is consistent with a random encounter of zoospores with the root, indicating that no long range signal is evidenced in our set-up.

biophysics↗

bioRxiv2021

Cooperation of two opposite flagella allows high-speed swimming and active turning in zoospore

Phytophthora species cause diseases in a large variety of plants and represent a serious agricultural threat, leading, every year, to multibillion dollar losses. Infection occurs when these biflagellated zoospores move across the soil at their characteristic high speed and reach the roots of a host plant. Despite the relevance of zoospore spreading in the epidemics of plant diseases, characteristics of individual swimming of zoospores have not been fully investigated. It remains unknown about the characteristics of two opposite beating flagella during translation and turning, and the roles of each flagellum on zoospore swimming. Here, combining experiments and modeling, we show how these two flagella contribute to generate thrust when beating together, and identify the mastigonemes-attached anterior flagellum as the main source of thrust. Furthermore, we find that turning involves a complex active process, in which the posterior flagellum temporarily stops, while the anterior flagellum keeps on beating and changes its pattern from sinusoidal waves to power and recovery strokes, similar to Chlamydomonass breaststroke, to reorient its body to a new direction. Our study is a fundamental step towards a better understanding of the spreading of plant pathogens motile forms, and shows that the motility pattern of these biflagellated zoospores represents a distinct eukaryotic version of the celebrated "run-and-tumble" motility class exhibited by peritrichous bacteria.

biophysics↗

bioRxiv2018

Guidance of zoospores by potassium gradient sensing mediates aggregation

Biflagellate zoospores of some phytopathogenic Phytophthora species spontaneously aggregate within few minutes in suspension. Depending on species auto-aggregate formation results from bioconvection or from a sequence bioconvection-positive chemotaxis. In this study, we show that P. parasitica zoospores may form aggregates upon application of a K+ gradient in particular geometric arrangements. Based on the use of macro- and microfluidic devices, in addition to time-lapse live imaging both in the vertical and horizontal planes, we defined (i) the spatiotemporal and concentration scale evolution within the gradient in correlation with (ii) cell distribution and (iii) metrics of zoospore motion (velocity, trajectory). The results indicated that K+-induced aggregates result from a single bi-phasic temporal sequence involving negative chemotaxis and then bioconvection in a K+ gradient concentration scale [0-17 mM]. Each K+-sensing cell undergoes a forward-to-backward movement within a threshold range of 1-4mM, thereby forming progressively a swarm. Once a critical population density is achieved zoospores form a plume which undergoes a downward migration leading to aggregate formation on the support surface. We discuss putative sources for K+ gradient generation in natural environment (zoospore population, microbiota, plant roots, soil particles), and implication for events preceding inoculum formation on host plant.

microbiology↗