Abstract

Simple SummaryThe microalga Haematococcus lacustris is a source of the natural colorant astaxanthin, a powerful antioxidant and key component of cosmetics and animal feed. Haematococcus is cultivated in photobioreactors. It can obtain energy just from a light illuminating photobioreactor and uses inorganic salts and CO2 as sources for chemical elements. The most widespread approach for Haematococcus cultivation is the two stage scheme. At the first stage, biomass accumulation under favorable growth conditions occurs. At the second stage, the cells are subjected to stress inducing astaxanthin synthesis. Generally, the culture of Haematococcus is not axenic. It exists in the form of a community with bacteria constituting its microbiome. The information on photobioreactor-cultivated Haematococcus microbiome is scarce. We analyzed its dynamic during astaxanthin production by DNA metabarcoding and microscopic observations. The main results of the work include the characterization of the daily dynamic of this microbiome and the revealing of contact between microalgae and bacteria. These findings are of potential significance for biotechnology. On one hand, they provide an insight into possible bacterial contamination of the harvested algal biomass. On the other hand, they reveal the presence of a core microbiome or bacteria essential for the growth of the microalga existing in all Haematococcus cultures.Haematococcus lacustris is a natural source of a valuable ketocarotenoid astaxanthin. Under autotrophic growth conditions, it exists in the form of a community with bacteria. The close coexistence of these microorganisms raises two questions: how broad their diversity is and how they interact with the microalga. Despite the importance these issues, little is known about microorganisms existing in Haematococcus cultures. For the first time, we characterize the dynamic of the H. lacustris microbiome of the microbiome of Haematococcus (a changeover of the bacterial associated species as function of the time) cultivated autotrophically in a photobioreactor based on 16S rRNA metabarcoding data. We found that Proteobacteria and Bacteroidetes are predominant phyla in the community. The Caulobacter bacterium became abundant during astaxanthin accumulation. These data were supported by microscopy. We discuss possible roles and interactions of the community members. These findings are of potential significance for biotechnology. They provide an insight into possible bacterial contamination in algal biomass and reveal the presence of bacteria essential for the algal growth.

Highlights

  • Haematococcus lacustris (Volvocales, Chlorophyceae) is a unicellular green alga; a biotechnological source of the ketocarotenoid astaxanthin and a natural pigment sought after in the market for use in cosmetics, functional food and animal feed [1,2,3,4,5,6,7]

  • Most of the bacteria visually discovered in the H. lacustris monoalgal cultures cultivated autotrophically in photobioreactors were rod-shaped with a typical structure of gram-negative cells, 0.2–0.6 μm wide and 1–5 μm long (Figure 2B)

  • Our microscopy observations showed that rod-shaped bacteria abundant in the H. lacustris cultures attached to the surface of H. lacustris vegetative cells by their apexes or lateral surface (Figure 2A)

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Summary

Introduction

Haematococcus lacustris (Volvocales, Chlorophyceae) is a unicellular green alga; a biotechnological source of the ketocarotenoid astaxanthin and a natural pigment sought after in the market for use in cosmetics, functional food and animal feed [1,2,3,4,5,6,7]. H. lacustris is cultivated at an industrial scale because it accumulates the pigment in up to 4–5% of dry cell weight [3,4,5,6,8]. H. lacustris can grow under autotrophic, mixotrophic and heterotrophic conditions [1,2,9,10,11,12,13] using acetate as an organic carbon source [2,9]. The “inductive phase”, the vegetative cells are subjected to stressful conditions inducing astaxanthin accumulation.The heterotrophic and mixotrophic cultivation of H. lacustris are carried out aseptically. The prokaryotes common for H. lacustris-based natural communities from the White Sea are dominated by Comamonadaceae and include representatives of the families Cytophagaceae, Xanthomonadaceae, Acetobacteraceae, Rhodobacteraceae and Rhodocyclaceae [26]. Isolation under laboratory conditions and the removal of the cyanobacterial and eukaryotic consorts changes the bacterial diversity of H. lacustris cultures and shifts the dominant taxa

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