An Antibiotic Protocol for Preparing Axenic Cell Cultures of Porphyra linearis

Abstract

A procedure for producing axenic cell suspension cultures of Porphyra is described. Bacteria from a gametophytic frond of P. linearis were evaluated for their sensitivity to different antibiotics. The most effective were ampicillin, carbenicillin, erythromycin, kanamycin, neomycin and ticarcillin. Cells derived from P. linearis clonal cell line CMAC-25, which were contaminated with bacteria, were treated in D-ll medium containing a mixture of these antibiotics at a concentration of 2mgmL~I of each. Next, an abalone enzyme mixture was applied to remove the antibiotic-treated cell walls that normally harbour bacteria. After regulating the osmolality of the protoplasts derived from the cells, single protoplasts were isolated and incubated in an antibiotic solution for 24 h before transferring to D-ll medium for propagation. Axenicity was confirmed in sterility testing media. A success rate of 85 -100% was achieved.

Formação de calos e regeneração de segmentos apicais de Hypnea musciformis (Wulfen) Lamouroux (Gigartinales, Rhodophyta): obtenção de culturas axênicas e efeitos da concentração do ágar

“…No entanto, uma das principais dificuldades encontradas nesses estudos em macroalgas é a fase de obtenção de culturas axênicas, livres de quaisquer tipos de contaminação (Collantes et al 1990). Com o intuito de otimizar essa fase, alguns trabalhos foram realizados por Saga & Sakai (1983), Garcia-Reina et al (1987), Lawlor et al (1987) e Chen & McCracken (1993). Apesar dessa dificuldade, estudos sobre micropropagação, particularmente nas rodofíceas, foram realizados, mais recentemente, em vários locais do mundo, enfocando o crescimento, a indução de calos e a regeneração dos talos por Gusev et al (1987), PolneFuller & Gibor (1987), Bradley & Cheney (1991), Robaina et al (1990), Liu & Kloareg (1991), Dawes & Koch (1991), Kaczyma & Megnet (1993), Yokoya et al (1993Yokoya et al ( , 1999Yokoya et al ( , 2003, Yokoya & Handro (1996, Huang & Fujita (1997), Garcia-Jimenez et al (1998) e Yokoya (2000.…”

Effect of polysaccharide admixtures on expression of multiple polysaccharide-degrading enzymes in Microbulbifer strain CMC-5

“…Gracilaria corticata collected from the Anjuna coast, Goa, India was used as test seaweed. Axenic culture of Gracilaria corticata was prepared in Provasali’s enriched sea water (ESP) medium according to Chen and McCracken (1993) [ 15 ]. Bacterial strain CMC-5 was grown in ASW medium containing 0.2% glucose for 24 h at 30 °C on an orbital shaker at 130 rpm.…”