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Cultivation of Green Microalgae in Bubble Column Photobioreactors and an Assay for Neutral Lipids

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An improved method to isolate lichen algae by gel filtration.

M J Pérez¹, C Vicente, M E Legaz

¹Department of Plant Physiology, The Lichen Team, Faculty of Biology, Complutense University, E-28040, Madrid, Spain.

Plant Cell Reports

|November 21, 2013

Summary

Researchers successfully isolated photobiont cells from the Evernia prunastri lichen using Sepharose 2B filtration. The absence of mannitol, a mycobiont-exclusive compound, confirmed successful separation.

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Area of Science:

Lichenology
Cell biology
Biochemistry

Background:

Lichens are symbiotic organisms comprising a fungal partner (mycobiont) and an algal or cyanobacterial partner (photobiont).
Efficient separation of these partners is crucial for studying their individual physiology and biochemistry.
Previous methods for photobiont isolation were often inefficient or lacked clear indicators of purity.

Purpose of the Study:

To develop and validate a reliable method for the complete separation of photobiont cells from the mycobiont in Evernia prunastri.
To establish biochemical markers for monitoring the isolation process and confirming the purity of the isolated photobionts.

Main Methods:

Photobiont cells were separated from the mycobiont using filtration through a Sepharose 2B column.
Mannitol and ribitol concentrations were quantified using gas-liquid chromatography (GLC) at various stages of the isolation.
The mycobiont-specific sugar alcohol, mannitol, was used as a key indicator of fungal contamination.

Main Results:

Complete separation of photobiont cells from the fungal partner was achieved using Sepharose 2B filtration.
Gas-liquid chromatography confirmed the presence of ribitol in both partners but the exclusive presence of mannitol in the mycobiont.
The absence of mannitol in the isolated fraction served as a definitive marker for pure photobiont populations.

Conclusions:

Sepharose 2B filtration is an effective method for isolating photobionts from Evernia prunastri.
Quantification of mannitol via GLC provides a robust method for assessing the purity of isolated photobiont cultures.
This technique facilitates further research into the physiology and biochemistry of lichen photobionts.