卵孢金孢藻(Chrysosporum ovalisporum)——中国水华蓝藻新记录

卵孢金孢藻(Chrysosporum ovalisporum(Forti)Zapomelováet al.)是一种在欧洲地中海地区和澳大利亚较常见的水华蓝藻,但我国目前还未有该物种分布的报道。本文对采自上海市滴水湖中的藻种进行分离培养,获得了一株纯化藻株CFWA01007,经光学显微镜观察,初步判定其为卵孢金孢藻,对其形态特征进行了详细描述并测定了其16S rRNA基因序列,结果与来自欧洲和北美的Chrysosporum ovalisporum(Forti)Zapomelováet al.相似度极高,故判断其即为卵孢金孢藻,这是该种在中国的首次发现。对该种的分类学和生态学特征进行了讨论。     

镇海水库拟柱孢藻的分离鉴定和氮磷对其生长的影响

以分离自广东省镇海水库的拟柱孢藻N8为对象, 探究其在不同磷浓度及氮磷浓度组合下的生长情况。结果表明, 拟柱孢藻N8对磷的适应范围很宽, 在0.025.12 mg/L磷浓度下均能生长, 最适生长磷浓度范围为0.165.12 mg/L, 磷浓度的升高能显著延长拟柱孢藻的对数生长期和提高生物量。动力学分析表明, 拟柱孢藻N8有较低的KSP值, 对磷元素的亲和性较高, 在磷营养贫乏的环境下更容易形成优势。在氮磷组合实验中, 低氮(0.5 mg/L)显著抑制拟柱孢藻的生长, 且这种生长抑制不受磷浓度的影响; 而在低磷(0.04 mg/L)条件下, 水体中氮浓度的增加会显著促进拟柱孢藻的生长, 拟柱孢藻在高氮中磷和高氮高磷下的生长显著优于其他氮磷组合条件。研究表明, 广东省水库拟柱孢藻的生长受磷的限制较弱, 氮是其生长的决定因子。       

光照-温度交互作用及不同氮源对拟柱孢藻生长的影响

为探讨不同环境因子对拟柱孢藻(Cylindrospermopsis raciborskii)生长的影响,对从广东省镇海水库分离的2株拟柱孢藻在不同的温度和光照组合,及不同氮源条件下的生长进行了研究。结果表明,在3种光强下,拟柱孢藻N1和N8藻株的生长随温度的上升而增加,均在28℃高光强下达到最大比生长速率,而N8藻株对低温的适应能力要高于N1藻株。拟柱孢藻N1和N8在各硝态氮浓度下均能正常生长,但仅能在中低浓度的铵态氮和尿素氮中生长,高浓度(128~247 mg L–1)的铵态氮和尿素氮会显著抑制藻细胞生长;在3种氮源下,N1藻株的比生长速率均显著大于N8藻株,这说明N1藻株对不同氮源的利用能力要高于N8藻株。因此,广东省的拟柱孢藻具株系多样性,喜好高温,适应较宽的光照范围,并可利用多种氮源用于生长,这可能是该地区拟柱孢藻水华频发的原因。     

大沙河水库拟柱孢藻昼夜垂直分布特征

为了解大沙河水库拟柱孢藻(Raphidiopsis raciborskii)昼夜垂直分布模式,于2018年9月20—21日(丰水期)和2018年12月31日(枯水期)对该水库拟柱孢藻进行昼夜定点分层采样,分析影响拟柱孢藻垂直分布的主要环境因子。用Morisita指数和平均滞留深度定量检验拟柱孢藻垂直分布格局;用方差分析检验拟柱孢藻是否存在昼夜垂直迁移现象,并采用多元回归方法分析拟柱孢藻垂直分布格局和环境因子的关系。结果表明:拟柱孢藻在丰水期呈现集群分布模式,主要分布于0.5～2 m水层,平均滞留深度为1.5 m;枯水期在6 m水深以上的水层呈现随机分布模式;丰水期和枯水期均无明显的昼夜垂直迁移现象;大沙河水库温跃层的出现是影响拟柱孢藻的昼夜垂直分布模式的主导因素。     

华南热带水库拟柱孢藻产毒基因型的低优势及影响因子分析——以珠海市水库为例

研究以珠海市20座水库两次(2018年和2022年)调查数据为基础,分析华南热带地区产毒拟柱孢藻的发生规律和拟柱孢藻毒素(CYN)的健康风险。基于rpoC1和cyrJ基因的荧光定量PCR分析发现,在两次采样的40个样品中均检到拟柱孢藻,在其中的33个样品中检到产毒拟柱孢藻。种群内产毒基因型的比例在0.046%—38.66%,表明产毒株广泛存在但不占优势。与2018年相比, 20座水库的总拟柱孢藻丰度均值在2022年增加近10倍,但两年的产毒拟柱孢藻丰度无显著差异;产毒基因型的比例明显下降,比例均值和最大值分别从2018年的14.86%和38.66%降至2022年的4.17%和17.24%; CYN浓度与产毒细胞比例具有类似的变化趋势,平均浓度也从0.56降至0.19μg/L。线性混合效应模型分析表明,非产毒基因型丰度随无机磷的升高及硝氮的下降而显著增加,而产毒拟柱孢藻比例随水温升高而增加。     

拟柱孢藻N8全基因组测序及磷吸收转运通路的比较分析

为从分子水平解析热带特征性蓝藻-拉氏拟柱孢藻(Cylindrospermopsis raciborskii, 简称拟柱孢藻)环境适应机制, 研究以广东省镇海水库中分离的拟柱孢藻藻株N8为材料, 采用PacBio单分子实时测序技术(Single Molecule Real-Time, SMRT)进行测序, 并初步进行全基因组特征的比较分析。结果显示拟柱孢藻N8全基因组大小为3.857 Mb, GC含量为40.13 %, 预测编码基因个数为3598个, 在COG、KEGG和GO数据库中注释到的基因数分别为2429、1664和2244个。N8藻株与美国国立生物技术信息中心(NCBI)上公布的27株拟柱孢藻基因组大小和GC含量基本一致, 但N8的编码基因数最多。基于拟柱孢藻全基因组单拷贝基因的系统进化树表明N8藻株与韩国藻株GIHE 2018的亲缘关系最近。N8藻株的基因组中未发现拟柱孢藻毒素(CYN)和石房蛤毒素(STX)合成酶基因簇, 表明该藻株不产蓝藻毒素CYN和STX。对N8藻株和其他7藻株的磷吸收转运通路比较分析表明, 这些藻株均拥有较为完整的磷吸收转运基因(双组分调节系统、低亲和力无机磷转运基因、高亲和力无机磷转运系统、有机磷酸盐转运复合体、C-P裂解酶和碱性磷酸酶), 表明拟柱孢藻具有灵活利用环境中不同形态磷源的潜能; 而不同藻株间的基因拷贝数和排列顺序存在差异, 如拟柱孢藻N8基因组中有2个C-P裂解酶复合体蛋白phnF和phnM, 其余7株拟柱孢藻则只有1个, 表明拟柱孢藻存在株系特异性。N8藻株为中国首株公开完成全基因组测序数据的拟柱孢藻, 对其基因组分析和磷吸收转运通路的解析将有助于阐明华南地区拟柱孢藻水华优势形成的分子机制。     

采自青海湖的刚毛藻属一新种——青海刚毛藻

对青海湖的丝状藻类进行了样品采集, 分析引起水华的藻类种类和组成。基于SSU-LSU的联合进化分析和基于ITS的系统进化分析显示: 形成青海湖水华的丝状藻为一种刚毛藻, 该种与史氏刚毛藻Cladophora stimpsonii和散束刚毛藻Cladophora vagabunda具有较近的亲缘关系, 属于海洋刚毛藻和淡水刚毛藻的中间过渡种类。和邻近种类的区别特征如下: 该种生长于咸水生境, 植物体纤细柔软, 呈绿色或浅绿色, 分枝旺盛, 具明显的顶端生长和居间生长。细胞直径较小。主轴细胞直径30.0—90.0 μm, 细胞长宽比2.4—8.0。分枝细胞直径25.0—70.0 μm, 长宽比4.0—12.3。顶端细胞直径25.0—50.0 μm, 长宽比3.6—12.0。基于形态学特征和分子系统发育分析, 将其定为一新种, 即青海刚毛藻Cladophora qinghaiensis sp. nov.。     

柱孢鱼腥藻的藻蓝蛋白亚基和F_(678)色素蛋白

柱孢鱼腥藻的藻蓝蛋白包含有两个亚基。β亚基具有578nm吸收峰和600nm荧光发射峰,分子量19.80±0.40KD,可表明β亚基仅有一个载色团。α亚基具有578nm和630nm吸收峰及645nm的荧光发射峰,分子量17.35±0.38,α亚基可能有两个载色团。别藻蓝蛋白有635nm和650nm吸收蜂及664nm的荧光发射峰。具藻胆体的类囊体膜从高盐浓度缓冲液移至低盐浓度缓冲液时表现678nm荧光发射峰,可能柱孢鱼腥藻存在的F_(678)色素蛋白相当于GLazer(1975)的别藻蓝蛋白B。     

卵孢金孢藻对不同磷化合物的利用及生长响应

为了解水华蓝藻卵孢金孢藻对水体不同磷化合物的利用能力,试验以磷酸氢二钾为对照,通过室内模拟的方法探究不同磷基质条件下卵孢金孢藻的生长响应.结果表明: 卵孢金孢藻能直接利用三聚磷酸钠和十水焦磷酸钠,且对三聚磷酸钠有极高的利用率.15天后,三聚磷酸钠处理组的藻细胞生物量和叶绿素a浓度最高,分别达到(426.96±47.42) mg·L^-1和(1852.34±116.60) μg·L^-1.2-氨基乙基膦酸和五水β-甘油磷酸钠处理组生物量与对照无明显差异,可溶性无机磷变化特征响应了碱性磷酸酶活性大小,表明卵孢金孢藻能通过酶水解利用这两种有机磷.而整个培养过程中,草甘膦处理组可溶性无机磷浓度趋近于0 mg·L^-1,藻细胞生物量、比生长速率、叶绿素a浓度及光合活力均显著低于对照,表明草甘膦不仅不利于藻细胞摄取磷,还对其生长产生抑制作用.本研究结果为了解卵孢金孢藻向不同水生态系统的扩散机理提供了新的思路,对我国新型蓝藻水华的防治具有一定的理论参考价值.     

南海长茎葡萄蕨藻共附生真菌的多样性

为了揭示南海长茎葡萄蕨藻Caulerpa lentillifera共附生真菌的群落结构,并分析自来水清洗对真菌群落多样性的影响,本研究以收集于深圳南海各月份长茎葡萄蕨藻为材料,利用Illumina MiSeq 2500测序平台进行真菌ITS1扩增子测序,分析自来水清洗前后真菌类群组成和多样性的差异。研究发现,从所有18个样品中总共获得914个OTUs,检测出真菌3门12纲32目50科74属106种,其中优势属为曲霉属Aspergillus、单胞瓶霉属Phialemonium、枝孢属Cladosporium、链座菌属Catenulostroma和茎点霉属Phoma;不同月份真菌Alpha多样性指数显示南海长茎葡萄蕨藻共附生真菌丰度和多样性均随着温度降低而降低;自来水清洗前后南海长茎葡萄蕨藻共附生真菌的多样性及群落结构存在一定差异,清洗后真菌OTUs和Alpha多样性指数均有所降低,其中Chao1和Ace指数为显著性下降,表明自来水清洗对真菌群落组成丰度及多样性影响显著,这对长茎葡萄蕨藻的食品安全具有积极意义。     

水华鱼腥藻类菌胞素氨基酸的分子鉴定和化学检测

类菌胞素氨基酸 (MAAs) 是一类具有吸收紫外线能力的物质, 蓝藻MAAs的生物合成及其分子机制的揭示为MAAs基因的快速检测提供了可能。研究采用分子生物学方法扩增了一株分离自太湖的水华鱼腥藻(Dolichospermum flos-aquae CHAB1629)编码脱氢醌合成酶(DHQS)基因的部分片段, 系统树分析发现其与报道的Anabaena sp. 90核酸序列相似度达99%, 而与Anabaena variabilis ATCC 29413仅为53.6%; 同时运用HPLC检测发现, 该株水华鱼腥藻MAAs的类型为shinorine。研究结果可为后续浮游类鱼腥藻MAAs的分子鉴定及野外适应性研究提供依据。       

Invasibility of a reservoir to exotic Daphnia lumholtzi: experimental assessment of diet selection and life history responses to cyanobacteria

SUMMARY 1. In reservoirs of the south-central United States, the exotic cladoceran Daphnia lumholtzi is common during warm midsummer conditions, when cyanobacteria are abundant and native Daphnia are rare. In the current study, we employed surveys, field experiments, and a life table experiment to investigate the role of food quality in explaining the distribution and phenology of D. lumholtzi , relative to two native species ( Daphnia parvula and Daphnia mendotae ).
2. During May–September 2000 in eutrophic McDaniel Lake, Missouri U.S.A., cyanobacteria (primarily Oscillatoria ) first appeared at 6-m depth and then became abundant throughout the epilimnion.
3. During the May field experiment, D. lumholtzi , D. parvula and D. mendotae all consumed a similar diet of algae, showing positive selection for small greens (chlorophytes and cryptophytes <20 μm). During the July experiment, when the epilimnion exceeded 25 °C and cyanobacteria were common in the lake, D. lumholtzi consumed significantly more total algae and more cyanobacteria than the two native species. Although the Daphnia selected against cyanobacteria, all three species consumed about 25% of this food in their diet.
4. A life table experiment compared the responses of D. lumholtzi and D. parvula with variation in density of high-quality food ( Ankistrodesmus ) and concentration of a toxic strain of cyanobacteria ( Anabaena flos-aquae ). Both Daphnia species showed reduced survivorship, fertility and intrinsic rates of increase in response to elevated concentrations of cyanobacteria, particularly at the higher food level.
5. The results suggest that D. lumholtzi shows similar inhibition from cyanobacteria as does the native Daphnia . However, their continued high in situ feeding rates imply that D. lumholtzi is less affected by midsummer conditions in warm-water reservoirs than are native Daphnia .     

Generational and dual-species exposures to invertebrate predators influence relative head size in Daphnia mendotae

Laboratory cyclomorphosis experiments with crustacean zooplanktonoften fail to produce morphological trait expression as extremeas that observed in field populations. A seven-generation laboratoryexperiment found that the maximum expression of relative headarea in a round-helmet morph of Daphnia mendotae was delayeduntil the third generation of non-lethal, chemical exposureto Leptodora kindtii. By the third generation and after, relativehead area remained high and comparable in degree to field specimens.The induction pattern from parents to offspring was statisticallypositive into the fourth generation. Published studies on kairomoneinduction of morphological trait expression extending only onegeneration may fail to measure transgenerational effects, whichcould account for mismatch between laboratory and field results.In a second laboratory experiment with a pointed-helmet morphof D. mendotae exposed to L. kindtii and Chaoborus punctipennis,we show that induction levels are not the same for all predatorsor combinations of predators. The maximum relative head areafor pointed morph individuals exposed to both predator cueswas still not as extreme in the laboratory as in the field.This is the first demonstration that delayed generational effectsof kairomone induction of helmet expression in the Cladoceracan reconcile laboratory experiments with the field observationsof cyclomorphosis.     

Detection of Microcystin-Producing Cyanobacteria Microcystis,Planktothrix, and Dolichospermum Using Multiprimer Amplification of the mcy Genes

Microbiology - A multiprimer PCR-based technique for the joint molecular detection of microcystin-producing planktonic cyanobacteria of the genera Microcystis, Planktothrix and Dolichospermum was...     

Genetic assessment of species boundaries in the North American Daphnia longispina complex (Crustacea: Daphniidae)

Species boundaries in the North American Daphnia longispina group have proved difficult to establish on the basis of morphology alone. This confusion may be due to hydridization, phenotypic plasticity or the existence of sibling species. We therefore used genetic analysis to delineate species boundaries by examining 27 North American populations belonging to the longispina complex for variation at 15–26 allozyme loci. The populations consisted of Daphnia thorata from two western sites and two eastern sites, Daphnia galeata mendotae from its type location and seven sites across its range, and Daphnia rosea from eight temperate and seven arctic sites. Two populations from the Eurasian longispina complex were also included for reference. Populations assigned to D. galeata mendotae formed a genetically cohesive group, whereas a genetic dichotomy was found between temperate and arctic D. rosea , suggesting that this taxon includes two species. Genetic analysis also confirmed the distinctness of western D. thorata from other members of the longispina group. Unexpectedly, eastern populations resembling D. thorata were genetically more similar to temperate D. rosea than to any helmeted species ( D. galeata, Daphnia hyalina or D. thorata ). Our results suggest that the helmet character is a poor indicator of phylogenetic relationships, as the genetic ability to produce this feature has been lost or acquired several times in the evolution of the longispina group.     

Alternative antipredator responses of two coexisting Daphnia species to negative size selection by YOY perch

Our study showed for the first time in nature that two coexistingDaphnia adopted alternative life history and behavioural strategiesto cope with negative size-selection predation by gape-limitedyoung-of-the-year (YOY) perch. We evaluated the phenotypic plasticityin life history and behavioural traits of two coexisting Daphniaspecies, D. pulicaria (2 mm) and D. galeata mendotae (1.4 mm),in response to seasonal changes in predation by YOY yellow perch(Perca flavescens) in a mesotrophic lake. We expected that thelarge-sized D. pulicaria, the most likely subjected to size-selectivepredation by YOY perch, will show stronger antipredator responsesthan the small-sized D. galeata mendotae. To test this hypothesis,we examined changes in life history and behavioural traits injuveniles and adults of both species during four YOY fish predationperiods that were selected based on the presence of YOY perchin the pelagic zone and the relative abundance of Daphnia preyin their gut contents. Our study supports the scenario of negativesize-selective predation by gape-limited YOY perch on both Daphniaspecies. The electivity index indicated that no daphnids witha body length > 1.75 mm were predated by YOY yellow perch.Coexisting Daphnia exhibited phenotypic plasticity in theirantipredator defenses based on their vulnerability to seasonalchanges in size-selective predation of YOY perch. Juvenile Daphniawere the targeted prey and they responded by a decreased bodylength. Behavioural defenses were the dominant strategy usedby both adult Daphnia populations to withstand high predation.A decreased size at maturity was not employed by Daphnia, exceptat the very end of the predation period. Behavioural defensesare short-term strategy adopted to avoid predation. Both antipredatordefenses became unnecessary expenses and were no longer sustainedafter the predation period.     

Phosphorus strategy in bloom-forming cyanobacteria (Dolichospermum and Microcystis) and its role in their succession

Dolichospermum (formerly Anabaena) and Microcystis cause harmful cyanobacterial blooms in freshwater ecosystems worldwide. Input reduction of both nitrogen (N) and phosphorus (P) are commonly recognized as basic ways of controlling blooms, but little is known about the roles of nutrients and their using strategy among cyanobacteria in triggering the succession of diazotrophic to non-diazotrophic cyanobacteria. In this study, we investigated in situ responses of cyanobactria to ambient P status during the transition from Dolichospermum flos-aquae to Microcystis spp. in Lake Taihu and Lake Chaohu. While dominant in phytoplankton community, D. flos-aquae experienced P deficiency as evidenced by qualitative detection of extracellular phosphatase via enzyme labeled fluorescence (ELF). The percentage of ELF-labelled D. flos-aquae cells was 33% when it dominated the phytoplankton community, and was 78% when it co-dominated with Microcystis spp., indicating an increase in P deficiency. Meanwhile, no ELF-labelled Microcystis cells were observed while polyphosphate body (PPB) were present, suggesting that Microcystis spp. were not P deficient. Additionally, the percentages of Microcystis cells containing PPB showed an inverted “U-shaped” relationship with concentrations on soluble reactive phosphorus (SRP). To validate the field observation, a laboratory study of the monocultures of the dominant cyanobacteria was conducted. Extracellular alkaline phosphatase activity (APA) and PPB accumulation were regulated by P availability in monocultures of D. flos-aquae. Interestingly, no cell bound extracellular phosphatase was found on Microcystis aeruginasa even in the culture without P supply. Consistently, the expressions of phosphatase encoding gene phoX showed no differences among the treatments. The way in which PPB accumulation occurred in Microcystis spp. in response to P availability in the cultures was similar to that observed in the field, demonstrating a strategy of energy conservation over P accumulation. The competitive advantage of Microcystis spp. was displayed at low P concentrations: where it could rapidly uptake and store inorganic P, which also increased the P deficiency of the coexisting phytoplankton species. Responses of P-transport gene pstS confirmed this hypothesis. The physiological and molecular mechanisms mentioned above enable Microcystis to survive and proliferate in environment with low available P supply more efficiently. In conclusion, different cyanobacterial species have distinct ways of responding to P availability, suggesting that the control of cyanobacterial blooms by targeted nutrient reduction is largely dependent upon the dominant species. P reduction is more effective in controlling diazotrophic cyanobacteria than non-diazotrophic cyanobacteria.     

中国蓝藻植物的新记录属-拟甲色球藻

拟甲色球藻(Chroococcidiopsis Geitler 1933)在全球分布广泛, 并在极端环境下多有发现, 但目前在中国尚无报道。研究在对太湖水体进行野外调查时观察到了水体中的Chroococcidiopsis, 并分离得到纯培养藻株, 编号为CHAB1690。拟甲色球藻属(Chroococcidiopsis)为我国新记录属, 本文描述了该属的特征。对16S rRNA基因进行测序分析表明: CHAB1690与拟甲色球藻属的模式种温泉拟甲色球藻(Chroococcidiopsis thermalis)基因序列相似度仅为91%, 暂未定种; CHAB1690与欧洲的两株未定种Chroococcidiopsis相似度较高, 并与大多数拟甲色球藻(Chroococcidiopsis)聚集在一起, 但仍有少数藻株聚集在距离较远的另一个类群。比较两个类群中藻株的最低相似率, 结果表明, 基于形态界定的拟甲色球藻属(Chroococcidiopsis)可能包含多个属。       

Biogeographically interesting planktonic Nostocales (Cyanobacteria) in the Czech Republic and their polyphasic evaluation resulting in taxonomic revisions of Anabaena bergii Ostenfeld 1908 (Chrysosporum gen. nov.) and A. tenericaulis Nygaard 1949 (Dolichospermum tenericaule comb. nova)

Questions of biogeography of freshwater cyanobacteria and their ability to colonize new areas have been recently discussed in connection with increasing occurrence of some formerly rare morphospecies in temperate zones. Nevertheless, the general knowledge about the distribution of cyanobacterial species is still fragmentary, and any new findings on cyanobacterial biogeography and spread are valuable. In this study, we provide updated information on the occurrence of Anabaena bergii, Raphidiopsis mediterranea, and Sphaerospermopsis aphanizomenoides in the Czech Republic. In addition, more nostocacean morphospecies are newly reported from the Czech Republic (A. fusca, A. tenericaulis, Dolichospermum curvum, D. mucosum, and S. reniformis). All of these morphospecies were characterized from a morphological point of view, and their phylogenetic affiliations were assessed on the basis of their 16S rRNA gene sequences. Based on these results, Anabaena bergii was reclassified into Chrysosporum gen. nov., and D. tenericaule comb. nova was established.