Early development of Costaria costata (C. Agardh) Saunders and cultivation trials

Costaria costata (C. Agardh) Saunders is one of common kelps distributed in many coastal areas worldwide; however, in China, no reports have been made on cultivation of the genus. To investigate potential cultivation of the species in the northern part of China, trials on isolation and preservation of the gametophytes were conducted using C. costata from Korea; growth and development of the gametophytes were observed. We showed that at 10±1°C, 60 μmol m⁻²s⁻¹ and 12:12 h (L:D), freshly released zoospores settled down within 1 hour, and then developed into the primary cell during the following 2 days. After a vegetative growth phase lasting 6–8 days, female gametophytes became 3–4 times larger in diameter than that of the primary cell, but still remained at a unicellular stage, while male gametophytes divided into 4–10 cells with only a slight change in size. Fertilization occurred within 10 days after the zoospores were released from the sporangia, and the apical and basal tissues of the juvenile sporophyte divided and differentiated into the blade and stipe. Temperature and irradiance influenced gametophytic vegetative growth and developmental patterns. Generally, low irradiance (15 μmol m⁻²s⁻¹ and 30 μmol m⁻²s⁻¹) was unfavorable to the induction of fertility, but it enhanced female gametophyte division. The optimal conditions for vegetative growth were 15°C and 30 μmol m⁻²s⁻¹. After transplantation of the juvenile seedlings and after eight months cultivation, the harvested mature blade reached 194 cm in length and 32.7 cm in width. Our study proves that it is feasible to implement propagation and large scale cultivation of C. costata in northern China.     

Cryopreservation of Gametophytes of Laminaria japonica （Phaeophyta） Using Encapsulation-Dehydration with Two-Step Cooling Method

Gametophytes of Laminaria japonica were cryopreserved in liquid nitrogen using encapsulation-dehydration with two-step cooling method. Gametophytes cultured at 10℃ and under continuous irradiance of 30 μmol m-2 s-1 for 3 weeks were encapsulated in calcium alginate beads. The beads were dehydrated in 0.4 molL-1 sucrose prepared with seawater for 6 h, desiccated in an incubator set at 10℃ and 70% relative humidity for 4 h, pre-frozen at either -40℃ or -60℃ for 30 min, and stored in liquid nitrogen for ＞24 h. As high as 43% of survival rate was observed when gametophytes were thawed by placing the beads in 40℃seawater and re-hydrated in 0.05 molL-1 citrate sodium prepared using 30‰ NaC1 7 d later. More cells of male gametophytes survived the whole procedure in comparison with female gametophytes. The cells of gametophytes surviving the preservation were able to grow asexually and produce morphologically normal sporophytes.     

Formation and early development of tetraspores of Polysiphonia urceolata （Rhodomelaceae, Rhodophyta）

Polysiphonia urceolata is one type of potential commercial red seaweeds used for breeding and cultivation, because of its significant biochemical and biomedical application. However, the information of breeding and seedling incubation for cultivation is limited, especially the early development. In this study, tetrasporohyte and gametophyte of P. urceolata were taken as the study materials in Huiquan Bay, Qingdao, China. The cleaned and sterilized tetrasporophytes and gametophytes were pre-cultured in sterilized seawater, then nurtured at 18°C, 25 μmol photons m⁻² s⁻¹ in 12:12 h (light:dark) photoperiod. Continuous observation under microscope showed that the early development consists of bipolar division stage and seedling stage. In the division stage, tetraspores germinate into bipolar sporelings that further differentiate into a colorless rhizoidal portion and a lightly pigmented upright shoot. The lightly pigmented rhizoidal cell develops to a rhizoid and the larger pigmented cell transforms to an erect axis. In the seedling stage, several quasi-protuberances appear on the erect axis and form juvenile seedlings. The results demonstrate the culture of P. urceolata from tetraspores under laboratory conditions. Supported by National Key Technology Support Program, Development Program of China (No.2006AA09Z21), National Natural Science Foundation of China (No. 40618001 and N_CUHK438/06) and Shandong Agricultural Seed Stock Breeding Project     

Optimization of conditions for tetraspore release and assessment of photosynthetic activities for different generation branches of Gracilaria lemaneiformis Bory

Gracilaria lemaneiformis Bory is an economically important alga that is primarily used for agar production. Although tetraspores are ideal seeds for the cultivation of G. lemaneiformis, the most popular culture method is currently based on vegetative fragments, which is labor-intensive and time-consuming. In this study, we optimized the conditions for tetraspore release and evaluated the photosynthetic activities of different colonies formed from the branches of G. lemaneiformis using a PAM (pulse-amplitude-modulated) measuring system. The results showed that variations in temperature and salinityhad significant effects on tetraspore yield. However, variations in the photon flux density (from 15 μmol m⁻² s⁻¹ to 480 μmol m⁻² s⁻¹) had no apparent effect on tetraspore yield. Moreover, the PAM-parameters Y(I), Y(II), ETR(I), ETR(II) and F _v/F _m of colonies formed from different branches showed the same trend: parameter values of first generation branches>second generation branches>third generation branches. These results suggest that the photosynthetic activities of different colonies of branches changed with the same trend. Furthermore, photosynthesis in G. lemaneiformis was found to be involved in vegetative reproduction and tetraspore formation. Finally, the first generation branches grew slowly, but accumulated organic compounds to form large numbers of tetraspores. Taken together, these results showed that the first generation branches are ideal materials for the release of tetraspores.     

Early development of Chondrus ocellatus Holm （Gigartinaceae, Rhodophyta）

1 INTRODUCTION Red algae C. ocellatus lives in temperate zone of coastal area, and inhabits at substrates from high tide to low tide regions. Its life history is composed of three phases: tetrasporophyte, gametophyte and carposporophyte. Chondrus is one o…     

Effect of temperature and irradiance on the growth and reproduction of Enteromorpha prolifera J. Ag. （Chlorophycophyta,Chlorophyceae）

Effect of temperature and irradiance on growth and reproduction of Enteromorpha prolifera that bloomed offshore along the Qingdao coast in summer 2008, was studied. It was showed that E. prolifera propagated mainly asexually with specific growth rate （SGR） of 10.47 at 25℃/40 μmol m^-2s^-1. Under this condition, gametes with two flagellate formed and released in 5 days. At the beginning of the development, the unicell gamete divided into two cells with heteropolarity, and then the apical cell developed into thalli primordial cells, whereas the basal cell developed into rhizoid primordial cells. In 8-day culture, the monoplast gamete developed into juvenile germling of 240 μm in length. Unreleased gametes can develop directly within the alga body. E. prolifera could either reproduce through lateral branching or fragmenting except apomixis revealed by Microscopic observation. On aged tissue of E. prolifera, although the degraded pigments partially remained in faded algal filaments, numerous vegetative cells could still divide actively in the algal tissues.     

Light demands of juvenileLaminaria japonica

The light demands of seaweeds is an interesting and rather complex phenomenon because they depend not only on the species but also on their different development stages. Even different parts of the same plant sometimes have different light demands. Light control is an important procedure at large scaleLaminaria nursery stations in China. Technicians and scientists have different viewpoints on the best method to regulate light. A culture study onLaminaria japonica starting from zoospores to several centimeter sporophytes to find the optimal and critical irradiance ranges for juvenileLaminaria at different development stages added more knowledge on this aspect. Experiment results show gametophytes can not tolerate irradiance of more than 150 μE m⁻²s⁻¹ while sporophytes can tolerate more than 519 μE m⁻²s⁻¹. This big difference starts from the very early stage of 1-to 2-celled sporophytes. The biological basis and mechanism of this phenomenon need further research. Contribution No. 1515 from the Institute of Oceanology, Academia Sinica Contribution No. 1 from the Experimental Marine Biological Laboratory     

Effects of salinity, light and temperature on growth rates of two species of Gracilaria （Rhodophyta）

Effects of temperature, salinity and light intensity on growth rates of Gracilaria lichenoides and G. tenuistipitata var. liui Zhang et Xia were tested. Eight to ten levels of each factor were first tested separately. The best growth rate was obtained under the conditions of 32°C, 30 and 240 μmol/(m²·s) for G. lichenoides, and 24°C, 20 and 200 μmol/(m²·s) for G. tenuistipitata, respectively. Then a uniform design was used to evaluate the optimal combinations of the three factors. The best conditions for the highest daily specific growth rates (% increase in wet weight) are determined to be 31.30°C, 32.10, and 287.23 μmol/(m²·s) for G. lichenoides (16.26%/d), and 25.38°C, 21.10, and 229.07 μmol/(m²·s) for G. tenuistipitata (14.83%/d), respectively. Supported by the 908 Special Program (908-02-04-07), the National Basic Research Program of China (973 Program, No. 2006CB400608), and K. C. Wong Magna Fund in Ningbo University     

Study on gametophyte vegetative growth ofUndaria pinnatifida and its applications

When cultured under certain environmental conditions (25°C, light intensity 80 μmol/m²·s, LD 12/12, in enriched seawater medium with 7×10⁻⁴ mol/L NO₃−N, 1.56×10⁻⁴ mol/L, PO₄−P and supplements of other elements like Mn, Fe, I, etc.), male and female gametophytes ofU. pinnatifida kept growing vegetatively and propagated fast at average daily fresh weight increase rate of about 20%. The empirical formulaG _m=G _o·3^m was established to estimate the output of vegetative gametophytes. Vigorous vegetative gametophyte cells began to form reproductive structures (oogonium and spermatangium, when the temperature was lower than 25°C and other environmental factors were kept optimal. The sufficient supply of gametophyte cells provided enough seeds for raisingUndaria sporelings on production scale. Controlled cross-breeding experiments using selected male and female gametophyte clones which increase their cell number by mitosis instead of meiosis were also carried out in vitro. Juvenile sporophytes from the cross-breeding had almost the same length and width increase rates as those of the control. The fact that vegetative gametophytes can be purposely selected, propagated quickly, cross-bred, with the resulting sporophytes having almost the same characteristics leads to a new way to select desiredUndaria strains for long time use without losing the desired economic characteristics. Contribution No. 2696 from the Institute of Oceanology, Chinese Academy of Sciences. Supported by National Natural Science Foundation of China.     

Effects of temperature, salinity, light intensity, and pH on the eicosapentaenoic acid production of Pinguiococcus pyrenoidosus

The effects of temperature, light intensity, salinity, and initial pH on the growth and fatty acid composition of Pinguiococcus pyrenoidosus 2078 were studied for eicosapentaenoic acid (EPA) production potential. The fatty acid composition was assayed by gas chromatography-mass spectrometry, which indicated that the main fatty acids were C14:0, C16:0 and EPA. The highest EPA percentage 20.83% of total fatty acids was obtained at 20℃ with the temperature being set at 20, 24, and 28℃. Under different salinities and light intensities, the highest percentages of total polyunsaturated fatty acids (PUFAs) and EPA were 17.82% and 31.37% of total fatty acids, respectively, which were achieved at salinity 30 and 100μmol photon m-2s-1 illumination. The highest percentages of total PUFAs and EPA were 38.75% and 23.13% of total fatty acids, respectively, which were reached at an initial pH of 6 with the test range being from 5.0 to 9.0.     

Molecular identification and culture trials of Eutreptiella gymnastica (Eutreptiales, Euglenophyceae)

During a survey for marine microalgal resources, we isolated a rare marine euglenoid from the coastal waters of Qingdao, China in 2009, and established a pure culture. Electron microscopic and molecular phylogenetic (18S rDNA and 16S rDNA sequences) analyses revealed a close affinity with Eutreptiella gymnastica, a bloom-forming species. Different culture conditions were monitored to understand optimal E. gymnastica growth characteristics. The optimal growth conditions in a batch culture of this isolate were 20°C, 160 μmol photons/(m2×s) of white light, and a salinity of 10-31. Nutrient experiments demonstrated that growth increased dramatically with a phosphorus concentration greater than 72 μmol/L. Understanding the effect of culture conditions on E. gymnastica may help understanding the blooming mechanism of this alga in its natural environment.     

Intraspecific genetic analysis, gamete release performance, and growth of Sargassum muticum （Fucales, Phaeophyta） from China

Sargassum muticum is one of the most abundant and conspicuous native macroalgae species on the northern coasts of China. It often forms large-scale seaweed beds in subtidal zones. This investigation was designed to understand the intraspecific genetic relationships of this alga based on samples collected from four northern coastal sites of China, and to evaluate gamete release and growth capacity in laboratory conditions. The nuclear internal transcribed spacer 2 （ITS2） sequences of 16 samples from four locations were identical. Based on cox3 gene and partial rbcLS operon sequences, intraspecific genetic variability was detected with three and two ribotypes, respectively. Temperature, not irradiance, was shown to significantly affect gamete release and fertilization. Elevated temperature and irradiance enhanced the growth of germlings and vegetative branchlets. Maximum growth rate of germlings was detected at 18-24℃and an irradiance of 60-100 μmol photons/（m^2.s）. Under ambient conditions （12-25℃ and 60-125 gmol photons/ （m2.s））, relative growth rate of young branchlets could reach 7.5%/d.     

Starvation metabolism of two common species of chironomids in Biandantang Lake

1 INTRODUCTION Chironomid larvae are main groups in most aquatic ecosystems, playing a crucial ecological role in decomposition of detritus and material ex- changes between water column and the sediment (Liang et al., 1995a, b; Chen, et al., 1982). They a…     

Early development of grateloupia turuturu (Halymeniaceae, Rhodophyta)

Grateloupia turuturu is a commercial red alga with potential value in nutraceuticals and pharmaceuticals. To supplement information on its life history and verify whether carpospores can be used for seedling culture, early development of G. turuturu was investigated under culture conditions (27°C, 10-13 mol/(m2.s) in irradiance, photoperiod 10:14 h L:D). Three physiological stages were recognized by continuous microscopic observation: division stage, discoid crust stage, and juvenile seedling stage. At the beginning of the division stage, the carpospores developed germ tubes into which the carpospore protoplasm was evacuated, and then the carpospore protoplasm in the germ tubes began to divide continuously until discoid crusts formed. Finally, upright thalli appeared on the discoid crusts and developed into juvenile seedlings. It took about 60 days for carpospores to develop into juvenile seedlings. The growth parameters, including germination rate for carpospores and discoid crust diameter, were recorded. These results contribute more information on the life cycle, and at the same time are of great significance in the scaling-up of artificial seedling cultures of G. turuturu.     

The South Pacific Subtropical Mode Water in the Tasman Sea

From the synopical CTD sections in the WOCE PR11 repeated cruises, the South Pacific Subtropical Mode Water (SPSTMW) has been identified in the region of the Tasman Front Extension (TFE) around 29?S to the east of Australia. In the depth range of 150-250 m, the SPSTMW appears as a thermostad with vertical temperature gradient lower than 1.6℃(100 m)-1 and a tem- perature range of 16.5-19.5℃ and as a pycnostad with PV lower than 2×10-10 m-1 s-1 and a potential density range of 25.4-26.0 kg m-3. Like the subtropical mode waters in the North Atlantic and North Pacific, the formation of the SPSTMW is associated with the convective mixing during the austral wintertime as manifested from the time series of the Argo floats. And cold water entrains into the mixed layer with the deepening mixed layer from September to the middle of October. During the wintertime formation process, mesoscale eddies prevailing in the TFE region play an important role in the SPSTMW formation, and have a great effect on the SPSTMW distribution in the next year. The deeper (shallower) mixed layer in wintertime, consistent with the depressed (uplifted) permanent thermocline, is formed by the anticyclonic (cyclonic) eddies, and the substantial mode water thicker than 50 m is mainly found in the region of the anticyclonic eddies where the permanent thermocline is deeper than 450 m.     

Molecular identification and culture observation on <Emphasis Type="Italic">Acrochaete leptochaete</Emphasis> (Chaetophoraceae,Chlorophyta) from China

Acrochaete leptochaete (Huber) Nielsen (Chaetophoraceae, Chlorophyta) was isolated from the macroalgae Chaetomorpha collected from intertidal pools in Rongcheng, Shandong, China. 18S rDNA combined with ITS regions were used to ascertain the morphological identification of the isolated material. Based on the unialgal culture, asexual reproduction and growth characteristics of A. leptochaete were investigated over wide ranges of temperature and irradiance. Results revealed that asexual reproduction of A. leptochaete could be realized by biflagellate zoospores. The zoospores germinated directly to give self- replicating generations. Zoospore germination was bipolar. A temperature range from 13-21°C and a lower irradiance of 36 μmol/(m2·s) were most favorable for the growth of A. leptochaete. Thallus organization, an important taxonomic criterion for the genus Acrochaete, was affected markedly by temperature and irradiance. Our results extend the knowledge about the species’ general biology and its morphological plasticity. For classification and identification of a simple microphytic algae like A. leptochaete, which are traditionally placed in the class Chaetophoraceae, we propose that molecular tools associated with culture observations are applied.     

NUTRIENTS NEAR THE ARCTIC POLE REGION

The distributions of SiO_3-Si, PO_4-P, NO_3-N and NO_2-N concentrations in the seawater, iceand snow from 82°45′21″W, 88°01′20″N to the North Pole are reported and the exchange of thenutrients among the seawater, ice and snow are discussed. The average concentrations of the nutrients inthe seawater near the North Pole were 4.9±2.4μmol/L for SiO_3-Si; 0.60±0.10μmol/L for PO_4-P,3.4±1.7μmol/L for NO_3-N and 0.13±0.04μmol/L for NO_2-N. The nutrient concentrations inthe ice and snow in the North Pole were 0.20 and 0.65μmol/L for SiO_3-Si; 0.22 and 0.25μmol/L forPO_4-P; 1.6 and 5.8μmol/L for NO_3-N, 0.19 and 0.15μmol/L for NO_2-N.     

Size-fractionated phytoplankton biomass in autumn of the Changjiang （Yangtze） River Estuary and its adjacent waters after the Three Gorges Dam construction

A cruise was undertaken from 3rd to 8th November 2004 in Changjiang （Yangtze） River Estuary and its adjacent waters to investigate the spatial biomass distribution and size composition of phytoplankton. Chlorophyll-a （Chl-a） concentration ranged 0.42-1.17 μg L^-1 and 0.41-10.43 μg L^-1 inside and outside the river mouth, with the mean value 0.73 μg L^-1 and 1.86 μg L^-1, respectively. Compared with the Chl-a concentration in summer of 2004, the mean value was much lower inside, and a little higher outside the river mouth. The maximal Chl-a was 10.43 μg L^-1 at station 18 （122.67°E, 31.25°N）, and the region of high Chl-a concentration was observed in the central survey area between 122.5°E and 123.0°E. In the stations located east of 122.5°E, Chl-a concentration was generally high in the upper layers above 5 m due to water stratification. In the survey area, the average Chl-a in sizes of 〉20 μm and 〈20 μm was 0.28 μg L^-1 and 1.40 μg L^-1, respectively. High Chl-a concentration of 〈20 μm size-fraction indicated that the nanophytoplankton and picophytoplankton contributed the most to the biomass of phytoplankton. Skeletonema costatum, Prorocentrum micans and Scrippsiella trochoidea were the dominant species in surface water. The spatial distribution of cell abundance of phytoplankton was patchy and did not agree well with that of Chl-a, as the cell abundance could not distinguish the differences in shape and size of phytoplankton cells. Nitrate and silicate behaved conservatively, but the former could probably be the limitation factor to algal biomass at offshore stations. The distribution of phosphate scattered considerably, and its relation to the phytoplankton biomass was complicated.     

THE DENSITIES OF CHANGJIANG ESTUARINE WATERS

The International Eq一lation of State of Seawater,1980 and the PraeticalSalinity Scale,1978 have been adoPted by theUNESCO江CES沼COR八APSOJoint Panel ono‘eanogral,hie Tables and Standards(JPOTS),and endorsed bythese organizati6ns(Miller‘〕and Poisson,1981;Uneseo,1981).Th已new equa-tion and the Praetieal Salinity Seale are to be used for all values Published fromJan .1,1982 .The new equation 15 aeeurate for use in all oeeanie surfaee waters,but eannot be aeeurately aPPlied to…     

Periodic exposure to ambient solar irradiance benefits the growth of juvenile seedlings of Hizikia fusiformis

In our trials, from 2007 to 2008, of mass production of seedlings of Hizikia fusiformis using synchronization techniques, problems of a "dark thalli" phenomenon and epiphytes contamination severely threatened the health of juvenile seedlings. In this investigation, we optimized conditions for improving the growth of juvenile seedlings. Seven string collectors were seeded with zygotes and a series of experiments were conducted including direct exposure to solar irradiance, co-culture with Ulva spp. and treatment with sodium hypochlorite. It was found that direct exposure to solar irradiance (maximum: 1 740 μmol photons/(m2?s)) for 2 h per day could efficiently enhance the growth of young seedlings and simultaneously inhibit the growth of epiphytic algae. In this treatment, 50-day old seedlings could reach an average of 0.44 cm in length and possess up to nine leaflets. However, a single treatment with 18-mmol/L sodium hypochlorite for 10 min severely harmed 15-day old seedlings. In comparison, weekly treatment with 2.2-mmol/L sodium hypochlorite for 10 min brought no apparent harm to seedlings and eliminated epiphytic algae efficiently. However, this treatment significantly increased the detachment rate of seedlings. Inoculating Ulva spp. onto the collector caused a dramatic decrease in the number of seedlings. However, the growth of the remaining seedlings appeared unhampered. All collectors except the control were daily sprayed with a high pressure water jet from the 8th day post fertilization. From the first day to 50th day, no "dark thallus" was observed on any of the seven collectors. We believe that well-timed daily exposure to solar irradiance would favor H. fusiformis in its early growing stages.