共球藻纲丝状藻中国新记录属种——洪泛拟寇丽藻

从甘肃省张掖市黑河流域分离了一株不分枝的丝状绿藻,对其进行纯培养并保藏于中国科学院淡水藻种库,编号FACHB 2451。形态观察表明,该藻株具有相对较长的营养细胞 (细胞长为宽的3~8倍)、双叶形的片状叶绿体、末端细胞顶端钝圆或略尖细等特征。结合形态比较和基于18S rDNA 基因序列的系统发育分析,确定该藻株为中国淡水共球藻纲新记录属种——洪泛拟寇丽藻 (Koliellopsis inundata Lokhorst)。分子系统发育研究结果表明,所分离的藻株与分离自比利时与荷兰边境农地的洪泛拟寇丽藻模式藻株序列相似度极高,且拟寇丽藻属与寇丽藻属、针丝藻属和拟针丝藻属等不分枝丝状拟寇丽藻科成员亲缘关系密切,可以较好地聚成一个亚支。     

中国淡水共球藻纲新记录属种——土佐牧野藻(Makinoella tosaensis Okada)

报道分别从湖北省武汉市内和云南省西双版纳小水池中分离培养的两株绿藻,对其进行了形态和18S r DNA基因序列分析,编号分别为FACHB-1783和FACHB-1784。这两株绿藻具独特的四边形群体形态,通常为4或16个细胞,细胞为宽椭圆形至不规则卵圆形、细胞壁两端无增厚,叶绿体多数、片状,具蛋白核。结合形态和分子系统发育分析,确定这两株绿藻为我国1种淡水共球藻纲新记录属种——土佐牧野藻(Makinoella tosaensis Okada)。基于18S r DNA基因的系统发育研究表明这两株绿藻与分离自韩国的土佐牧野藻基因序列相似度可达99.6%~99.9%,并且以较高的支持值与土佐牧野藻聚在一起。     

我国北方岩溶泉域刚毛藻的系统发育及形态学研究

脆弱刚毛藻(Cladophora fracta)是一种大型丝状绿藻,生境分布广泛。然而,对于岩溶泉域分布的刚毛藻研究较少,它们的遗传多样性、生物地理亲缘性和生理特性都有待于深入研究。该研究对我国北方地区五个典型岩溶泉域的50个脆弱刚毛藻样本进行了形态学和分子系统学描述。主要研究目标:(1)对我国北方地区五个典型岩溶泉的刚毛藻生境进行描述;(2)根据形态学特征和分子序列对藻体进行鉴定;(3)探究生境对藻体生理特性的影响。结果表明:基于SSU和LSU序列的结果,发现所分析的50株刚毛藻个体为同一种,同时还发现了13个不同的核糖体基因型。基于SSU和LSU的系统发育树,刚毛藻属均未能形成单系分支,分布在三个不同的分支上。13个样本基因型在SSU和LSU树中的位置相似,与Cladophora vagabunda有很高的序列同源性,但是形态特征却差异很大。从显微结构结果来看,五个岩溶泉域采集到的刚毛藻在细胞直径上无显著差异,藻体的形态特征与脆弱刚毛藻相一致。但是,岩溶泉域采集的藻体细胞直径比文献报道中在湖泊和河流中采集的脆弱刚毛藻直径要大。另外,仅在两个地点(XA和ST)采集的标本中发现有假根状分枝。因此,基于形态学和分子序列的结果,将这五个泉域的刚毛藻鉴定为脆弱刚毛藻(Cladophora fracta)。     

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。     

氮源类型和水平对3株球状绿藻生长、油脂和花生四烯酸积累的影响

【背景】缺刻叶球藻(Lobosphaera incisa Reisigl)是一种单细胞球状绿藻,是已知花生四烯酸(Arachidonic acid,AA)含量最丰富的植物资源之一。然而目前其分类和命名仍然较为混乱。【目的】明确3株球状绿藻(SAG2468、SAG2043、H4301)的分类地位及在不同氮源(硝酸钠、尿素、碳酸氢铵、碳酸铵、硝酸铵、氯化铵和硫酸铵)和氮浓度(18mmol/L,3mmol/L)条件下油脂和AA积累的特性。【方法】通过分子系统学和形态观察的方法对3株球状绿藻进行分类界定;采用干重法、重量法和气相色谱分析的方法分别对其生物量、油脂含量、脂肪酸组成及AA含量进行测定。【结果】3株球状绿藻均隶属于叶球藻属(Lobosphaera),SAG2468原定名为缺刻缘绿藻(Parietochloris incisa),现修订为缺刻叶球藻(Lobosphaera incisa),与H4301为缺刻叶球藻的不同地理株系。SAG2043原定名为双隔蚁形藻(Myrmecia bisecta),现修订为双隔叶球藻(Lobosphaera bisecta)。3株微藻在高氮(18 mmol/L)和低氮(3 mmol/L)浓度的硝酸钠和尿素培养条件下均可良好生长,铵盐对藻细胞生长普遍有抑制作用,且浓度越高抑制作用越显著。低氮胁迫能显著促进油脂和AA的积累(P0.05),SAG2043在3 mmol/L硝酸钠条件下油脂和AA产率最高,分别为142.15mg/(L·d)和35.51mg/(L·d),明显高于另外2株微藻(P0.05)。此时SAG2043对应获得的生物量为4.9 g/L,油脂含量为43.49%,AA含量高达干重的10.86%,占总脂肪酸含量的31.75%。【结论】SAG2043是一株更具AA开发潜力的微藻。     

重离子诱变创制高产油微拟球藻新品种

以具有产业化前景的微拟球藻Nannochloropsis oceanica OZ-1为实验材料,利用碳重离子进行诱变育种,采用Imaging-PAM和酶标仪进行大规模筛选,最终获得两株高生长速率微拟球藻突变藻株(HP-1和HP-2),进一步分析显示两株突变藻株(HP-1和HP-2)生物量积累较野生型藻株大幅提高,在18d培养末期生物量分别提高了18％和26％,两株突变藻株油脂产率分别为295 mg/(L·d)和275 mg/(L·d),而野生型藻株为247 mg/(L·d).所获两株突变藻株生长速度快、油脂产率高,较野生型藻株优势明显.     

绿球藻目的几个中国新记录属种

6个生境比较特殊而有趣的绿球藻目物种首次在我国发现,它们分别是生于土壤的香喇网绿藻(Dictyochloris fragrans Vischer ex Start),寄生于眼子菜叶片的浮萍绿点藻(Chlorchytrium lemmae Cohn.),气生种类—生长在樟树皮上的分层胶囊藻(Gloeocystis polydermatica(Kutz.) Hindak),与真菌共生形成地衣的土著共球藻(Trebouxia aboricola de Puymaly)和团集共球藻(Trebouscia glomerata(Waren) Ahmadjian),还有附生于一种刚毛藻(Cladophora sp.)细胞表面的伊乐外生藻(Ectogeron elodeae Dangeard)。除胶囊藻属(Gloeocystis)外,其它属均为中国新记录属。     

云南石蕊共生藻形态及分子系统学研究

该研究以2016年采自云南凤凰山的云南石蕊(Cladonia yunnan)为材料,在无菌条件下,采用微量微管吸法和直接法分离纯化培养云南石蕊共生藻,利用光学显微镜观察共生藻细胞的形态结构特征,并对该共生藻进行分子系统学分析。结果表明：(1)从云南石蕊中仅分离到1种共生藻,该藻细胞大小约2.5~6 μm,球形或椭球形,单细胞,不运动,色素体片状,无性繁殖,含有中央蛋白核和淀粉粒,且3~5个细胞包裹在粘液状的胶壳中,细胞核不可见。(2)该共生藻主要形态学特征与自由生活的胶球藻高度相似,根据采集地及形态特征将其命名为云南胶球藻(Coccomyxa sp. YN)。(3)基于对云南石蕊分子数据(rbcL, ITS)构建的云南石蕊共生藻的系统发育关系显示,云南胶球藻与胶球藻聚为一枝; ITS2 rRNA二级结构分析发现,云南胶球藻与胶球藻具有几乎一致的ITS2 rRNA二级结构。该研究首次揭示了胶球藻属(Coccomyxa)藻细胞与石蕊科地衣共生,且云南胶球藻与胶球藻具有高度一致的亲缘关系,而“地衣化”使两者细胞形态特征有明显差异,推测共生藻细胞可能是由自由生活的藻细胞进化而来。     

中国青岛沿岸潮间带海藻中的可培养真菌多样性（英文）

从中国青岛沿岸潮间带采集9种海藻,包括3种褐藻、4种绿藻和2种红藻,通过传统培养方法共分离获得92株真菌。根据形态学特征和真菌转录间隔区(ITS)r DNA序列分析对所得菌株进行鉴定,结果显示:92株真菌隶属子囊菌门56种,担子菌门1种及接合菌门1种;优势属为枝顶孢属、曲霉属、枝孢属、青霉属和帚霉属;无性型真菌40种,占总菌株数量的75%;绿藻门刺松藻的真菌多样性最高,其次为褐藻门鼠尾藻,红藻门珊瑚藻的真菌多样性最低。本研究首次对中国沿岸潮间带海藻中的真菌多样性进行报道,为进一步研究藻生真菌生态功能提供参考。     

细胞周期蛋白依赖性激酶抑制剂Roscovitine对3种蓝藻生长和活性的影响

为了明确蓝藻中丝氨酸/苏氨酸激酶的功能是否与调控细胞的生长分裂相关,以丝状鱼腥藻7120、单细胞集胞藻6803和聚球藻7002为对象,利用OD750光吸收测定和MTT方法研究了不同浓度丝氨酸苏氨酸激酶抑制剂roscovitine对其生长和脱氢酶活性的影响。结果表明:4 h roscovitine处理后对鱼腥藻7120和集胞藻6803生长量影响不大,对聚球藻7002的生长有促进作用。4 h roscovitine的处理对鱼腥藻7120有浓度依赖的显著抑制活性,对集胞藻6803的活性无影响,但是却促进聚球藻7002的活性。药物作用4 d后,7120的生长和活性均显著降低,并有浓度效应;6803的生长量较对照减少,但活性变化不明显;聚球藻7002的生长和活性均未受影响。显微观察结果显示,roscovitine对3种细胞形态没有影响,但药物作用4 d后的7120藻丝体较短。结果表明丝氨酸/苏氨酸抑制剂roscovitine影响丝状藻7120的生长和活性。     

冬季赤水河流域刚毛藻多样性调查及系统发育分析

研究于2019年冬季对赤水河流域开展刚毛藻多样性调查,共设计采样点38个,覆盖赤水河上、中、下游。调查发现:21个采样位点分布有刚毛藻目藻类,其中19个位点有刚毛藻分布。基于核糖体小亚基(SSU rDNA)、核糖体大亚基(LSU rDNA)和内转录间隔区(ITS)对采集样品进行系统发育分析,结果显示:(1)赤水河流域刚毛藻多样性较高,且该流域上、中、下游均有刚毛藻分布;(2)目前淡水刚毛藻类群包含至少10个支系,赤水河流域采集到的刚毛藻覆盖其中6个支系(分别是clade 1、clade 2、clade 4、clade 7、clade 9和clade 10);(3)相比基于SSU+LSU双分子标记构建的系统进化树,基于SSU+ITS+LSU三分子标记构建的进化树各支系支持度更高,可以较有效地将淡水刚毛藻不同支系区分开来。研究较好地展示了冬季赤水河流域刚毛藻的广泛分布及其分子多样性,丰富了中国淡水丝状绿藻的分类研究,也为赤水河段的水生态环境保护提供了基础数据支持。     

Cladophora (Chlorophyta) spp. harbor human bacterial pathogens in nearshore water of Lake Michigan

Cladophora glomerata, a macrophytic green alga, is commonly found in the Great Lakes, and significant accumulations occur along shorelines during the summer months. Recently, Cladophora has been shown to harbor high densities of the fecal indicator bacteria Escherichia coli and enterococci. Cladophora may also harbor human pathogens; however, until now, no studies to address this question have been performed. In the present study, we determined whether attached Cladophora, obtained from the Lake Michigan and Burns Ditch (Little Calumet River, Indiana) sides of a breakwater during the summers of 2004 and 2005, harbored the bacterial pathogens Shiga toxin-producing Escherichia coli (STEC), Salmonella, Shigella, and Campylobacter. The presence of potential pathogens and numbers of organisms were determined by using cultural methods and by using conventional PCR, most-probable-number PCR (MPN-PCR), and quantitative PCR (QPCR) performed with genus- and toxin-specific primers and probes. While Shigella and STEC were detected in 100% and 25%, respectively, of the algal samples obtained near Burns Ditch in 2004, the same pathogens were not detected in samples collected in 2005. MPN-PCR and QPCR allowed enumeration of Salmonella in 40 to 80% of the ditch- and lakeside samples, respectively, and the densities were up to 1.6 x 10(3) cells per g Cladophora. Similarly, these PCR methods allowed enumeration of up to 5.4 x 10(2) Campylobacter cells/g Cladophora in 60 to 100% of lake- and ditchside samples. The Campylobacter densities were significantly higher (P < 0.05) in the lakeside Cladophora samples than in the ditchside Cladophora samples. DNA fingerprint analyses indicated that genotypically identical Salmonella isolates were associated with geographically and temporally distinct Cladophora samples. However, Campylobacter isolates were genetically diverse. Since animal hosts are thought to be the primary habitat for Campylobacter and Salmonella species, our results suggest that Cladophora is a likely secondary habitat for pathogenic bacteria in Lake Michigan and that the association of these bacteria with Cladophora warrants additional studies to assess the potential health impact on beach users.     

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—90.0μm,细胞长宽比2.4—8.0。分枝细胞直径25.0—70.0μm,长宽比4.0—12.3。顶端细胞直径25.0—50.0μm,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.。     

MOLECULAR PHYLOGENY AND TAXONOMY OF THE AEGAGROPILA CLADE (CLADOPHORALES,ULVOPHYCEAE), INCLUDING THE DESCRIPTION OF AEGAGROPILOPSIS GEN. NOV. AND PSEUDOCLADOPHORA GEN. NOV.1

The Aegagropila clade represents a unique group of cladophoralean green algae occurring mainly in brackish and freshwater environments. The clade is sister to the species‐rich and primarily marine Cladophora and Siphonocladus lineages. Phylogenetic analyses of partial LSU and SSU nrDNA sequences reveal four main lineages within the Aegagropila clade, and allow a taxonomic reassessment. One lineage consists of two marine ‘Cladophora’ species, for which the new genus Pseudocladophora and the new family Pseudocladophoraceae are proposed. For the other lineages, the family name Pithophoraceae is reinstated. Within the Pithophoraceae, the earliest diverging lineage includes Wittrockiella and Cladophorella calcicola, occurring mainly in brackish and subaerial habitats. The two other lineages are restricted to freshwater. One of them shows a strong tendency for epizoism, and consists of Basicladia species and Arnoldiella conchophila. The other lineage includes Aegagropila, Pithophora and a small number of tropical ‘Cladophora’ species. The latter are transferred to the new genus Aegagropilopsis. Previously, polypyramidal pyrenoids had been suggested to be apomorphous for this clade, but we report the finding of both polypyramidal and bilenticular pyrenoids in members of the Pithophoraceae, and thus show that this character has no diagnostic value.     

Occurrence of Escherichia coli and enterococci in Cladophora (Chlorophyta) in nearshore water and beach sand of Lake Michigan

Each summer, the nuisance green alga Cladophora (mostly Cladophora glomerata) amasses along Lake Michigan beaches, creating nearshore anoxia and unsightly, malodorous mats that can attract problem animals and detract from visitor enjoyment. Traditionally, elevated counts of Escherichia coli are presumed to indicate the presence of sewage, mostly derived from nearby point sources. The relationship between fecal indicator bacteria and Cladophora remains essentially unstudied. This investigation describes the local and regional density of Escherichia coli and enterococci in Cladophora mats along beaches in the four states (Wisconsin, Illinois, Indiana, and Michigan) bordering Lake Michigan. Samples of Cladophora strands collected from 10 beaches (n = 41) were assayed for concentrations of E. coli and enterococci during the summer of 2002. Both E. coli and enterococci were ubiquitous (up to 97% occurrence), with overall log mean densities (+/- standard errors) of 5.3 (+/- 4.8) and 4.8 (+/- 4.5) per g (dry weight). E. coli and enterococci were strongly correlated in southern Lake Michigan beaches (P < 0.001, R(2) = 0.73, n = 17) but not in northern beaches (P = 0.892, n = 16). Both E. coli and enterococci survived for over 6 months in sun-dried Cladophora mats stored at 4 degrees C; the residual bacteria in the dried alga readily grew upon rehydration. These findings suggest that Cladophora amassing along the beaches of Lake Michigan may be an important environmental source of indicator bacteria and call into question the reliability of E. coli and enterococci as indicators of water quality for freshwater recreational beaches.     

PHYLOGENETIC RELATIONSHIPS WITHIN CLADOPHORALES (ULVOPHYCEAE,CHLOROPHYTA) INFERRED FROM 18S rRNA GENE SEQUENCES,WITH SPECIAL REFERENCE TO AEGAGROPILA LINNAEI 1

The phylogenetic position of a freshwater green alga, Aegagropila linnaei (Cladophorales, Ulvophyceae), was investigated using nuclear 18S rRNA gene sequences. This alga has usually been called Cladophora aegagropila (L.) Rabenhorst or Cladophora sauteri (Nees ex Kütz.) Kütz. Based on morphology, it was formerly classified into the section Aegagropila or into the subgenus Aegagropila, together with several marine species of the genus Cladophora. This classification is not supported by the present phylogenetic analyses in which two very distinct Cladophorales clades are recognized. Aegagropila linnaei groups together in a well‐supported clade with Cladophora sp., Pithophora sp., Chaetomorpha okamurae, Arnoldiella conchophila, Wittrockiella lyallii, and Cladophora conchopheria. Aegagropila linnaei and its closely related species share some ultrastructural and biochemical characteristics, like pyrenoid structure, carotenoid composition, and cell wall composition. Freshwater species, included in the analysis, were located in two distantly related lineages, indicating that adaptation from a marine to a freshwater habitat has happened at least twice independently in the Cladophorales.     

Animal associations with the dominant species of shallow water macrophytes along the coastline of the Vestfold Hills,Antarctica

Benthic grab samples, taken through core holes in sea-ice 1983, returned eight species of marine macrophytes which had associated fauna on at least one sample. A total of 24 animal taxa were found on these eight macrophytes. Phyllophora antarctica had a significantly greater biomass of attached fauna per unit of macrophyte biomass than any of the other seven species. It also had the greatest variety of epiphytic fauna (21 taxa). Phyllophora antarctica was the only macrophyte collected from comparatively protected Ellis Fjord, apart from filamentous algae, including Cladophora subsimplex and Geminocarpus austrogeorgiae, which were found in the most landward basin. It therefore played an important role in the ecology of the fjord. Palmaria decipiens was commonly collected at more exposed coastal sites and was dominant in Long Fjord, characterized here as intermediate in terms of water movement. Sixteen animal taxa were collected from this macrophyte with the greatest numbers being found in July and August. Only four animal taxa were collected from Iridaea cordata. The other five macrophyte species from which epiphytic animals were collected (Porphyra endiviifolium, Plocamium cartilagineum, Desmarestia menziesii, Himantothallus grandifolius and Cladophora subsimplex had animals associated with them on less than half the occasions when they were found.     

The palaeolimnology of Lake Mývatn, northern Iceland: plant and animal microfossils in the sediment

SUMMARY. Sediment from a precursor of the 2300-year old Lake Mývatn was traced by diatoms in pseudocrater scoria. This lake (Mývatn I) was deeper than the present Mývatn but similar in size. Tephrochronology confirms an equal age of the Ytrlflói and Sydrifloi basins of the present Mývatn. Diatom succession in the sediment of the Sydriflói basin was divided into four zones, of which the lowest corresponds to inherited and highly disturbed Mývatn I sediment. At 90 cm depth, i.e. around 1600 AD, in the Sydriflói sediment a simultaneous increase occurred in Cladophora aegagropila (Chlorophyta), Eurycercus lamellatus (Cladocera) and three diatom species epiphytic on Cladophora. The density of Cladocera also followed this trend. The increase in Cladophora was perhaps caused by improved light conditions on the bottom, either because of a decrease in water depth due to sedimentation, or a decrease in Anabaena blooms, or both.     

Isolation of chlorophylls and carotenoids from freshwater algae using different extraction methods

Usually marine algae are an excellent source of pigments for different commercial sectors. Freshwater macroalgae can be exploited as a good source of biologically active compounds provided an appropriate extraction method is developed. The efficiency of four methods, like microwave‐assisted (MAE), ultrasound‐assisted extraction (UAE), supercritical fluid extraction (SFE) with ethanol as a co‐solvent, as well as conventional Soxhlet extraction were studied in the same conditions (time, solvent and temperature) for the recovery of chlorophylls and carotenoids from three freshwater green algae species: Cladophora glomerata, Cladophora rivularis and Ulva flexuosa. UV‐Vis spectrophotometry was used to determine chlorophyll a, chlorophyll b and total carotenoid content in obtained extracts. The results of this study showed that the advantages of novel extraction techniques (MAE and UAE) include higher yield and, in consequence, lower costs compared to traditional solvent extraction techniques. These methods were much more efficient in freshwater green algae pigment recovery than the classic Soxhlet extraction as well as SFE.