Study of light signal receptor of Stephanopyxis palmeriana during sexual reproduction

We collected centric diatom Stephanopyxis palmeriana samples in coastal waters of Xiamen for characteristic red light/far red light (R/FR) phytochrome reactions to identify its photoreceptor in the course of sexual reproduction. The result showed that pre-illumination of 2-3h red light before darkness could induce sexualization of S.palmeriana, while the follow-up illumination of far red light could reverse the effect of red light, which is a featured reaction of phytochrome. The Southern Dot Blot was carried out to identify the type of phytochrome that induces the sexualization. The result also showed high homogeneity of DNA fragment of S. palmeriana with phyB, but phyA. This means the photoreceptor in the process of sexual reproduction of S. palmeriana is phytochrome B (phyB).     

Effects of blue light on gametophyte development of Laminaria japonica (Laminariales, Phaeophyta)

Laminaria gametophyte was greatly influenced by light in its growth and development. Using light-emitting diodes (LED) as blue and red light sources, we analyzed the light effect on gametophytes development ofLaminaria japonica Aresch. The gametophytes were obtained from zoospores collected in April, May, July, 2003 and September, 2004. We found that the growth of gametophytes was stimulated by increasing intensity of blue light (BL) and red light (RL) illumination, of which BL was obviously stronger than that of RL. The fertilization of gametophytes depended largely on BL, and only sufficient BL illumination could take the reproductive effect. In addition, we noticed that there was a significant difference in light responses for gametophytes developed from zoospore collected in different times. For zoospores released in April, under BL₁ (73.90 μmol photons/m·s), the unicellular female gametophytes and multi-cellular male gametophytes produced eggs and sperms respectively, and further developed towards sporophytes. However, for gametophytes developed in May, July or September, they became multi-cellular and never formed oogonia or antheridia. It is believed that theLaminaria sporangium maturation stage could affect the gametophytes reaction to BL under laboratory culture conditions. Therefore, cryptochrome- or phototropin-like BL photoreceptors is probably involved in BL-induced development ofLaminaria gametophytes. This research was supported by the NSFC (No.40376049) and Japan Society for the Promotion of Science (The Invitation Fellowship for Research in Japan 2002).     

Effect of germanium dioxide on growth of Spirulina platensis

This study on the effect of different concentrations of germanium dioxide (GeO₂) on the specific growth rate (SGR), pigment contents, protein content and amino acid composition ofSpirulina platensis showed that Ge was not the essential element of this alga; that GeO₂ could speed up growth and raise protein content ofS. platensis, and could possibly influence the photosynthesis system. The concentration range of GeO₂ beneficial to growth ofS. platensis is from 5–100mg/l. GeO₂ is proposed to be utilized to remove contamination byChlorella spp. usually occurring in the cultivation ofSpirulina.     

Unpalatable weed Stellera chamaejasme L. provides biotic refuge for neighboring species and conserves plant diversity in overgrazing alpine meadows on the Tibetan Plateau in China

Steller chamaejasme L. (S. chamaejasme for short) is one of the most noxious unpalatable weeds in China, which has been frequently reported its negative interaction (i.e. competition and allelopathy) with other herbaceous species in grasslands. This study compared species diversity, biomass and sexual reproduction of herbaceous plants in meadows with S. chamaejasme and in open meadows without S. chamaejasme in overgrazing meadows on the Tibetan Plateau in China to determine whether positive facilitation exist between S. chamaejasme and other herbaceous species under livestock’s overgrazing. The results showed that there are more herbaceous species in meadows with S. chamaejasme than those in open meadows (35s and 30s, respectively). Diversity index and above-ground biomass were also significantly higher in meadows with S. chamaejasme. There were 39% (11/28) of all species with sexual reproduction found in meadows with S. chamaejasme, which was 7 times more than those in open meadows. Our study showed that S. chamaejasme could provide biotic refuge for neighboring plants and preserve plant diversity from livestock’s overgrazing in alpine meadows on the Tibetan Plateau. It also suggested that inter-specific facilitation between S. chamaejasme and other herbaceous species may play a key role in overgrazing alpine meadows.     

Promotive effect of se on the growth and antioxidation of a blue-green algaSpirulina maxima

Cultures of a blue-green algaSpirulina maxima (Setch. et Gard.) Geitler with various concentrations of Se in Zarrouk's medium showed that not higher than 40 mg/L Se could promote its growth. The present experiments showed thatS. maxima grown under normal conditions, has an oxidant stress defence system for hydrogen peroxide (H₂O₂) removal, which is the Halliwell-Asada pathway. When 4 to 20 mg/L Se was added to the algal medium, this pathway was replaced by a so-called Sestressed pathway containing GSH peroxidase (GSH-POD). As a result of the occurrence of both higher activity of GSH-POD and lower levels of hydroxyl radical (OH·), the Se-stressed pathway scavenged H₂O₂ so effectively that the growth ofS. maxima was promoted by 4 to 20 mg/L Se. While GSH-POD activity of the alga disappeared at 40 mg/L Se, the recovery of ascorbate peroxidase was observed. The lower levels of ascorbic acid and GSH made the Halliwell-Asada pathway for scavenging H₂O₂ less effective, while the highest activity of catalase might be responsible in part for the H₂O₂ removal, causing the level of OH· inS. maxima grown at 40 mg/L Se to be much higher than the OH· level in this alga grown at 4 to 20 mg/L Se, but lower than that in the control. The OH· level changes caused the growth ofS. maxima cultured at 40 mg/L Se to increase slightly to close to that of the control.     

Excitation energy transfer in vitro between phycobiliproteins and thylakoid photosystem II of higher plants

The excitation energy transfer from phycobiliproteins to thylakoid PSII of higher plants was investigated. When incubated with spinach thylakoids, phycobiliproteins isolated from red and blue-green algae transferred light energy absorbed to spinach PSII. The efficiency of energy transfer was dependent on the kind of phycobiliproteins used. If spinach thylakoids were replaced by the thylakoids ofBrassica chinensis, R-phycoerythin or C-phycocyanin did not transfer their excitation energy to PSII ofBrassica chinensis unless allophycocyanin was present. Contribution No. 69 from the Experimental Marine Biology Laboratory IOAS.     

The effects of nitrogen,phosphorus, vitamins and trace metals on the growth of the red tide organismProrocentrum Micans

Laboratory culture experiments showed that <100μ mol/L nitrate, amonium or mixture of amino acids promote the growth of the red tide organismProrocentrum micans Ehrenb, but that >100μmol/L of ammonium, or mixture of glycine and glutamate was harmful to growth, and that orthophosphate wasP. micans’ main phosphorous source in the ocean. Presence of 80μ mol/L EDTA, 0.5 to 1 μmol/L Fe³⁺, 1.0 to 20.0 μ mol/L Mn²⁺ 0.1 to 0.4 μmol/L Co²⁺ in the culture medium could improve the growth ofP. micans. Vitamin B₁ promoted growth, but vitamin B₁₂ and biotin did not. The estimated minimum cell quotas (q _o) for nitrogen and phosphorus being 0.74 pmole/cell and 0.045 pmole/cell show that phosphorus (more than nitrogen) limits the growth ofP. micans in the study area. This project was supported by the Natural Science Foundation of Zhejiang Province.     

Serum immunoglobulin of the mandarin fish, Siniperca chuatsi with development of polyclonal antibody

Serum immunoglobulin from the mandarin fish, or the so-called Chinese perch,Siniperca chuatsi (Basilewsky), was successfully purified using affinity chromatography. Heavy and light chains were detected on electrophoresis gel, with molecular weights being estimated at 72 and 29 kDa, respectively. The tetrameric IgM ofS. chuatsi was calculated to be 808 kDa. The rabbit polyclonal antisera against the purifed immunoglobulin were developed and tested by Western blot analysis. The antisera reacted strongly with the heavy chains ofS. chuatsi immunoglobulin. Humoral immune responses of the mandarin fish can then be examined using the developed polyclonal antibody. Project No. 30025035 supported by NSFC, State Key Laboratory of Freshwater Ecology and Biotechnology (2000FB02), and the Chinese Academy of Sciences (KZ9511-B1-111).     

Main nutritional contents of 30 Dalian coastal microalgae species

This paper reports results of study on the contents of proteins, amino acids, polysaccharose and uronic acids in 30 species of macroalgae from Shicao, Heishijiao, Shimiao, and Xiaofujiazhuang in the vicinity of Dalian City, N.E.China. The results showed that the protein contents of the 30 algae from highest (112.55μg/ml) to the lowest (0.24 p.g/ml) was in the descending order of Dictyopteris ndalata, Gelidium vagum,Gymnogongrus japonican, Ectocarpus confervoides, Tinocladia crassa, Sargassum thunberii. In general, the protein content in red algae was higher than that in brown algae. The content of free ami no acids showed nosignificent differences from 7.44μg/ml-4.96μg/ml in all these algae, in the descending order of Gymnogongrusrus japonican, Sargassum confusum, Undoria pinnatifida, Laminaria japonica and Ectocarpus confervoides. The content of polysaccharose varied from 168.2 gg/ml-22.15 p.g/ml in the descending order of Symphyocladiaia tiuscula, Scytosiphon lomentarius, Desmarestia viridis., Tinocladia crassa, Gracilaria asiatica and Porphyra yezoensis. The content of uronic acids is from 196.24μg/ml-20.77μg/ml in the descending order of Ulva lactuca,Symphyoclaldia latiuscula, Scytosiphon lomentarius, Ceramimum kondoi, Gracilaria vemucosa and Porphyra yezoensis. The fatty acids in 30 species of algae belong to Rhodophyta, Chlorophyta and Phaeophyta. Most phaeophytes have many (4-12) types of fatty acids.     

Studies on color type variants from mutagenized protoplasts ofPorphyra haitanensis Chang et Zheng &P. Yezoensis ueda (rhodophycease)

Isolated protoplasts from thalli ofPorphyra haitanensis andPorphyra yezoensis were treated with colchicine or irradiated by ultraviolet (UV). Several types of color variants were observed among the protoplast offspring. After treatment with colchicine: (1) 0.04–0.09% of red type variants inP. haitanensis were obtained; (2) The rate of red type variants and the variegated chimeral thalli composed of red type and wild type of sectors were 0.31–1.11% inP. yezoensis. After irradiation with UV: (1) 3.5–10.5% of red type variants inP. yezoensis were obtained: (2) 0.5–2.0% of red type variants and the variegated chimeral thalli composed of red type and wild type of sectors were obtained inP. haitanensis. Colchicine and UV’s mutangenic effects onP. yezoensis protoplasts were stronger than those onP. haitanensis protoplasts. The most efficient concentration of colchicine was 0.05%. The optimal length of UV-radiation was 1/2 min (radiation distance 5 cm). The red type variants induced by colchicine treatment grew faster than the wild type thalli. The clones of vegetative propagation from protoplasts of red type variants were still red type thalli. The red type variants will be good materials for genetic studies and improvement ofPorphyra strains.     

Production of DMS and DMSP in different physiological stages and salinity conditions in two marine algae

Dimethylsulfide(DMS) and dimethylsulfoniopropionate(DMSP) production by Scrippsiella trochoidea and Prorocentrum minimum was investigated to characterize the effects of physiological stage and salinity on DMS and DMSP pools of these two marine phytoplankton species.Axenic laboratory cultures of the two marine algae were tested for DMSP production and its conversion into DMS.The results demonstrated that both algal species could produce DMS,but the average concentration of DMS per cell in S.trochoidea(12.63 fmol/L) was about six times that in P.minimum(2.01 fmol/L).DMS and DMSP concentrations in algal cultures varied significantly at different growth stages,with high release during the late stationary growth phase and the senescent phase.DMS production induced by three salinities(22,28,34) showed that the DMS concentrations per cell in the two algal cultures increased with increasing salinity,which might result from intra-cellular DMSP up-regulation with the change of osmotic stress.Our study specifies the distinctive contributions of different physiological stages of marine phytoplankton on DMSP and DMS production,and clarifies the influence of salinity conditions on the release of DMS and DMSP.As S.trochoidea and P.minimum are harmful algal bloom species with high DMS production,they might play an additional significant role in the sulfur cycle when a red tide occurs.     

Isolation,purification and characterization of the hydrogen evolution promoting factor of hydrogenase ofSpirulina platensis

A component (s-factor) with obvious promoting effect on hydrogen evolution of hydrogenase has been isolated and extracted from a cell-free preparation ofSpirulina platensis. The effect of the s-factor in the reaction system is similar to that of Na₂S₂O₄, but is coupled with light. The s-factor has the maximum absorption peak at 620 nm in the oxidized state, at 590 nm in the reduced state. The partially purified s-factor showed two bands by SDS-PAGE and is distinctly different from phycocyanin, which has no change of oxidized state and reduced state absorption spectra, and also has no promoting effect on hydrogenase ofSpirulina platensis under the light. The research supported by the NSFC.     

Growth,reproduction and population growth ofCeriodaphnia cornuta sars and comparison of 7-day fecundity withCeriodaphnia dubia richard

Biology of parthenogenetic females ofCeriodaphnia cornuta Sars was studied under laboratory conditions. The mean length of neonates (first-young instars) ofCeriodaphnia cornuta was found to be 0.27 mm at 25±1°C and that of the first adult instars 0.50 mm. A maximum length of 0.69 mm was attained at the 8th adult instar. The mean longevity was found to be 19.0 days for 3 preadult and 8 adult instars. Sex maturity was reached in 4.26 days. The mean number of young per brood was 6.21. The total production for each adult reached 49.7. The maximum reproduction period occurred during the 4th adult instar. At this age, the animals were 10.25 days old, and had a body length of 0.65 mm. The innate capacity for increase (r _m) was 0.40 per female per day, the net reproduction rate (R ₀) was 33.601 per generation, the mean length of a generation (T) was 8.786 days, and the finite rate for increase (λ) was 1.492 per female per day. Comparisons using the t-test indicated that the number of eggs produced and number of broods ofC. cornuta andC. dubia were not significantly different (p>0.05) after 7 days.     

Simultaneous estimates ofsynechococcus spp. Growth and grazing mortality rates in the English Channel

The marine chroococooid phycoerythrin-containingSynechococcus spp. cyanobacterium has been implicated as a substantial component of the photosynthetic picoplankton in the ocean. Although its importance as food source for heterotrophic nanoplankton is now recognized, information about the cycling ofSynechococcus biomass and its diel pattern is limited and study methodology varies among authors. The selective metabolic inhibitor method was used to simultaneously estimate growth and grazing disappearance rates ofSynechococcus in the English Channel where growth rates ranged from 0.25 to 0.72/d (mean ±SD=0.51±0.17/d) and grazing mortality rates ranged from 0.19 to 0.64/d (mean ±SD=0.48±0.17/d). Size-fractionated experiments demonstrated that up to 70% ofSynechococcus disappearance could be attributed to grazers going through a 2 μm Nuclepore filter.Synechococcus grazing mortality rates (mean=0.74 ±0.25/d) during the day were always higher than that (mean=0.2±0.20/d) during the night, while growth rates showed no clear diel pattern. A positive correlation was observed between growth rates andin situ temperature ranging from 9 to 17°C, while in contrast grazing was independent of temperature. The close similatiry between average growth and grazing rates suggests a rapid recycling ofSynechococcus biomass in English Channel coastal waters.     

PHYLOGENETIC RELATIONSHIPS AMONG PUFFERFISH OF GENUS TAKIFUGU BY RAPD ANALYSIS

AmpLifications with 300 10-base primers under predetermined optimal reaction yielded 2762 reproducible amplified fragments ranging from 200 to 3000 bp. Genetic distances among five species of Takifugu and Lagocephalus spadiceus as outgreup were calculated based on the presence/absence of the amplified fragments. Topological phylogenie trees of the 5 species of Takgfugu and the outgroup were gen-erated by Parsimony and Neighbor-Joining analysis based on RAPD data set. The genetic distance between T. rubripes and T. pseudommus was almost the same as that between individuals within each species, but was much smaller than that among T. rubripes, T. pseudommus and the other species. The molecu-lar data from RAPD analysis convincingly showed that T. rubripes and T. pseudommus should be the same species.     

Growth,reproduction & population structure of the freshwater crabSinopotamon yangtsekiense bott, 1967, from Zhejiang,China

Monthly investigations were mae on the population of Chinese freshwater crab,Sinopotamon yangtsekiense Bott, 1967 from April, 1984 to March, 1985. The data on 4413 specimens show that the growth was affected mainly by temperature. During the April to November growth period, the crabs' major development occurred from June through October. One year was required for a fine white oocyte to develop into a mature egg. The reproduction period was June–October. Females bearing eggs were taken from June–August, and crabs with young were found from July–October. The females reproduced once a year but could for more than one year. The number of eggs carried by a female varied greatly according to the size of the crab, ranging from 30 to 100 eggs. New-born crabs become mature after 1–2 years. The sex ratio was approximately 1∶1 in the overall population. However, the larger crabs are predominantly male. The age distribution ofS. yangtsekinese was estimated from size frequency histograms. There were more adult crabs (over 70%) from June to October and more immature crabs (over 50%) from November to May.     

Study on sori formation inLaminaria japonica aresch

Sori formation inLaminaria japonica was studied from the ecological and physiological point of view. Cutting through the medulla ofLaminaria japonica improved the formation of sori. Sori formation under different water temperature, photoperiod, and light intensity was investigated and total nitrogen and reducible sugar content determined. The result showed that sori ofL. japonica can be formed in 1–22°C water in Dalian. No sori were found in water over 23°C. High temperature inhibits the formation of sori, but low temperature has no effect. Apparently, light intensity affects the formation of sori, but photoperiod does not. In shallow waterL. japonica grows slower but stores dry material faster and forms sori rapidly. In deeper water it grows faster in length but stores dry material slower and the sori formation is slow. Analysis of dry material shows that during sori formation the water content decreases but total nitrogen and total reducible sugar increase. Only when the storage of sugar reaches a certain level can sori begin to form. Cutting through the medulla interrupts the passage of nutrients to the growth area. Storage of carbohydrates and nitrogen is the main factor affecting sori formation.     

Cell cycles and proliferation patterns in Haematococcus pluvialis

Most studies on Haematococcus pluvialis have been focused on cell growth and astaxanthin accumulation;far less attention has been paid to cell cycles and proliferation patterns.The purpose of this study was to clarify cell cycles and proliferation patterns in H.pluvialis microscopically using a camera and video recorder system.The complicated life history of H.pluvial i s can be divided into two stages:the motile stage and the non-motile stage.All the cells can be classifi ed into forms as follows:motile cell,nonmotile cell,zoospore and aplanospore.The main cell proliferation,both in the motile phase and non-motile phase in H.pluvialis,is by asexual reproduction.Under normal growth conditions,a motile cell usually produces two,sometimes four,and exceptionally eight zoospores.Under unfavorable conditions,the motile cell loses its fl agella and transforms into a non-motile cell,and the non-motile cell usually produces 2,4 or 8 aplanospores,and occasionally 20–32 aplanospores,which further develop into non-motile cells.Under suitable conditions,the non-motile cell is also able to release zoospores.The larger non-motile cells produce more than 16 zoospores,and the smaller ones produce 4 or 8 zoospores.Vegetative reproduction is by direct cell division in the motile phase and by occasional cell budding in the non-motile phase.There is,as yet,no convincing direct evidence for sexual reproduction.     

Life strategy traits of the liana Sericostachys scandens spreading in the montane forests in the Kahuzi-Biega national park (DR Congo)

Sericostachys scandens is a monocarpic and heliophilous liana, native in tropical African forests. In the montane forests of the Kahuzi-Biega National Park (KBNP) (East of DR Congo), it has been expanding very strongly for a decade, and is currently considered as having negative impacts on biodiversity conservation. In this paper, we test if S. scandens differs from three co-occurring, native, non spreading lianas (Gouania longispicata, Tacazzea apiculata and Adenia bequaertii) for functional traits which might influence plant expansion. For leaf traits (SLA, dry matter content, nitrogen concentration), S. scandens did not show extreme values compared to those of the three other lianas. In contrast, S. scandens had much higher biomass allocation to sexual reproduction. It also differs from the three other lianas for its reproductive strategy that combines both vegetative propagation and sexual reproduction, and propagule dispersal by wind. Moreover, S. scandens has larger leaves and a greater number of lateral branches per unit stem length. It is argued that the particular combination of functional traits exhibited by S.scandens may in part explain its propensity to behave as an opportunistic weed in the disturbed areas in the montane forests of Kahuzi-Biega.     

Comparative studies of inorganic carbon utilization inEmiliania huxleyi and some non-calcifying marine microalgae

Inorganic carbon utilization in the non-calcifying marine microalgae,Nannochloropsis oculata, Phaeodactylum tricornutum andPorphyridium purpureum was compared with high- and low-calcifying strains ofEmiliania huxleyi grown in artificial seawater medium aerated with either air (0.03% V/V CO₂) or CO₂-free air. For high-calcifying strain ofE. oculata andP. tricornutem, similar growth patterns were observed in air-and CO₂-free air-grown cultures.P. purpureum showed a less final cell density in CO₂-free air than in air-grown culture. However, low-calcifying strain ofE. huxleyi was able to grow only in air-grown culture, but not in CO₂-free air-grown culture. Measurements of alkalinity, pH, concentration of dissolved inorganic carbon (DIC) and free CO₂ showed different patterns of DIC utilization. WithN. oculata, P. tricornutum andP. purpureum the pattern of DIC utilization was characterized by an increase of pH and a decrease of DIC but a constant alkalinity in the cultures aerated with air or CO₂-free air, suggesting that bicarbonate utilization was concomitant with an efflux of OH⁻. Both alkalinity and pH were maintained rather constant in air-grown culture of low-calcifying strain ofE. huxleyi, suggesting that diffusive entry of CO₂ could meet the requirement of DIC for its photosynthesis and growth. High-calcifying strain ofE. huxleyi, however, showed a pattern of decrease of alkalinity and DIC but an almost constant pH, indicating that bicarbonate was the major form of inorganic carbon utilised by this organism and bicarbonate uptake is unlikely to be accompanied by an efflux of OH⁻. The final pH values reached byN. oculata, P. tricornutum andP. purpureum in a closed system were 10.75, 10.60 and 9.85 respectively, showing that bicarbonate utilisation is concomitant with an efflux of OH⁻. While the final pH of 8.4 in high-calcifyingE. huxleyi suggests that bicarbonate utilization was not accompanied by an efflux of OH⁻. Contribution No. 3557 from the Institue of Oceanology, Chinese Academy of Sciences. This work was supported by NERC grant GRE3/7853 U. K. and partly supported by Bio-Engineering Center, SSTC 96-C01-05-01.