鄱阳湖的中国水华蓝藻新记录属气丝藻属

2012年4月和10月在江西省鄱阳湖进行野外调查时, 发现一种浮游的丝状蓝藻。通过分离纯培养, 获得了6个纯化藻株。根据藻株主要形态学特征及其16S rRNA基因序列与比较, 鄱阳湖这些藻株与老挝的2株Aerosakkonema funiforme较为相近, 其中16S rRNA基因序列的相似度达到98%。鉴于这些藻株不同于颤藻目中其他属的藻株, 因此确定为我国一水华蓝藻新记录属气丝藻属Aerosakkonema Nanda Watanabe 2012, 模式种为索状气丝藻Aerosakkonema funiforme Nanda Watanabe 2012。       

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。     

汾河太原河段浮游植物多样性及微囊藻产异味物质成因

2012年7月和10月,对汾河太原河段浮游植物多样性及微囊藻产异味物质进行了调查研究,并对其水质进行了评价。结果显示:(1)共鉴定出浮游植物6门65属126种,其中绿藻门种类最多,有25属57种,其次为硅藻门,有26属40种,蓝藻门有9属20种,其它门种类相对较少,包括裸藻门2属5种,甲藻门2属2种,隐藻门1属2种;(2)2012年7月份的优势种为蓝藻门的微小平裂藻(Merismopedia tenuissima)、不定微囊藻(Microcystis incerta),硅藻门的尖针杆藻(Synedra acus),绿藻门的四尾栅藻(Scenedesmus quadricauda)、美丽网球藻(Dictyosphaerium pulchellum)和甲藻门的微小多甲藻(Peridinium pusillum),10月份的优势种为蓝藻门的微小平裂藻、阿氏浮丝藻(Planktothrix agardhii)、两栖颤藻(Oscillatoria amphibia),硅藻门的尖针杆藻,绿藻门的四尾栅藻和隐藻门的啮蚀隐藻(Cryptomonas erosa);(3)2012年7月份和10月份迎泽桥段浮游植物平均细胞密度分别为188.28×106个/L和58.66×106个/L,南内环桥段浮游植物平均细胞密度分别为83.78×106个/L和65.99×106个/L,表明水体为富营养型;(4)Margalef多样性指数(D)为2.04—2.68,Shannon-Wiener多样性指数(H)为0.49—1.00,Pielou均匀度指数(J)为0.13—0.26,数据分析显示汾河太原河段水质为中度污染至重污染;(5)通过对分离纯化的微囊藻挥发性异味物质的检测,8株微囊藻中有6株可产生明显的异味。经过嗅味判断及与常见异味种类标准品的比对,确定这6株微囊藻产生的异味物质主要为β-环柠檬醛(β-cyclocitral)。     

晋阳湖丝状蓝藻的形态学分析与分子鉴定北大核心CSCD

蓝藻是地球上最古老的生物之一,其形态结构较为简单,为产氧型光合作用的原核生物。山西省晋阳湖为华北地区最大的人工湖,该研究以采自晋阳湖水体及岸边附着的蓝藻为材料,采用经典毛细管法分离纯化出5株丝状蓝藻,利用光学显微镜观察其形态结构特征(如细胞形状、藻丝体宽度、是否有鞘)和显微结构,并采用16S rRNA序列分析其系统发育关系,以明确晋阳湖的蓝藻种类,为预防湖泊蓝藻水华的发生、维护水资源环境稳定与生态平衡提供理论数据。结果显示:(1)所分离纯化的5株丝状蓝藻依形态特征归属于3个科,其中2株(JYH005和JYH012)为细鞘丝藻亚科(Leptolyngbyaceae),2株(JYH008和JYH022)为伪鱼腥藻科(Pseudanabaenaceae),1株(JYH010)为沙丝藻科(Desertifilaceae)。(2)基于16S rRNA序列构建的系统发育树显示,5株丝状蓝藻中JYH005为结丝藻属(Nodosilinea)的一种;JYH008可归为Arthronema,该株蓝藻在培养条件下观察到不同的形态特征,可能为新物种;JYH010为沙丝藻属(Desertifilum)的一种;JYH012可归为细鞘丝藻属(Leptolyngbya);JYH022与伪鱼腥藻科聚为一支,由于与该科其他藻相似度低于90%,且不能聚为一支,因此只能归为伪鱼腥藻科。研究表明,基于16S rRNA序列系统发育分析与形态学鉴定结果相一致。该研究结果丰富了山西省晋阳湖丝状蓝藻的多样性,为该湖的资源利用和环境保护提供了一定的科学依据。     

阿氏浮丝藻mcyT基因序列多样性研究

为研究我国浮丝藻（Planktothrix Anagnostidis et Komrek）的毒素相关基因,选取分离自我国不同省份水体的13株阿氏浮丝藻,通过PCR检测其微囊藻毒素合成酶基因mcyA、mcyE及mcyT研究其毒素基因特性。PCR结果表明除mcyT之外其他引物检测均无扩增产物,说明这13株浮丝藻不具备产微囊藻毒素的能力。通过克隆测序得到mcyT序列,并进行分子系统分析,构建了关于mcyT序列的Neighbor-Joining系统树,结果表明mcyT序列可以将产毒与不产毒浮丝藻分为两大独立的分支,两个分支之间的最低序列相似度分别为98.5%和99.1%。研究结果可为后续研究我国浮丝藻的微囊藻毒素合成相关基因的多样性以及分子监测提供参考。       

基于激光共聚焦扫描显微术和16SrRNA基因序列探讨一株水华蓝藻的分类地位

首次从福州市一重要供水水库中分离到一株体型微小的丝状蓝藻,该藻喜粘附在微囊藻胶被内外。为了确定该蓝藻的种类和分类学地位,通过形态特征分析和16S rRNA序列分析的方法鉴定其种类,并利用激光共聚焦扫描显微镜的荧光光谱以及色素提取物的吸收光谱分析其色素组成。结果表明,该藻株为粘伪鱼腥藻(Pseudanabaena mucicola)(Gen Bank序列登录号为KR912197),黏伪鱼腥藻不仅含有藻蓝蛋白,同时也含藻红蛋白。此结果为该藻株分类位置的确定提供了有利佐证。     

大亚湾水域并基拟菱形藻的种类鉴定和产毒特征分析

为了丰富我国海域拟菱形藻(Pseudo-nitzschia Peragallo)的物种多样性, 并澄清其产毒特征, 研究从广东大亚湾海域分离并建立了一株拟菱形藻的单克隆培养株系MC298, 通过光学显微镜下的群体特征和透射电镜下的超微形态特征观察, 结合基于核糖体转录间隔区(Internal Transcribed Spacer, ITS)的分子系统学数据, 以及基于ITS2转录RNA的二级结构分析, 鉴定到我国拟菱形藻属的1个新记录种: 并基拟菱形藻P. decipiens Lundholm & Moestrup。研究对其形态学特征进行了较为详细地描述, 并与相似种进行了比较, 还对其ITS2-RNA的特有标志结构进行了阐述。同时, 利用高效液相色谱-质谱联用法(Liquid chromatography tandem mass spectrometry, LC-MS/MS)对该藻株的产毒特征进行了检测, 结果未检测到DA的存在。研究不仅丰富了我国拟菱形藻属的物种多样性, 也可为拟菱形藻的产毒特征研究提供基础数据。     

洱海的浮游蓝藻布氏常丝藻及其分类学的讨论

常丝藻(Tychonema)是1988年由Anagnostidis和Komárek从颤藻属分离出来而新成立的蓝藻属,以纤细常丝藻(T.tenue)为模式种类。目前确认的常丝藻有纤细常丝藻、博多常丝藻(T.bornetii)和布氏常丝藻(T.bourrellyi)三个种类。我国已经有博多常丝藻的纪录,但是对模式种纤细常丝藻和它的相似种类布氏常丝藻却没有报道。在洱海中采集到布氏常丝藻(T.bourrellyi),研究并描述了该藻的藻丝颜色、藻细胞内含物的结构、藻体形态特征等。同时,通过藻种的分离培养技术,得到了布氏常丝藻的纯培养藻株,编号为CHAB663,并且测定了藻株的16S rRNA基因序列。测序结果表明,该藻株与T.bouurrellyi/T.tenue聚为一族。洱海分布的布氏常丝藻,是在欧洲以外首次发现此种藻类,也是我国的新纪录种。研究说明布氏常丝藻不仅仅分布在温带欧洲较为寒冷的水体中,在亚热带的水体中也可以存在。布氏常丝藻被认为是出现在轻微富营养化湖泊中,而我国洱海也被认为是富营养化的初级阶段的水体,这也表明,布氏常丝藻的出现对洱海的水环境状况起到了指示作用。此外,研究还对常丝藻属分类学问题进行了讨论。     

一株虾池来源的螺旋拟柱孢藻藻株的分离鉴定及重金属离子Cu~(2+)、Cd~(2+)和Pb~(2+)对其生长的影响

【目的】探讨凡纳滨对虾养殖水体中入侵蓝藻拟柱孢藻的生长生理特性。【方法】从汕头澄海人工对虾养殖池分离纯化藻株,通过形态及其16SrRNA基因鉴定,之后在CT与BG11两种蓝藻通用培养基的基础上优化最佳培养条件,最后分析了不同浓度的3种重金属离子即Cu~(2+)(0–0.8 mg/L)、Cd~(2+)(0–4 mg/L)和Pb~(2+)(0–80 mg/L)对藻株生长的影响。【结果】澄海虾池来源的分离纯化藻株形态呈卷曲螺旋型,16S rRNA基因序列与多株其他来源的拟柱孢藻相似度均达98%以上。实验室培养,藻株最佳生长状态的培养条件是在BG11培养基的基础上调整氮浓度及氮磷比分别为N 62 mg/L,N︰P=9︰1,在此条件下,藻丝生物量可达(0.632±0.170)×107/L,藻丝比平均生长速率最高为(0.063±0.001)/d。本分离藻株活体对重金属Cu~(2+)、Cd~(2+)和Pb~(2+)具有一定的耐受性,其耐受浓度范围分别为0–0.2、0–0.5和1–40 mg/L,其中,Cu~(2+)和Cd~(2+)对藻的生长具有抑制作用,而且此抑制作用随着金属离子剂量的增加及作用时间的延长更加显著,Cu~(2+)和Cd~(2+)对藻体的半数抑制浓度(96 h EC50)分别为0.125和0.551 mg/L;而浓度范围为0–80 mg/L的Pb~(2+)对藻体的生长则表现为低剂量(≤40 mg/L)呈促进,高剂量(≥80 mg/L)则抑制。【结论】从凡纳滨对虾养殖池中分离鉴定出一株形态呈螺旋型的拟柱孢藻,命名为螺旋拟柱孢藻(Cylindrospermopsis raciborskii helix),本藻株活体能够在一定浓度的Cu~(2+)、Cd~(2+)和Pb~(2+)中生长,为螺旋拟柱孢藻活藻生物吸附重金属离子而改善虾池水体环境提供了可能性。     

中国沿海新报道的三种产毒拟菱形藻

为了明确我国海域拟菱形藻属(Pseudo-nitzschia)物种的产毒特征, 从中国沿海建立了15个拟菱形藻单克隆培养株系, 利用高效液相色谱-质谱联用法对其多莫酸特征进行检测, 在10个株系中检测到多莫酸。结合光学显微镜下的群体特征和透射电镜下的超微形态学特征, 以及基于核糖体转录间隔区的分子系统学数据, 确认上述10个产毒株系分别隶属于3个物种:尖细拟菱形藻P. cuspidata、伪柔弱拟菱形藻P. pseudodelicatissima、伪善拟菱形藻P. fraudulenta, 其中伪善拟菱形藻是我国的新记录种。建立尖细拟菱形藻的11个尖细拟菱形藻株系, 其中3个株系未检出多莫酸, 其余8个株系有检出, 单细胞产毒水平为0.4—5.5 fg。建立伪柔弱拟菱形藻株系2个, 1个未检出多莫酸, 另1个株系的单细胞产毒量为1 fg。建立伪善拟菱形藻株系2个, 纯种培养株系均未检出多莫酸。利用卤虫(Artemia salina)对部分藻株进行混培诱导, 其中尖细拟菱形藻(MC4049)和伪柔弱拟菱形藻(MC3015)的产毒水平略有下降, 单细胞产毒水平分别由2、1 fg降至0.2、0.4 fg, 而伪善拟菱形藻(MC4074)的产毒能力则有显著改变, 单细胞产毒水平由未检出上升至17.5 fg。研究丰富了我国产毒拟菱形藻的物种多样性, 明确了其物种信息和产毒水平, 可为后续深入研究提供基础数据。     

Identification of two cyanobacterial strains isolated from the Kotel'nikovskii hot spring of the Baikal rift

Two cyanobacterial strains, Pseudanabaena sp. 0411 and Synechococcus sp. 0431, were isolated from a sample collected in the Kotel'nikovskii hot spring of the Baikal rift. According to the results of light and transmission electron microscopy, as well as of the phylogenetic analysis of the 16S rRNA gene, these cyanobacteria were classified as Pseudanabaena sp. nov. and Synechococcus bigranulatus Skuja. The constructed phylogenetic tree shows that the studied strains are positioned in the clades of cyanobacteria isolated from hydrothermal vents of Asia and New Zealand, separately from marine and freshwater members of these genera, including those isolated from Lake Baikal.     

Adaptation to sulfide and to the underwater light field in three cyanobacterial isolates from Lake Arcas (Spain)

Abstract: Three strains of cyanobacteria isolated from karstic Lake Arcas were tested for photosynthetic adaptations to soluble sulfide. One of them, AO11, was identified as Oscillatoria cf. ornata , and forms dense populations in the sulfide-rich anoxic hypolimnion of this lake. This cyanobacterium was able to perform sulfide-dependent anoxygenic photosynthesis and its oxygenic photosynthesis was relatively insensitive to sulfide. The other strains studied were AP1 and AO21, identified respectively as Pseudanabaena sp. and Oscillatoria cf. tenuis , populations of which were present only in epilimnetic waters at low population densities. Pseudanabaena sp. also carried out anoxygenic photosynthesis, but oxygenic photosynthesis was totally inhibited by 0.5 mM sulfide. Oscillatoria cf. tenuis lost most of its oxygenic photosynthetic capacity when submitted to 0.1 mM sulfide and anoxygenic photosynthesis accounted for less than 20% of sulfide-free controls. In addition to different photosynthetic capabilities, the three cyanobacteria exhibited differences in light-harvesting photosynthetic accessory pigments. Pigment analysis of cultures grown under different light conditions showed the capacity of Oscillatoria cf. ornata AO11 to produce phycoerythrin under low light intensity or under predominantly green light, while neither Pseudanabaena sp. AP1 nor Oscillatoria cf. tenuis AO21 produced this pigment. The complementary chromatic adaptation of Oscillatoria cf. ornata correlates well with its summertime distribution under the dim light field of the hypolimnion. The distribution and abundance of specific cyanobacterial populations in Lake Arcas can thus be explained by the interplay of light regime and presence of sulfide as some of the most determinant ecological parameters.     

Culture temperature affects gene expression and metabolic pathways in the 2-methylisoborneol-producing cyanobacterium Pseudanabaena galeata

A volatile metabolite, 2-methylisoborneol (2-MIB), causes an unpleasant taste and odor in tap water. Some filamentous cyanobacteria produce 2-MIB via a two-step biosynthetic pathway: methylation of geranyl diphosphate (GPP) by methyl transferase (GPPMT), followed by the cyclization of methyl-GPP by monoterpene cyclase (MIBS). We isolated the genes encoding GPPMT and MIBS from Pseudanabaena galeata, a filamentous cyanobacterium known to be a major causal organism of 2-MIB production in Japanese lakes. The predicted amino acid sequence showed high similarity with that of Pseudanabaena limnetica (96% identity in GPPMT and 97% identity in MIBS). P. galeata was cultured at different temperatures to examine the effect of growth conditions on the production of 2-MIB and major metabolites. Gas chromatograph–mass spectrometry (GC–MS) measurements showed higher accumulation of 2-MIB at 30 °C than at 4 °C or 20 °C after 24 h of culture. Real-time-RT PCR analysis showed that the expression levels of the genes encoding GPPMT and MIBS decreased at 4 °C and increased at 30 °C, compared with at 20 °C. Furthermore, metabolite analysis showed dramatic changes in primary metabolite concentrations in cyanobacteria grown at different temperatures. The data indicate that changes in carbon flow in the TCA cycle affect 2-MIB biosynthesis at higher temperatures.     

An early origin of plastids within the cyanobacterial divergence is suggested by evolutionary trees based on complete 16S rRNA sequences

It is generally accepted that the plastids arose from a cyanobacterial ancestor, but the exact phylogenetic relationships between cyanobacteria and plastids are still controversial. Most studies based on partial 16S rRNA sequences suggested a relatively late origin of plastids within the cyanobacterial divergence. In order to clarify the exact relationship and divergence order of cyanobacteria and plastids, we studied their phylogeny on the basis of nearly complete 16S rRNA gene sequences. The data set comprised 15 strains of cyanobacteria from different morphological groups, 1 prochlorophyte, and plastids belonging to 8 species of plants and 12 species of diverse algae. This set included three cyanobacterial sequences determined in this study. This is the most comprehensive set of complete cyanobacterial and plastidial 16S rRNA sequences used so far. Phylogenetic trees were constructed using neighbor joining and maximum parsimony, and the reliability of the tree topologies was tested by different methods. Our results suggest an early origin of plastids within the cyanobacterial divergence, preceded only by the divergence of two cyanobacterial genera, Gloeobacter and Pseudanabaena.      

Phylogeography of the Microcoleus vaginatus (Cyanobacteria) from three continents--a spatial and temporal characterization

It has long been assumed that cyanobacteria have, as with other free-living microorganisms, a ubiquitous occurrence. Neither the geographical dispersal barriers nor allopatric speciation has been taken into account. We endeavoured to examine the spatial and temporal patterns of global distribution within populations of the cyanobacterium Microcoleus vaginatus, originated from three continents, and to evaluate the role of dispersal barriers in the evolution of free-living cyanobacteria. Complex phylogeographical approach was applied to assess the dispersal and evolutionary patterns in the cyanobacterium Microcoleus vaginatus (Oscillatoriales). We compared the 16S rRNA and 16S-23S ITS sequences of strains which had originated from three continents (North America, Europe, and Asia). The spatial distribution was investigated using a phylogenetic tree, network, as well as principal coordinate analysis (PCoA). A temporal characterization was inferred using molecular clocks, calibrated from fossil DNA. Data analysis revealed broad genetic diversity within M. vaginatus. Based on the phylogenetic tree, network, and PCoA analysis, the strains isolated in Europe were spatially separated from those which originated from Asia and North America. A chronogram showed a temporal limitation of dispersal barriers on the continental scale. Dispersal barriers and allopatric speciation had an important role in the evolution of M. vaginatus. However, these dispersal barriers did not have a permanent character; therefore, the genetic flow among populations on a continental scale was only temporarily present. Furthermore, M. vaginatus is a recently evolved species, which has been going through substantial evolutionary changes.     

热带直流型水库蓝藻季节变化特征

于2004年调查了广东省飞来峡和深圳两座直流型水库的蓝藻季节分布特征。共检到蓝藻17属,绝大多数为丝状体种类。常见属有假鱼腥藻(Pseudanabaena)、微囊藻(Microcystis)和蓝纤维藻(Dactylococcopsis)。蓝藻细胞密度为5-2052个细胞ml-1。深圳水库最高蓝藻细胞密度出现在6月,其中丝状蓝藻的相对丰度在75%以上;飞来峡水库最高蓝藻细胞密度出现在12月,丝状蓝藻的相对丰度达到90%。水动力学条件是限制蓝藻丰度和种类的主要环境因子。由于飞来峡水库的水力滞留时间变化较大,所以蓝藻季节性变化受水动力学条件的影响比深圳水库的大。假鱼腥藻等丝状蓝藻更适合在水库中生长,特别是在氮磷营养水平较高,水力滞留时间明显增加时,其细胞密度将大幅度上升。     

古尔班通古特沙漠生物结皮微鞘藻分类学研究

实验研究了从古尔班通古特沙漠生物土壤结皮中分离纯化培养出的11株与微鞘藻(Microcoleus)形态接近的丝状蓝藻,通过形态特征、16S rRNA和ITS二级结构相结合的多相分析方法对其进行分类学研究。研究结果表明,实验藻株隶属于微鞘藻科(Microcoleaceae)的微鞘藻属(Microcoleus)和束脉藻属(Symplocastrum),其中包括2个中国新记录种:斯坦微鞘藻(Microcoleus steenstrupii)和细长束脉藻(Symplocastrum flechtnerii),另外还有具鞘微鞘藻(Microcoleus vaginatus)和类似斯坦微鞘藻的存疑物种。藻丝多少与排列方式、细胞大小与末端细胞形状,以及16S rRNA系统发育位置是确定微鞘藻(Microcoleus)与束脉藻(Symplocastrum)属于不同物种的关键依据, ITS二级结构是区分属内不同物种的重要参考。     

Host re-identification of cyanophage PP and its implications for host range and specificity

Dear Editor, In this study,we re-identified the hosts of cyanophage PP.Twenty-three candidate algal strains were tested,and the results indicated that seven strains belonging to Plectonema and Phormidium are the hosts of cyanophage PP,including two previously reported filamentous cyanobacteria,Plectonema boryanum and Phormidium foveolarum (Zhao et al.2002).However,several species or strains within the two genera were found not to be hosts of cyanophage PP,implying that the host range is relatively specific.