前言

<正>在人类活动和气候变化共同影响下,我国近海的赤潮问题呈现出致灾危害加剧、致灾藻种增多的变化趋势。例如2012年,福建近海暴发有毒米氏凯伦藻(Kareniamikimotoi)赤潮,造成大量皱纹盘鲍死亡,经济损失超20亿元; 2016年,秦皇岛近海发生因有毒赤潮引起的中毒事件,导致消费者死亡。特别值得注意的是, 2015年广西近海暴发球形棕囊藻赤潮,堵塞滨海核电冷源系统,为国内外首例。赤潮灾害已成为威胁滨海核电冷源安全的新隐患,引起国家领导人及相关部门的高度关注。综观我国近海赤潮问题所带来的危害效应,有两类致灾赤潮最令人关注:一类影响核电冷源安全;另一类危害养殖业发展与人类健康。前者以球形棕囊藻(Phaeocystis globosa)赤潮为代表,后者以米氏凯伦藻等有毒赤     

北部湾海域微型、微微型浮游植物类群季节变化及其与棕囊藻赤潮的关系初探

近年来,北部湾海域连年暴发球形棕囊藻（Phaeocystis globosa）赤潮,对近海生态环境构成了潜在威胁。为了解北部湾海域球形棕囊藻赤潮生消过程及其与其他浮游植物类群的关系,自2016年9月至2017年8月期间,在北部湾海域开展了9个航次的调查,并应用流式细胞仪分析了微型真核藻类、微微型真核藻类、原绿球藻（Prochlorococcus spp.）和聚球藻（Synechococcus spp.）等4类浮游植物类群的变化情况。结果表明,北部湾海域各浮游植物类群主要分布在调查区北部钦州湾外侧海域和雷州半岛西侧近岸海域,其丰度有明显的季节变化。微微型真核藻类在春、秋两季丰度较高,而冬、夏季较低。微型真核藻类、聚球藻和原绿球藻的丰度均呈现夏季高、冬季低的特点,其中聚球藻丰度的季节变化最为显著,与水温变化密切相关。结合对球形棕囊藻赤潮生消过程以及调查海域水文条件的分析可以看出,北部湾海域球形棕囊藻赤潮发生过程中,海水温度下降至20℃左右,并且聚球藻和原绿球藻丰度具有明显变化,有望作为棕囊藻赤潮的预警指标,为该海域棕囊藻赤潮的预测和防控提供依据。     

中国沿海球形棕囊藻(Phaeocystis globosa)的分类、分布及其藻华

球形棕囊藻(Phaeocystis globosa Scherffel)是全球海洋广泛分布的有害藻华种类。1997年10月,中国东南沿海首次暴发了此种藻类的大规模藻华,其后陆续在福建、广东、广西、海南、河北及天津等省市沿海暴发多起同种藻华。中国近海的球形棕囊藻藻华呈现两大独有的特点,即藻类囊体较大(可达3厘米),以及藻华可毒害养殖业。历经20多年,球形棕囊藻在中国沿海已从一个"藻华新记录种"变成了"藻华常见种"。值得注意的是, 2014年以来,广西北部湾海域棕囊藻藻华肆虐,威胁核电冷源安全,受到了社会高度关注,也对球形棕囊藻藻华的研究提出了新的挑战与要求。针对这一生态灾害的最新发展趋势,本文总结了20年来中国球形棕囊藻及其藻华灾害的发生与发展状况,分别就棕囊藻的分类、生活史特征、营养特性、藻华形成的环境驱动因素、生态毒理等诸多方面开展简要综述,冀望为棕囊藻藻华的研究及防治提供基础资料及思路。     

改性黏土法消除球形棕囊藻赤潮的现场实验与效果评估

球形棕囊藻是一种具有复杂异型生活史的有毒有害赤潮生物,近年来在我国近海频繁暴发成灾,形成的巨大囊体为国内外罕见。迄今还未见到有效消除囊体型球形棕囊藻赤潮的方法报道。本文通过海上围隔实验和现场赤潮消除工程跟踪监测,考察了喷洒改性黏土消除囊体型球形棕囊藻赤潮的可行性与效率,并分析了改性黏土法治理赤潮时对水体及沉积环境的可能影响。围隔实验结果表明,喷洒改性黏土可以有效消除水体中过量的微藻细胞,其中以少量多次喷洒方法的效果最好,生物量(chl a)去除率90%。在对2016年2月广西防城港附近海域球形棕囊藻赤潮消除时的跟踪监测结果表明,改性黏土法适用于工程化消除赤潮作业,能够快速消除水体中的大量球形棕囊藻囊体,随改性黏土絮凝体沉入海底的赤潮藻可以快速分解而失去活性。喷洒改性黏土对于赤潮水体的主要理化指标影响不显著,所监测的水体COD、pH和不同形态氮、磷、硅等生源要素浓度都在原有水质水平范围内波动。     

近10年珠江口海域赤潮发生特征及原因初探

对2000-2009年珠江口海域发生的各种赤潮灾害资料进行统计分析,研究珠江口海域赤潮发生的时空分布规律、赤潮种类的变化规律、海水营养限制的变化状况.结果表明,珠江口赤潮多发区为深圳湾海区,近年珠江口西部珠海沿岸海域频发大面积赤潮;赤潮灾害发生的持续时间呈上升趋势;赤潮多发种为中肋骨条藻,多发于深圳湾;定鞭藻类原球形棕囊藻引发的有毒赤潮明显增多且影响范围较大.研究发现,珠海沿岸海域频发大面积赤潮和深圳湾春季频发赤潮可能与出现潜在硅酸盐营养限制有关,冬季频发球形棕囊藻与季节骤然升温有关.     

北部湾海域球形棕囊藻遗传多样性分析

南海北部湾海域近年来连年暴发球形棕囊藻赤潮,严重威胁近海生态安全,亟待探明赤潮成因。球形棕囊藻是一种广泛分布于温带至热带海域的有害藻华微藻,具有明显的种下遗传分化。为了解北部湾海域球形棕囊藻的遗传多样性及其对赤潮藻种来源的指示意义,本研究针对分离于北部湾海域赤潮发生期间的4株棕囊藻,以核糖体28S大亚基rRNA基因（28S rDNA）D1-D2区和转录间隔区（ITS区）为靶区构建克隆文库,通过测序分析了北部湾海域球形棕囊藻的遗传多样性。结果表明,从北部湾分离的4株棕囊藻均为球形棕囊藻,但不同年份分离的球形棕囊藻藻株之间存在明显的遗传差异。部分球形棕囊藻藻株（PG2015和PG2017）的不同克隆间也存在遗传差异,这种遗传差异在由单一藻细胞建立的克隆培养系中仍存在,表明遗传差异是来自细胞内多拷贝的基因序列,可能源自不同地理种群球形棕囊藻之间的基因交流。与ITS区相比,28S rDNA D1-D2区能更好地反映球形棕囊藻遗传多样性状况,具有作为分子标记指示球形棕囊藻地理种群的潜力,但仍需深入研究。研究结果深化了对北部湾球形棕囊藻遗传多样性的认识,有望为进一步解析北部湾海域球形棕囊藻赤潮原因种来源提供判据。     

XX核电厂附近海域生态环境及潜在致灾生物研究

为了了解XX核电厂周边海域的生态环境状况,筛选潜在威胁核电厂冷源安全的致灾生物,分析影响致灾生物时空分布的主要环境因素,本研究于2018年进行了四个季节的生态调查。结果表明,研究海域浮游植物密度秋季最高,春季优势种为东海原甲藻,其他季节优势种为中肋骨条藻等硅藻。浮游动物密度夏季最高,以桡足类为主,春、夏季优势种还包括球型侧腕水母和薮枝螅水母。底栖动物的密度和生物量在秋季最高,优势种主要为豆形短眼蟹、长吻沙蚕和丝异须虫。浮游植物、浮游动物和底栖动物群落均在夏季最为稳定,群落多样性水平和物种丰度较高,且分布较为均匀。浮游植物群落的细胞丰度与悬浮物和磷酸盐的浓度正相关性最高。浮游动物的密度受温度和盐度的影响较大。底栖动物的群落分布主要受悬浮物和无机营养盐的浓度影响。本研究共筛选出17种威胁XX核电厂冷源安全的潜在致灾生物。海地瓜、棘刺锚参、海葵和球型侧腕水母的风险等级较高,其分布主要受温度和盐度的影响。后续工作中,XX核电厂应根据潜在致灾生物与环境因子的关系加强暴发机制研究、提升监测预警能力,并优化驱赶消杀技术和干扰体系,为核电厂冷源安全提供理论和技术保障。     

青岛近海球形棕囊藻的宏条形码分析

自1997年球形棕囊藻(Phaeocystis globosa)赤潮首次在我国南海海域暴发以来,该赤潮在我国南海和渤海的多个海域频繁暴发,表明球形棕囊藻在我国海域的分布比较广泛。尽管如此,在此之前开展的分类和生态调查研究均未发现我国海域球形棕囊藻的存在。近年来基于分子标记扩增和高通量测序的宏条形码分析检测到了球形棕囊藻,显示出宏条形码分析对球形棕囊藻鉴定的优势。然而,由于包括球形棕囊藻在内的定鞭藻的18S rDNA序列具有较高的G(Guanine,鸟嘌呤)C(Cytosine,胞嘧啶)含量等特点,导致针对球形棕囊藻的宏条形码分析结果在很大程度上依赖分子标记的选择或PCR扩增引物的选择。以青岛栈桥海域样本为研究对象,评估了扩增通用分子标记18S rDNA V4的两套常用PCR引物(即Stoeck引物和Song引物)对球形棕囊藻的宏条形码分析结果的影响,并利用Song引物对青岛近海2021年冬季突发性球形棕囊藻赤潮样本开展了宏条形码分析。分析发现不同引物对定鞭藻门物种的分析结果具有显著影响。利用Stoeck引物的宏条形码分析没有获得任何对应的定鞭藻的ASV(ampliconsequence...     

北部湾棕囊藻藻华原因种分析

近年来, 棕囊藻已成为我国北部湾海域藻华发生的主要原因种, 严重影响北部湾海洋生态环境、水产增养殖业和防城港核电安全, 急切需要对棕囊藻藻华原因种作进一步分析确认。2017年2月和3月在钦州湾和涠洲岛藻华发生海域分离得到9株棕囊藻, 采用光学显微镜进行形态学初步鉴定, 并通过核糖体小亚基序列(SSU rDNA)进行系统进化分析。研究结果表明, 北部湾9株棕囊藻与球形棕囊藻形态特征基本符合, 系统发育树与不同海域来源的球形棕囊藻聚在同一大分支上, 与其他海域来源的球形棕囊藻遗传距离为0.0000~0.0071, 其中北部湾WZS3-1株与墨西哥湾株CCMP627、苏里南株CCMP628、厄瓜多尔株CCMP1528、南非株P162、中国海株RCC:K1398及汕头株Santou97的球形棕囊藻亲缘关系最近, 遗传距离为0.0000, 而北部湾WZS1、WZS2、WZS4、WZS5株与香港株SKLMP_T005和渤海株BOHAI1亲缘关系最近, 遗传距离为0.0000。球形棕囊藻的种内遗传距离(0.0000~0.0073)明显小于种间遗传距离(0.0084~0.0440), 因此可确定来自北部湾不同海域的9株棕囊藻均为球形棕囊藻。此外, 球形棕囊藻各藻株间的亲缘关系远近与其地理位置分布不能完全对应。本研究结果将为我国棕囊藻藻华藻种库构建、棕囊藻北部湾株生理生态研究和北部湾棕囊藻藻华发生机理研究等提供参考。     

磷限制对球形棕囊藻生长及TEP形成的影响

广西北部湾近岸海域虽常年为磷限制状态,但球形棕囊藻（Phaeocystis Globosa）赤潮灾害频繁发生。球形棕囊藻暴发时主要以囊体形式存在,赤潮衰退后,囊体破裂产生大量的透明胞外聚合颗粒物（Transparent Exopolymer Particles,TEP）,在海洋动力作用下形成高碳、高黏性泡沫,影响渔业和核电取水安全。本文通过设置氮磷比为32∶1和64∶1的半连续培养实验,探讨磷限制对球形棕囊藻的生长,以及透明胞外聚合颗粒物产量的影响。结果表明：在氮磷比为32∶1和64∶1的磷限制条件下,球形棕囊藻的细胞密度、叶绿素a浓度、囊体密度及囊体表面细胞数,在培养的后半期都受到了抑制。与控制组相比,两个磷限制的处理组在囊体直径上并没有表现出显著的差异（显著性水平p> 0.05）。在氮磷比为32∶1的条件下,球形棕囊藻的TEP生产量与控制组相比没有显著差异（p> 0.05）,在氮磷比为64∶1的条件下,TEP的生产量显著降低（p <0.05）,磷限制程度的加剧对球形棕囊藻细胞分泌TEP产生影响。研究结果为进一步研究环境胁迫下球形棕囊藻TEP形成机制提供了依据。     

Molecular identification of bloom-forming species Phaeocystis globosa (Pryninesiophyta) and its dispersal based on rDNA ITS sequence analysis

The colony-forming Phaeocystis species are causative agents of dense bloom occurrences in coastal waters worldwide. It is difficult to separate them because of the different morphologies associated with their colonial stages. In this study we applied molecular approaches to analyze the genetic variation of Phaeocystis globosa and Phaeocystis pouchetii from several geographic regions, and to assist in tracing the dispersal of bloom-forming Phaeocystis species in coastal waters of China. The sequences of the internal transcribed spacers (ITS1 and ITS2) of rDNA and the 5.8S ribosomal RNA gene of Phaeocystis strains were determined. Sequence comparison shows that P.globosa was the most divergent to P. pouchetii, exhibiting sequence divergence higher than 0.08. However, lower genetic divergences existed between strains of P.globosa. The sequence comparison of the Phaeocystis rDNA ITS clearly shows that the species isolated from the southeast coast of China is identified as P.globosa rather than P. cf. pouchetii or other species. Furthermore, the significance of rDNA variation in distinct global populations of P.globosa suggested it might have had sufficient time to accumulate detectable mutations at the rDNA locus, supporting the hypothesis of ancient dispersal of P.globosa to many areas, meaning that P.globosa blooms in the coastal waters of China are endemic rather than a newly introduced species or a foreign source. Finally, based on the high divergent region of rDNA ITS, a pair of species-specific primers for P.globosa were designed, they could be useful to detect the presence of this species in mixed plankton assemblages or flagellate stages that are recognized with diffculties by means of conventional microscopy.     

某核电厂附近海域生态环境特征及潜在致灾生物研究

为了了解某核电厂周边海域的生态环境状况,筛选潜在威胁核电厂冷源安全的致灾生物,分析影响致灾生物时空分布的主要环境因素,本研究于2018年进行了4个季节的生态调查。结果表明,研究海域浮游植物密度秋季最高,春季优势种为东海原甲藻(Prorocentrumdonghaiense),其他季节优势种为中肋骨条藻(Skeletonema costatum)等硅藻。浮游动物密度夏季最高,以桡足类为主,春、夏季优势种还包括球型侧腕水母(Pleurobrachiaglobosa)和薮枝螅水母(Obeliasp.)。底栖动物的密度和生物量在秋季最高,优势种主要为豆形短眼蟹(Xenophthalmus pinnotheroides)、长吻沙蚕(Glycera chirori)和丝异须虫(Heteromastusfiliforms)。浮游植物、浮游动物和底栖动物群落均在夏季最为稳定,群落多样性水平和物种丰度较高,且分布较为均匀。浮游植物群落的细胞丰度与悬浮物和磷酸盐的浓度正相关性最高。浮游动物的密度受温度和盐度的影响较大。底栖动物的群落分布主要受悬浮物和无机营养盐的浓度影响。本研究共筛选出17种威胁该核电厂冷...     

Mesoscale and microscale spatial variability of bacteria and viruses during a Phaeocystis globosa bloom in the Eastern English Channel

Sampling was conducted within inshore and offshore sites, characterized by highly dissimilar hydrodynamic and hydrobiological conditions, in the Eastern English Channel. The eutrophic inshore site was dominated by the influence of a dense bloom of the Prymnesiophyceae phytoplankton species Phaeocystis globosa, while the offshore site was characterized by more oceanic conditions. Within each site the microscale distributions of chlorophyll a and several flow cytometrically-defined subpopulations of heterotrophic bacteria and viruses were measured at a spatial resolution of 5 cm. The inshore site was characterized by comparatively high levels of microscale spatial variability, with concentrations of chlorophyll a, heterotrophic bacteria, and viruses varying by 8, 11 and 3.5-fold respectively across distances of several centimeters. Within the offshore site, microscale distributions of chlorophyll a and bacteria were markedly less variable than within the inshore site, although viruses exhibited slightly higher levels of heterogeneity. Significant mesoscale variability was also observed when mean microbial parameters were compared between the inshore and offshore sites. However, when the extent of change (max/min and coefficient of variation) was compared between meso- and microscales, the variability observed at the microscale, particularly in the inshore site, was substantially greater. This pattern suggests that microscale processes associated with Phaeocystis globosa bloom dynamics can generate heterogeneity amongst microbial communities to a greater degree than large scale oceanographic discontinuities.     

An inventory of factors that affect polysaccharide production by Phaeocystis globosa

Phaeocystis material contains polysaccharides that are built from at least eight different monosaccharides. Differences have been reported between the carbohydrate composition of different Phaeocystis species, and also between samples taken from Phaeocystis globosa blooms in different areas. In order to elucidate factors that could play a role in determining variation in carbohydrate composition and production, a number of Phaeocystis globosa strains were studied under laboratory conditions. Although there was a clear distinction of a northern and a southern cluster in the Phaeocystis globosa strains based on RAPD analysis, the differences in the composition of the mucopolysaccharides were relatively small. The contribution of glucose, however, ranged from 7–85% of total sugars. A strain that was cultured in seawaters of diverse origin produced polysaccharides of a different composition, suggesting the effect of environmental factors. The presence of bacteria affected neither the amount, nor the composition of the carbohydrates that were produced by Phaeocystis globosa. Glucose is part of both the intracellular polysaccharide pool and of the mucopolysaccharides in the colony matrix. Using specific digestion of the intracellular chrysolaminaran by laminarinase, the distribution of polysaccharides over different pools could be assessed. During growth of an axenic, mucus-producing strain, the portion of glucose present as chrysolaminaran appeared to increase. The polyglucose that was not digested by laminarinase remains unidentified. This study shows that environmental factors rather than strain differences determine differences in the sugar composition of Phaeocystis globosa, especially with respect to the glucose content of the material. A difference in the contribution of glucose could be correlated to the portion of cells in the culture that are not in the colonies. Our study emphasises that for studying polysaccharide dynamics in Phaeocystis globosa it is important to be able to discriminate between the different polysaccharide pools. Preliminary results of an enzymatic approach were promising     

中国新记录定鞭藻物种冠状棕囊藻的遗传进化分析

棕囊藻属(Phaeocystis)分类学上位于定鞭藻门(Haptophyta)定鞭藻纲(Haptophyceae)棕囊藻目(Phaeocystales)棕囊藻科(Phaeocystaceae)。迄今在我国海域仅分离鉴定到一种棕囊藻属物种,即自1997年以来在我国海域频繁形成有害藻华的球形棕囊藻(Phaeocystis globosa)。作者从中国南海分离到一个单细胞鞭毛类株系CNS01077,结合形态特征观察及基于18S rDNA序列的系统发育分析,将其鉴定为冠状棕囊藻(Phaeocystis rex)。这是该棕囊藻物种在我国海域的首次报道。该研究构建了该物种的首个叶绿体基因组序列和首个线粒体基因组序列。与球形棕囊藻和南极棕囊藻的细胞器基因组比较分析发现冠状棕囊藻的细胞器基因组发生了显著的结构重排和序列变异。该物种在我国海域的发现及细胞器基因组的构建,将为棕囊藻的生物多样性组成和地理分布研究提供支撑。     

Mesozooplankton grazing during the Phaeocystis globosa bloom in the southern bight of the North Sea

During spring blooms 1998 and 1999, three complementary methods were used to evaluate the in situ feeding activities of the dominant copepod species of the Belgian coastal zone: gut pigment content analysis using HPLC, the ¹⁴C tracer method, and cell count experiments. The results obtained by all three methods consistently showed that Phaeocystis globosa is not an adequate food source for the spring copepods in the Belgian coastal zone. Our results demonstrated that, among the potential prey, copepods strongly selected diatoms and microzooplankton, and that these types of prey accounted for the major part of the ingested carbon. However, diatoms and microzooplankton ingestion did not always seem sufficient in terms of carbon to avoid food limitation. Comparison of clearance rates exerted on different potential prey types during the P. globosa peak with those before and after the P. globosa peak showed that the copepods' feeding pressure on diatoms was reduced during the P. globosa peak while that on microzooplankton was not. The low grazing pressure on P. globosa, together with the preferential grazing on diatoms, which reduces the competition for nutrients, and the predation on microzooplankton organisms, which reduces the microzooplankton grazing pressure on P. globosa cells, are likely to favour the P. globosa bloom in the Southern Bight of the North Sea.     

Effects of irradiance on pigment signatures of harmful algae during growth process

Three harmful algal bloom (HAB) species, Phaeocystis globosa, Thalassiosira rotula, and Proro-centrum donghaiense were isolated from the coast of China and cultured in batches at three light intensities (40, 70 and 150 μmol photons·m^-2·s^-1). The variation patterns of cell numbers and growth rates with light intensity during growth process were different among species. In P. globosa and T. rotula, maximum growth rates were found at 150 μmol photons·m^-2·s^-1 and ranged from 0.60 divisions per day in T. rotula, to 1.17 divisions per day in P. globosa. The highest growth rate of P. donghaiense, however, was found at 70 μmol photons·m^-2·s^-1 (0.36 divisions per day). In general, all the three HAB species showed adaptation to increasing light intensity by decreas-ing cellular concentrations of chlorophyll a (Chl a), but the variation patterns during the growth process were species-specific. The cellular concentrations of Chl a in P. donghaiense and T. rotula increased gradually with incubation time, but the opposite trend was found in P. globosa. Most of the pigment ratios and pigment indices of these three species were nearly constant during the growth process and showed small changes at different light intensities illustrating the applicability of chemotaxonomy during the initial and developing stages of HAB events, which is very important to study the ecological issues related to HAB species. Ratios of photoprotective carotenoids, such as diadinoxanthin, diatoxanthin and β,β-carotene to total chlorophylls a (Tchl a) showed the trend of increasing with the increase of light intensity during growth process. The species-specific and pigment-specific variations in pigment ratios/indices at different light intensities during growth pro-cess probably reflected the differences in the pigment composition as well as the adaption capabilities of different species to the changes of physical conditions.