A Rapid and Inexpensive Method for Surface Sterilisation of Ecklonia radiata (Phaeophyta) for Tissue Culture

Abstract

Sections of the thallus of the brown alga Ecklonia radiata (C. Ag.) J. Ag. were surface sterilised for tissue culture by dipping in 70% ethanol for 30 s, followed by sterile deionised water for 30 min. A high percentage of aseptic viable explants could be obtained using this treatment, and growth in vitro was initiated.

Decontamination treatments to eliminate problem biota from macroalgal tank cultures of Osmundea pinnatifida, Palmaria palmata and Ulva lactuca

“…KI has been used successfully before in the disinfection of kelp (Yan 1984); however, a slightly high concentration of 1.5 % for 10 s was found to be lethal to Ecklonia radiata (Lawlor et al 1991). In addition, KI can lead to iodine staining of the tissue (Kientz et al 2012), which may affect the suitability and value of the treated tissue for food.…”

“…For instance, Lawlor et al (1991) dipped E. radiata into 70 % ethanol for 30 s, Kawashima and Tokuda (1990) dipped into 70 % ethanol for 30–60 s and Kientz et al (2012) recommend 30–60 s in 40–50 % ethanol and 1 % NaClO. Such short exposure times could be an effective treatments against the epiphytes reported.…”

A new method for removing microflora from macroalgal surfaces: an important step for natural product discovery

“…Despite the importance of the topic, few publications have addressed the issue of disinfection of macroalgal surfaces prior to metabolite extraction (Fu et al 1999, Schulz et al 2008. Nevertheless, development of axenic cultures has focused on these aspects (Gusev et al 1984, Xue-wu and Gordon 1987, Lawlor et al 1991, Garcia-Jiménez et al 1998, Choi et al 2002, Rorrer and Cheney 2004, Harrison and Berges 2005 and it has been shown that disinfection of macroalgae is difficult as they lack a thick surface cuticle and can be easily damaged (Reddy et al 2008, Baweja et al 2009). Methods developed for aquaculture purposes often combine physical surface cleaning of macroepiphytes (using brushes, razors, sonication or osmotic shocks) (Xue-wu and Gordon 1987) and chemical treatment for microflora elimination (surfactant, alcohols, oxidizing agents, iodine and antibiotics) (Baweja et al 2009).…”

“…Harrison and Berges (2005) published protocols based on the use of antibiotics. Various techniques have been developed in which algae are submerged in different bacterial disinfectants including ethanol (Gusev et al 1987, Lawlor et al 1991, Schulz et al 1998 and sodium hypochlorite (Garcia-Jiménez et al 1998). Lawlor et al (1991) demonstrated that a treatment with 70% ethanol for 30 s combined with 30 min in deionized water was efficient in producing axenic tissue cultures from Ecklonia radiata (C. Agardh) J. Agardh (Phaeophyta).…”

“…Various techniques have been developed in which algae are submerged in different bacterial disinfectants including ethanol (Gusev et al 1987, Lawlor et al 1991, Schulz et al 1998 and sodium hypochlorite (Garcia-Jiménez et al 1998). Lawlor et al (1991) demonstrated that a treatment with 70% ethanol for 30 s combined with 30 min in deionized water was efficient in producing axenic tissue cultures from Ecklonia radiata (C. Agardh) J. Agardh (Phaeophyta). Similarly, Gusev et al (1984Gusev et al ( , 1987 employed 70% ethanol and 1% chlorhexidine bigluconate for 15 min to produce axenic marine algae.…”