Abstract
Paraphysoderma sedebokerense (P. sedebokerense) (Blastocladiomycota) is a facultative pathogenic chytrid that causes irreversible damage to some green microalgae. Specific attacks leading to culture collapse under different conditions have only been described in the lucrative microalga Haematococcus pluvialis (H. pluvialis), while generating biomass for ketocarotenoid astaxanthin production, both indoors and outdoors. In order to manage the infection, parasite propagules (zoospores/amoeboid swarmers), the initiators of the disease, must be studied. Until now, no report on isolated P. sedebokerense propagules has been published. Here, we report on a reproducible method for the stimulation of P. sedebokerense propagule release and their isolation from fungal cultures in synthetic media and infected H. pluvialis cultures, and we further studied their development under different conditions. The isolated propagules featured different spore morphotypes, with coatless spherical spores and amoeboid swarmers being the most dominant in the first pulse of propagule release in both cultures. Inoculating the pure propagules with the host, in both the presence and absence of nitrogen, resulted in epidemic development in both green and red cells; however, in red cells, the epidemic developed more quickly in the presence of nitrogen. Biologically non-active autoclaved host cells were used to distinguish the initial stages of recognition from more progressive stages of the epidemics; on these cells, propagules encysted but did not develop further. These results prove the existence of heat-stable recognition sites on the host and an obligatory signal transduction from the host to support fungal cyst development. The propagule isolation method described herein is a breakthrough that will enable researchers to study the influence of different substances on the propagules, specifically as the initiators of the infection, and thus assist in the management of chytrid diseases. Moreover, it will be useful in studying host-parasite recognition and, therefore, will increase our understanding of the multiple chytrid infections found in nature.
Highlights
The parasite Paraphysoderma sedebokerense (P. sedebokerense) from Blastocladiomycota, isolated and initially characterized in our laboratory by Hoffman et al (2008), has gained much attention because of its host-specific infection of Haematococcus pluvialis (H. pluvialis), which results in the collapse of the algal cultures and the ultimate decline in the production of industrially important red ketocarotenoid astaxanthin
An axenic blastoclad monoculture was grown in an enriched chytrid growth medium (Gutman et al, 2009), with glucose replaced by sucrose, called the blastoclad growth medium (BGM)
We found that replacing distilled water with the modified BG-11 (mBG11) medium for plate flooding resulted in higher propagule production, with even higher production when mBG11 without NO3 was used
Summary
The parasite Paraphysoderma sedebokerense (P. sedebokerense) from Blastocladiomycota, isolated and initially characterized in our laboratory by Hoffman et al (2008), has gained much attention because of its host-specific infection of Haematococcus pluvialis (H. pluvialis), which results in the collapse of the algal cultures and the ultimate decline in the production of industrially important red ketocarotenoid astaxanthin. The macroscopic symptoms of this parasite in H. pluvialis include flocculation at the initial stages of infection, followed by a change of algal cells’ color from green to brown. Most parasites of freshwater algae are zoosporic true fungi and fungi-like organisms, belonging to the taxonomical groups of Chytridiomycota, Blastocladiomycota, Cryptomycota, Oomycota, Alveolata, and Labyrinthulomycota (Carney and Lane, 2014). Based on their motility, zoospores of true fungi (Chytridiomycota, Blastocladiomycota) are the primary dispersive and infective propagules in aquatic habitats, originating from sporangium production after cyst vegetative growth. Blastocladiomycota zoospores are capable of locomotion, either by a single posteriorly located whiplash flagellum in a water environment, or a wall-less propagule capable of an amoeboid motion on the surface of a substrate, such as a host or a nutrient agar (Strittmatter et al, 2016), as well as on a microscope slide
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