An improved enzyme preparation for rapid mass production of protoplasts as seed stock for aquaculture of macrophytic marine green algae

Differential response of varying salinity and temperature on zoospore induction, regeneration and daily growth rate in Ulva fasciata (Chlorophyta, Ulvales)

“…The unipolar germination pattern demarcating the elongated rhizoidal cell after first cell division favors firm attachment prior to development of the erect shoot. The formation of the extensive rhizoidal system in U. fasciata makes it suitable for open sea mariculture.The highest regeneration in U. fasciata (78.53±10.05%) at 25°C and 30 psu and at 15 μmol photons m −2 s −1 with a 12:12 h light:dark photoperiod is in agreement with observations on several other tropical seaweeds Reddy et al (2006). reported protoplast regeneration in several species of Ulvaceae at 21°C and 32 psu at 15 μmol photons m −2 s −1 and a 12:12 h light: dark photoperiod.…”

“…Furthermore, due to contamination of intertidal coastal waters by sewage and other domestic and industrial effluent, the spores collected in the field are highly susceptible to microbial attack. In view of these bottlenecks, other methods such as isolation and seeding of protoplasts (Reddy et al 2006), germling clusters (Hiraoka and Oka 2008), and dark light treatment (Mairh et al 1986) have been proposed for artificial seeding. However, the utility of these techniques for large-scale commercial farming is yet to be assessed.…”

Isolation of seaweed-associated bacteria and their morphogenesis-inducing capability in axenic cultures of the green alga Ulva fasciata

“…The flasks with algal pieces were then maintained in a Multi Thermo Incubator (MTI-202; Eyela) at 25 ± 1°C temperature under daylight white fluorescent lamps at 15 µmol photon m -2 s -1 irradiance with a 12:12 h light:dark photoperiod. After 2 to 3 d of acclimatization, fronds were given sequential treatment with different chemicals to obtain an axenic algal culture as described by Reddy et al (2006). The axenicity of the algal culture was tested by incubating randomly selected algal tissue on Zobell agar medium over a week at 37 ± 1°C in an incubator.…”

Production and regeneration of protoplasts from Grateloupia turuturu Yamada (Rhodophyta)

“…Even if variations in protoplast yields have been generally attributed to the physiological, biochemical, and seasonal changes of algal material used (Cheney et al 1986;Reddy et al 2006), the effects of the concentration and the composition of enzymes, the incubation period, and ionic and osmotic strengths, which have been reported to influence protoplast yields (Reddy et al 2006), were evaluated to optimize the isolation protocol. A maximum yield of 1.5×10 7 protoplasts g −1 fresh weight was obtained after tissue has been incubated in an enzyme mixture with 2% cellulase Onozuka R-10, 0.5% macerozyme R-10, and 2% crude extract of H. tuberculata for 4 h. Generally, the incubation times in mixture enzyme for producing Rhodophyta protoplasts are variable according to the protocols for a period ranging from 1 to 48 h. Enzyme incubation periods are only 1-1.5 h for Palmaria palmata (Liu et al 1992), 2 h for Porphyra yezoensis (Liu et al 2004), 2 to 4 h for Gracilaria (Cheney et al 1986;Yeong et al 2008), 18 h for G. sparsa and G. filicina (Chen and Chiang 1994), and until 48 h for Kappaphyccus alvarezii (Salvador and Serrano 2005).…”