The influence of Fe~(2+) on growth and development of cells enzy-matically isolated from Porphyra yezoensis blades

1 INTRODUCTION Porphyra (also called nori in Japanese) is one of the most important algae in aquaculture in the world. In the early 1980’s, basic studies on cell and protoplast isolation, cultivation and “seedling” breeding of Porphyra were conducted in laboratory. Zhao and Zhang (1981) isolated and cultured isolated cells by grinding the nori thallus, and Lu (1983) isolated cells by digesting bacteria. An enzyme isolated from a sea snail, was used to isolate cells and protoplasts (T…     

Effect of cytokinins and auxins on the growth of free-living conchocelis ofPorphyra yezoensis

IAA 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 ofPorphyra yezoensis which showed that its growth in medium with cytokinins, IAA and NAA was more rapid than that in medium with non—phytohormones; that the optimal concentrations for promoting growth were 10 μg/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, IAA and cytokinins, NAA/ZA 1–1000/1 μg/L, NAA/BA 10/1-1000 μg/L, NAA/KIN 1/1-1000 μg/L promoted growth. IAA/ZA 0.1-1/0.1-1 μg/L; IAA/BA 0.1-1/0.1-10 μg/L IAA/KIN 1/0.1-1000 μg/L also promoted growth. Contribution No. 2335 from the Institute of Oceanology, Chinese Academy of Sciences. Contribution No. 94 from the Experimental Marine Biology Lab Qingdao, China. The research was supported by a grant from the Chinese Academy of Sciences.     

Gamma-rays induction of mutation in conchocelis ofPorphyra yezoensis

Free-living conchocelis ofPorphyra yezoensis Ueda (Bangiales, Rhodophyta) were treated with⁶⁰Co-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.     

Cloning and Analysis of Calmodulin Gene from Porphyra yezoensis Ueda （Bangiales, Rhodophyta）

In order to understand the mechanisms of signal transduction and anti-desiccation mechanisms of Porphyra yezoensis, cDNA and its genomic sequence of Calmodulin gene (CaM) was cloned by the technique of polymerase chain reaction (PCR) based on the analysis of P. yezoensis ESTs from dbEST database. The result shows that the full-length cDNA of CaM consists of 603 bps including an ORF encoding for 151 amino acids and a terminate codon UGA, while the length of genomic sequence is 1231 bps including 2 exons and 1 intron. The average GC content of the coding region is 58.77%, while the GC content of the third position of this gene is as high as 82.23%. Four Ca²⁺ binding sites (EF-hand) are found in this gene. The predicted molecular mass of the deduced peptide is 16688.72 Da and the pI is 4.222. By aligning with known CaM genes, the similarity of CaM gene sequence with homologous genes in Chlamydomonas incerta and Chlamydomonas reinhardtii is 72.7% and 72.2% respectively, and the similarity of the deduced amino acid sequence of CaM gene with homologous genes in C. incerta and C. reinhardtii are both 71.5%. This is the first report on CaM from a species of Rhodophyta.     

Toxic effects of Zn2+, Cu2+, Cd2+ and NH3 on Chinese prawn

This study deals with the toxic effects of Zn²⁺, Cu²⁺, Cd²⁺ and NH₃ on Chinese prawn (Penaeus orientalis). The median lethal concentrations of the toxic matters on Chinese prawn were determined. Cd²⁺>Cu²⁺>Zn²⁺>NH₃-Nm is the toxic order of these chemicals to Chinese prawn. The toxic order of concentrated Zn²⁺ and Cd²⁺ in tissues and organs of Chinese prawn is in the order. gill>shell+appendage>viscera>muscle.     

Comparative studies on structural feature of agar polysaccharides fromPorphyra haitanensis grown in south and north China

The structural feature of agar polysaccharides fromPorphyra haitanensis grown in south China and transplanted to the north was investigated by fractionation on DEAE—Sephadex A 50, chemical analysis, and infrared and¹³C-NMR spectroscopy. The agars composed mainly of charged molecules were eluted from DEAE—Sephadex A 50 with 1.0 mol/L NaCl solution from the southernP. haitanensis and with 0.5 mol/L NaCl from the northern one. The¹³C-NMR spectra showed that agarobiose [(1→3)-β-D-galactopyranosyl-(1→4)-3,6-anhydro-α-L-galactopyranose] and the biological precursor of agarobiose [(1→3)-β-D-galactopyra nosyl-(1→4)-6-sulfate-α-L-galactopyranose] were the major disaccharide repeating units in the charged fractions. The content of the biological precursor in the agar polysaccharides from southernP. haitanensis was higher than that in the northern one, the content of the biological precursor extracted from cold water was higher than that from hot water, and the content of 6-OMe-D-galactose in the southernP. haitanensis polysaccharides was higher than in the northern one. This distinct difference will be of significance for further study of the physiological characters ofP. haitanensis. Contribution No. 1849 from the Institute of Oceanology, Academia Sinica.     

Bioaccumulation of Pb<Superscript>2+</Superscript> and its effects on growth,morphology and pigment contents of <Emphasis Type="Italic">Spirulina</Emphasis> (<Emphasis Type="Italic">Arthrospira</Emphasis>) <Emphasis Type="Italic">platensis</Emphasis>

A laboratory experiment was conducted to assess the bioaccumulation of Pb²⁺ and its effects on growth, morphology and pigment contents of Spirulina (Arthrospira) platensis. The specimen cultured in Zarrouk liquid medium was treated with various initial metal concentrations (0, 5, 10, 30, 50 and 100 μg mL⁻¹). The growth of S. platensis was adversely affected by Pb²⁺ at high concentrations (30, 50 and 100 μg mL⁻¹). However, at low concentrations (5 μg mL⁻¹), Pb²⁺ could stimulate its growth slightly. The pigment contents (chlorophyll α and β carotene) were decreased in a dose-dependent manner. The highest reductions (67% and 53% respectively in chlorophyll α and β carotene) were observed in 100 μg mL⁻¹ treatment group. The LC₅₀ (96 h) of Pb²⁺ was measured as 75.34 μg mL⁻¹. Apart from a few cases of filament breakages at elevated concentrations (50 and 100 μg mL⁻¹), morphological abnormalities are not specific. Metal bioaccumulation increased with Pb²⁺ concentrations, but decreased with exposure time. The maximum accumulated amount was 188 mg g⁻¹ dry weight. The bioconcentration factor (BCF) reached to a peak at day 2, followed by a gradual reduction for all the exposure concentrations. S. platensis is able to tolerate considerably high Pb²⁺ concentrations. Consequently it can be used as a potential species to remove heavy metal from contaminated waters.     

Effects of Na+/K+ ratio of groundwaters on the gill ion-transport enzyme activity, plasma osmolality and growth of Cynoglossus semilaevis juveniles

The effects of environmental Na⁺/K⁺ ratio on the gill ion-transport enzyme activity, plasma osmolality and growth of Cynoglossus semilaevis juveniles were investigated. The results showed that, plasma osmolality was similar among flounder adapted to different Na⁺/K⁺ ratios of saline groundwaters (P>0.05), while the growth, gill Na⁺, K⁺-ATPase and HCO₃ ⁻-ATPase activities were affected by Na⁺/K⁺ ratio significantly (P<0.05). The gill Na⁺, K⁺-ATPase activity reached its maximum on day 3, then decreased gradually from day 3 to day 9 and remained constant from day 9 to day 15. The peak values of gill Na⁺, K⁺-ATPase activity were detected on day 3 for all Na⁺/K⁺ ratios of saline groundwaters, then the enzyme activities descended, and on day 9 the enzyme activities achieved steady state, and the gill HCO₃ ⁻-ATPase activity increased rapidly and achieved steady state after one day. During steady state, the gill Na⁺, K⁺-ATPase and HCO₃ ⁻-ATPase activity of Na⁺/K⁺ ratios 20 and 40 treatments were significantly higher than those in the control group (Na⁺/K⁺ ratio 27.5), while there were no significant differences between the Na⁺/K⁺ ratio 30 treatment and the control group; the gill Na⁺, K⁺-ATPase activity of Na⁺/K⁺ ratio 20 and 40 treatments were significantly higher than that for ratio 30 treatment, but there were no significant differences of gill HCO₃ ⁻-ATPase activity among these treatments. At the end of the 15-day experiment, the weight gain (%) and specific growth rate (SGR) of flounders maintained in seawater were significantly higher than those in groundwaters; significant differences also occurred among the treatments; Na⁺/K⁺ ratio 30 treatment had the highest values (33.7% and 1.94 respectively), which were significantly higher than those under Na⁺/K⁺ ratios 20 and 40 treatments. Therefore, for the saline groundwater used in this experiment, it is suggested that the Na⁺/K⁺ ratio be adjusted to approximately 30, i.e., as close to that of natural seawater as possible in the culture of flounder.     

The ultrastructure of separated and cultured cell of Porphyra yezoensis

There are many reports that cells (protoplasts) separated from the thallus ofPorphyra by enzyme can develop to normal leafy thalli in the same way as monospores. But there are few investigations on the subcellular structure of the isolated vegetative cell for comparison with the subcellular structure of monospores. To clarify whether the separated and cultured cells undergo the same or similar ultrastructure changes during culture and germination as monospores undergo in their formation and germination, we observed their ultrastructure, compared them with those of the monospore and found that the ultrastructure of separated and cultured cells did not have the characteristic feature as that of monospore formation, such as production of small and large fibrous vesicles, but was accompanied by vacuolation and starch mobilization like that in monospore germination. The paper also discusses the relations between monospores and separated and cultured cells. Contribution No. 3875 from the Institute of Oceanology, Chinese Academy of Sciences. Project 39770593 supported by NSFC; Project 96-C01-05-01 of the 9th Five-Year Plan supported by Science and Technology Commision of China.     

EFFECTS OF TEMPERATURE AND Ca^2＋ ON THE LARVAL DEVELOPMENT OF THE DECAPODA CRUSTACEAN ： ERIOCHEIR SINENSIS

With increasing demand in China for the mitten crab larvae, understanding its survival mechanism gets more important. This research focused on the effects of temperature and Ca^2 on the larval growth and development. Eriocheir sinensis larvae were reared in laboratory under 21 different combinations of temperature (15, 20, 25℃) and Ca^2 content (120, 130, 140, 150, 160, 170, 180mg/l) and constant salinity (20) and pH (8), The results suggested that the survival rate increases with temperature and Ca^2 content. These combinations of temperature and Ca2 content maximized survival rate inour study and it may be the optimum water environmental conditions for culturing the larvae. To predict surviving larvae number under different water environmental conditions, 21 dynamic mathematical models were developed. This for the first time observation of the zoeal Ⅵ larvae of the Changjiang River E.sinensis population showed that they occurred under stressed water environmental conditions: temperature of 15℃ and Ca^2 content of 120,130mg/l.