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How endo- is endo-? Surface sterilization of delicate samples: A Bryopsis (Bryopsidales, Chlorophyta) case study
Hollants, J.; Leliaert, F.; De Clerck, O.; Willems, A. (2010). How endo- is endo-? Surface sterilization of delicate samples: A Bryopsis (Bryopsidales, Chlorophyta) case study. Symbiosis 51(1): 131-138. dx.doi.org/10.1007/s13199-010-0068-0
In: Symbiosis. Springer: Rehovot. ISSN 0334-5114, more
Peer reviewed article

Available in Authors 
    VLIZ: Open Repository 215665 [ OMA ]

Keywords
    Bacteria [WoRMS]; Bryopsis J.V.Lamouroux, 1809 [WoRMS]; Chlorophyta [WoRMS]; Marine
Author keywords
    Bryopsis ; Endosymbionts; Endosymbiotic bacteria; Marine green algae; Surface sterilization

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Abstract
    In the search for endosymbiotic bacteria, elimination of ectosymbionts is a key point of attention. Commonly, the surface of the host itself or the symbiotic structures are sterilized with aggressive substances such as chlorine or mercury derivatives. Although these disinfectants are adequate to treat many species, they are not suitable for surface sterilization of delicate samples. In order to study the bacterial endosymbionts in the marine green alga Bryopsis, the host plant's cell wall was mechanically, chemically and enzymatically cleaned. Merely a chemical and enzymatic approach proved to be highly effective. Bryopsis thalli treated with cetyltrimethylammonium bromide (CTAB) lysis buffer, proteinase K and bactericidal cleanser Umonium Master showed no bacterial growth on agar plates or bacterial fluorescence when stained with a DNA fluorochrome. Moreover, the algal cells were intact after sterilization, suggesting endophytic DNA is still present within these algae. This new surface sterilization procedure opens the way to explore endosymbiotic microbial communities of other, even difficult to handle, samples.

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