DEVELOPMENT OF PROTOPLASTS OFULVA FASCIATA(CHLOROPHYTA) FOR ALGAL SEED STOCK

Abstract

The aim of this study was to isolate and cultivate protoplasts of the green alga Ulva fasciata Delile and subsequently induce them to form a microthallus suspension for algal seed stock. The protoplasts were covered with secreted mucilage following 6 h of culture when viewed with SEM. The mucilage fused to form thick layers during day 1 of culture. Microfibrillar cell walls were deposited into the thick layers of mucilage on the 5th day of culture. An average of about 10% of the freshly isolated protoplasts began to divide at 6-14 days. These protoplasts subsequently developed varied morphologies, depending on the time of collection during the year. Protoplasts isolated from U. fasciata collected in March to June developed frond thalli or microthalli when they were cultured in low or high densities (cells/area), respectively. The microthallus suspension was cultured for more than two years at 10-40 μmol·m ·s . Frond thalli formed when the suspension was cultivated at 100-160 μmol·m ·s . Therefore, microthallus suspension can serve as a seed stock of U. fasciata.

Optimization of protoplast yields from the red algae Gracilaria dura (C. Agardh) J. Agardh and G. verrucosa (Huds.) Papenfuss

“…2006). The variations in protoplast yields from Gracilariales, especially with enzymatic methods, have been attributed partly to physiological and biochemical as well as seasonal changes of the material being investigated (Cheney et al 1986;Kloareg et al 1989;Chen and Shih 2000). Moreover, the activity of crude enzymes also varies from batch to batch, leading to inconsistent protoplast yield from time to time.…”

Seaweed protoplasts: status, biotechnological perspectives and needs

“…The yield, viability and regeneration rate of isolated protoplasts is dependent on several factors, including the enzyme constituents and their concentrations, pH, osmotic conditions and ionic strength of protoplast isolation medium, incubation temperature, physiological state and age of donor plant, and protoplast culture medium and its culture conditions Cheney et al 1986;Bjork et al 1990;Butler et al 1990;Chen 1998;Krishna Kumar et al 1999;Chen and Shih 2000;Matsumura et al 2000;Shikh et al 2005;Reddy et al 2006). Pretreatment of thallus with proteases (protease or papain) or plasmolytic solutions was also carried out prior to enzymatic treatment to obtain enhanced yields of viable protoplasts from some members of Chlorophyta and Rhodophyta (Kawashima et al 1989;Yamaguchi et al 1989;Fujita and Saito 1990;Bjork et al 1992;Amano and Noda 1992;Chen and Chiang 1994a, b;Wakabayashi et al 1999).…”

“…In order to provide continuous seed stock for cultivation of green seaweeds, Chen (1998) and Chen and Shih (2000) developed protoplast-based methods for producing stocks of seedlings (micro thalli) with survival abilities of many years in an incubator while maintaining their potential to develop into leafy thalli of Ulva and Monostroma. Protoplasts from Monostroma and Porphyra have been successfully tested for their seeding and regeneration in laboratory conditions (Reddy et al 2006;Shikh et al 2005).…”

Quantification of DNA content in freshwater microalgae using flow cytometry: a modified protocol for selected green microalgae.

“…There exist actually differences in cell wall composition within different algal taxa (Stace 1991), but the concentrations of the main cell wall polymers of land plants are similar to that of Streptophyta (eder et al 2008(eder et al , anderSOn & kinG 1961. We modified the enzyme mixture used for the genera Cucumis and Cucurbita (GajdOVá et al 2007;Ondřej et al 2009) and marine green macroalgae (reddy et al 2006;uPPalaPati & Fujita 2002;chen & Shih 2000). Flow cytometry and DNA amount estimation FC is the optimal method for nuclear genome size estimation.…”