MOLECULAR PHYLOGENY OF SELECTED MEMBERS OF THE ORDER THALASSIOSIRALES (BACILLARIOPHYTA) AND EVOLUTION OF THE FULTOPORTULA1

Recent phylogenetic studies of the diatoms indicate that members of the order Thalassiosirales occupy an interesting position in the diatom evolutionary tree. Despite their radial morphology and scaly auxospores, they are consistently recovered in molecular analyses as a member of subdivision Bacillariophytina and a sister clade to non‐fultoportulate and non‐radial lithodesmioids. This study included 46 species from nine traditionally accepted extant genera, and analyzed 43 nuclear small subunit (SSU) rRNA sequences in parallel with a survey of the variation in fultoportula structure. Three possible scenarios leading to the evolution of the fultoportula are discussed in the context of molecular and morphological similarities between the examined Thalassiosirales and their SSU rRNA sister clade Lithodesmiales. We speculate that the fultoportula might be derived by a modification of either a cribrum in an areola (fultoportula within an areola), or structures similar to marginal ridges now seen in lithodesmioids around a cluster of poroids (fultoportula in a tube), or finally, that the central fultoportula may have an origin different from the marginal fultoportulae. Our data confirm that fultoportula‐bearing diatoms constitute a natural phylogenetic group. The families Thalassiosiraceae, Skeletonemaceae, and Stephanodiscaceae and the genus Thalassiosira Cleve were unexpectedly found to be paraphyletic. Further, Cyclotella Kutz. and Stephanodiscus Ehr. may not be closely related and some species of these genera are more closely allied to other species of Thalassiosira. The generitype, T. nordenskioeldii, is embedded within a large poorly structured cluster of species that includes several members of Thalassiosira, Planktoniella sol, Minidiscus trioculatus, and two members of Stephanodiscus. An emendment of the order Lithodesmiales and the family Lauderiaceae are proposed.     

采自广东大亚湾水域的海链藻属一新种——扇形海链藻

为澄清广东大亚湾水域海链藻属的物种多样性, 研究建立了它的单克隆培养株系。结合光镜和扫描电镜下的形态学特征, 以及基于核糖体大、小亚基序列的分子系统发育分析, 报道了中国海链藻属的1个新种, 即扇形海链藻Thalassiosira flabellata sp. nov. X.H. Guo, Y.Q. Guo & Y. Li。该种以短链状群体或单细胞生活, 壳面具有一圈壳缘支持突、一个中央支持突和一个壳缘唇形突。壳面孔纹束状排列, 是此种的典型特征。此种与帕尔海链藻T. partheneia较为相似, 区别特征如下: 此种的壳面支持突和唇形突均具有外管, 而帕尔海链藻的突起没有外管, 只有内管; 其次, 此种的壳面较平, 而帕尔海链藻的壳面隆起呈鼓状; 第三, 此种的中央支持突具有3个围孔, 而帕尔海链藻具有4—5个围孔。     

THALASSIOSIRA ANDAMANICA SP. NOV. (BACILLARIOPHYCEAE), A NEW DIATOM FROM THE ANDAMAN SEA (THAILAND)

A new marine diatom, Thalassiosira andamanica, is described from light and electron microscopy. The specimens were collected in the vicinity of Phuket Marine Biological Center, Thailand, and later brought into clonal culture. Thalassiosira andamanica possess a rimoportula with a pronounced outer extension, one marginal ring of fultoportulae, and three rings of fultoportulae on the valve face. Cells are united into colonies by a single thread secreted through a central fultoportula. Marginal fultoportulae extensions are shortest on the inside of the valve. The areolae are arranged in sectors, and the valve margin is ribbed with approximately 38 ribs in 10 μm. The valvocopula and copula have rows of pores, four to six pores in 1 μm. Apparently, the pleurae are hyaline. Experiments with a clonal culture isolated at Phuket, Thailand, showed that growth (cell divisions·24 h⁻¹) was reduced for cultures grown at 14° C compared to those grown at 19°, 24°, or 30° C. The maximum growth rate (2.2 divisions·24 h⁻¹) was at 30° C. Thalassiosira andamanica is compared with morphologically similar taxa. On the basis of morphological features and the response to different temperature regimens, it is concluded that this taxon must be recognized as a new warm-water species. In addition, T. andamanica does not clearly belong to any of the two subgroups of species of Thalassiosira. To accommodate the morphological characteristics of T. andamanica, the establishment of a possible third subgroup is discussed.     

Description of Conticribra tricircularis,a new genus and species of Thalassiosirales,with a discussion on its relationship to other continuous cribra species of Thalassiosira Cleve (Bacillariophyta) and its freshwater origin

A new diatom genus Conticribra is erected to accommodate C. tricircularis, described from a freshwater Pliocene deposit in Trout Creek, Oregon (USA). The genus accommodates species possessing: (i) loculate areolae with (semi-) continuous cribra; (ii) non-plicated valve face; (iii) rimoportula located on the valve mantle, replacing a fultoportula. Conticribra tricircularis has no valve face fultoportulae and can easily be distinguished by its marginal fultoportulae with four satellite pores arranged in three rings. Three species are transferred to the new genus from Thalassiosira sensu lato. Using evidence from the fossil record and recent molecular data, a hypothesis concerning the freshwater origin of Conticribra is discussed.     

Thalassiocyclus pankensis sp. nov., a new diatom from the Panke Swamp, northern Japan (Bacillariophyta)

Thalassiocyclus pankensis sp. nov. is described from Panke Swamp (Panke-numa) located in Horonobe Town, Hokkaido, northern Japan. The species is characterized by a strong transversely undulate central area of the valve face with one to three fultoportulae on the elevated part and one fultoportula on the depressed part. The external valve face has costate radial interfascicles and very wide fascicles on the marginal area. Thickened openings of mantle fultoportulae are located at the end of interfascicles with roofed board spines. The cingulum consists of five bands. The new species is compared to the only other species belonging to genus Thalassiocyclus , T. lucens (Hustedt) Håkansson & Mahood. Thalassiocyclus pankensis is the second species in the genus Thalassiocyclus and also the first report of a Thalassiocyclus species in Japan.     

Morphology and molecular phylogeny of Thalassiosira sinica sp. nov. (Bacillariophyta) with delicate areolae and fultoportulae pattern

Thalassiosira Cleve is one of the most species-rich marine diatom genera. Previous studies have mainly focused on polar and temperate areas, but recent studies on material from Asian waters suggested that a high and undescribed species diversity of Thalassiosira occurs in Asia. On the basis of plankton samples collected from the South China Sea, a new species, T. sinica sp. nov. Y. Li & Y. Q. Guo is described. The morphology of the cells was examined by light and scanning electron microscopy. The hypervariable region of the nuclear large-subunit ribosomal DNA and the relatively conserved region of the nuclear small-subunit ribosomal DNA were sequenced for information on phylogenetic relationships. The living cells are usually solitary and drum-shaped. The areolae on the valve are delicate, small and arranged in fascicles. In addition to a regular ring of marginal fultoportulae on the valve edge, T. sinica possesses one central fultoportula and a number of fultoportulae arranged into 2–3 irregular rings on the valve face. A rimoportula located inside the ring of marginal fultoportulae possesses a long and strong external tube. The valvocopula and the copulae have rows of pores, but the pores on the valvocopula are larger than those on the copulae. Thalassiosira sinica appears to be included in subgroup C sensu Gedde because of a rimoportula with a distinct external tube located on the valve face. The molecular phylogeny, inferred from both SSU and LSU sequences, does, however, not support the validity of subgroup C, as the closest allies of T. sinica here turned out to be T. diporocyclus and T. lundiana, species in which the rimoportulae are located on the valve margin.     

Cyclotella orientalis sp. nov. (Bacillariophyceae) from South Korea

Cyclotella ortentalis sp. nov, is described from an epilithic sample taken in Lake Imha, located in Kyungsangbuk-do. South Korea. The species is characterized by a conspicuous and short costa regularly extending from each marginal fultoportula to the valve margin. It can be included in the Cyclotella stelligeroid group. The new species can be separated from the most similar species C. stelligera Cleve et Grunov var. stelligera by the more distant position of marginal fultoportulae from the valve margin, the external long tubes of fultoportulae and having short costae. It differs from C. stelligera var. robusta Must, and var. hyalina Hust., in having much denser marginal striae; from C. glomerata H. Bachm., C. pseudostelligera Hust., C. stelligeroides Must, and C. woltereckii Hust., in its much larger valve, coarser marginal striae and short costae with fultoportulae.     

海链藻属重要形态学特征的二次研究——以艾伦海链藻为例

海链藻属（Thalassiosira Cleve）是硅藻门中物种多样性最为丰富的大属之一,广泛分布于全球各类水体。目前沿用的海链藻属分类体系是基于形态特征建立的,尤其是电镜下的超微特征具有重要的分类价值。然而,也有研究认为某些形态特征存在不稳定或变化的情况,应对其开展二次研究和重新评估。故此,本文选择全球近岸广布种艾伦海链藻（T.allen Takano）作为典型代表,基于建立的单克隆培养株系,结合传统形态学方法和现代分子生物学技术,对其重要形态特征进行了二次研究。结果显示：（1）艾伦海链藻壳面孔纹的排列方式存在一定的变化,呈切线形或偏心状;壳缘唇形突的位置多有变化,占据一个壳缘支持突的位置、或位于两个壳缘支持突正中间、或靠近其中一个。艾伦海链藻这两个特征并不具有较高的稳定性,其分类学意义或应降低;（2）壳缘不具有肋纹,此特征相对稳定,可作为艾伦海链藻的识别特征之一;（3）艾伦海链藻具有较保守的遗传多样性,尚未发现（拟）隐形种。结合其它海链藻的形态学报道,我们认为孔纹排列方式易受环境的影响,稳定性较差,在分类时应慎重应用;壳缘唇形突的位置在少数海链藻属物种中易发生变化,分类时应视具体物种而针对性应用;支持突等突起特征较为稳定,在海链藻属物种的分类体系中仍可作为重要依据;艾伦海链藻中壳缘肋纹具有较好的稳定性,可作为分类依据,但在其它海链藻属物种中是否也具有很好的分类价值,还需要更多的研究加以验证;分子信息在形态特征上的二次研究和评价中具有重要应用价值,但目前相关分子数据的缺乏和不足限制了对其进一步的研究。本研究对重新评估艾伦海链藻形态特征的分类学价值、完善现存的分类学体系提供了参考。     

我国沿海双唇形突海链藻的形态学研究

唇形突是海链藻中常见的一类突起, 其形态学特征和排列方式是重要的分类学依据。绝大多数海链藻只有一个唇形突, 两个唇形突的现象仅报道于极少数海链藻种类。我国关于双唇形突海链藻的报道较少。研究利用扫描电镜(SEM)技术, 对采自我国沿海海域的自然水样, 以及分离获得的单克隆培养藻株进行了形态学观察, 针对其中的海链藻属Thalassiosira Cleve种类开展了形态学的专题研究。报道了4个具有双唇形突的海链藻种类, 分别是有翼海链藻T. bipartita (Rattray) Hallegraeff、脆弱海链藻T. fragilis Fryxell、亨氏海链藻T. hendeyi Hasle &; Fryxell和维斯吉思海链藻T. visurgis Hustedt, 其中有3个为我国新记录种类: 脆弱海链藻、亨氏海链藻和维斯吉思海链藻。对每个种类的形态学特征、生活习性和生态分布进行了描述, 提供了扫描电镜照片, 对相似种类的形态学特征进行了比较研究, 分析了唇形突的变化类型及其分类学价值。     

小字海链藻双卫变种,海链藻属一新变种

采用毛细管复洗技术建立海链藻属(Thalassiosira)物种的单克隆培养株系,结合形态学和分子系统学研究,对小字海链藻原变种(Thalassiosira minuscula var.minuscula Krasske)的形态特征进行观察,并报道了1个新变种:小字海链藻双卫变种(T.minuscula var.bicustodis X.H.Guo,Y.Q.GuoY.Li)。双卫变种与原变种的形态特征基本一致,区别仅在于唇形突的伴生支持突数目不同,双卫变种具有2个,原变种只有1个。基于对多个株系生活史的连续观察,确认其伴生支持突的数目稳定,具有分类学意义。     

DIVERSITY IN THE GENUS SKELETONEMA (BACILLARIOPHYCEAE): III. PHYLOGENETIC POSITION AND MORPHOLOGICAL VARIABILITY OF SKELETONEMA COSTATUM AND SKELETONEMA GREVILLEI,WITH THE DESCRIPTION OF SKELETONEMA ARDENS SP. NOV.1

Skeletonema costatum (Grev.) Cleve emend. Zingone et Sarno and S. grevillei Sarno et Zingone were known only from the type material collected from Hong Kong waters more than a century ago. Both species have now been collected as live material, and their morphology and phylogenetic position are investigated in this study. Eight Skeletonema strains isolated from Florida, USA; Uruguay; and Brazil are attributed to S. costatum, while one strain from Oman is ascribed to S. grevillei based on morphological similarity to the type material of these species. In addition, a new Skeletonema species, S. ardens Sarno et Zingone, is described for a strain from Singapore and two from northern Australian waters. Skeletonema ardens has terminal fultoportula processes ending in a tapered, undulate protrusion and long intercalary fultoportulae with 1:1 junctions. The rimoportula of terminal valves is located at the margin of the valve face. No major morphological variations were observed within S. grevillei and S. ardens along a salinity gradient, whereas in S. costatum, the processes shortened and the valves came into close contact at low salinities, as already described for S. subsalsum (Cleve) Bethge. Consistent with their morphology, Skeletonema costatum and Skeletonema subsalsum also had similar rDNA sequences. Skeletonema grevillei and S. ardens were distinct in the large subunit (LSU) phylogeny. Skeletonema ardens exhibited consistent intraspecific genetic differences in both the LSU and small subunit (SSU) rDNA.     

TRENDS IN THE EVOLUTION OF THE EUGLENID PELLICLE

Abstract Trends in the evolution of the euglenid pellicle were described using phylogenetic methods on 18S rDNA, morphological, and combined data from 25 mostly phototrophic taxa. The tree topology from a total‐evidence analysis formed a template for a synthetic tree that took into account conflicting results derived from the partitioned datasets. Pellicle character states that can only be observed with the assistance of transmission and scanning electron microscopy were phylogenetically mapped onto the synthetic tree to test a set of previously established homology statements (inferences made independently from a cladogram). The results permitted us to more confidently infer the ancestral‐derived polarities of character state transformations and provided a framework for understanding the key cytoskeletal innovations associated with the evolution of phototrophic euglenids. We specifically addressed the character evolution of (1) the maximum number of pellicle strips around the cell periphery; (2) the patterns of terminating strips near the cell posterior end; (3) the substructural morphology of pellicle strips; (4) the morphology of the cell posterior tip; and (5) patterns of pellicle pores on the cell surface.     

THE INTERACTION OF BEHAVIORAL AND MORPHOLOGICAL CHANGE IN THE EVOLUTION OF A NOVEL LOCOMOTOR TYPE: “FLYING” FROGS

“Flying” frogs have evolved independently several times among anurans. In all cases flyers are distinguished from their nonflying arboreal relatives by a unique set of morphological features and behavioral postures. Using both live animal field tests and wind tunnel models, this study examines the importance of this characteristic morphology and limb position on five aerial performance variables: horizontal traveling distance, minimum glide speed, maximum time aloft, maneuverability, and stability. Comparison of relative performance between a model frog with a generalized nonflying morphology and limb position and a model frog with flying morphology and limb position reveals that the morphological and positional features associated with “flying” actually decrease horizontal traveling distance but improve maneuverability. This finding suggests that maneuverability rather than horizontal travel may be the key performance parameter in the evolution of “flying” frogs. More generally, this study illustrates that (1) derived morphological and postural features do not necessarily change a suite of performance variables in the same way, and (2) the performance consequences of postural shifts are a function of morphology. These findings indicate that the potential complexity of morphological and behavioral interactions in the evolution of new adaptive types is much greater than previously considered.     

Phylum Tardigrada: an “individual” approach

Phylum Tardigrada consists of ～1000 tiny, hardy metazoan species distributed throughout terrestrial, limno‐terrestrial and oceanic habitats. Their phylogenetic status has been debated, with current evidence placing them in the Ecdysozoa. Although there have been efforts to explore tardigrade phylogeny using both morphological and molecular data, limitations such as their few morphological characters and low genomic DNA concentrations have resulted in restricted taxonomic coverage. Using a protocol that allows us to identify and extract DNA from individuals, we have sequenced 18S rDNA from 343 tardigrades from across the globe. Using maximum parsimony and Bayesian analyses we have found support for dividing Order Parachela into three super‐families and further evidence that indicates the traditional taxonomic perspective of families in the class Eutardigrada are nonmonophyletic and require re‐working. It appears that conserved morphology within Tardigrada has resulted in conservative taxonomy as we have found cases of several discrete lineages grouped into single genera. Although this work substantially adds to the understanding of the evolution and taxonomy of the phylum, we highlight that inferences gained from this work are likely to be refined with the inclusion of further taxa—specifically representatives of the nine families yet to be sampled. © The Willi Hennig Society 2008.     

THE EVOLUTION OF THERMOTOLERANCE IN HOT SPRING CYANOBACTERIA OF THE GENUS SYNECHOCOCCUS

The extension of ecological tolerance limits may be an important mechanism by which microorganisms adapt to novel environments, but it may come at the evolutionary cost of reduced performance under ancestral conditions. We combined a comparative physiological approach with phylogenetic analyses to study the evolution of thermotolerance in hot spring cyanobacteria of the genus Synechococcus. Among the twenty laboratory clones of Synechococcus isolated from collections made along an Oregon hot spring thermal gradient, four different 16S ribosomal RNA gene sequences were identified. Phylogenies constructed using these sequence data indicated that the clones were polyphyletic but also that three of the four sequence groups formed a clade. Differences in thermotolerance were observed for clones with different 16S rRNA gene sequences, and comparison of these physiological differences within a phylogenetic framework provided evidence that more thermotolerant lineages of Synechococcus evolved from less thermotolerant ancestors. The extension of the thermal limit in these bacteria was correlated with a reduction in thermal niche breadth, which may have implications for the geographic distributions of these organisms. This study illustrates the utility of using phylogenetic comparative methods to investigate how evolutionary processes have shaped historical patterns of ecological diversification in microorganisms.     

COMMENT ON MARDEN (2013): “REANALYSIS AND EXPERIMENTAL EVIDENCE INDICATE THAT THE EARLIEST TRACE FOSSIL OF A WINGED INSECT WAS A SURFACE SKIMMING NEOPTERAN”

Marden's (2013) reanalysis of Knecht et al. (2011) suggesting that specimen SEMC‐F97 is the result of the skimming behavior of a neopteran insect and, more importantly, fossil evidence of “… surface skimming as a precursor to the evolution of flight in insects” (Marden 2013) is found to be deficient on three fronts: (1) the principal specimen was never viewed firsthand which led to significant morphological misinterpretations; (2) poorly designed and executed neoichnological experiments led to incredulous results; and (3) the assumption that this specimen is fossil evidence supporting the surface skimming hypothesis of the origin of insect flight despite the fact that since its induction into the literature that hypothesis has been refuted based on significant paleontological, phylogenetic, genetic, and developmental evidence.     

Detecting the historical signature of key innovations using stochastic models of character evolution and cladogenesis

Phylogenetic evidence for biological traits that increase the net diversification rate of lineages (key innovations) is most commonly drawn from comparisons of clade size. This can work well for ancient, unreversed traits and for correlating multiple trait origins with higher diversification rates, but it is less suitable for unique events, recently evolved innovations, and traits that exhibit homoplasy. Here I present a new method for detecting the phylogenetic signature of key innovations that tests whether the evolutionary history of the candidate trait is associated with shorter waiting times between cladogenesis events. The method employs stochastic models of character evolution and cladogenesis and integrates well into a Bayesian framework in which uncertainty in historical inferences (such as phylogenetic relationships) is allowed. Applied to a well-known example in plants, nectar spurs in columbines, the method gives much stronger support to the key innovation hypothesis than previous tests.     

SPECIES BOUNDARIES IN THE TOXIC DINOFLAGELLATE PROROCENTRUM LIMA (DINOPHYCEAE,PROROCENTRALES), BASED ON MORPHOLOGICAL AND PHYLOGENETIC CHARACTERS1

Wild and cultured specimens of Prorocentrum lima (Ehrenb.) F. Stein from 26 widely different areas in 13 countries were examined in order to determine consistent characters for delimiting species boundaries in this taxon. The morphological characters valve shape, valve size, valve ornamentation, number and shape of valve pores, number and shape of marginal pores, and periflagellar platelets were observed using LM and SEM, and two molecular genetic regions were sequenced. We identified stable morphological characters that were consistent among wild specimens and all cultures, which were valve shape, valve ornamentation, and number and arrangement of periflagellar platelets. All cultures of P. lima identified by these characters formed a monophyletic group in phylogenetic analyses based on the two genes, which, however, included the species Prorocentrum arenarium. P. arenarium was determined to be within the range of morphological variation of P. lima, and therefore we synonymize the two taxa. Within this monophyletic group, P. lima was divided into several subclades in the all phylogenetic analyses. There were no morphological characters specifically related to any one subclade. The subclades appeared to correlate broadly to sample collection regions, suggesting that geographically separated populations may have become genetically distinct within this epi‐benthic species. We have emended species boundaries in P. lima.     

A multi-locus analysis of phylogenetic relationships within cheilostome bryozoans supports multiple origins of ascophoran frontal shields

Phylogenetic relationships within the bryozoan order Cheilostomata are currently uncertain, with many morphological hypotheses proposed but scarcely tested by independent means of molecular analysis. This research uses DNA sequence data across five loci of both mitochondrial and nuclear origin from 91 species of cheilostome Bryozoa (34 species newly sequenced). This vastly improved the taxonomic coverage and number of loci used in a molecular analysis of this order and allowed a more in-depth look into the evolutionary history of Cheilostomata. Maximum likelihood and Bayesian analyses of individual loci were carried out along with a partitioned multi-locus approach, plus a range of topology tests based on morphological hypotheses. Together, these provide a comprehensive set of phylogenetic analyses of the order Cheilostomata. From these results inferences are made about the evolutionary history of this order and proposed morphological hypotheses are discussed in light of the independent evidence gained from the molecular data.Infraorder Ascophorina was demonstrated to be non-monophyletic, and there appears to be multiple origins of the ascus and associated structures involved in lophophore extension. This was further supported by the lack of monophyly within each of the four ascophoran grades (acanthostegomorph/spinocystal, hippothoomorph/gymnocystal, umbonulomorph/umbonuloid, lepraliomorph/lepralioid) defined by frontal-shield morphology. Chorizopora, currently classified in the ascophoran grade Hippothoomorpha, is phylogenetically distinct from Hippothoidae, providing strong evidence for multiple origins of the gymnocystal frontal shield type. Further evidence is produced to support the morphological hypothesis of multiple umbonuloid origins of lepralioid frontal shields, using a step-wise set of topological hypothesis tests combined with examination of multi-locus phylogenies.