Details zur Publikation

Referenztyp Zeitschriften
DOI / URL Link
Titel (primär) Recovery of soil unicellular eukaryotes: an efficiency and activity analysis on the single cell level
Autor Lentendu, G.; Hübschmann, T.; Müller, S.; Dunker, S.; Buscot, F.; Wilhelm, C.;
Journal / Serie Journal of Microbiological Methods
Erscheinungsjahr 2013
Department UMB; BOOEK; iDiv;
Band/Volume 95
Heft 3
Sprache englisch;
POF III (gesamt) T11;
Keywords Soil unicellular eukaryotes; Cell recovery; Flow cytometry; Cell sorting; Low abundant microorganisms
UFZ Querschnittsthemen RU1
Abstract

Eukaryotic unicellular organisms are an important part of the soil microbial community, but they are often neglected in soil functional microbial diversity analysis, principally due to the absence of specific investigation methods in the special soil environment. In this study we used a method based on high-density centrifugation to specifically isolate intact algal and yeast cells, with the aim to analyze them with flow cytometry and sort them for further molecular analysis such as deep sequencing. Recovery efficiency was tested at low abundance levels that fit those in natural environments (104 to 106 cells per g soil). Five algae and five yeast morphospecies isolated from soil were used for the testing. Recovery efficiency was between 1.5 to 43.16% and 2 to 30.2%, respectively, and was dependent on soil type for three of the algae. Control treatments without soil showed that the majority of cells were lost due to the method itself (58% and 55.8% respectively). However, the cell extraction technique did not much compromise cell vitality because a fluorescein di-acetate assay indicated high viability percentages (73.3% and 97.2% of cells, respectively). The low abundant algae and yeast morphospecies recovered from soil were cytometrically analyzed and sorted. Following, their DNA was isolated and amplified using specific primers. The developed workflow enables isolation and enrichment of intact autotrophic and heterotrophic soil unicellular eukaryotes from natural environments for subsequent application of deep sequencing technologies.


ID 13771
dauerhafte UFZ-Verlinkung http://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=13771
Lentendu, G., Hübschmann, T., Müller, S., Dunker, S., Buscot, F., Wilhelm, C. (2013):
Recovery of soil unicellular eukaryotes: an efficiency and activity analysis on the single cell level
J. Microbiol. Methods 95 (3), 463 - 469