海洋生物固氮作用研究进展

近年来,海洋生物固氮作用研究已成为海洋氮循环研究热点之一,因为它补充了海洋中的结合态氮,影响着海洋氮储库的收支平衡,进而调控海洋的初级生产力,并因此与海洋颗粒有机碳的迁出以及海洋对大气CO2的吸收密切相关.海洋生物固氮作用的研究主要是在近30a开展起来的,现有观点认为全球海洋生物固氮速率为100～200 Tg/a.15↑N2示踪法和乙炔还原法是最重要的生物固氮速率实测方法,而硝酸盐异常指数法、15↑N同位素收支平衡法、卫星遥测法等则是重要的间接估测方法.束毛藻曾被认为是全球海洋最重要的固氮生物,但近年来也发现了固氮生物在微微（pi-co）级和微（nano）级的分布,表明以往基于束毛藻的固氮速率可能是被低估的.海洋生物固氮作用营养盐限制的研究主要集中在铁和磷这2个元素的作用,但目前尚无定论.在中国边缘海生物固氮作用的研究还很匮乏,少数研究表明除黑潮区外,由束毛藻所支持的固氮速率并不高.就海洋固氮作用研究的未来发展,指出今后必须在更大时空尺度上开展海洋生物固氮作用研究,以弥补现有实测数据的不足,并获取固氮微生物种类组成的准确信息,在确定海洋生物固氮作用控制因素的同时,追踪新固定氮的流向,进而为海洋氮循环提供准确信息.     

海洋蓝藻固氮基因

氮是海洋浮游植物必需的大量元素,又是海洋浮游植物生长的主要限制因子之一,研究氮元素的补充机制对了解海洋生态系统有重要意义。海区氮营养盐的补充主要通过新生氮实现,其主要补充机制为径流输入、大气干湿沉降、水体混合、上升流及生物固氮作用。生物固氮是指固氮生物将大气中的氮气还原成氨的过程,对地球上氮循环和氮平衡具有不可替代的作用。海洋生物固氮作用是维持海洋初级生产力和新生产力的一个重要生态反应。海洋中固氮生物种类较多,主要包括固氮蓝藻、异养细菌和光合细菌。     

一株嗜碱兼性好氧反硝化菌Marinobacter sp.B3的分离鉴定及脱氮性能研究

为获得反硝化脱氮效率较好的菌株,实验从海水螺旋藻培养体系中分离获得一株嗜碱兼性好氧反硝化菌, 通过观察细菌形态以及16S rRNA基因序列的同源性分析, 鉴定该菌株为海杆菌属, 命名为Marinobacter sp. B3。为明确该海杆菌的反硝化性能及氮转化途径, 研究开展了溶解氧(DO), 碳氮摩尔比(C/N), pH和温度等不同单因素对反硝化性能影响实验和氮平衡实验。单因素影响实验结果表明, 当硝酸钾(KNO₃)作为唯一氮源, NO₃^--N的初始浓度为100 mg/L, 盐度32, 振荡速度为150 r/min (初始DO质量浓度是5.6 mg/L), C/N=10, pH=8.0±0.2, 温度为35 °C时, 可获得最大脱氮效果。氮平衡实验结果得出, 在好氧环境下, 有20.11%的NO₃^--N转化为胞内氮, 5.58 mg/L的NO₃^--N转化为其他形态(NO₂^--N、NO₄⁺-N和有机氮), 74.72%转化为N₂释放; 厌氧环境下, 有26.65%的NO₃^--N转化为胞内氮, 72.86%的NO₃^--N转化为气态产物释放。最终实验结果表明, Marinobactersp. B3在好氧和厌氧条件下, 48 h对NO₃^--N的去除率分别为99.89%和93.80%, 具有较好的反硝化脱氮能力, 且在好氧条件下NO₃^--N去除效率更高, 在海水工厂化循环水养殖尾水处理方面具有良好的应用前景。     

养马岛附近海域藻华期间有色溶解有机质的生物可利用性研究

本研究利用吸收光谱和荧光激发-发射矩阵光谱-平行因子分析(EEMs-PARAFAC),研究了养马岛附近海域海水中有色溶解有机质(CDOM)的浓度、组成、来源和生物可利用性,并估算了浮游植物生长繁殖对CDOM及具有生物可利用性CDOM的贡献。结果表明,表、底层海水中CDOM浓度(以吸收系数a₃₅₀计)平均值分别为1.62±0.42 m^-1和1.30±0.47 m^-1,光谱斜率(S_275-295)平均值分别为0.022±0.003 nm^-1和0.023±0.003 nm^-1。利用PARAFAC模型识别出4种荧光组分,分别为陆源类腐殖酸C1、类色氨酸C2、类酪氨酸C3和微生物源类腐殖酸C4。荧光指数(FIX)、腐殖化指数(HIX)和生物指数(BIX)显示,CDOM受陆源输入和海洋自生源的综合影响。降解实验结果显示,表、底层海水中生物可利用性CDOM百分比(%△a₃₅₀)平均值分别为(23.36%±17.94%)和(8.93%±20.30%)。C1、C2和C4组分的荧光强度在培养之后降低,而C3组分的荧光强度上升。各荧光组分生物可利用性依次递减的顺序为:%△C1(23.75%±8.96%)＞%△C4(20.83%±11.71%)＞%△C2(11.67%±38.87%)＞%△C3(-29.61%±39.90%),显示培养之后CDOM的平均分子量和腐殖化程度降低。表层海水中a₃₅₀、%△a350与Chl a之间存在显著线性相关关系,据此可以估算出浮游植物生长繁殖对CDOM的贡献为36.9%,对具有生物可利用性CDOM的贡献为85.0%。     

在流水系统中浒苔对营养盐吸收和转化的模拟实验

在实验室内选用不进行营养盐加富的流水系统模拟研究了浒苔（Ulva prolifera）对海水营养盐的吸收,测定了浒苔体内不同形态的碳、氮、磷的含量变化,探讨了浒苔对海水中碳、氮、磷3种生源要素的转化作用。结果显示,流动海水的实验体系是一个能给藻体创造稳定环境的模拟装置,可以不断地给藻体更新海水和补充营养盐。浒苔对海水中溶解无机氮（DIN）、溶解有机氮（DON）、溶解无机磷（DIP）和溶解有机磷（DOP）都有吸收作用,其平均吸收速率分别为10.87 μmol·g^-1·d^-1、2.41 μmol·g^-1·d^-1、0.183 μmol·g^-1·d^-1和0.023 μmol·g^-1·d^-1。光照时段的DTN和DTP的平均吸收速率与无光照时段的平均吸收速率没有显著差异（P>0.05）。浒苔对海水中无机形态的营养盐的吸收量高于对该元素的有机形态的吸收量;而浒苔的体内有机形态的成分均远高于无机形态的成分。浒苔对无机形态的氮、磷和碳转化为自身有机形态的转化效率分别为：97.33%,99.99%和96.84%。以上结果表明,浒苔能快速吸收无机形态的生源要素并转化为有机形态,能够加快生源物质进入生态系统的物质循环。     

富油微藻富碳培养下油脂积累及固碳研究

微藻固碳是一种新型节能减排技术,具有长期可持续发展的潜力。本文对两株富油微藻（球等鞭金藻和微拟球藻）进行了富碳培养下生长特性及中性脂积累特性的研究。两株富油微藻的最佳培养条件为10%CO₂浓度和f培养基。本研究对两株富油微藻的最大生物量产率、总脂含量、最大油脂产率、微藻的C含量和CO₂固定率进行了测定。球等鞭金藻的各参数指标分别为：142.42±4.58g/（m²·d）,39.95%±0.77%,84.47±1.56g/（m²·d）,45.98%±1.75%和33.74±1.65g/（m²·d）。微拟球藻的各参数指标分别为：149.92±1.80g/（m²·d）,37.91%±0.58%,89.90±1.98g/（m²·d）,46.88%±2.01%和34.08±1.32g/（m²·d）。实验结果显示,两株海洋微藻均属于高固碳优良藻株,适合应用于微藻烟气减排技术开发,具备用于海洋生物质能耦合CO₂减排开发的潜力。     

光谱法测定海水中溶解态无机氮的研究进展

溶解态无机氮（dissolved inorganic nitrogen, DIN）主要由亚硝酸盐-氮（NO^-₂-N）、硝酸盐-氮（NO^-₃-N）和铵氮（NH⁺₄-N）组成,它们在海洋的生物地球化学循环过程中起重要作用。但人类活动向海洋输入了大量无机氮,导致一系列环境问题。为了更好地开展海洋氮循环研究和环境污染管理,需对海水中的DIN进行测定。在众多分析方法中,光谱法因其通用性好、适用范围广、所需设备简单,成为测定海水DIN的首选。本文总结了近10年来基于光谱法测定海水DIN的研究进展,包括紫外分光光度法测定NO^-₃-N、萘乙二胺分光光度法测定NO^-₂-N和NO^-₃-N、次溴酸盐氧化-分光光度法测定NH⁺₄-N、靛酚蓝分光光度法测定NH⁺₄-N、酸碱指示剂-分光光度法测定NH⁺₄-N、荧光法和化学发光法测定DIN等,比较了各分析方法的特点,并展望了光谱法测定海水DIN的发展趋势。总的来说,在分析方法上,新试剂的使用以及一些新合成材料的出现,丰富了DIN的分析手段;在分析仪器上,以流动分析技术为基础的分析仪器在DIN的实验室及现场分析中得到了广泛应用。DIN的分析方法均朝着简单便捷、全自动化、分析速度快、精确度高、可适用范围广的方向发展。     

球形棕囊藻同化吸收硝酸盐的氮氧稳定同位素分馏研究

球形棕囊藻(Phaeocystis globosa)赤潮是一种全球范围的生态灾害,而硝酸盐稳定同位素技术是研究海洋富营养化与赤潮暴发机制的前沿技术。为将该技术应用于球形棕囊藻赤潮暴发机制方面的研究,首先需了解其同化吸收硝酸盐的稳定同位素分馏特征。为此开展了球形棕囊藻室内培养实验,获取培养过程中氮、磷、硅等营养盐的浓度及硝酸盐氮、氧稳定同位素(δ¹⁵N-NO₃^-、δ¹⁸O-NO₃^-)等参数的变化特征,计算球形棕囊藻同化吸收硝酸盐的稳定同位素分馏系数。结果显示,NO₃^-和PO₄^3-浓度随培养时间均呈现先明显下降后稳定的特征,同时伴随直径2~3mm的囊体出现并生长至2~3cm。NH₄⁺浓度先后两次出现升高,推测可能是受到有机氮矿化过程的补充所致。随NO₃^-浓度降低,δ¹⁵N和δ¹⁸O的值分别在第13天和第7天达到相对峰值。经计算,球形棕囊藻同化吸收硝酸盐过程δ¹⁵N和δ¹⁸O分馏系数分别为3.32‰±0.38‰和3.12‰±0.59‰,而前者的分馏系数呈逐渐降低的特点,原因可能是随球形棕囊藻生长,发生酶还原的NO₃^-较参与跨膜运输的NO₃^-比例逐渐升高。研究首次给出了球形棕囊藻同化吸收硝酸盐过程的氮、氧稳定同位素分馏系数及其变化特征,补充了海洋微藻同位素分馏数据库,为稳定同位素技术研究球形棕囊藻赤潮暴发的营养机制提供了重要的基础数据。     

双层反铁磁体K3 Cu2 F7 中轨道序驱动的自旋二聚化

基于自旋-轨道-晶格Hamilton量,应用团簇自洽场方法,研究了双层钙钛矿结构材料K₃Cu₂F₇基态的晶格、磁及轨道结构,发现近孤立的双层的对称破缺和Jahn-Teller晶格畸变使得Cu²⁺离子在每层内交替占据 z²-x²〉/ z²-y²〉轨道,进而导致双层的层间表现为强的反铁磁耦合,层内为弱的铁磁耦合.强反铁磁耦合导致层间     

离子色谱法测定海水中氯离子和硫酸根离子

氯离子和硫酸根离子是海水中重要的无机阴离子,在研究海洋生态变化、海洋循环作用过程与海洋全球气候变化等领域具有重要的指示意义。其测定方法较多,但缺少相应的测试方法。本文对测定海水中Cl^-,SO₄^2-的离子色谱方法进行了优化,选用IonPacAS14碳酸盐选择性离子色谱柱,以3.5 mmol/L Na₂CO₃+1 mmol/L NaHCO₃为流动相,可消除海水样品中碳酸盐及其他阴离子的干扰。该方法对Cl^-检出限为0.29 mg/L,线性相关系数r²=0.999 2,对SO₄^2-检出限为0.42 mg/L,线性相关系数r²=0.997 9。样品的加标回收率在95%~102%,Cl^-和SO₄^2-的相对标准偏差分别为1.92%和4.18%。该方法简便、迅速、灵敏、准确度高,可满足批量海水样品中Cl^-与SO₄^2-的准确测试。     

N2 fixation variability in the oligotrophic Sulu Sea, western equatorial Pacific region over the past 83 kyr

N₂ fixation is an important biological process that adds new nitrogen to oceans and plays a key role in modulating the oceanic nitrate inventory. However, it is not known how, when, and where N₂ fixation rates have varied in response to past climate changes. This study presents a new record of nitrogen isotopic composition (δ¹⁵N) over the last 83 kyr from a sediment core (KH02-4 SUP8) taken in the Sulu Sea in the western equatorial Pacific region; data allow the N₂ fixation variability in the sea to be reconstructed. Sediments, sinking, and suspended particulate organic matter (POM) all have lighter isotopic values compared to the δ¹⁵N values of substrate nitrate (av. 5.8‰) in North Pacific Intermediate Water. These lighter δ¹⁵N values are regarded as reflecting N₂ fixation in the Sulu Sea surface water. A δ¹⁵N mass balance model shows that N₂ fixation rates were significantly enhanced during 54–34 kyr in MIS-3 and MIS-2. It has been speculated that higher interglacial denitrification rates in the Arabian Sea and the eastern tropical Pacific would have markedly decreased the global oceanic N inventory and contributed to the increase in N₂ fixation in oligotrophic regions, but such a model was not revealed by our study. It is possible that changes in N₂ fixation rates in the Sulu Sea were regional response, and accumulation of phosphate in the surface waters due to enhanced monsoon-driven mixing is thought to have stimulated enhancements of N₂ fixation during MIS-3 and MIS-2.     

The relationship between dissolved hydrogen and nitrogen fixation in ocean waters

Fixed nitrogen is a key nutrient involved in regulating global marine productivity and hence the global oceanic carbon cycle. Oceanic nitrogen (N₂) fixation is estimated to supply 8×10¹² moles N y^?1 to the ocean, approximately equal to current riverine and the atmospheric inputs of fixed N, and between 50 and 100% of current estimates of oceanic denitrification. However, the spatial and temporal variability of N₂ fixation remains uncertain, mostly because of the normal low resolution sampling for diazotroph distribution and fixation rates. It is well established that N₂ fixation, mediated by the enzyme nitrogenase, is a source of hydrogen (H₂), but the extent to which it leads to supersaturation of H₂ in oceanic waters is unresolved. Here, we present simultaneous measurements of upper ocean dissolved H₂ concentration (nmol L^?1), and rates of N₂ fixation (μmol N m^?3 d^?1), determined using ¹⁵N₂ tracer techniques (at 7 or 15 m), on a transect from Fiji to Hawaii. We find a significant correlation (r=0.98) between dissolved H₂ and rates of N₂ fixation, with the greatest supersaturation of H₂ and highest rates of N₂ fixation being observed in the subtropical gyres at the southern (～18°S) and northern (18°N) reaches of the transect. The lowest H₂ saturation and N₂ fixation were observed in the equatorial region between 8°S and 14°N. We propose that an empirical relationship between H₂ supersaturations and N₂ fixation measurements could be used to guide sampling for ¹⁵N fixation measurements or to aid the spatial interpolation of such measurements.     

Long‐term nitrogen and phosphorus fertilization effects on N2 fixation rates and nifH gene community patterns in mangrove sediments

The bioavailability of nutrients is important in controlling ecological processes and nitrogen cycling in oligotrophic mangrove forests, yet the variation of diazotrophic community structure and activity with nutrient availability in sediments remains largely unexplored. To investigate for the first time how nutrients in sediments affect spatial and temporal patterns of diazotrophic community structure and activity, the sedimentary environment of Twin Cays, Belize, was examined with respect to the effects of long‐term fertilization [treatments: control (Ctrl), nitrogen (N), and phosphorus (P)] on N₂ fixation rates and nifH gene community structure. We found that N₂ fixation rates were significantly higher at the P‐treatment, intermediate at the Ctrl‐treatment and lower in the N‐treatment (P: 4.2 ± 0.5, Crtl: 0.8 ± 0.1, N: 0.4 ± 0.1 nmol·N·g^?1·h^?1; P < 0.001) with spatial (Ctrl‐ and P‐treatments) and temporal (only P‐treatment) variability positively correlated with live root abundance (r² = 0.473, P < 0.001) and concentration (r² = 0.458, P < 0.0001). The community structure of diazotrophs showed larger spatial and temporal variability in the fertilized treatments than in the Ctrl‐treatment, with the relative abundance of OTUs (nifH operational taxonomic units) at the fertilized treatments inversely related to live root abundance. Overall, long‐term fertilization (with either N or P) affects not only nutrient levels in mangrove sediments directly, but also spatial and temporal patterns of both community structure and activity and likely plant‐microbe interactions as well. Our findings suggest that the maintenance of natural nutrient conditions in mangrove sediments is important to ensure the stability of microbial functional groups like diazotrophs.     

The relative contributions of unicellular and filamentous diazotrophs to N2 fixation in the South China Sea and the upstream Kuroshio

We studied the seasonal, diel, and vertical distribution of phytoplankton N₂ fixation to understand the relative contributions of unicellular and filamentous nitrogen fixers (diazotrophs) to N₂ fixation and nitrogen recycling in the northern South China Sea (SCS) and the neighboring upstream Kuroshio. N₂-fixation rates were measured by the ¹⁵N₂ tracer technique (addition by bubble) on unicellular (<10 or 20 µm) and the filamentous diazotrophs (>10 or 20 µm, mostly Trichodesmium and Richelia) fractionated by 10- or 20-µm mesh sizes. The mean depth-integrated total (unicellular+filamentous) N₂-fixation rates in the SCS (51.7±6.2 µmol N m⁻² d⁻¹) averaged 1/3 of that in the Kuroshio (142.7±29.6 µmol N m⁻² d⁻¹), with higher rates in the winter than in other seasons in the SCS and the opposite seasonal pattern in the Kuroshio. Unicellular diazotrophs contributed 65% of the total N₂ fixation in the SCS, which were negatively correlated with surface temperature and, as for total N₂ fixation, were higher in the winter when Trichodesmium spp. were scarce. In comparison, the unicellular diazotrophs contributed 50% of total N₂ fixation in the Kuroshio, and their contributions were not significantly correlated with surface temperature. In both the SCS and the Kuroshio, the unicellular N₂ fixation was more important during the night than during the day, and in the deep euphotic layer than in the surface layer, even in the daytime. Our results show that the unicellular diazotrophs were important N₂ fixers and contributed significantly to N₂ fixation in the tropical marginal seas, more so in the SCS than the Kuroshio.     

Assessment of excess nitrate development in the subtropical North Atlantic

Geochemical estimates of N₂ fixation in the North Atlantic often serve as a foundation for estimating global marine diazotrophy. Yet despite being well-studied, estimations of nitrogen fixation rates in this basin vary widely. Here we investigate the variability in published estimates of excess nitrogen accumulation rates in the main thermocline of the subtropical North Atlantic, testing the assumptions and choices made in the analyses. Employing one of these previously described methods, modified here with improved estimates of excess N spatial gradients and ventilation rates of the main thermocline, we determine a total excess N accumulation rate of 7.8 ± 1.7 × 10¹¹ mol N yr^− 1. Contributions to excess N development include atmospheric deposition of high N:P nutrients (adding excess N at a rate of 3.0 ± 0.9 × 10¹¹ mol N yr^− 1 for  38% of the total), high N:P dissolved organic matter advected into and mineralized in the main thermocline (adding excess N at 2.2 ± 1.1 × 10¹¹ mol N yr^− 1 for  28% of the total), and, calculated by mass balance of the excess N field, N₂ fixation (adding excess N at 2.6 ± 2.2 × 10¹¹ mol N yr^− 1 for  33% of the total). Assuming an N:P of 40 and this rate of excess N accumulation due to the process, N₂ fixation in the North Atlantic subtropical gyre is estimated at  4 × 10¹¹ mol N yr^− 1. This relatively low rate of N₂ fixation suggests that i) the rate of N₂ fixation in the North Atlantic is greatly overestimated in some previous analyses, ii) the main thermocline is not the primary repository of N fixed by diazotrophs, and/or iii) the N:P ratio of exported diazotrophic organic matter is much lower than generally assumed. It is this last possibility, and our uncertainty in the N:P ratios of exported material supporting excess N development, that greatly lessens our confidence in geochemical measures of N₂ fixation.     

Top-down control on major groups of global marine diazotrophs

Dinitrogen (N₂) fixed by a group of prokaryotes (diazotrophs) is the dominant process adding bioavailable nitrogen into the ocean. Although it has been intensively studied how N₂ fixation is controlled by resources (bottom-up factors), it is unclear whether the grazing (top-down control) effectively impacts growth and distribution of different diazotroph groups. In this study, we evaluate this question by conducting log-log regression of diazotroph biomass onto corresponding N₂ fixation rates in the global ocean. The slope of the regression for Trichodesmium is ~0.8, indicating that a small portion of the increase in N₂ fixation does not accumulate as its biomass. This leads to a conclusion that Trichodesmium is under a substantial top-down control, although bottom-up control still dominates. We also analyze the residuals of the regression in the North Atlantic, concluding that free trichomes of Trichodesmium are subject to stronger top-down control than its colonies. The weak correlation between the biomass and N₂ fixation of unicellular cyanobacterial diazotrophs indicates that the degree of top-down control on this type of diazotrophs varies greatly. The analyses obtain unrealistic results for diatom-diazotroph assemblages due to complicated nitrogen sources of these symbioses. Our study reveals the variability of top-down control among different diazotroph groups across time and space, suggesting its importance in improving our understandings of ecology of diazotrophs and predictions of N₂ fixation in biogeochemical models. Measurements of size-specific N₂ fixation rates and growth rates of different diazotroph groups can be useful to more reliably analyze the top-down control on these key organisms in the global ocean.     

生物固氮速率测定中15N2同位素示踪剂的引入

生物固氮作用是一个重要的海洋新氮来源,在海洋生物地球化学循环中扮演着不可替代的角色。基于稳定同位素（15N₂）示踪吸收法,是目前直接测定海洋生物固氮速率最有效的手段。其中,高效、洁净地将15N₂引入海水培养体系,并准确定量培养体系底物的同位素示踪水平,是同位素示踪吸收法准确获取固氮速率的关键。本研究针对15N₂同位素示踪剂引入这一关键环节进行了探讨,确认改进气泡法是将15N₂引入海水培养体系的首选操作。在对培养体系造成的较小扰动的情况下,可将培养体系氮气底物的15N原子丰度提升至10%以上,相对于另一种导入同位素示踪剂的手段——预溶解海水法,改进气泡法将培养瓶中氮气底物的15N原子丰度提升了近200%。此外,改进气泡法还具有最小化痕量金属沾污、操作简便等优点。将改进气泡法结合与稳定同位素比值质谱测定结合,是准确测定水体生物固氮速率的推荐方法。     

~(15)N示踪法测定海洋反硝化速率中~(29)N_2浓度的准确定量

利用15 N示踪法实测南海水体反硝化速率的研究发现,培养水样在长时间密闭放置过程中也会受到外界空气的污染,且其29N2/28N2比值恒定为0.007 35。根据空气背景中29N2/28N2比值恒定的特征,提出基于质量平衡关系校正空气N2污染的方法,通过将样品实测29N2浓度扣除由外界空气贡献的29N2浓度,可获得由生物反硝化作用所产生的29N2准确浓度,进而可计算出准确的反硝化速率。经空气29N2背景校正后,29N2浓度的偏差明显小于未经校正的结果,且29N2浓度与培养时间之间的线性相关性显著加强,凸显出空气29N2背景校正是获取准确反硝化速率的关键。鉴于15 N示踪法已被广泛应用于海洋水体与沉积物反硝化速率的测定中,所提出的空气29N2背景校正方法具有重要的意义。     

Enteromorpha and the cycling of nitrogen in a small estuary

The nitrogen relations of Enteromorpha spp. growing on intertidal mud flats have been examined over a twelve-month period. Nitrogen assimilation rates using ¹⁵N have been used to calculate the production of the alga and were between 0·046 and 0·217 mg $\text{NH}{}_4{}^{\mathrm{+}}\text{N}$ (g dry wt alga)^?1 h^?1 A considerable quantity of the alga was buried beneath the sediment over the growth season and was calculated to be equivalent to an input of up to 9·52 g N m^?2 per month and 32 g N m^?2 over one complete growth season. Based on carbon, this latter value represented an input of approximately 320 g C m^?2 annually. Low rates of nitrogenase activity (acetylene reduction) were found to be associated with the Enteromorpha. The organisms responsible for the nitrogenase activity were probably heterotrophic bacteria but they did not contribute significant quantities of nitrogen to the alga.