ABSTRACT
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.
Authors’ contributions
EG and XN designed experiments.
EG and CE carried droplet and millifluidic analyses.
XN, CC and PT carried out microfluidic analyses.
EG, XN, CC, PT wrote the manuscript.
Acknowledgements
The authors thank the Microscopy Platform-Sophia Agrobiotech Institut-INRA 1355-UNS-CNRS 7254-INRA PACA Sophia Antipolis for access to instruments and technical advice. The authors would like to thank Fernando Peruani, Emiliano Perez Ipiña (UAD, Nice) and Laurent Counillon (LP2M, Nice) for fruitful discussions.
Footnotes
EMAIL ADRESSES: Eric Galiana, eric.galiana{at}inra.fr, Catherine Etienne, catherine.mura{at}inra.fr, Xavier Noblin, xavier.noblin{at}unice.fr, Celine Cohen, celine.cohen{at}unice.fr, Philippe Thomen, Philippe.THOMEN{at}unice.fr
Competing interests: The authors declare that they have no competing interests.
Funding: This work has been supported by the French government, through the UCAJEDI Investments in the Future project managed by the National Research Agency (ANR) with the reference number ANR-15-IDEX-01; through the “Credits Scientifiques Incitatifs” of the University of Nice Sophia-Antipolis and the “Action Recherche” of the INRA Plant Health and Environment Division.