Establishment and maintenance of an axenic culture of Ettlia sp. using a species-specific approach

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Title
Establishment and maintenance of an axenic culture of Ettlia sp. using a species-specific approach
Author(s)
Hyung Gwan Lee; Sang Yoon Shin; L Jin; Chan Yoo; A Srivastava; Hyun Joon La; Chi-Yong AhnHee-Sik KimHee-Mock Oh
Bibliographic Citation
Biotechnology and Bioprocess Engineering, vol. 20, no. 6, pp. 1056-1063
Publication Year
2015
Abstract
The establishment of an axenic culture of microalgae is essential step in understanding its physiology, genetics, and ecology. However, culturing of microalgae is usually accompanied by complex and variable associated prokaryotic and eukaryotic microorganisms. Conventional approaches used for obtaining axenic cultures of microalgae are time-consuming and often involve difficulties in maintaining and preserving axenicity. In this study, we developed a procedure for establishing an axenic culture of Ettlia sp. YC001 and demonstrate that we maintained the axenic culture through subculture in the long term. Three sequential treatments, an antibiotic cocktail, serial dilution, and plate spreading, were applied to strain YC001 and we confirmed axenicity using molecular and physiological methods. The bacterial community associated with strain YC001 was investigated to select antibiotics for their specific elimination. The xenic culture (1 × 106 cells/mL) was treated with the antibiotic cocktail-5 (AC-5), carbendazim, chloramphenicol, imipenem, rifampicin, and tetracycline for 3 days, followed by serial dilution up to 1 × 102 cells and spreading on agar plates. The pure colonies were analyzed using denaturing gradient gel electrophoresis (DGGE), fluorescence-activated cell sorting (FACS), and scanning electron microscopy (SEM). The procedure we developed can be applied to other strains of microalgae for the establishment of axenic cultures.
Keyword
antibiotic cocktailaxenic cultureEttlia sp. YC001plate spreadingserial dilution
ISSN
1226-8372
Publisher
South Korea
DOI
http://dx.doi.org/10.1007/s12257-015-0289-4
Type
Article
Appears in Collections:
Division of Biomaterials Research > Cell Factory Research Center > 1. Journal Articles
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