Effects of Light Quality on Growth and Physiological Characteristics of Neopyropia yezoensis Free Living Conchocelis

[Objectives]To study the differences of growth rate,morphology,ultrastructure,pigment content and antioxidant enzyme activity of free-living conchocelis of cultivated type of Neopyropia yezoensis under different light qualities(white,red,blue,and green light).[Methods]The study was carried out through light quality design and culture,growth rate determination,microstructure and ultrastructure observation,chlorophyll a content and carotenoid content determination,phycobiliprotein content determination,malondialdehyde(MDA)content determination,superoxide dismutase(SOD)activity determination.[Results]After 21 d of culture,the specific growth rate(SGR)and chlorophyll a content of free-living conchocelis of N.yezoensis were significantly increased by white light(WL),followed by red light(RL)and green light(GL),and they were the lowest under blue light(BL).Compared with the WL group,the BL group had the highest content of phycoerythrin(PE),and the RL and GL groups had the highest content of phycocyanin(PC).The algal body of WL group was normal black brown,and the cell wall was the thickest.In RL and GL groups,the algal bodies were green,and their diameters and cell wall thicknesses were similar to those in WL group.In BL group,the algal body was bright red,the diameter was the smallest,the cell wall was the thinnest,and the ultrastructure showed that the number of plastoglobulus on the thylakoid was the largest.After BL irradiation,the highest MDA content and the lowest SOD activity were observed.The results revealed that WL is the most beneficial to the growth of free-living conchocelis,followed by RL and GL,while BL has adverse effects.[Conclusions]This study explored the most suitable light quality conditions for the propagation of free-living conchocelis.It is expected to provide germplasm guarantee for the production and seedling of N.yezoensis.

GAMMA-RAYS INDUCTION OF MUTATION IN CONCHOCELIS OF PORPHYRA YEZOENSIS

Free living conchocelis of Porphyra yezoensis Ueda (Bangiales, Rhodophyta) were treated with 6 0Co γ rays of different doses (ranging from 100 Gy to 1000 Gy) to induce mutation. Most of the conchocelis maintained the capability of penetrating into shells, growing and forming colonies in shells, but their vitality was seriously impaired by the irradiation of γ rays. A few conchocelis pigments were mutagenized directly into different color pigment mutants whose progeny conchospores and foliose thalli had the same colors. However, some irradiated conchocelis did not show the change in color at the conchocelis stage. The pigment mutation could be observed only after the conchospores of these conchocelis had germinated into young foliose thalli. Irradiation of low dose (100 Gy) promoted the growth of thallus and many with altered morphology were observed. Conchospores of the irradiated conchocelis attached to the culture nets were cultured in the sea, and growth of these progenies was observed and measured.

Study on Cryopreservation of Porphyra yezoensis Conchocelis

Cryopreservation of Porphyra yezoensis conchocelis was conducted with cryoprotectants and a proposed pretreatment procedure and thawing methods explored. Six cryoprotectants combined by DMSO with ethylene glycol(EG),propylene glycol(PEG),sorbitol and sucrose were developed. The effect of prefreezing at - 40℃ or -20℃ for different time durations was compared and the thawing methods were screened. It was shown that the cryoprotectant including 10% DMSO with 0.5 molL-1 sorbitol exhib-ited the optimal effect. The ideal pretreatment was that conchocelis segments were stayed for 20 min at - 40℃ before stored in liquid nitrogen,and 40℃ water bath was proper for quick thawing. The highest recovery rate of cryopreserved P. yezoensis conchocelis reached 89.41%.

AXENIC CULTURE OF FREE-LIVING CONCHOCELIS OF PORPHYRA YEZOENSIS AND PORPHYRA HAITANENSIS

After discarding marine microorganisms from conchocelis of Porphyra yezoensis and Porphyra haitanensis, their axenic cultures were obtained through treatment with antibiotics. Antibiotic disc tests were carried out to determine the effectiveness of each antibiotic in eliminating contaminating microorganisms. Five of 12 antibiotics tested were selected and used to produce the axenic cultures in this study, which showed that 200 μg/mL streptomycin, 250 μg/mL penicillin, 252 μg/mL kanamycin, 30 μg/mL neomycin, 200 μg/mL chloramphenicol were effective concentrations for eliminating microorganisms from conchocelis when antibiotics were added singly step by step; whereas simultaneous combination of 150 μg/mL streptomycin, 250 (or 350) μg/mL penicillin, 150 (or 250) μg/mL kanamycin, 70 μg/mL neomycin and 200 μg/mL chloramphenicol was also effective for producing the axenic cultures. However, it seemed that the treatments with antibiotics applied individually were more feasible than those with all antibiotics added at the same time. This may be due to the combined inhibiting effect of antibiotics on the growth and development of conchocelis.

EFFECT OF CYTOKININS AND AUXINS ON THE GROWTH OF FREE-LIVING CONCHOCELIS OF PORPHYRA YEZOENSIS

1AA 3-Indolylacetic acid, NAA a-Naphthylacetic acid and cytokinins in PESI culture medium were used in a study on the effects of plant hormones on the growth of free-living conchocelis of Porphyra yezoensis which showed that its growth in medium with cytokinins, 1AA and NAA was more rapid than that in medium with non-phytohormones; that the optimal concentrations for promoting growth were 10μg/L for IAA and ZA (Zeatin), and 0.1 μg/L for BA 6-Benzyl amino purine and KIN 6-Furfurylamino- purine. Mix use of NAA, 1AA and cytokinins, NAA/ZA 1-1000/1μg/L, NAA/BA 10/1-1000 μg/L, NAA/KIN 1/1-1000 μg/L promoted growth. 1AA/ZA 0.1-1/0.1-1μg/L; 1AA/BA 0.1-1/0.1-10 μg/L IAA/KIN 1/0.1-1000μg/L also promoted growth.

Formation of primary pit connection during conchocelis phase of Porphyra yezoensis (Bangiophyceae, Rhodophyta)

The formation of pit connection during conchocelis phase of Porphyra yezoensis Ueda was observed and examined with transmission electron microscope (TEM) and epifluorence microscope. It is indicated that the pit connection was formed in late stage of conchocelis phase and the early stages of conchosporangial cell development, and disappeared in bispore stage. The pit connection contained a thin membrane layer at outer pit plug. Stained with 4’, 6’-diamidino-2-phenylidole dihydrochloride hydrate (DAPI), transferring of DNA or RNA between adjacent cells were observed in late stage of conchocelis development, it was deduced that pit connection might serve as a channel for signal transduction and ge- netic substance transportation in conchocelis phase.

DEVELOPMENT OF SUSPENDED CONCHOCELIS OF PORPHYRA HAITANENSIS (BANGIALES, RHODOPHYTA)

Ths Mar. 1993 to Aug. 1994 study on suspended conchocelis of Porphyra haitanensisshowed tha there were three pattems for developmnt of vegetative filaments: filaments to filaments by"budding"; filaments to sporangial branchlets by "budding", or cell swelling. There were also three pat-terns for sporangial branchlet development: vegetatively propagting, changing into conbhospores, or dy-ing. Each developmntal stage had one or mere different developmntal directions between vegetative fila-ments and Sporangial branchlets. Developmnts from conchosporangial branchlets to conchospores were se-quential and irreversible. Although sporangial brachlets formed at 29℃ could give rise to filaments, theycould not propagae as healthily under the same condihons as those formed at 25℃ did. Probaby the cru-cial peried of plant cell differenhation is in the late stage of spompal branchlets. In line with the devel-opmental directions of different stages, the authors reghated the developmnt of conchocelis to get idealdifferent developmental smps materials to obtain very develoPmentally homogeneous stages, including fil-aments and sporanglal branchiets.

STUDIES ON INTERCROPPING OF PORPHYRA HAITANENSIS T.J.CHANG ET B.F.ZHENG WITH BAY SCALLOP ARGOPECTEN IRRADIANS LAMARCK

Results of the experiment of northward transplantation of Porphyra haitanensis showed thatintercrop of P. haitanensis with Bay Scallop is feasible because the growth duration of both of them is about the same in August - December.Full use of cultivating area, lowering its cost and increasing its product per unit area as well as improving the ecological envronment are realized during intercrop.