STOICHIOMETRIC RESPONSES OF PHYTOPLANKTON SPECIES TO THE INTERACTIVE EFFECT OF NUTRIENT SUPPLY RATIOS AND GROWTH RATES1

Three species of phytoplankton, Rhodomonas sp., Phaeodactylum tricornutum Bohlin, and Isochrysis galbana Parke, were cultivated in semicontinuous culture to analyze the response of carbon (C):nitrogen (N):phosphorus (P) stoichiometry to the interactive effect of five N:P supply ratios and four growth rates (dilution rates). The relationship between cellular N and P quotas and growth rates fits well to both the Droop and Ågren’s functions for all species. We observed excess uptake of both N and P in the three species. N:P biomass ratios showed a significant positive relationship with N:P supply ratios across the entire range of growth rates, and N:P biomass ratios converged to an intermediate value independent of N:P supply ratios at higher growth rates. The effect of growth rates on N:P biomass ratios was positive at lower N:P supply ratios, but negative at higher N:P supply ratios for both Rhodomonas sp. and I. galbana, while for P. tricornutum this effect was negative at all N:P supply ratios. A significant interactive effect of N:P supply ratios and growth rates on N:P biomass ratios was found in both Rhodomonas sp. and P. tricornutum, but not in I. galbana. Our results suggest that Ågren’s functions may explain the underlying biochemical principle for the Droop model. The parameters in the Droop and Ågren’s functions can be useful indications of algal succession in the phytoplankton community in changing oceans.     

澳门湿地浮游植物群落特征

2007年7月、10月和2008年1月, 对澳门的湿地——筷子基湾、白鹭林、莲花大桥滩涂和南湾湖的浮游植物群落特征进行了调查。3次采样共检测到76种(属), 其中, 蓝藻门藻类13种(属), 绿藻门30种(属), 硅藻门25种(属), 甲藻门2种, 裸藻门4种, 金藻门和隐藻门各1种(属)。浮游植物丰度的最高值出现在莲花大桥, 为3 922.33×10⁴ cells·L^-1; 最小值出现在南湾湖, 为1.58×10⁴ cells·L^-1。浮游植物主要是由蓝藻、绿藻、裸藻和硅藻组成。2008年1月硅藻的含量最高, 而2007年7月蓝藻的含量最高。浮游植物多样性指数和均匀度指数值均指示出: 筷子基湾2008年1月、莲花大桥2007年7月和10月污染最严重, 其他属于中度污染或无污染。透明度对澳门湿地浮游植物的影响较大, 与绿藻(r=0.683, p<0.05)、甲藻(r=0.715, p<0.05)、金藻(r=0.707, p<0.05)和隐藻(r=0.701, p<0.05)都存在较高的正相关关系。硅藻与pH值存在较强的负相关关系(r=–0.674, p<0.05), 与总氮(r=0.895, p<0.05)、总磷(r=0.920, p<0.05)和正磷酸(r=0.668, p<0.05)存在较强的正相关关系。     

GROWTH RATE VARIATION IN THE N:P REQUIREMENT RATIO OF PHYTOPLANKTON1

Theoretical considerations predict that the cell N:P ratio at transition from nitrogen limitation to phosphorus limitation of phytoplankton growth (critical ratio, R_c) varies, as a function of population growth rate. This prediction is confirmed by experimental, data from the literature along with new experimental data for the marine, prymnesiophyte Pavlova lutheri (Droop) Green. R_c passes through a maximum at intermediate growth rates for the three phytoplankton species for which data, are available, but there is significant interspecific variability in its value. There is no theoretical or experimental evidence to support the idea that the ratio of subsistence N and P cell quotas is equal to R_c over the range of growth rates, or that the subsistence quota ratio equals the ratio of the N and P cell quotas minus a storage fraction. Examination of N:P composition ratios can be used to determine which nutrient is limiting, but cannot be used to determine relative growth rates or competitive advantage between species limited by the same nutrient. Growth rates are determined by environmental conditions and by the cell quota of the limiting nutrient, without reference to the cell quota of the non-limiting nutrient.     

CONTRIBUTION OF THE CLASS CRYPTOPHYCEAE TO PHYTOPLANKTON STRUCTURE IN THE GERMAN BIGHT1

The class Cryptophyceae (Division Cryptophyta) contains ecologically relevant species, which are widespread in aquatic environments. However, classification, identification, and enumeration of cryptophytes are challenged by a morphology that must be usually examined with EM to permit species identification. The quantitative importance of this group has been revealed by HPLC data. But ecological information assessing the occurrence or seasonality of cryptophytes in the marine environment is still scarce. Molecular techniques allow for a refined assessment of taxonomically challenging taxa, such as the cryptophytes. In our laboratory, a Phylochip was developed to facilitate and refine the assessment of cryptophyte microalgae. Here, we present the results of an environmental study that took advantage of the Phylochip. The study was designed to elucidate the seasonality and contribution of cryptophytes to phytoplankton structure in the German Bight. The occurrence of cryptophytes in total plankton versus the picoplankton fraction was assessed with the Phylochip between the years 2004 and 2006. Our data indicate that cryptophytes are an important and constant contributor to phytoplankton structure of the German Bight, especially in the picoplankton fraction.     

KINETICS OF NUTRIENT UPTAKE AND GROWTH IN PHYTOPLANKTON1

Microscopic algae ran grow rapidly in natural waters that are extremely low in essential macro and micro nutrients. Yet, their nutrient uptake systems exhibit only mediocre nutrient affinities, the saturation constants being often 10–1000 times the (estimated) ambient concentrations. The large difference which exists between the saturation constants for growth (K_μ) and short term uptake (K_ρ) are due to the acclimation capabilities of the organisms. Over the acclimation range, K_μ to K_ρ, the algae can maintain maximum growth rate by modulating both their internal nutrient quotas (Q) and their maximum short term nutrient uptake rates (ρ_max) in response to variations in external nutrient concentrations. The commonly assumed hyperbolic relationships for steady growth and uptake (viz “chemostat theory”) are coherent with a hyperbolic expression for short term uptake including a variable maximum (ρ_max). The ratio of the saturation constants for growth and uptake is then directly related to the extreme in quotas and maximum uptake rates: K_μ/K_ρ= Q_min/Q_max·ρ^lo_max/ρ^hi_max. This result is applicable even when the exact hyperbolic laws are not. Published data on Fe, Mn, P and N limitation in algae are generally in accord with the theory and demonstrate a wider acclimation range for trace than for major nutrients.     

KINETICS OF NUTRIENT UPTAKE AND GROWTH IN PHYTOPLANKTON1

Microscopic algae can grow rapidly in natural waters that are extremely low in essential macro and micro nutrients. Yet, their nutrient uptake systems exhibit only mediocre nutrient affinities, the saturation constants being often 10–1000 times the (estimated) ambient concentrations. The large difference which exists between the saturation constants for growth (K_u) and short term uptake (K_p) are due to the acclimation capabilities of the organisms. Over the acclimation range, K_u, to K_p the algae can maintain maximum growth rate by modulating both their internal nutrient quotas (Q) and their maximum short term nutrient uptake rates (P_max) in response to variations in external nutrient concentrations. The commonly assumed hyperbolic relationships for steady growth and uptake (viz “chemostat theory”) are coherent with a hyperbolic expression for short term uptake including a variable maximum (P_max). The ratio of the saturation constants for growth and uptake is then directly related to the extreme in quotas and maximum uptake rates: K^μ/K^ρ= Q^min/Q^maxρ^max/ρQ^max. This result is applicable even when the exact hyperbolic laws are not. Published data on Fe, Mn, P and N limitation in algae are generally in accord with the theory and demonstrate a wider acclimation range for trace than for major nutrients.     

COMBINING NEW TECHNOLOGIES FOR DETERMINATION OF PHYTOPLANKTON COMMUNITY STRUCTURE IN THE NORTHERN GULF OF MEXICO 1

In situ analysis of phytoplankton community structure was determined at five stations along the Texas Gulf coast using two instruments, the Fluoroprobe and FlowCAM. Results were compared with traditional methods to determine community structure (pigment analysis and microscopy). Diatoms and small nanoplankton (most likely haptophytes) dominated the phytoplankton community at all stations. Estimated chl concentrations for diatoms+dinoflagellates obtained via the Fluoroprobe were not significantly different for three of the five stations sampled when compared with HPLC‐chemical taxonomy analysis, whereas the concentrations of green algal and cryptophyte chl were overestimated. The FlowCAM estimates of overall nanoplankton and microplankton cell abundance were not significantly different when compared with epifluorescence microscopy, and recorded images of phytoplankton cells provided a representative population of the phytoplankton community at each station. The Fluoroprobe and FlowCAM, when used in tandem, are potentially capable of determining the general characteristics of phytoplankton community structure in situ and could be an important addition to biological observing systems in the coastal ocean.     

NUTRIENT-LIMITED GROWTH OF AUREOUMBRA LAGUNENSIS (PELAGOPHYCEAE), WITH IMPLICATIONS FOR ITS CAPABILITY TO OUTGROW OTHER PHYTOPLANKTON SPECIES IN PHOSPHATE-LIMITED ENVIRONMENTS

Growth and chemical compositional characteristics of the brown tide-forming alga, Aureoumbra lagunensis Stockwell, DeYoe, Hargraves et Johnson, were studied through a series of nitrogen-limited and phosphate-limited continuous cultures over a range of growth rates. The specific growth rate of A. lagunensis was hyperbolically related to the cell quota of the limiting nutrient in ammonium-limited cultures. In phosphate-limited cultures, the relationship was best described by a straight line. The N cell quota of A. lagunensis ranges from about 20 fmol at zero growth rate under N-limited conditions to a high of roughly 85 fmol under N-replete conditions. Similarly, the P cell quota of A. lagunensis ranges from about 0.15 fmol at zero growth rate under P-limited conditions to a high of 2 fmol under P-replete conditions. Aureoumbra lagunensis has a very high N:P critical ratio (>100). The high N:P critical ratio, as well as the organism's apparent ability to use forms of phosphorus other than phosphate under severe phosphate deficiency, may partially explain its success in P-limited environments, such as the Laguna Madre. In addition, a uniqe quadratic relationship between the productivity index (PI) and growth rate was discovered. Such a relationship supports an earlier argument that PI may not be a good indicator of nutritional status.     

GROWTH RESPONSES OF SEVERAL DIATOM SPECIES ISOLATED FROM VARIOUS ENVIRONMENTS TO TEMPERATURE

Eight diatom species (Chaetoceros pseudocurvisetus Mang ., Stephanodiscus hantzschii Grun ., Skeletonema costatum ( Grev.) Cleve , Asterionella formosa Hass ., Thalassiosira nordenskioeldii Cleve , Detonula confervacea ( Cleve) Gran , Chaetoceros sp., and Nitzschia frigida Grun.) were isolated from various temperature environments ranging from temperate to the Arctic, and their growth responses to temperature were determined. Each species grew over a different temperature range. The lower and upper limits of each species varied from −1.8° to 20° C and from 2° to 30° C, respectively. The width of the growth range of each species. also varied from 3.8° to 25° C, and the growth of these species was observed, as a whole, between a wide temperature range from −1.8° to 30° C .
Within the growth temperature ranges, the growth rate of each species increased with temperature until reaching a maximum, which was followed by a steep decrease up to the upper limit of the growth range. As a result, each species showed a maximum rate at the temperature very near to the upper limit, which was generally higher than the isolation temperature. The specific growth rates were compared among the eight species. The interspecific maximum rate at each temperature exhibited an exponential increase with a Q₁₀ = 1.48. The relative growth rates of each species were calculated by normalizing the specific growth rates with the interspecific maximum rate at each respective temperature. The higher relative growth rates tended to occur at the isolation temperature of each species, suggesting that temperature is a significant control on species distributions in nature .     

南亚热带贫营养水库春季浮游植物群落结构与动态

2005年1~6月,通过每两周一次的高频率采样,对南亚热带贫营养水库——梅溪水库的水文、营养盐和浮游植物进行了调查,并计算水体浮游植物生物量。主要结果如下:梅溪水库浮游植物具有物种少,生物量低,以飞燕角甲藻(Ceratium hirundinella)和多甲藻(Peridinium sp.)为优势藻的特征。12次采样24个样品共检测到浮游植物42种。浮游植物在早春(1~3月)和晚春(4~6月)有显著的差别,其中每次采样浮游植物早春平均13种,晚春平均21种。浮游植物总的细胞丰度为31~273 cells·ml^-1,总生物量为0.176~2.024 mg·L^-1之间。晚春浮游植物平均生物量明显高于早春。低营养盐和弱酸性水体有利于能够垂直迁移获得营养的鞭毛藻类和其它藻类之间竞争,而使其成为整个春季优势类群。在晚春,随着水温显著增加,浮游植物丰度和生物量也明显增加,但是降雨的增加降低了水体的透明度,大大减缓了由水温上升导致生物量增长的趋势。水温是梅溪水库浮游植物变化的主要限制因子,但是降雨有明显的干扰作用。     

PHOTOACCLIMATION AND GROWTH RATE RESPONSES OF ULVA ROTUNDATA (CHLOROPHYTA) TO INTRADAY VARIATIONS IN GROWTH IRRADIANCE1

A vegetative clone of the chlorophyte macroalga Ulva rotundata was maintained in an outdoor continuous flow system under nutrient sufficient conditions and various light regimes. Step changes between 9 and 100% incident irradiance (I^o) were employed to simulate cloud passage. Temporary (1–4 h) midday (I^o)perturbations evoked net changes in growth rate (μ) and chlorophyll (chl) content. Under I_o alternating at various periodicities from 15 min to 7 h, net μ was the average of the μ under steady state 9 and 100% I^o, regardless of periodicity. However, the μ in alternating light was considerably less than μ under steady state 55% I^o(? 9%+ 100%/2), as expected based on the nonlinear shape of the μ-I relationship. Unlike μ, chl content depended primarily on the total daily irradiance, probably clue to the slower response of chl compared to photosynthetic rate. On time scales ≥ one day, chl was linearly correlated with light-regulated daily μ under both steady state and intraday fluctuating irradiance, consistent with photosynthetic feedback regulation of chl concentration.     

GROWTH RATE RELATIONSHIPS TO PHYSIOLOGICAL INDICES OF NUTRIENT STATUS IN MARINE DIATOMS

The growth of two species of marine diatom, Thalassiosira weissflogii (Grunow) and Thalassiosira pseudonana (Hustedt), was followed in batch cultures at four concentrations of dissolved inorganic carbon from N- and C-replete lag phase into N- and/or C-deplete stationary phase. Results describe the relationship between carbon-specific growth rate (μ_C) and chl a :carbon (chl a :C) and glutamine:glutamate (gln:glu) ratios with changes in the cells' nutritional status (N:C), during the utilization of either NO₃ ⁻ or NH₄ ⁺ . The use of the gln:glu ratio as an index of N:C requires further clarification. For both species and N sources, N stress resulted in a decrease in μ_C, chl a :C, and N:C relative to μ_Cmax values, whereas C stress resulted in a decrease in μ_C and an increase in chl a :C and N:C relative to μ_Cmax values. Both species attained a chl a :C ratio of approximately 15 μg·g ^{− 1} at μ_Cmax using either N source. However, this value was not necessarily an indicator of maximal growth rate. NC colimitation resulted in decreased μ_C to values less than 20% of μ_Cmax with only minor changes in chl a :C and N:C relative to μ_Cmax values. Chl a :C results suggest a similarity between the light stress and C stress responses of marine diatoms. The potential for C stress in the marine environment needs to be addressed.     

STEADY STATE GROWTH OF PHYTOPLANKTON IN CONTINUOUS CULTURE: COMPARISON OF INTERNAL AND EXTERNAL NUTRIENT EQUATIONS1,2

Previously, there have been conflicts over whether external or internal nutrient concentrations control phytoplankton growth rates at steady state in continuous culture. To experimentally demonstrate that both equations equally describe steady state growth, continuous culture studies with phosphorus-limited growth of the chrysophyte Monochrysis lutheri Droop were carried out over the entire growth rate region up to biomass washout. Data were examined using both the Monod and Droop equations, and, even though there were significant variations in the yield coefficient with growth rate, the data fit both equations reasonably well. Because of their relative simplicity, the Droop equation and an equation combining both the Monod and Droop equations are better suited for expressing kinetic data than the Monod equation. It is crucial, though, that the criteria necessary to achieve steady state be fulfilled.     

七子花土壤微生物数量及呼吸速率的季节动态*

研究濒危植物七子花群落土壤微生物的生长及其呼吸速率的季节动态,结果表明：根际土壤细菌、放线菌,根围土壤细菌、真菌、放线菌的数量和土壤总呼吸速率具有相似的季节变化规律：全年呈单峰曲线变化,最大值均出现在9月份,但根际土壤真菌数量的最大值出现在10月份。土壤微生物数量受土壤含水量和温度的影响较大;土壤总呼吸速率不仅与土壤含水量和温度存在密切关系,而且与土壤微生物数量显相关,土壤微生物是土壤总呼吸的重要承担。     

INTERACTING EFFECTS OF LIGHT AND NUTRIENT LIMITATION ON THE GROWTH RATE OF SYNECHOCOCCUS LINEARIS (CYANOPHYCEAE)1

The blue-green alga Synechococcus linearis (Naeg.) Kom. was grown in P- and N-limited chemostats over a range of potentially limiting irradiances in order to determine the combined effects of light and nutrient limitation on some aspects of the composition and metabolism of this alga. Over a narrow range of low irradiances, simultaneous limitation of growth rate by light and either N or P was shown. This simultaneous limitation of growth rate by a nutrient and a physical factor can be explained by the ability of an increased supply of one to compensate in part for a decreased supply of the other. At all irradiances, the internal concentration of the limiting nutrient increased with increasing dilution rate, and the results could be fitted to the Droop relationship. With decreasing irradiance, the internal concentration of the limiting nutrient increased. There appeared to be little or no effect of light on the minimum internal concentration of P but that of N increased with decreasing light. Both chlorophyll a and biliprotein per unit particulate C increased with increasing dilution rate and decreasing irradiance. The critical N/P ratio increased with decreasing light as the N requirement of N-limited cells increased faster than did the P requirement of P-limited cells. The composition of exponentially growing cells in complete medium varied much less with light. Neither dilution rate nor irradiance during growth had a great effect on saturated rates of P or N uptake or alkaline phosphatase activity. Calculated assimilation ratios increased with light and dilution rate. The role of the flexibility of nutrient composition in adaptation to adverse conditions and the implications of the results for the use of physiological indicators of nutrient status are discussed.     

MERCURY AND TEMPERATURE INTERACTIONS ON THE GROWTH RATES OF THREE SPECIES OF FRESHWATER PHYTOPLANKTON1,2

Cultures of freshwater algae, representing three algal divisions, Synura petersenii Korshikov; Chlamydomonas sp.: and Nitzschia sp., were subjected to four different mercury-temperature shock interactions to demonstrate synergistic effects between mercury and temperature. Algal growth, measured by temporal changes in vivo fluorescence of chlorophyll was used to ascertain the effects. Mercury addition and temperature shock had various inhibitory effects on algal growth. Prior environmental conditions influence the effect of subsequent treatments. Growth of S. petersenii was severely inhibited by mercury in all experiments. Control cultures would not grow at 30 C and died when shacked at this temperature. Chlamydomonas sp. cultures initially inhibited by mercury were able to recover under most conditions after a period of reduced growth. Nitzschia sp. was resistant to mercury except when simultaneously shocked with temperature. Mercury analyses showed that Nitzschia cells at 25 C and 30 C contained a high percentage of the mercury initially added to the cultures. There was a significant loss of mercury at the end of each experiment from all cultures, probably due to volatilization.     

PHYTOPLANKTON COMMUNITY COMPOSITION AND GENE EXPRESSION OF FUNCTIONAL GENES INVOLVED IN CARBON AND NITROGEN ASSIMILATION1

A functional gene microarray was developed and used to investigate phytoplankton community composition and gene expression in the English Channel. Genes encoding the CO₂‐fixation enzyme RUBISCO (rbcL) and the nitrate assimilation enzyme nitrate reductase (NR) representing several major groups of phytoplankton were included as oligonucleotide probes on the “phytoarray.” Five major groups of eukaryotic phytoplankton that possess the Type 1D rbcL gene were detected, both in terms of presence (DNA) and activity (rbcL gene expression). Changes in relative signal intensity among the Type 1D rbcL probes indicated a shift from diatom dominance in the spring bloom to dominance by haptophytes and flagellates later in the summer. Because of the limitations of a smaller database, NR probes detected fewer groups, but due to the greater diversity among known NR sequences, NR probes provided higher phylogenetic resolution than did rbcL probes and identified two uncultivated diatom phylotypes as the most abundant (DNA) and active (NR gene expression) in field samples. Unidentified chlorophytes and the diatom Phaeodactylum tricornutum Bohlin were detected at both the DNA and cDNA (gene expression) levels. The reproducibility of the array was evaluated in several ways, and future directions for further improvement of probe development and sensitivity are outlined. The phytoarray provides a relatively high‐resolution, high‐throughput approach to assessing phytoplankton community composition in marine environments.     

闽南-台湾浅滩近岸上升流区浮游植物碳同化速率的研究

本文采用~(14)C同位素示踪法,测定了浮游植物光合作用速率,结合浮游植物细胞含碳量,计算了不同季节、不同深度下浮游植物碳同化速率常数,讨论了不同环境条件对浮游植物碳同化速率常数的影响。结果表明,闽南-台湾浅滩近岸上升流的形成,是该海域高生产力的主要原因,上升流期间浮游植物复制时间要比非上升流期间浮游植物复制时间缩短1.8倍。同时还表明,温度,光照强度、营养盐是控制浮游植物生长的主要因子。上升流期间营养盐始终保持在较高的水平。是上升流区具有较大的浮游植物碳同化速率常数(3.3d~(-1))的原因之一。适宜该海域浮游植物生长的光照强度在3 000—15 000lx之间,温度的影响可用Goldman和Carpenter模式近似表示。