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。     

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。     

淡水藻种库(FACHB)库藏产2-MIB蓝藻的鉴定及其产嗅特征研究

为认识产二甲基异莰醇(2-MIB)蓝藻的形态和产嗅特征,从国家水生生物种质资源库淡水藻种库中筛选出24株可产2-MIB藻株,描述了这些藻株的形态特征和生境分布。结合形态和16S rRNA基因分析对藻株进行物种鉴定复核,修订了部分库藏藻株物种名称,例如发现库藏产2-MIB的浮丝藻属种类应当被重新鉴定为拉氏拟浮丝藻或索状气丝藻。基于mic基因系统发育树分析,显示蓝藻mic基因形成5个分支。通过2-MIB含量检测发现,不同藻株间单个细胞总2-MIB含量为6—2549 fg/cell,其含量通常为拉氏拟浮丝藻>索状气丝藻>灰假鱼腥藻。研究提供了产2-MIB蓝藻的形态、分子、生态和产嗅特性等的基础数据,首次报道气丝藻、沙丝藻、苏打丝藻种类可产2-MIB,并在国内首次报道了产2-MIB的微鞘藻,为进一步研究产2-MIB蓝藻生理生态特性提供重要的实验材料和科学依据。     

陕北黄土高原生物结皮中7种微藻的形态与分子生物学鉴定

陕北黄土高原实施退耕还林后,生物结皮成为其典型的地表覆盖类型,含有丰富的土生藻类,对固定土壤和促进养分循环具有极其重要的作用。该研究通过平板法与水滴稀释法对陕北黄土高原生物结皮土生藻类进行分离培养,采用光学显微镜观察结皮微藻的形态特征,并对单藻种进行分子鉴定,为黄土高原生物结皮藻类的研究奠定基础。结果显示:(1)共纯化获得7种结皮藻类,经光学显微镜初步确定,其中5株为绿藻、2株为蓝藻。(2)5株绿藻SM-2-1、DB-2-1、DB-2-2、SD-1和SD-2的序列长度分别为664 bp、663 bp、662 bp、589 bp和688 bp,GC含量分别为33.43%、49.47%、50.15%、50.76%和51.01%;2株蓝藻YJ-3、YJ-2的序列长度分别为570 bp和465 bp,GC含量分别为46.31%和49.03%。(3)序列比对并构建系统树分析发现,5株结皮绿藻可分为4个分支,分别为栅藻科(Scenedesmaceae)2株(DB-2-1和SM-2-1)、衣藻目(Chlamydomonadales)、环藻科(Sphaeropleaceae)、真眼点藻科(Eustigmataceae),但5株绿藻的5.8S+ITS2序列在属内差异小、非常保守,极易确定到属;2株蓝藻YJ-3和YJ-2聚在同一大类分支的伪鱼腥藻科(Pseudanabaenaceae)中,但分别归于不同的属。研究认为:5株结皮绿藻中DB-2-1藻株是Scotiellopsis属的一种、SM-2-1归于尖带藻属(Acutodesmus)、DB-2-2可能是红球藻属(Haematococcus)的一物种、SD-1是Ankyra属的一物种、SD-2归于真眼点藻属(Eustigmatos);2株生物结皮蓝藻中YJ-3可能是伪鱼腥藻科的一新物种、YJ-2可能是细鞘丝藻属(Leptolyngbya)亲缘关系较近的一新物种。     

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。       

洱海鱼腥藻优势种的形态鉴定与16S rRNA 基因序列分析

分别在2004年、2005年和2006年洱海鱼腥藻水华暴发时期,分离优势种,获得藻株EH-A、EH-B和EH-C,通过形态学特征和16S rRNA基因序列分析鉴定了藻株的种类.选用藻丝的形态、气囊的存在与否、异形胞和孢子的位置、各种细胞的形状以及营养细胞、异形胞和孢子的大小等传统的分类特征描述藻株的形态.依据形态特征,初步判断这3个藻株可能为卷曲鱼腥藻(Anabaena circinalis)或 A.crassa株系成员.利用16S rRNA基因序列构建邻接树分析了藻株间的系统进化关系,分析表明:藻株EH-A、EH-B和EH-C序列的同源性达到100%,且与A.circicular 和A.crassa藻株组成一个群(cluster),其藻株间的序列相似度高达100%,进一步说明藻株EH-A、EH-B和EH-C为相同的物种,且均为卷曲鱼腥藻(A.circinalis)或A.crassa.     

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。     

贵州中部埃迪卡拉系—寒武系留茶坡组的颤藻化石*

在我国贵州省麻江县埃迪卡拉纪—寒武纪过渡时期的留茶坡组硅质岩中,发现了丰富的丝状蓝藻化石。通过观察岩石薄片共鉴定出蓝藻门颤藻科的6个种,包括Calyptothrix perfecta,Calyptothrix major,Megathrix longus,Obruchevellasp.,Palaeolyngbya barghoorniana,以及Palaeolyngbyasp.。此外,还发现一种未定名的丝状微体化石,经观察与对比,认为该丝状化石形态接近于现代鞘丝藻Lyngbya,由于其胶鞘具有独特的规则环纹,可能为鞘丝藻属的一个新种,在本组硅质岩中为首次发现。该发现丰富了留茶坡组的微体生物化石资料,为埃迪卡拉纪—早寒武世的生物演化提供新的化石材料。     

淡水刚毛藻目一新组合种及其系统发育分析

作者于2012-2014年先后在湖北省采集到4株刚毛藻目的丝状绿藻。经鉴定,其形态特征和莫拉瓦刚毛藻Cladophora moravica Gardavsk(1986)非常相似;但相较于刚毛藻属各个种的形态,4株藻体更接近黑孢藻科的分类特征。基于SSU和LSU rDNA序列构建的系统发育树显示,4株刚毛藻目的丝状绿藻均属于黑孢藻科拟湖球藻属(Aegagropilopsis)。据此,作者建议将该种名称作为一个新组合处理,即莫拉瓦拟湖球藻Aegagropilopsis moravica(Dvoˇrák)Zhao et Liu com.nov。拟湖球藻属仅含有3个种:莫拉瓦拟湖球藻、硬枝拟湖球藻和棒状拟湖球藻,其形态特征与湖球藻相近且易混淆,故采用核基因组序列分析方法区分二者是非常有效的。     

鞘丝菌属的一个新种

自同氮蓝藻藻种培养池的土壤中分离得到一株具有透明薄鞘、宽度均一、长度不等、可届挠滑行运动的丝状菌。随菌龄增长或培养条件的改变,丝状体内分隔形成单列杆状细胞链。此种杆状细胞可由鞘内滑出形成新的丝状体。产生内生芽孢。革兰氏染色阳性,但不稳定。30—320c生长良好,45℃亦可生长,并形成大量不规则卷曲的螺旋状丝状体。在含铁蛋白胨琼脂上出现膨大的葫芦状细胞。初次分离物对固氮蓝藻细胞有明显的溶解能力。经鉴定为鞘丝菌属(Coleomitus)的一个新种,命名为直孢鞘丝菌(Coleomitus rectisporus sp．nov．)。     

Genotypic and phenotypic diversity of cyanobacteria assigned to the genus <Emphasis Type="Italic">Phormidium</Emphasis> (Oscillatoriales) from different habitats and geographical sites

In this study, 30 strains of filamentous, non-heterocystous cyanobacteria from different habitats and different geographical regions assigned to diverse oscillatorian genera but here collectively referred to as members of the Phormidium group have been characterized using a polyphasic approach by comparing phenotypic and molecular characteristics. The phenotypic analysis dealt with cell and filament morphology, ultrastructure, phycoerythrin content, and complementary chromatic adaptation. The molecular phylogenetic analyses were based on sequences of the 16S rRNA gene and the adjacent intergenic transcribed spacer (ITS). The sequences were located on multiple branches of the inferred cyanobacterial 16S rRNA tree. For some, but not all, strains with identical 16S rDNA sequences, a higher level of discrimination was achieved by analyses of the less conserved ITS sequences. As shown for other cyanobacteria, no correlation was found between position of the strains in the phylogenetic tree and their geographic origin. Genetically similar strains originated from distant sites while other strains isolated from the same sampling site were in different phylogenetic clusters. Also the presence of phycoerythrin was not correlated with the strains’ position in the phylogenetic trees. In contrast, there was some correlation among inferred phylogenetic relationship, original environmental habitat, and morphology. Closely related strains came from similar ecosystems and shared the same morphological and ultrastructural features. Nevertheless, structural properties are insufficient in themselves for identification at the genus or species level since some phylogenetically distant members also showed similar morphological traits. Our results reconfirm that the Phormidium group is not phylogenetically coherent and requires revision.     

Natronolimnobius baerhuensis gen. nov., sp. nov. and Natronolimnobius innermongolicus sp. nov., novel haloalkaliphilic archaea isolated from soda lakes in Inner Mongolia, China

Three novel isolates of haloalkaliphilic archaea, strains IHC-005^T, IHC-010, and N-1311^T, from soda lakes in Inner Mongolia, China, were characterized to elucidate their taxonomic positions. The three strains were aerobic, Gram-negative chemoorganotrophs growing optimally at 37–45°C, pH 9.0–9.5, and 15–20% NaCl. Cells of strains IHC-005^T/IHC-010 were motile rods, while those of strain N-1311^T were non-motile pleomorphic flats or cocci. The three strains contained diphytanyl and phytanyl-sesterterpanyl diether derivatives of phosphatidylglycerol and phosphatidylglycerophosphate methyl ester. No glycolipids were detected. On phylogenetic analysis of 16S rRNA gene sequences, they formed an independent cluster in the Natro group of the family Halobacteriaceae. Comparison of their morphological, physiological, and biochemical properties, DNA G+C content and 16S rRNA gene sequences, and DNA-DNA hybridization study support the view that strains IHC-005^T/IHC-010 and strain N-1311^T represent separate species. Therefore, we propose Natronolimnobius baerhuensis gen. nov., sp. nov. for strains IHC-005^T (=CGMCC 1.3597^T =JCM 12253^T)/IHC-010 (=CGMCC 1.3598=JCM 12254) and Natronolimnobius innermongolicus sp. nov. for N-1311^T (=CGMCC 1.2124^T =JCM 12255^T).     

Diversity of Freshwater <Emphasis Type="Italic">Thioploca</Emphasis> Species and Their Specific Association with Filamentous Bacteria of the Phylum <Emphasis Type="Italic">Chloroflexi</Emphasis>

Phylogenetic diversity among filamentous sulfur-oxidizing bacteria of the genus Thioploca inhabiting freshwater/brackish environments was analyzed in detail. The 16S rRNA gene sequence of Thioploca found in a freshwater lake in Japan, Lake Okotanpe, was identical to that of Thioploca from Lake Ogawara, a brackish lake. The samples of the two lakes could be differentiated by the sequences of their 23S rRNA genes and 16S–23S rRNA internal transcribed spacer (ITS) regions. The 23S rRNA-based phylogenetic relationships between Thioploca samples from four lakes (Lake Okotanpe, Lake Ogawara, Lake Biwa, and Lake Constance) were similar to those based on the 16S rRNA gene sequences. In addition, multiple types of the ITS sequences were obtained from Thioploca inhabiting Lake Okotanpe and Lake Constance. Variations within respective Thioploca populations were also observed in the analysis of the soxB gene, involved in sulfur oxidation. As major members of the sheath-associated microbial community, bacteria of the phylum Chloroflexi were consistently detected in the samples from different lakes. Fluorescence in situ hybridization revealed that they were filamentous and abundantly distributed within the sheaths of Thioploca.     

16S rRNA GENE HETEROGENEITY IN THE FILAMENTOUS MARINE CYANOBACTERIAL GENUS LYNGBYA1

The SSU (16S) rRNA gene was used to investigate the phylogeny of the cyanobacterial genus Lyngbya as well as examined for its capacity to discriminate between different marine species of Lyngbya. We show that Lyngbya forms a polyphyletic genus composed of a marine lineage and a halophilic/brackish/freshwater lineage. In addition, we found morphological and genetic evidence that Lyngbya spp. often grow in association with other microorganisms, in particular smaller filamentous cyanobacteria such as Oscillatoria, and propose that these associated microorganisms have led to extensive phylogenetic confusion in identification of Lyngbya spp. At the species level, the phylogenetic diversity obtained from the comparison of 16S rRNA genes exceeded morphological diversity in Lyngbya. However, the expectation that this improved phylogeny would be useful to species and subspecies identification was eliminated by the fact that phylogenetic species did not correlate in any respect with the species obtained from current taxonomic systems. In addition, phylogenetic identification was adversely affected by the presence of multiple gene copies within individual Lyngbya colonies. Analysis of clonal Lyngbya cultures and multiple displacement amplified (MDA) single‐cell genomes revealed that Lyngbya genomes contain two 16S rRNA gene copies, and that these typically are of variable sequence. Furthermore, intragenomic and interspecies 16S rRNA gene heterogeneity was approximately of the same magnitude. Hence, the intragenomic heterogeneity of the 16S rRNA gene overestimates the microdiversity of different strains and does not accurately reflect speciation within cyanobacteria, including the genus Lyngbya.     

Epilithic cyanobacterial communities of a marine tropical beach rock (Heron Island, Great Barrier Reef): diversity and diazotrophy

The diversity and nitrogenase activity of epilithic marine microbes in a Holocene beach rock (Heron Island, Great Barrier Reef, Australia) with a proposed biological calcification "microbialite" origin were examined. Partial 16S rRNA gene sequences from the dominant mat (a coherent and layered pink-pigmented community spread over the beach rock) and biofilms (nonstratified, differently pigmented microbial communities of small shallow depressions) were retrieved using denaturing gradient gel electrophoresis (DGGE), and a clone library was retrieved from the dominant mat. The 16S rRNA gene sequences and morphological analyses revealed heterogeneity in the cyanobacterial distribution patterns. The nonheterocystous filamentous genus Blennothrix sp., phylogenetically related to Lyngbya, dominated the mat together with unidentified nonheterocystous filaments of members of the Pseudanabaenaceae and the unicellular genus Chroococcidiopsis. The dominance and three-dimensional intertwined distribution of these organisms were confirmed by nonintrusive scanning microscopy. In contrast, the less pronounced biofilms were dominated by the heterocystous cyanobacterial genus Calothrix, two unicellular Entophysalis morphotypes, Lyngbya spp., and members of the Pseudanabaenaceae family. Cytophaga-Flavobacterium-Bacteroides and Alphaproteobacteria phylotypes were also retrieved from the beach rock. The microbial diversity of the dominant mat was accompanied by high nocturnal nitrogenase activities (as determined by in situ acetylene reduction assays). A new DGGE nifH gene optimization approach for cyanobacterial nitrogen fixers showed that the sequences retrieved from the dominant mat were related to nonheterocystous uncultured cyanobacterial phylotypes, only distantly related to sequences of nitrogen-fixing cultured cyanobacteria. These data stress the occurrence and importance of nonheterocystous epilithic cyanobacteria, and it is hypothesized that such epilithic cyanobacteria are the principal nitrogen fixers of the Heron Island beach rock.     

When Is A Lineage A Species? A Case Study In Myxacorys gen. nov. (Synechococcales: Cyanobacteria) With The Description of Two New Species From The Americas

Soil cyanobacteria are crucial components of biological soil crusts and carry out many functions in dryland ecosystems. Despite this importance, their taxonomy and population genetics remain poorly known. We isolated 42 strains of simple filamentous cyanobacteria previously identified as Pseudophormidium hollerbachianum from 26 desert locations in the North and South America and characterized these strains using a total evidence approach, that is, using both morphological and molecular data to arrive at taxonomic decisions. Based on a phylogenetic analysis of 16S rRNA gene sequences, we propose and characterize Myxacorys gen. nov. with two new species Myxacorys chilensis, the generitype, and M. californica. We also found distinct 16S‐23S ITS sequence variability within species in our dataset. Especially interesting was the presence of two distinct lineages of M. californica obtained from locations in close spatial proximity (within a few meters to kilometers from each other) suggesting niche differentiation. The detection of such unrecognized lineage‐level variability in soil cyanobacteria has important implications for biocrust restoration practices and conservation efforts.     

Relationships between the Arctic and the Antarctic cyanobacteria; three Phormidium-like strains evaluated by a polyphasic approach

Selected strains of filamentous Phormidium-like cyanobacteria isolated from two Arctic regions (Ellesmere Island, High Canadian Arctic and Svalbard) and from Antarctica (Antarctic peninsula, South Shetland Islands and South Orkney Islands) were studied. The polyphasic approach used included phenotypic observations of morphological features and genotypic analyses (restriction fragment length polymorphism of 16S rRNA gene, internal transcribed space, 16S rRNA gene sequence analysis). Although genotypes generally correspond to observed morphotypes, the genetic analyses revealed a high degree of biodiversity that could not be unveiled using solely morphological evaluations. According to the phylogenetic analysis, the three clones were divided into two major clades, indicating that the phylogenetic distance between Arct-Ph5/Ant-Ph68 and Ant-Ph58 was so large they belonged to different genera. The polyphyletic position of strains of the genus Phormidium was confirmed by this study, attesting the need to entirely revise classification in this taxon in the future.     

Preliminary studies of cyanobacteria, picoplankton, and virioplankton in the Salton Sea with special attention to phylogenetic diversity among eight strains of filamentous cyanobacteria

Cyanobacterial diversity in the Salton Sea, a high-salinity, eutrophic lake in Southern California, was investigated using a combination of molecular and morphological approaches. Representatives of a total of 10 described genera (Oscillatoria, Spirulina, Arthrospira, Geitlerinema, Lyngbya, Leptolyngbya, Calothrix, Rivularia, Synechococcus, Synechocystis) were identified in the samples; additionally, the morphology of two cultured strains do not conform to any genus recognized at present by the bacteriological system. Genetic analysis, based on partial 16S rRNA sequences suggested considerable cryptic genetic variability among filamentous strains of similar or identical morphology and showed members of the form-genus Geitlerinema to be distributed among three major phylogenetic clades of cyanobacteria. Cyanobacterial mats, previously described from the Sea were, in fact, composed of both filamentous cyanobacteria and a roughly equivalent biomass of the sulfur-oxidizing bacterium Beggiatoa, indicating their formation in sulfide rich regions of the lake. Flow cytometric analysis of the water samples showed three striking differences between samples from the Salton Sea and representative marine waters: (1) phycoerythrin-containing unicells, while abundant, were much less abundant in the Salton Sea than they were in typical continental shelf waters, (2) Prochlorococcus appears to be completely absent, and (3) small (3–5 m) eukaryotic algae were more abundant in the Salton Sea than in typical neritic waters by one-to-two orders-of-magnitude. Based on flow cytometric analysis, heterotrophic bacteria were more than an order of magnitude more abundant in the Salton Sea than in seawater collected from continental shelf environments. Virus particles were more abundant in the Salton Sea than in typical neritic waters, but did not show increases proportionate with the increase in bacteria, picocyanobacteria, or eukaryotic algae.