Nutrient requirements for cell differentiation progression in Batrachochytrium dendrobatidis

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Abstract

Batrachochytrium dendrobatidis(Bd) is the causative agent of the deadly amphibian disease chytridiomycosis, which has decimated amphibian populations across the globe.Bdreproduces via a multistage asexual pathway. TheBdlife cycle begins with a motile zoospore. Upon contact with a surface, the zoospore anchors itself and forms a cyst. The cyst then develops into a germling that produces rhizoid structures. Finally, the germling matures into a zoosporangium which then releases multiple new zoospores.

In this study, we investigated the effects of nutrients on the growth and differentiation ofBd. The presence of carbohydrates, vitamins, and trace minerals had a negligible effect onBd’sability to progress through its life cycle. In contrast, the amino acids L-leucine and L-isoleucine (and to a lesser extent L-arginine) were required for cysts to germinate into germlings. However, none of these amino acids were able to sustain further cellular development. Furthermore, the presence of nitrate or ammonium aided progression from germlings into immature zoosporangia that were not able to produce zoospores. The progression to mature zoosporangia was blocked by nitrite or the combined action of calcium ions and bicarbonate. The addition of soluble and insoluble protein sources alone also allowedBdto develop into immature zoosporangia and affected the rate of development. Finally, protein sources in combination with L-amino acids allowed the completion of theBdcycle by allowing development from zoospores to mature zoosporangia that were able to release new zoospores.

Importance

Even though the cell differentiation cycle ofBdhas been microscopically described, not much is known about the requirements forBdto progress throughout its life stages. The identification of the molecular cues that allowBdcell cycle progression may be crucial to disease mitigation efforts. As a corollary, we have identified conditions that allow for the arrest ofBdat various stages of its life cycle. By changing the metabolite availability, we can efficiently obtain cultures enriched for zoospores, cysts, germlings, immature zoosporangia, or spore-releasing zoosporangia. The ability to temporarily freeze and then restartBdat different points of its life cycle, can allow for determining the mechanisms behind individual transitions between cell types.

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