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.     

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 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).     

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.     

Effects of temperature and irradiance on early development of Chondrus ocellatus Holm (Gigartinaceae, Rhodophyta)

Chondrus is a type of commercially produced red seaweed that widely used for food and carrageen extraction. Although the natural life history of the alga had been well understood, the factors influencing development of the tetraspore and carpospore remain poorly understood. In the perspective of seedling resources, the regulation of early development is crucial for the seedling nursing; therefore, it is necessary to understand the physiological influences during its early development. In this study, we studied the effects of temperature and irradiance on the early development of Chondrus ocellatus Holm under laboratory conditions. The released tetraspores and carpospores were cultivated at different temperatures (10–28°C) and irradiances (10, 60 μmol photons m⁻²s⁻¹) with a photoperiod of 12L:12D. The results indicate that both tetraspores and carpospores are tolerant to temperatures of 10–25°C, and have the highest relative growth rate at 20°C. Irradiance variances influenced the growth of the discoid crusts, and the influence was more significant with increasing temperature; 60 μmol photons m⁻²s⁻¹ was more suitable than 10 μmol photons m⁻²s⁻¹. The optimum temperature and irradiance for the development of seedlings was 20°C and 60 μmol photons m⁻²s⁻¹, respectively.     

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.     

Intraspecific crossings ofUndaria pinnatifida (Harv.) Sur.—A possible time-saving way of strain selection

Unicellular gametophytes ofUndaria pinnatifida (Harv.) Sur. were isolated in Qingdao, P. R. China in April 1993 and in Tokushima, southern Japan in March 1995. Different intraspecific crossings by using unicellular male and female gametophytes were successfully undertaken in Sept. of 1995 in Qingdao. Sporophytes were transplanted to two different locations for open sea cultivation. One was at Zhanqiao (ZQ) Bay where the water current was slower than that of another location—Taipingjiao(TPJ). A total of 218 adult sporophytes were harvested on January 12 and 18 from TPJ and ZQ respectively. For eacn combination, 10 sporophytes were cultivated. Analysis of the morphological characteristics of adult sporophytes indicated that the longest length between two bases of the serration of pinnate blades (W2) is a morphological characteristic that can be transferred from the parent plant to the next generations regardless of environmental variations. There was evidence that W2 was apparently determined by sex-linked factors, i.e., by male parental gametophyte. Sporophytes from certain crossing combinations showed more vigorous growth than those from other crossing combinations. It is therefore possible to select gametophyte strains which can be used as parental gametophytes for the seedling production of sporophytes with more vigorous growth within shorter cultivation period. The morphology of hybrids from a Qingdao strain and a Tokushima cultivated strain resembled that of both parental plants in frond features (wrinkled or smooth) and W2. Sporophyll formation also varied with strains. The fact that adult sporophytes resulting from the same crossing combinations have identical morphological characteristics under the same environmental conditions indicates the possibility of a new way to select strains which are expected to be ideal for commerical production by purposely selecting, propagating, and seeding unicellular gametophytes for sporeling production through freeliving techniques of gametophytes. Project 39400102 supported by NSFC, also supported by National Climbing Plan B (PD-B642); and Bioengineering Center, SSTC.     

A preliminary comparative study of the productivity of three economic seaweeds

The rates of photosynthesis and respiration ofPorphyra, Laminaria andUndaria were determined with an oxygenmeter at different temperatures in an enclosed continuous-flow system. The temperature coefficients of respiration and the net primary productivities for the three species were calculated. The spectacular rates of primary production were found to be as high as 270 g C m⁻² forP. yezoensis, 2200 g C m⁻² forL. japonica and 160 g C m⁻² forU. pinnatifida annually. The low respiratory rates of these three species as well as low light compensation points especially at low temperatures are considered as a reason to save heat energy, resulting in a high productivity, especially in case ofLaminaria. Contribution No. 945 from the Institute of Oceanology, Academia Sinica. The paper was presented at the International Phycological Society Meeting, 19–22 Aug. 1980, Glasgow, UK.     

DYNAMIC CHANGES OF INORGANIC NITROGEN AND ASTAXANTI—IIN ACCUMULATION IN HAEMATOCOCCUS PLUVIALIS

This study on dynamic changes of culture color,astaxanthin and chlorophylls,inorganic N including N-NO3^-,N-NO2^- and N-NH4^ in batch culture of Haematococcus pluvialis exposed to different additive nitrate concentration showed(1)ast/chl ratio was over 0.8 for brown and red algae,but was usually less than 0.5 for green and yellow algae;(2)N-NO3^-,in general,was unstable and decreased,except for a small unexpected increase in nitrate enriched treatment groups;(3)measurable amounts of N-NO2^- and N-NH4^ were observed respectively with three change modes although no extemal nitrite and ammonia were added into the culture;(4)a non-linear correlation between ast/chl ratio(or color)changes and the levels of N-NO3^-,N-NO2^-,N-NH4^ in H.pluvialis culture;(5)up and down variation of the ast/chl ratio occurred simultaneously with a perceptible color change from yellow to brown(or red)when N-NO3^-,N-NO2^- and N-NH4^ fluctuated around 30,5,5μmol/L respectively;(6)existence of three dynamic modes of N-NO3^-,N-NO2^- and N-NH4^ changes,obviously associated with initial extemal nitrate;(7)the key level of total inorganic N concentration regulating the above physiological changes during indoor cultivation was about 50 μmol/L;and(8)0.5-10mmol/L of nitrate was theoretically conducive to cell growth in batch culture.     

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     

Abundance and production ofBranchiura sowerbyi (Oligochaeta: Tubificidae) in two typical shallow lakes (Hubei, China)

An April 1996 to March 1997 comparative study on the abundance and secondary production ofBranchiura sowerbyi Beddard, 1892 in two typical shallow lakes showed that in Houhu, an algae-dominated lake, the worm density (68 ind·m⁻²) peaked in July, biomass (1.930 g·m⁻²) peaked in June, while in Biandantang, a macrophyte-dominated lake, standing stock (density: 60 ind·m⁻²; biomass: 1.019 g·m⁻² in wet weight) peaked in December. Secondary production of the animal in Houhu Lake was 3.413 g wet wt m⁻² a⁻¹, a little more than that (2.675 g wet wt m⁻² a⁻¹) in Biandantang Lake. Their turnover rates (P/B ratios) were 4.0 and 5.0, respectively. Project (39600019 and 39430101) supported by NSFC.     

Standing crop and primary production of benthic microalgae on the tidal flats in Yueqing Bay

The standing stock and primary production of benthic microalgae on tidal flats were measured seasonally at 3 transects (Puqing, Dahengchuang and Puqi) in Yueqing Bay during 2002 2003. The results showed that the integral chlorophyll a (Chl a) concentration in tidal flat mud exhibited a seasonal variation with the order of magnitude: winter (14.0 4.2 mg m-2) > spring (13.0 6.3 mg m-2) > autumn (7.7 5.9 mg m-2) > summer (4.6 3.2 mg m-2). The primary production showed an order of magnitude: spring (270.5 224.9 mgC m-2 d-1)>winter (238.7 225.5 mgC m-2 d-1)>autumn (214.1 56.2 mgC m-2 d-1)>summer (71.6 44.6 mgC m-2 d-1). Both chlorophyll a and primary production showed maximum values in the surface layer of sediment, and decreased rapidly with increasing depth due to sun light limitation. The results of variance analysis indicated that seasonal variation and tidal flat condition affected Chl a greatly, but had no significant effect on primary production. The annual primary production of benthic microalgae on tidal flats in Yueqing Bay was estimated at 16143 tons carbon, which is sufficient to support 1.02×105 tons shellfish production. The environmental factors affecting chlorophyll and primary production on the tidal flats in Yueqing Bay were discussed. By comparing with other bays on China’s coast, it was observed that Yueqing Bay is a region with high benthic microalgae standing crop and primary production, which may be related to the type of its sediment.     

Mixing in Shallow Waters： Measurements, Processing, and Applications

Microstructure profiling measurements taken on a shallow Black Sea shelf and in Lake Banyoles and Boadella reservoir (Both in Spain) are analyzed to investigate the influence of boundary-layer-induced turbulence of various sources on mixing in the water interior. The state of turbulence in shallow waters is examined and details of microstructure data processing and error analysis are discussed. The dependence between averaged activity parameter AG and buoyancy Reynolds number Reb for the shelf turbulence indicates that for Reb < 1 the state of turbulence can be described by the fossil turbulence model, which postulates AG-Reb1/2. For Reb> 1, however, the influence of Reb on AG is weak, signifying that the buoyancy Reynolds number can no longer serve as the governing parameter for active turbulent mixing. The generation of turbulence by a one-minute long wind bursts (the Boadella reservoir) increases the averaged dissipation rate (e) of the surface mixed layer by more than 5 times (up to 3×10-6 W kg-1). The influence of the wind bursts was also traced below the ther-mocline, where turbulent patches with     

<Emphasis Type="Italic">p</Emphasis>CO<Subscript>2</Subscript> and carbon fluxes across sea-air interface in the Changjiang Estuary and Hangzhou Bay

Partial pressure of CO2 （pCO2） was investigated in the Changjiang （Yangtze River） Estuary, Hangzhou Bay and their adjacent areas during a cruise in August 2004, China. The data show that pCO2 in surface waters of the studied area was higher than that in the atmosphere with only exception of a patch east of Zhoushan Archipelago. The pCO2 varied from 168 to 2 264 μatm, which fell in the low range compared with those of other estuaries in the world. The calculated sea-air CO2 fluxes decreased offshore and varied from -10.0 to 88.1 mmol m^-2 d^-1 in average of 24.4 ± 16.5 mmol m^-2 d^-1. Although the area studied was estimated only 2 × 10^4 km^2, it emitted （5.9 ± 4.0） × 10^3 tons of carbon to the atmosphere every day. The estuaries and their plumes must be further studied for better understanding the role of coastal seas playing in the global oceanic carbon cycle.     

Effects of NO and Different Media on the Growth of Prorocentrum micans

1Introduction NOisadiffusible,gaseousfreeradicalthatplaysimportantrolesinanimalsandplants.Itfunctionsin diseaseresistance,abioticstress,celldeath,respira tion,senescence,rootdevelopments,germinationandhormoneresponses.Itisanunusualsignalinthatitis areactive,lipophilicandvolatilefreeradicalthatcanbecytotoxic.ThewidespreadbiologicalsignificanceofnitricoxidewasrecognizedbySciencein1992whichnamedthefreeradicalNO‘Moleculeoftheyear’,andin1998theNobelPrizeinPhysiologyandMedicinewasawardedforwor…     

Transpiration rates of Carex Meyeriana in relation to micrometerological factors in a mountain valley wetland

In order to reveal transpiration rates of wetland plants and its relationships to micro- meterological factors in a mountain valley wetland, relative humidity, air temperature, leaf temperature, soil temperature, photo flux density and transpiration rates were measured once two hours in a Carex meyeriana wetland of the Changbai Mountain valley in dry （July） and wet （September） of 2003, respectively. Results showed that the tendency of ＂decreasing after increasing＂ was obvious in wet season. However, a relatively stable trend was observed for the transpiration in dry season.. Generally, the photon flux density of Carex meyeriana was higher in wet season than that in dry season. However, the variabilities of leaf temperature, air temperature and relative humidity were similar in both seasons. Higher transpiration rates of Carex meyeriana leaves were observed in July （varied from 40 to 150 mol.m^-2.s^-1） compared to those （varied from 7 to 14 mol.m^-2.s^-1） in September. Transpiration rates were significantly correlated with air temperature （P〈0.01）, leaf temperature （P〈0.01）, and wind speed （P〈0.05）, but correlationship between relative humidity and photo flux density was not significant （P〈0.05）.     

Effects of nutritional factors on the growth and heterotrophic eicosapentaenoic acid production of diatom <Emphasis Type="Italic">Nitzschia laevis</Emphasis>

The effects of several nutritional factors on the growth and eicosapentaenoic acid (EPA) production of diatom Nitzschia laevis were studied. 4 LDM (quadrupled concentration of the nutrient salt) was the optimal concentration of nutrient salt for the growth and EPA production of N. laevis. The growth of N. laevis was inhibited when the glucose concentration was either lower than 10 gL⁻¹ or higher than 15 gL⁻¹. Both sodium nitrate and urea were good nitrogen sources for the growth and EPA production, while ammonium chloride seriously decreased the dry cell weight (DW) and the EPA content. Silicate seriously influenced the growth of N. laevis. The maximum DW of 2.34 gL⁻¹ was obtained in the presence of 150 mgL⁻¹ Na₂SiO₃·9H₂O. The EPA content remained almost the same when the silicate concentration was lower than 150 mgL⁻¹; however, higher silicate concentrations resulted in a steady decrease of EPA content. Low medium salinity (⩽29) did not seem to influence the DW of N. laevis, and high salinity resulted in a decrease of DW. The highest EPA content (4.08%) and yield (110 mgL⁻¹) were observed at the salinity of 36 and 29, respectively.