PHOSPHORUS,NITROGEN AND THE GROWTH OF ALGAE IN LAKE KINNERET1

The intracellular concentrations of carbon, nitrogen, phosphorus and chlorophyll a of phytoplankton and zooplankton in Lake Kinneret were determined from 1969 to 1973. The ratios C:P, C:N, chlorophyll a:P, chlorophyll a:N of the algae showed fluctuations which could be related to the nutrient conditions that influence the annual pattern of phytoplankton development, especially in respect to the dinoflagellate bloom of Peridinium cinctum (OFM) Ehr. f. westii (Lemm.) Lef. Relatively high intracellurar P values at the start of the bloom indicated adequate availability of this nutrient and luxury consumption over a short period of time. Later, Peridinium continued to grow despite unusually high C:P ratios (> 300:1). In most years, phosphorus may have eventually limited growth, however, in 1970, the bloom censed despite comparatively high intracellular P concentrations. These observations, together with supplementary evidence from nutrient addition experiments and determinations of specific alkaline phosphatase levels, indicated that, for most of the growth phase of the bloom, Peridinium cells were not directly limited by P. The decline of the bloom usually, but not always (e.g., in 1970), was marked by very high C:P ratios. Thus, a shortage of P may often be a contributory factor to the cessation of the Peridinium bloom and may be limiting phytoplankton growth in the fall. Over the years 1969–73, possibly due to an overall drop in salinity, there appears to be a trend to lower levels of biologically bound phosphorus in Lake Kinneret, without a concomitant decrease in carbon biomass.     

大型丝状绿藻对N、P去除效果研究

当前,N、P过剩引发水体富营养化现象仍十分严重,探索有效实用、易于推广的N、P去除技术和寻求对N、P具有较强吸收和降解能力的生物物种,始终为环保学者们所关注,利用藻类去除N、P的研究始于20世纪50年代,迄今已得到广泛应用并取得显著成效。为解决早期藻类生物技术中过量单细胞藻类不易收获之不足,自20世纪90年代中期,人们开始尝试利用大型丝状绿藻治理N、P污染。陈汉辉利用水网藻净化水源水质、王朝晖等人利用水网藻去除富营养化水体中N、P营养的研究结果均表明,大型丝状绿藻能在污水中正常生长代谢,并对水体中的N、P养分具有较高的转化率和去除率。       

CARBON,NITROGEN, AND PHOSPHORUS AND THE EUTROPHICATION OF FRESHWATER LAKES1

The question of nutrients responsible for eutrophication of freshwater lakes is reviewed, and recent additions to the literature on nutrient limitation are discussed. The paper by Lange is criticized on several grounds, including the facts that utilization of HCO₃^? by phytoplankton and the invasion of lake waters by atmospheric CO₂ are ignored as sources of photosynthetic carbon. The phosphorus and nitrogen concentrations used in Lange's experiments are far higher than values published by others for Lakes Erie and Ontario. Preliminary results of fertilizing a small oligotrophic lake with nitrogen and phosphorus are described. The standing crop of phytoplankton increased by 30–50 ×, while the P:N:C ratio in seston did not change from ratios found in unfertilized lakes. Other experiments done in water columns isolated with polyethylene film showed that addition of carbon did not increase the phytoplankton standing crop. Since the fertilized lake was initially lower in total CO₂ than any other recorded in the literature, it is concluded that carbon is unlikely to limit the standing crop of phytoplankton in almost any situation. Measurements of invasion of atmospheric gases to the fertilized lake by the Rn²²² technique were compared with phytoplankton production measurements, revealing that atmospheric invasion of CO₂ is sufficient to support the high phytoplankton standing crop in the epilimnion of the lake. Possible errors in interpretation of culture and bottle-bioassay experiments with respect to eutrophication are discussed.     

N、P营养盐胁迫对两株布朗葡萄藻生长的影响

采用批次培养方法,在光照强度60、110mol/m2s下分别设置了7个不同的氮、磷浓度(N:0-3500g/L,P:15-775g/L),研究两株布朗葡萄藻(Botryococcus braunii)对氮、磷胁迫的敏感性差异,筛选高营养利用效率的优良藻株。结果表明:两株藻对氮磷营养胁迫的耐受性存在差异,B.braunii764株对氮胁迫具有较高耐受性,而B.braunii765株对磷胁迫具有较高耐受性。光照强度110mol/m2s,不同氮浓度下B.braunii764株其平均生长速率均显著高于其他各处理组;不同磷浓度下B.braunii765株其平均生长速率显著高于B.braunii764株。在试验设定的光照强度条件下,适当增加光照强度能够显著降低氮胁迫对布朗葡萄藻生长的抑制效应。在光照强度110mol/m2s下,氮浓度3500g/L时两株布朗葡萄藻平均生长速率与在正常Chu-10培养基条件下无显著差异。磷浓度775g/L时两株布朗葡萄藻的平均生长速率均显著低于正常Chu-10培养基条件,增加光照强度对磷胁迫下藻细胞的生长无显著作用。两株布朗葡萄藻在第2天时磷吸收与初始磷浓度呈正相关关系,氮吸收在3500g/L时出现饱和现象。布朗葡萄藻的生长更容易受到培养基中磷营养胁迫的影响。       

EFFECTS OF NITROGEN AND PHOSPHORUS AVAILABILITY ON THE EXPRESSION OF THE COCCOLITH‐VESICLE V‐ATPASE (SUBUNIT C) Of PLEUROCHRYSIS (HAPTOPHYTA)1

The coccolithophores Emiliania and Pleurochrysis demonstrate increased coccolith production when growth is reduced by nitrate or phosphate limitation. The function of enhanced coccolith production under these conditions and its regulation have not been resolved. Studies at the molecular level are ideally suited to determine the exact relationship between calcification and other cellular functions. In a previous study we provided evidence for the presence of a vacuolar H⁺‐ATPase on coccolith vesicle membranes of P. carterae. These trans‐Golgi–derived vesicles are the sites of coccolith production, with each vesicle containing one coccolith. In this study, expression of the vap gene, encoding subunit c of the vacuolar H⁺‐ATPase, was investigated. Our objective was to explore potential relationships between vap expression, nutrient‐dependent growth, and calcification. Specifically, we monitored vap expression relative to two genes, fcp and pcna, whose expression was previously shown to vary with growth conditions; fcp encodes a fucoxanthin chl a/c‐binding protein, and pcna encodes the proliferating cell nuclear antigen. Relative to the expression of pcna and fcp, vap expression was highest at nutrient concentrations where growth curves and chl a patterns indicated arrest of cell division. Our results indicate that the level of vap expression does not decrease when cell growth diminishes.     

DETECTION OF LIMITING OR SURPLUS NITROGEN IN ALGAE AND AQUATIC WEEDS1

The rate of NH₄+-N absorption by algae and aquatic weeds in the dark has been shown to be 4-5 times greater for plants which are N-limited as compared to plants with sufficient available N. Eight species of green algae, 2 blue-green algae, 2 diatoms, and 3 aquatic weeds were used to demonstrate the usefulness of the test in determining if available N was in surplus or limited supply in a particular environment. The test was shown not to differentiate between blue-green algae capable of fixing N (4 species) from media with NO₃-N or without combined nitrogen. The factors influencing the results of NH4⁺-N absorption tests have been investigated. In order to differentiate between plants with sufficient available N and those which are N limited, the rate of NH₄⁺-N absorption (0.1 mg N) over 1-hr incubation in the dark by 10-20 mg of algae or aquatic weed tissues is measured. The relatively simple analysis for NH₄⁺-N in the samples makes it very easy to follow the changing N nutrition of plants in cultures with a limited N supply or in the presence of possible N sources.     

保安湖沉积物和间隙水中氮和磷的含量及其分布

保安湖沉积物中氮(N)的含量平均为5.20mg/g,平面分布以桥墩湖区含量最高。沉积物中磷(P)的含量平均为0.75mg/g,平面分布以扁担塘湖区含量最高。沉积物间隙水中总氮的平均含量为3.63mg/L,无机氮中以氨氮的含量最高,占总氮的57.2%;间隙水中总磷的含量平均为0.098mg/L,磷酸盐占总磷的50.0%.间隙水中氮和磷含量的平面分布差异不明显。间隙水中氮和磷的含量比湖水中含量高,但除氨氮外,一般不超过5倍,表明湖水和沉积物间隙水之间营养交换十分强烈。       

EXTRACTIVE AND ENZYMATIC ANALYSES FOR LIMITING OR SURPLUS PHOSPHORUS IN ALGAE

An extractive procedure for detection of surplus-stored phosphorus (luxury consumption) in algae and an enzymatic analysis for conditions of P-limited growth in algae have been evaluated. A simple 60-min boiling water extraction of algae known to contain surplus P separates essential P compounds and surplus-stored P compounds. Surplus P compounds can be measured in the extract as orthophosphate. Extracts of algae limited in their growth by the amount of available P contain little or no orthophosphate. Limitation of algal growth by P supply induces the enzyme alkaline phosphatase. The activity of this enzyme can be measured at pH 9 using p-nitro-phenylphosphate as substrate. Algae which were P-limited and contained no extractable orthophosphate have as much as 25 times more alkaline phosphatase activity than algae with surplus available P.     

EXTRACTIVE AND ENZYMATIC ANALYSES FOR LIMITING OR SURPLUS PHOSPHORUS IN ALGAE

An extractive Procedure for detection of surplusstored phosphorus (luxury consumption) in algae and an enzymatic analysis for conditions of P-limited growth in algae have been evaluated. A simple 60-min boiling water extraction of algae known to contain surplus P separates essential P compounds and surplus-stored P compounds. Surplus P compounds can be measured in the extract as orthophosphate. Extracts of algae limited in their growth by the amount of available P contain little or no orthophosphate. Limitation of algal growth by P supply induces the enzyme alkaline phosphatase. The activity of this enzyme can be measured at pH 9 using p-nitro-phenylphosphate as substrate. Algae which were P-limited and contained no extractable orthophosphate have as much as 25 times more alkaline phosphatase activity than algae with surplus available P .     

RESPONSES OF FRESHWATER ALGAE TO INHIBITORY VANADIUM CONCENTRATIONS: THE ROLE OF PHOSPHORUS1

We found that species-specific differences exist among a variety of freshwater algae and cyanobacteria in the extent to which growth and photosynthesis are inhibited by vanadium. A major factor controlling the degree of inhibition by vanadium was the phosphorus state (P-sufficient vs. P-deficient) of the organisms. In P-sufficient cultures, vanadium was inhibitory when the vanadium concentration exceeded the phosphate concentration. In P-deficient cultures, the depression of photosynthesis by vanadium increased with increasing phosphorus deficiency. Our conclusion that vanadium competed with phosphate for uptake sites was supported by the following three observations: 1) the decreased influx of ³²P-PO ₄ into P-deficient cells in the presence of vanadium, 2) the amelioration of vanadium inhibition of photosynthesis by the addition of phosphate, and 3) the accumulation of vanadium by cells. At vanadium concentrations that severely inhibited growth, the cells of Scenedesmus obliquus (Turp.) Kruger were larger than normal and contained more vacuoles, lipid, and starch bodies than normal cells. Four-celled coenobia were replaced by unicells. Scenedesmus acutusf: alternans Hortobagyi cells from vanadium-inhibited cultures had 7.5 times more vanadium per cell than control cultures and contained numerous granules that did not stain for polyphosphate and may be composed of condensed vanadate molecules. The cellular P quota and turnover time of PO₄in the medium are important regulators of the extent of inhibition by vanadium.     

EFFECTS OF NITROGEN AND PHOSPHORUS ON THE ENDOSYMBIONT LOAD OF RHOPALODIA GIBBA AND EPITHEMIA TURGIDA (BACILLARIOPHYCEAE)1

Diatoms of the family Epithemiaceae possess a unicellular nitrogen-fixing cyanobacterial endosymbiont. We investigated the potential of extracellular nitrogen and phosphorus concentrations to affect the endosymbiont load of Rhopalodia gibba O. Müll, and Epithemia turgida Ehr. in field and culture populations. In a growth chamber experiment, monoclonal cultures of R. gibba were exposed to three levels of nitrate-nitrogen. Nutrient-diffusing substrates were used in a lake environment to create nine microhabitats of varying nitrogen and phosphorus ratios for natural populations of R. gibba and E. turgida. The number of endosymbionts per diatom increased as ambient nitrogen became limiting; mean endosymbiont volume increased as nitrogen increased. The mean endosymbiont surface area: volume ratio decreased with increasing nitrogen. Total endosymbiont volume per diatom (the product of the number of endosymbionts per diatom and their individual biovolumes) did not have a simple response to increasing nitrogen. Phosphorus limitation uncoupled the relationship between endosymbiont load and nitrogen. We suspect that flexibility of the endosymbiont load can reduce the metabolic cost to the diatom if the endosymbionts are dependent on the diatom for a resource.