Naslov
Isolation of DNA and proteins from marine microbial epiphytes

Autori
Markovski, Marsej ; Korlević, Marino ; Zhao, Zihao ; Herndl, Gerhard J. ; Najdek, Mirjana

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
FEMS Conference on Microbiology : Electronic abstract book / - Belgrade, 2022, 547-548

ISBN
978-86-914897-8-6

Skup
2nd FEMS Conference on Microbiology

Mjesto i datum
Beograd, Srbija, 30.06.2022. - 02.07.2022

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
DNA and protein isolation ; marine macrophytes ; Cymodocea nodosa ; Caulerpa cylindracea

Sažetak
Background: Marine macrophyte surfaces are inhabited by diverse microbial communities. When culture-independent methods such as 16S rRNA sequencing, metagenomics, and metaproteomics are used in the study of these communities, it is advisable to perform selective isolation of DNA and proteins prior to analysis. Methods for separating microbial cells from host tissue, such as shaking, scraping, and ultrasonication, do not result in complete cell detachment and, in the case of ultrasonication, may result in host tissue disruption. Objectives: Our objective was to adapt protocols for selective DNA and protein isolation from microbial communities inhabiting two marine macrophyte species, the seagrass Cymodocea nodosa and the macroalga Caulerpa cylindracea, and to evaluate the suitability of the recovered material for culture-independent molecular methods. Methods: The developed protocols use universally available laboratory chemicals and are based on direct cell lysis on surfaces of marine macrophytes. The removal efficiency of the procedures was evaluated using confocal microscopy, while the recovered DNA and proteins were tested by 16S rRNA sequencing, metagenomics and metaproteomics. Results: The adapted protocols allowed almost complete removal of epiphytic microbial cells, regardless of host species. The obtained DNA was suitable for metagenomic and 16S rRNA sequencing, while isolated proteins were identified by mass spectrometry. The low presence of host DNA and proteins observed in the isolated samples indicates a high specificity of the protocols for epiphytic communities.

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Interdisciplinarne prirodne znanosti

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