游离单细胞和囊体细胞在球形棕囊藻囊体形成中的作用

球形棕囊藻是我国沿海常见有害藻华物种, 游离单细胞和囊体细胞是其异型生活史中主要的细胞形态。该研究通过分离游离单细胞和囊体细胞分别引发囊体的再次形成, 以阐明两种类型细胞在球形棕囊藻生活史转化过程中的作用。游离单细胞可诱导形成更高的生物量和更多的囊体, 但是两者诱导产生的囊体体积和囊体细胞数量并无显著性差异。游离单细胞诱导产生的囊体在单位表面积上的细胞密度更高, 排列更加紧密, 因此结构更加紧致。同棕囊藻藻华发生时以囊体为优势形态不同, 该研究发现只有不到15%的细胞以囊体细胞的形态存在。游离单细胞和囊体细胞均可引发囊体形成, 有利于球形棕囊藻藻华的形成以及规模的维持。     

球形棕囊藻的生长、囊体形态以及囊体细胞的分布

球形棕囊藻(Phaeocystis globosa)是中国近海海区常见有害藻华原因种, 其异型生活史中包含单细胞和球形囊体两种形态。游离单细胞直径一般为几微米, 囊体最大直径可达3 cm, 巨大的体积可能导致囊体具有特殊的结构和细胞分布。以球形棕囊藻汕头株为研究对象, 测定了囊体直径、囊体细胞丰度和游离单细胞丰度, 并探讨球形棕囊藻囊体形态与细胞分布的关系。研究结果表明, 囊体形态在其异型生活史中占优势, 囊体对生物量的贡献介于38%–95%之间, 在对数生长期的后期和稳定期, 囊体细胞与单细胞相比占绝对优势。囊体细胞数量与囊体直径的对数呈线性相关, 回归线斜率为1.34, 该值显著低于世界海区其它球形棕囊藻株系的研究结果, 表明汕头株单位囊体表面上分布的细胞数更少。中国海区的球形棕囊藻囊体结构和细胞分布与其它株系不同, 在爆发球形棕囊藻的海区, 巨大的囊体能够有效地抵御摄食, 可能对区域海洋食物链结构和功能有重要影响。     

球形棕囊藻囊体形成中光照、营养盐和共存硅藻的影响

在不同光照和营养盐结构条件下半连续培养球形棕囊藻和3种硅藻,研究光照、营养盐限制和硅藻竞争对球形棕囊藻囊体形成的影响。结果表明:高光照显著促进了藻类的生长,球形棕囊藻在低光环境下几乎不形成囊体。球形棕囊藻和3种硅藻对光限制和P限制更加敏感,而在N限制环境中均具有相对较高的生物量。粒径较小的球形棕囊藻游离单细胞和中肋骨条藻在营养盐和光限制条件下比粒径较大的细胞具有更强的竞争能力。硝酸盐是球形棕囊藻囊体形成的营养基础,但是营养盐结构并未改变棕囊藻囊体形态。具有两种生活史状态有利于球形棕囊藻度过资源限制的环境,从而有利于球形棕囊藻在硅藻藻华之后再次形成藻华。     

充气和搅动对球形棕囊藻生长及囊体形成的影响

球形棕囊藻生活史中包含游离单细胞和球形囊体两种生活形态,但是实验室中培养的球形棕囊藻经常无法形成囊体。研究通过向培养基中泵入过滤空气,以及给培养基提供不同程度的搅动,研究了充气和搅动对球形棕囊藻生长及囊体形成的影响。充气和搅动均显著提高了囊体的数量,并且提高了囊体内细胞的生长速率。但是充气对于囊体直径及囊体内细胞密度并无显著影响。搅动则明显的提高了囊体直径和囊体内细胞数量。然而,尽管充气以及搅动有利于球形棕囊藻囊体的形成,但是培养的囊体直径依然小于自然海区中囊体的大小。     

球形棕囊藻游离单细胞的密度与囊体形成的关系研究

球形棕囊藻(Phaeocystis globosa Scherffel)主要以囊体形态形成赤潮,由单细胞向囊体形态的转变是赤潮爆发的关键.本研究推测囊体形成的前提是游离单细胞达到一定密度阈值,当密度低于该阈值时,囊体无法形成.基于此,本文探究了不同条件(温度、营养充气搅动、摄食压力、初始密度)下囊体形成时游离单细胞的密...     

摄食信息对球形棕囊藻和布氏双尾藻竞争结果的影响

浮游动物的摄食信息能增大棕囊藻囊体体积,囊体形成被认为是棕囊藻的诱导性防御机制。利用桡足类火腿伪镖水蚤和异养甲藻海洋尖尾藻释放的摄食信息,研究了诱导性防御对球形棕囊藻和布氏双尾藻的竞争的影响。结果表明,球形棕囊藻接收了火腿伪镖水蚤和海洋尖尾藻释放的摄食信息之后形成更大的囊体。防御启动后的球形棕囊藻比未接收摄食信息的球形棕囊藻更快地形成囊体,且囊体维持的时间更长。对照组和火腿伪镖水蚤摄食信息诱导的球形棕囊藻的生物体积比布氏双尾藻更高,且球形棕囊藻在竞争中占优势;而海洋尖尾藻摄食信息诱导的球形棕囊藻生物体积低于布氏双尾藻,且球形棕囊藻相对布氏双尾藻的竞争力下降。微型浮游动物海洋尖尾藻摄食信息导致球形棕囊藻相对硅藻布氏双尾藻的竞争力的下降,有利于解释硅藻先于棕囊藻发生藻华。     

球状棕囊藻的形态结构、生活史及生态意义研究进展

球状棕囊藻生活史独特,在两种完全不同的生活状态--游离单细胞和粘液质的囊体之间相互转换.生活史中共出现4 种不同类型的单细胞,存在有性和无性两种生殖方式.不同的细胞能够适应不同的环境,占据不同的生态位,有利于棕囊藻细胞的生长.球状棕囊藻的囊体直径最大可达几个厘米,外被坚韧、紧密并且具有弹性,为内部的单细胞提供了一个良好的栖息和保护场所,并且储存着能量可供细胞在营养盐限制条件下继续生长.由于囊体粒径较大,浮游动物难以摄食,并且有效的抵御了细菌和病毒的侵蚀,从而降低了棕囊藻的死亡率.与其它浮游植物相比,球状棕囊藻的竞争策略更加优越,提高了球状棕囊藻在海洋生态系统中的竞争能力.     

球形棕囊藻的生长特性及生活史研究

对球形棕囊藻不同的藻株(香港株HK和汕头株ST)在实验室条件下分别进行了形态学,生活史及生长曲线的研究,结果表明:球形棕囊藻具有一个复杂的异型生活史,具有两种不同的生活形态:群体和游离的单细胞,香港株和汕头株二者单细胞呈球形或近球形,直径大多为3-9μm;群体呈中空的球形囊泡,细胞包埋在胶质囊中较均匀分布,但二者群体大小有较大差异。当培养达到一定阶段,群体衰亡破裂释放大量单细胞。批次培养中球形棕囊藻的生长周期约为20-30d,香港株最适生长温度接近25℃,最大比生长率为038;汕头株的最适生长温度接近30℃,最大比生长率为042,这说明温度是重要的生长限制因子之一。       

涠洲岛海域球形棕囊藻藻华过程中营养盐及浮游植物群落组成分析

球形棕囊藻(Phaeocystis globosa)是近年来北部湾常见的有害藻华原因种,涠洲岛海域作为广西沿海藻华的高发区之一,目前仍缺乏关于该海域球形棕囊藻藻华过程的相关研究。为探究涠洲岛海域球形棕囊藻藻华过程中营养盐的状况及藻华的发生对浮游植物群落组成的影响,对2017年3月下旬涠洲岛周边发生的球形棕囊藻藻华区域进行取样调查。结果表明,藻华期间水体中溶解有机磷含量与细胞及囊体密度呈显著正相关。本次藻华主要受水体中溶解态无机氮含量影响,多个调查站点的无机氮/无机磷(DIN/DIP)<10,且硅酸盐/无机氮(SiO₃^2–/DIN)>1,表明这些站点存在氮限制现象,而氮限制可能是本次藻华消亡的主要因素。调查区域共鉴定出浮游植物4门58种,球形棕囊藻细胞密度最高可达1.04×10⁸ cells/L,占浮游植物总细胞密度的98.28%。球形棕囊藻细胞密度与物种多样性指数存在显著的负相关关系。在营养缺乏条件下,有利于球形棕囊藻成囊的链状硅藻种类具有相对较大的细胞密度。     

棕囊藻北部湾株的18S rDNA分子鉴定

为研究北部湾棕囊藻(Phaeocystis)藻华的成因,采用PCR克隆了棕囊藻北部湾株核糖体18S r DNA序列。结果表明,棕囊藻北部湾株具有游动单细胞与群体结构两种形态;其18S r DNA序列和NCBI基因库中球形棕囊藻(Phaeocystis globosa)的同源性为99%~100%,在系统进化树上与不同海域来源的球形棕囊藻聚在一大分支上,且与球形棕囊藻间的遗传距离均小于其他种。首次从分子生物学上确定棕囊藻北部湾株为球形棕囊藻。     

Grazing and colony size development in Phaeocystis globosa (Prymnesiophyceae): the role of a chemical signal

The bloom-forming prymnesiophyte Phaeocystis globosa forms hollow,spherical, mucilaginous colonies that vary from micrometresto millimetres in size. A recent paper gave the first empiricalevidence that colony size increase in P. globosa is a defensiveresponse against grazers, and knowing the signalling mechanism(s)behind this response will thus be a key to understanding thetrophodynamics in systems dominated by this species. I conductedexperiments with specially designed diffusion incubators, eachof which consists of a non-grazing chamber (with P. globosaonly) and a grazing chamber (grazers + phytoplankton) connectedby 2 µm polycarbonate membrane filters. The results showedthat physical contact with grazers was not required to initiatethe defensive response; instead, P. globosa colony size increasewas found to be stimulated by dissolved chemicals generatedby ambient grazing activities. This signal was non-species specific,such that various combinations of three species of grazers andfour species of phytoplankton in the grazing chambers all resultedin significant, but different extents of colony enlargementin P. globosa in the non-grazing chambers (30–300% largerthan the ‘grazer-free’ control). High concentrationsof ambient solitary P. globosa cells and other phytoplanktonseemed to suppress colony enlargement in P. globosa, and grazerswould help reduce this inhibition by removing the ambient solitaryP. globosa cells and other phytoplankton. These non-species-specificmechanisms would allow P. globosa to regulate colony size developmentand defend itself in diverse planktonic systems, which may helpto explain the global success of this species.     

COLONY SIZE OF PHAEOCYSTIS ANTARCTICA (PRYMNESIOPHYCEAE) AS INFLUENCED BY ZOOPLANKTON GRAZERS1

The haptophyte Phaeocystis antarctica G. Karst. is a dominant phytoplankton species in the Ross Sea, Antarctica, and exists as solitary cells and mucilaginous colonies that differ by several orders of magnitude in size. Recent studies with Phaeocystis globosa suggest that colony formation and enlargement are defense mechanisms against small grazers. To test if a similar grazer‐induced morphological response exists in P. antarctica, we conducted incubation experiments during the austral summer using natural P. antarctica and zooplankton assemblages. Dialysis bags that allowed exchange of dissolved chemicals were used to separate P. antarctica and zooplankton during incubations. Geometric mean colony size decreased by 35% in the control, but increased by 30% in the presence of grazers (even without physical contact) over the 15 d incubation. The estimated colonial‐to‐solitary cell carbon ratio was significantly higher in the grazing treatment. These results suggest that P. antarctica colonies would grow larger in the presence of indigenous zooplankton and skew the carbon partitioning significantly toward the colonial phase. While these observations show that the colony size of P. antarctica was affected by a chemical signal related to grazers, the detailed nature and ecological significance of this signal remain unknown.     

An examination of the role of colonial <Emphasis Type="Italic">Phaeocystis antarctica</Emphasis> in the microbial food web of the Ross Sea

The extensive buildup of phytoplankton biomass in the Ross Sea conflicts with the view that high rates of herbivory occur in all regions of the Southern Ocean. Nano and microplanktonic consumers comprise a significant fraction of total plankton biomass; however, the importance of grazing remains uncertain in the Ross Sea. Microzooplankton ingestion of solitary and colonial cells of Phaeocystis antarctica were calculated using a novel live-staining fluorescently-labeled algae method. Different morphotypes of P. antarctica were stained different colors, mixed, and observed inside Euplotes to determine their feeding preference. The blue (7-aminocoumarin) (CMAC) stain was used on the colonies and the green (CMFDA) CellTracker Probe was used on solitary cells. Both morphotypes can be seen inside the food vacuoles of the ciliate, supporting the idea that microzooplankton are capable of ingesting cells within the colonial matrix. This suggests that P. antarctica colonies enter the microbial loop in the Ross Sea before sedimentation.     

How does eutrophication affect the role of grazers in harmful algal bloom dynamics?

Population dynamics of harmful algal bloom species are regulated both from the “bottom-up” by factors that affect their growth rate and from the “top-down” by factors that affect their loss rates. While it might seem apparent that eutrophication would have the greatest impact on factors affecting growth rates of phytoplankton (nutrient supply, light availability) the roles of top-down controls, including grazers and pathogens, cannot be ignored in studies of harmful bloom dynamics. Lags between the growth of phytoplankton and zooplankton populations, or disruption of zooplankton populations by adverse environmental conditions may be important factors in the initiation of plankton blooms under eutrophic conditions. Grazers that avoid feeding on harmful species and actively graze on competing species may also play important roles in bloom initiation. Grazers that are not affected by phytoplankton toxins and have growth rates comparable to phytoplankton (e.g. protozoan grazers) may have the potential to control the initiation of blooms. If the inhibition of grazers varies with cell density for blooms of toxic phytoplankton, eutrophication may increase the chances of blooms reaching threshold densities for grazer inhibition. In addition, secondary effects of eutrophication, including hypoxia and change in pH may adversely affect grazer populations, and further release HAB species from top-down control. The Texas brown tide (Aureoumbra lagunensis) blooms provide evidence for the role of grazer disruption in bloom initiation and the importance of high densities of brown tide cells in continued suppression of grazers.     

Isolation and phylogenetic analysis of novel viruses infecting the phytoplankton Phaeocystis globosa (Prymnesiophyceae)

Viruses infecting the harmful bloom-causing alga Phaeocystis globosa (Prymnesiophyceae) were readily isolated from Dutch coastal waters (southern North Sea) in 2000 and 2001. Our data show a large increase in the abundance of putative P. globosa viruses during blooms of P. globosa, suggesting that viruses are an important source of mortality for this alga. In order to examine genetic relatedness among viruses infecting P. globosa and other phytoplankton, DNA polymerase gene (pol) fragments were amplified and the inferred amino acid sequences were phylogenetically analyzed. The results demonstrated that viruses infecting P. globosa formed a closely related monophyletic group within the family Phycodnaviridae, with at least 96.9% similarity to each other. The sequences grouped most closely with others from viruses that infect the prymnesiophyte algae Chrysochromulina brevifilum and Chrysochromulina strobilus. Whether the P. globosa viruses belong to the genus Prymnesiovirus or form a separate group needs further study. Our data suggest that, like their phytoplankton hosts, the Chrysochromulina and Phaeocystis viruses share a common ancestor and that these prymnesioviruses and their algal host have coevolved.     

Variability in protist grazing and growth on different marine Synechococcus isolates

Grazing mortality of the marine phytoplankton Synechococcus is dominated by planktonic protists, yet rates of consumption and factors regulating grazer-Synechococcus interactions are poorly understood. One aspect of predator-prey interactions for which little is known are the mechanisms by which Synechococcus avoids or resists predation and, in turn, how this relates to the ability of Synechococcus to support growth of protist grazer populations. Grazing experiments conducted with the raptorial dinoflagellate Oxyrrhis marina and phylogenetically diverse Synechococcus isolates (strains WH8102, CC9605, CC9311, and CC9902) revealed marked differences in grazing rates-specifically that WH8102 was grazed at significantly lower rates than all other isolates. Additional experiments using the heterotrophic nanoflagellate Goniomonas pacifica and the filter-feeding tintinnid ciliate Eutintinnis sp. revealed that this pattern in grazing susceptibility among the isolates transcended feeding guilds and grazer taxon. Synechococcus cell size, elemental ratios, and motility were not able to explain differences in grazing rates, indicating that other features play a primary role in grazing resistance. Growth of heterotrophic protists was poorly coupled to prey ingestion and was influenced by the strain of Synechococcus being consumed. Although Synechococcus was generally a poor-quality food source, it tended to support higher growth and survival of G. pacifica and O. marina relative to Eutintinnis sp., indicating that suitability of Synechococcus varies among grazer taxa and may be a more suitable food source for the smaller protist grazers. This work has developed tractable model systems for further studies of grazer-Synechococcus interactions in marine microbial food webs.     

Zooplankton feeding ecology: does a diet of Phaeocystis support good copepod grazing, survival, egg production and egg hatching success?

The bloom-forming alga Phaeocystis is ingested by a varietyof zooplankton grazers, but is thought to be a poor source offood. We examined copepod grazing on solitary Phaeocystis cellsby adult females of Temora stylifera, and survival, fecal pelletproduction, egg production and egg hatching success in Calanushelgolandicus and T. stylifera over periods of 15 consecutivedays. Phaeocystis cell concentrations were high (1.2–3.6x 10⁴ cells ml^–1 for C. helgolandicus and 2.5–7.9x 10⁴ cells ml^–1 for T. stylifera), but within the rangeof maxima recorded for natural blooms. Both copepods survivedwell and continuously produced fecal pellets (indicating continuousgrazing) on a diet of Phaeocystis. However, egg production ratesfor both copepods were low, even though hatching success ofthe few eggs produced was high. Clearance rates for T. styliferawere higher than for most previous measurements of other copepodsfeeding on Phaeocystis solitary cells at lower cell concentrations.We conclude that even though copepods feed well upon Phaeocystis,resulting poor fecundity on this diet may inhibit copepod populationincreases during blooms, thereby contributing to the perpetuationof blooms. However, the high egg hatching success on this dietargues against Phaeocystis containing chemical compounds thatact as mitotic inhibitors reducing copepod egg viability, suchas those found in some other phytoplankters.     

The effect of grazer size manipulation on periphyton communities

Summary We examined the effect of grazer size on periphyton biomass, size structure, and species compostion by removing the largest invertebrate grazers on artificial macrophytes planted in the littoral of Lake Memphremagog (Que-Vt). A series of exclosures with increasingly fine mesh prevented colonization by large invertebrates but allowed in smaller grazers. Oligochaetes, chironomids, and cladocerans effectively replaced snails so that total grazer biomass in the various treatments was not significantly different from the controls. With one exception, algal biomass, measured as chlorophyll a, did not differ significantly among the various treatments. However algal size and taxonomy were affected because the dominance of large blue-green colonies was apperantly related to the presence of large grazers. The results of the size manipulations were qualitatively similar to those induced in phytoplankton communities by size selective zooplankton grazing and are consistent with models based on general allometric equations.Contribution 181 of the Lake Memphremagog Project, Limnology Research Centre