Novel approach for the development of axenic microalgal cultures from environmental samples

We demonstrated a comprehensive approach for development of axenic cultures of microalgae from environmental samples. A combination of ultrasonication, fluorescence‐activated cell sorting (FACS), and micropicking was used to isolate axenic cultures of Chlorella vulgaris Beyerinck (Beijerinck) and Ch...

Full description

Saved in:
Bibliographic Details
Published in:Journal of phycology 2013-08, Vol.49 (4), p.802-810
Main Authors: Cho, Dae-Hyun, Ramanan, Rishiram, Kim, Byung-Hyuk, Lee, Jimin, Kim, Sora, Yoo, Chan, Choi, Gang-Guk, Oh, Hee-Mock, Kim, Hee-Sik
Format: Article
Language:English
Subjects:
Algae
Axenic culture
Chlorella sorokiniana
Chlorella sp
Chlorella vulgaris
FACS
Microalgae
Microbiology
Micropicking
Scenedesmus
Scenedesmus sp
Ultrasonication
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:We demonstrated a comprehensive approach for development of axenic cultures of microalgae from environmental samples. A combination of ultrasonication, fluorescence‐activated cell sorting (FACS), and micropicking was used to isolate axenic cultures of Chlorella vulgaris Beyerinck (Beijerinck) and Chlorella sorokiniana Shihira & R.W. Krauss from swine wastewater, and Scenedesmus sp. YC001 from an open pond. Ultrasonication dispersed microorganisms attached to microalgae and reduced the bacterial population by 70%, and when followed by cell sorting yielded 99.5% pure microalgal strains. The strains were rendered axenic by the novel method of micropicking and were tested for purity in both solid and liquid media under different trophic states. Denaturing gradient gel electrophoresis (DGGE) of 16S rRNA gene confirmed the absence of unculturable bacteria, whereas fluorescence microscopy and scanning electron microscopy (SEM) further confirmed the axenicity. This is the most comprehensive approach developed to date for obtaining axenic microalgal strains without the use of antibiotics and repetitive subculturing.
ISSN:0022-3646
1529-8817
DOI:10.1111/jpy.12091