Cultivation of the brown alga <Emphasis Type="Italic">Hizikia fusiformis</Emphasis> (Harvey) Okamura: stress resistance of artificially raised young seedlings revealed by chlorophyll fluorescence measurement

Commercial farming of the intertidal brown alga Hizikia fusiformis (Harvey) Okamura in China and South Korea in the sea depends on three sources of seedlings: holdfast-derived regenerated seedlings, young plants from wild population and zygote-derived seedlings. Like many successfully farmed seaweed species, the sustainable development of Hizikia farming will rely on a stable supply of artificial seedlings via sexual reproduction under controlled conditions. However, the high rate of detachment of seedlings after transfer to open sea is one of the main obstacles, and has limited large-scale application of zygote-derived seedlings. To seek the optimal condition for growing seedlings on substratum in land-based tanks for avoidance of detachment in this investigation, young seedlings were grown in both outdoor tanks exposed directly to sunlight and in indoor raceway tanks in reduced, filtered sunlight. Results showed that young seedlings, immediately after fertilization, could withstand a daily fluctuation of direct solar irradiance up to a level of 1800 μmol photons m⁻² s⁻¹, and maintained a faster growth rate than seedlings grown in indoor tanks. Detailed experiments by use of chlorophyll fluorescence measurements further demonstrated that the overnight (12 h) recovery of optimal fluorescence quantum yield (F_v/F_m) of seedlings after 1 h treatment at 40°C was 98%, and the 48 h recovery of F_v/F_m of seedlings after 1 h exposure to 1800 μmol m⁻² s⁻¹ was 92%. Forty-one-day-old seedlings showed no significant decrease of optimal fluorescence quantum yield at salinity ranging from 30 to 5 ppt for a treatment up to 17 h. Six-hour desiccation treatment did not have any influence on the optimal fluorescence quantum yield. Exposure to 18 mmol L⁻¹ sodium hypochlorite for 10 min did not damage the PSII efficiency, and thus could be used to remove epiphytic algae. The strong tolerance of young seedlings to high temperature, high irradiance, low salinity and desiccation found in this investigation supports the view that mass production of Hizikia seedlings should be performed in ambient light and temperature instead of in shaded greenhouse tanks.     

大气CO2升高和紫外辐射相互作用对羊栖菜生理特性的影响

为了研究经济海藻羊栖菜对大气CO2浓度增加与紫外辐射(UVR)相互作用的响应,设置两个CO2浓度(380μL/L和800μL/L)以及两种辐射处理,即PAR处理(滤除UV-A、UV-B,藻体仅接受可见光,400—700nm)和PAB处理(全波长辐射280—700nm)培养海藻,探讨了羊栖菜生长、光合作用、呼吸作用、光合色素含量、可溶性糖和蛋白以及硝酸还原酶活性的变化情况。结果表明高浓度CO2显著提高羊栖菜藻体的相对生长速率,并且紫外辐射的负面效应在高CO2处理下表现不显著。高CO2降低了藻体的光合作用速率,而UVR的负面效应和生长体现为一致性,但是羊栖菜的呼吸作用没有受到环境变化的明显影响。羊栖菜的光合色素叶绿素a和类胡萝卜素在高浓度CO2处理下明显降低,而UVR没有明显影响。环境因子对羊栖菜的可溶性糖没有影响,但是在高CO2和全波长辐射处理下,藻体可溶性蛋白的含量显著增加。同时高CO2明显提高了硝酸还原酶的活性,并且仅在高浓度CO2处理下藻体中UVR对其活性有抑制作用。CO2和UVR对羊栖菜的大多数生理特性存在明显的交互作用,在未来CO2浓度进一步增加的情况下,UVR的负面效应将会得到一定程度的缓解,这样有利于羊栖菜在养殖海区获得更高的产量。     

Growth and photosynthesis in seedlings of Hizikia fusiformis (Harvey) Okamura (Sargassaceae, Phaeophyta) cultured at two different temperatures

This study examined temperature acclimation, growth, and photosynthetic characteristics of the zygote-derived seedlings of Hizikia fusiformis (Harvey) Okamura (Sargassaceae). The seedlings were cultured at 15°C or 25°C for 4 weeks. The average relative growth rate was significantly higher in seedlings acclimated at 25°C. The photosynthetic rate measured at 15°C was much higher in seedlings grown at 15°C than those grown at 25°C, indicating photosynthetic acclimation to a lower temperature. At 35°C, the photosynthetic rate of 15°C-grown seedlings was drastically decreased, whereas that of 25°C-grown seedlings was significantly increased. The maximum relative electron transport rate (rETR_max) measured at the respective growth temperature was significantly higher in seedlings grown at 25°C than at 15°C. At a measuring temperature of 35°C, the rETR_max in both 15°C- and 25°C-grown seedlings were considerably reduced with regard to those measured at 15°C or 25°C. Our results suggested that, compared with the seedlings grown at 25°C, those acclimated at a lower temperature could be disadvantaged under adverse conditions such as increased temperatures.     

Regulation of gamete release in the economic brown seaweed <Emphasis Type="Italic">Hizikia fusiforme</Emphasis> (Phaeophyta)

Gamete release is an essential event in artificial seeding of the economic brown seaweed, Hizikia fusiforme. Mass egg release occurred in the dark, with few eggs being discharged in the light. Release of eggs was elicited with eight practical salinity units (one PSU ≡ 1 g sea salts l⁻¹) and was inhibited by salinity levels >32 PSU. Egg release was optimal at 23 °C, and was decreased by 72% in agitated seawater compared to unstirred seawater. Inhibitors of photosynthesis and ions channels suppressed egg release, indicating that this process was physiologically associated with photosynthetic activity and ion transport.     

Seasonal Pattern of Reproduction Of Hizikia Fusiformis (Sargassaceae, Phaeophyta) from Nanao Island, Shantou, China

The maturation pattern of sexual reproduction in Hizikia fusiformis (Harvey) Okamura (Sargassaceae, Phaeaophyta) was examined in 2003 at Yunao Bay, Nanao Island, Shantou, China. Maturation began in mid-April (seawater temperature 19–21 ^∘C), reached the peak in mid-May (maturation rate ca. 70%, and seawater temperature 23.5–25 ^∘C) and finished in late-June (seawater temperature 27.5–30 ^∘C). The Hizikia plants continued to gain the length from the beginning of maturation season to reach a maximum mean length of 34.8 cm in mid-May, after which the mean length was reduced drastically due to the senescence and rupture of the larger plants in size. The major portion of the mature plants belonged to the larger plants between April and May, but to the smaller ones in June. It is suggested that the plant must achieve a critical size before reproductive maturation occurred. There was a positive relationship between the number of receptacles (NR), as well as the reproductive allocation (RA), and the plant size of Hizikia population, with the recorded maximum values of NR and RA being 1220 and 64.3% respectively, for a single plant.     

Cultivation of the brown alga Sargassum horneri: sexual reproduction and seedling production in tank culture under reduced solar irradiance in ambient temperature

As a large conspicuous intertidal brown alga, individuals of Sargassum horneri can reach a length of more than 7 m with a fresh weight of 3 kg along the coasts of the Eastern China Sea. The biomass of this alga as a vital component in coastal water ecology has been well documented. In recent years, a steady disappearance of the algal biomass along the once densely populated coastal areas of the Eastern China Sea has drawn attention in China. Efforts have been made to reconstruct the subtidal algal flora or even to grow the alga by use of long-lines. As part of the efforts to establish an efficient technique for producing seedlings of S. horneri, in this investigation a series of culture experiments were carried out in indoor raceway and rectangular tanks under reduced solar irradiance at ambient temperature in 2007–2008. The investigation demonstrated that: (1) sexual reproduction of S. horneri could be accelerated in elevated temperature and light climates, at least 3 months earlier than in the wild; (2) eggs of S. horneri had the potential to be fertilized up to 48 h, much longer than that of known related species; (3) suspension and fixed culture methods were both effective in growing the seedlings to the long-line cultivation stage; and (4) the life cycle of S. horneri in culture could be shortened to 4.5 months, thus establishing this alga as an appropriate model for investigating sexual reproduction in dieocious species of this genus.     

Effect of temperature on the filtration rate and percentage of assimilation ofSalpa fusiformis Cuvier (Tunicata: Thaliacea)

The effect of temperature on filtration and ingestion rates ofSalpa fusiformis Cuvier was determined over the temperature range 13–22°C at a constant algal concentration. Filtration and ingestion rates increased with temperature up to an optimum (19.7 to 20.2°C) beyond which they decreased. Food assimilation for this salp species was determined while fed onPhaeodactylum tricornutum Bohlin orHymenomonas elongata Stein at different algal concentrations. The percent assimilation varied between 28 and 39% (mean: 32%) with the diatomP. tricornutum as food, and between 39 to 81% (mean: 64%) with the flagellateH. elongata. Assimilation did not seem to depend on the algal concentration but it was greater with the flagellate than with the diatom, the latter having a higher ash content.     

Microscopic observations on tissue calcium distribution in the brown alga,Undaria pinnatifida (Harvey) Suringar

The distribution of Ca²⁺ in thallus tissues of the brown algaUndaria pinnatifida was investigated by microspectro-photometric analysis after fixation with Carnoy's reagent and staining with an alizarine sulfonate dye. Ca²⁺ accumulation in the young thallus was much higher in mucilage cells of the basal than the apical region; there was least in the pith of the basal region. A roughly uniform pattern was found in the stipe from the central part of the young thallus, while a highly irregular pattern occurred in both stipe and blade from the corresponding area of the mature thallus; the Ca²⁺ content was much lower than in the cortex, mucilage cells and vacuoles of the mature stipe than in the corresponding area of the young stipe. The contents of all regions of the mature blade were higher than those in the young blade; they were particularly high in mucilage cells.     

Canopy characteristics of the brown alga Sargassum muticum (Fucales,Phaeophyta) in Lake Grevelingen,southwest Netherlands

The present distribution of the invasive brown alga Sargassum muticum in the southwest Netherlands is updated. Populations of the alga were found to remain at their 1985 level in Lake Grevelingen, with a small eastward expansion into the Eastern Scheldt estuary. A new population for the brackish, non-tidal Lake Veere is reported. Within Lake Grevelingen S. muticum forms a persistent, extensive canopy of 100% cover (4,442.5 ± 525.6 g fresh wt m^–2, 640.3 ± 75.8 g dry wt m^–2) that has a marked effect upon the penetration of photosynthetically active radiation (PAR) (reduced by 97% at 0.1 m). Surface sea water temperatures can be elevated by 2.7 °C above water not associated with a Sargassum canopy; furthermore, the dense canopy shades and hence reduces water temperatures below 0.1 m depth. Productivity studies indicate that assimilation occurs in the upper levels of the canopy (57.09 µg C mg dry wt^–1 m^–2 at a mean PAR rate of 106.7 J cm^–2 h^–1). Self-shading and a resultant decrease in the rate of assimilation was evident below the canopy.     

Cultivation of Gracilaria in outdoor tanks and ponds

This review deals with the major problems of unattached Gracilaria intensive cultivation in outdoor tanks and ponds. These problems are presented through the main variables affecting the Gracilaria annual yield and the updated solutions evolved. The physical variables include tank and pond structure, seawater characteristics such as velocity, agitation practice, exchange rate, and salinity, light characteristics such as quantity and quality, and temperature modelling. The chemical variables include nutrient composition and regime of application, and inorganic carbon supply with the pH changes involved. The biological variables include seaweed density, epiphyte competition, grazer damage, bacterial disintegration, integrated mariculture and strain selection. The experience gained in the Israeli research on Gracilaria cultivation is discussed in view of other Gracilaria and seaweed intensive cultivation research.     

Protoplast isolation in the marine brown alga Dictyopteris prolifera (Dictyotales)

Protoplasts were isolated from thalli of Dictyopteris prolifera using a mixture of crude enzymes from vicera of live oysters (Crassostrea gigas) and the following commercial enzymes: an abalone enzyme, cellulase, polygalacturonase and hemicellulase. The enzyme mixtures produced up to 3.3 × 10⁷ cells per l g of tissue fresh weight. The conversion to protoplasts of the cells was about 100% using the oyster enzyme or the abalone enzyme alone. The optimum pH for protoplast isolation was 6.0 and 20 hours were required for conversion to protoplasts.     

Cultivation of Macrocystis integrifolia (Laminariales,Phaeophyta) in southeastern Alaskan waters

The artificial culture of Macrocystis integrifolia in southeastern Alaskan waters has been initiated as a first step in a project to augment the existing herring roe-on-kelp fishery in the state. The growing of Macrocystis in Prince William Sound has the potential of considerably enhancing this fishery by eliminating the costly importation of this kelp from southeast Alaska. In an ongoing feasibility study, Macrocystis has been cultured under laboratory conditions through the embryonic sporophyte stage and outplanted in waters near Sitka, Alaska (57° N). Growth of the outplanted kelps has been monitored as a function of the time and depth of the outplanting. Preliminary results suggest that light is limiting for growth in winter and that the optimal depth for outplanting will vary with the season.     

Characterization of two circular plasmids from the marine diatom Cylindrotheca fusiformis: plasmids hybridize to chloroplast and nuclear DNA.

Summary This paper reports the discovery and initial characterization of two small plasmids, pCfl and pCf2, in the marine diatomCylindrotheca fusiformis. Extracted diatom DNA separates into two bands in CsCI-Hoechst 33258 dye gradients. Upon agarose gel electrophoresis of a sample of the upper band of the gradient we observed, in addition to high molecular weight (genomic) chloroplast and mitochondrial DNA, pairs of lower molecular weight bands. These bands contained two species of circular plasmid DNA molecules, as shown by electron microscopy. The nucleotide composition of the plasmids, and chloroplast and mitochondrial DNAs is similar, as indicated by their co-banding in the gradients. They were cloned, and their restriction maps determined, showing that pCfl is 4.27 and pCf2 4.08 kb in size. By hybridization analysis, we showed that pCfl and pCf2 share regions of similarity, but not identity. Neither plasmid hybridizes with mitochondrial DNA. Both plasmids hybridize with chloroplast DNA, and pCf2 also hybridizes with nuclear DNA.     

The introduced alga Undaria pinnatifida (Laminariales,Alariaceae) in the lagoon of Venice

Since its appearance in 1992 in the lagoon of Venice, the brown algae Undaria pinnatifida (kelp) has gradually expanded along the banks of canals both at Chioggia and Venice, becoming the dominant species in the local algal community chiefly from February to July. In Chioggia Island, where another brown seaweed (Sargassum muticum) is present since 1992, the spreading of Undaria reaches a plateau. In Venice Island, instead, the colonisation process is still in progress. During 1999, the kelp has colonised the main canals (e.g. The Grand Canal) and, subsequently, the small inner ones. In order to evaluate the dynamics of substrate re-colonisation by Undaria, two mechanical eradications were carried out during (March) and after the fertile period (July) of the algae. Such experimental manipulations provided evidence of the fast re-colonisation potential of the algae mainly attributable to its efficient reproductive system. Eradication made during the fertile period, in fact, permitted the kelp development during the following year, while re-colonisation has started 2 years later where eradication was performed after the reproductive period. A significant decrease in the surface covered by other species has been observed both in shallow (Ulva rigida, Enteromorpha spp., Antithamnion pectinatum, Chondracanthus acicularis) and deeper areas (Rhodymenia ardissonei) during the period of maximum development of U. pinnatifida. On the basis of the results of this study, the following conclusions can be drawn: (1) the alga U. pinnatifida is continuously expanding in lagoon environments of Venice; (2) in order to limit its spreading, mechanical eradications would be done on a large spatial scale and before the zoospores release; (3) there is suggestive evidence of competition between Undaria and the remaining indigenous algae.     

Presence and development of populations of the introduced brown alga Sargassum muticum in the southwest Netherlands

Of the three Brachionus species used in aquaculture, Brachionus rubens, B. calycilorus and B. plicatilis, the latter is most widely used in raising marine fish and shrimp larvae due to its tolerance to the marine environment. In freshwater aquaculture the use of B. rubens and B. calycilorus is limited, probably because inert food products are readily available as feed for freshwater larvae.The rotifer Brachionus plicatilis is used in large numbers as the first food organism in intensive cultures of marine fish and shrimp larvae. An adequate supply of these rotifers relies on mass cultures. The reproductive rate of rotifers in these cultures depends on food quality and quantity, salinity, temperature and pH of the medium. Removal of waste products from culture tanks leads to higher and more efficient production of rotifers over extended periods of time. Rotifers have to be enriched with polyunsaturated fatty acids, which are essential for proper development and survival of marine fish and shrimp larvae.The future use of preserved rotifers and their resting eggs may help to overcome unforeseen failures of live cultures and may lead to more efficient use of these organisms in raising freshwater and marine fish and shrimp larvae.     

Relation between size and age of holdfasts of Ecklonia stolonifera Okamura (Laminariales,Phaeophyta) in northern Honshu,Japan

Ecklonia stolonifera is distributed along the coast facing the Sea of Japan. The size of various parts of the shoot (blade length and width and stipe length and diameter) and the age were determined at Ooma, Aomori Prefecture. The smaller the holdfast, the higher the percentage of one-year-old shoots. Holdfasts 10 cm in diameter seemed to be three years old, whereas holdfasts 40 cm in diameter seemed to be five or more years old. Zoosporangial sori were observed on blades three or more years old. Ecklonia stolonifera holdfast diameter expands only vegetatively by stoloniferous rhizoids. Zoospores, formed on shoots three or more years old, serve for the formation of new populations.     

Changes in M:G ratios of extracted and residual alginate fractions on boiling with water the dried brown alga Kjellmaniella crassifolia (Laminariales,Phaeophyta)

Kjellmaniella crassifolia, the edible macro-brown alga in Japan contained nearly 27% of alginates of which nearly 7% was extractable from the fronds with boiling water for 6 h and the residual alginates in the frond were almost exhaustively extracted with a dilute alkali at 60 °C for 6 h. The alginates dissolved in all these extracts with both boiling water and dilute alkali were purified by fractionation with MgCl₂ and alcohol.The content of MM blocks in the boiling water-soluble alginate sample increased remarkably during heating for 6 h while that of GG blocks from the same sample decreased. In contrast, MM blocks in the alkali-soluble alginate sample decreased during 6 h heating while GG blocks continued to increase. Since the amounts of MG blocks showed slight fluctation, the M:G ratio of alginates extracted with boiling water increased towards the end of extraction whereas the reverse is true for the alkali-soluble alginates.