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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.
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.
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