GENETIC ISOLATION BETWEEN THREE CLOSELY RELATED TAXA: FUCUS VESICULOSUS,F. SPIRALIS,AND F. CERANOIDES (PHAOPHYCEAE)1

All traditional markers, both phenotypic and phylogenetic, have failed to discriminate between the taxa composing the Fucus vesiculosus L., F. spiralis L., and F. ceranoides L. species complex, particularly in Brittany (France), so we used five microsatellite markers to compare the allelic frequencies of populations of the three taxa in this region. The aim of this study was to assess whether the different populations were grouped according to their geographical location, their habitat (open coast versus estuary), or their a priori taxonomic assignment. Species‐specific alleles were identified at one locus, demonstrating the utility of microsatellite markers for recognizing the three taxa in Brittany. Moreover, our results clearly support the separation of F. vesiculosus, F. spiralis, and F. ceranoides into distinct species, independently of geography. We also identified genetic differentiation between estuarine and coastal populations of F. vesiculosus.     

PHENOTYPIC VARIATION OF GROWTH TRAJECTORIES IN FINCHES

Evolution cannot proceed without phenotypic variation for selection to act on. This is particularly true of ontogenetic parameters because it is changes in these parameters that give rise to new phenotypes. I analyzed the amount and dimensionality of phenotypic variation on growth trajectories in early ontogeny in three species of finches (Fringillidae) using the recently developed infinite-dimensional model. For two species, eight traits were analyzed, and for a third, six traits. Growth data were analyzed only up to 6 d of age in two species and 8 d of age in the third. The results were very similar for all species and traits. A very large proportion of the phenotypic variation in growth trajectories was confined to a single dimension. This dimension corresponded to a simultaneous increase/decrease at all ages in early ontogeny. The eigenfunctions, each describing a family of similar-shaped growth trajectories, were highly collinear among traits. A high covariance existed among traits at the same and different ages. If some part of the phenotypic variation has an additive genetic basis, then any selection for a change in size at one age in one trait will lead to a response in a size at subsequent ages and in the other traits. This in turn suggests that morphological evolution frequently will move along a multivariate size axis, as has indeed been found in several taxa.     

MORPHOLOGICAL VARIABILITIES OF THREE CLOSELY RELATED MATING GROUPS OF CLOSTERIUM EHRENBERGII MENEGHINI (CHLOROPHYTA)1

The ranges of morphological variabilities in vegetative cells of three closely related mating groups of Closterium ehrenbergii Meneghini were statistically analyzed, having been grown under standard and uniform culture conditions, using Group A clones from Japan and Australia, Group B clones from Japan and Taiwan and Group H clones from Nepal. Significant differences in the morphological characters were not recognized between the two complementary mating types in any of the three groups. It has been shown that cells of Group A are smallest (mean width 50 μm and mean length 250 μm) and cells of Group B are largest (mean width 67 μm and mean length 404 μm), while cells of Group H are intermediate (mean width 57 μm and mean length 333 μm). There are considerable differences in the mean cell size between the three mating groups, although some intergrading clones were recognized. Degrees of the intergrading overlap were shown to be small between the sympatric groups (A and B) and large between the two pairs of allopatric groups (A and H & B and H). It has been shown that cells of Group A are shorter and much more curved than cells of the other two mating groups. Cells of Groups B and H are slender and less curved. It has also been shown that the ranges in cell size of each mating group are smaller than those currently accepted for C. ehrenbergii.     

TAXONOMIC REEXAMINATION OF 17 SPECIES OF NITELLA SUBGENUS TIEFFALLENIA (CHARALES,CHAROPHYCEAE) BASED ON INTERNAL MORPHOLOGY OF THE OOSPORE WALL AND MULTIPLE DNA MARKER SEQUENCES1

In an attempt to reconstruct the natural taxonomic system for Nitella, 17 species of Nitella subgenus Tieffallenia were reexamined using SEM observations of the internal morphology of the oospore wall (IMOW) and phylogenetic analyses of 4553 base pairs from multiple DNA markers (atpB, rbcL, psaB, and ITS‐5.8S rRNA genes). Our SEM observations identified three types of IMOW: homogeneous (HG), weakly spongy (W‐SG), and strongly spongy (S‐SG) types. Based on differences in the IMOW, species with reticulate or tuberculate oospore wall ornamentation in the external morphology of the oospore wall (EMOW) were subdivided into two distinct groups (characterized by the HG or S‐SG types of IMOW, respectively), which were robustly separated from each other in our molecular phylogenetic analyses. In our molecular phylogeny, the subgenus Tieffallenia consisted of four robust monophyletic groups—three clades of the HG type and a spongy (S‐SG and W‐SG) type clade—that were characterized by differences in the IMOW and EMOW. In addition, our SEM observations and sequence data verified the distinct status of five species (N. japonica Allen, N. oligospira A. Braun, N. vieillardii stat. nov., N. imperialis stat. nov., and N. morongii Allen) that R. D. Wood had assigned as infraspecific taxa. Moreover, our SEM observations of the IMOW also suggested that N. megaspora (J. Groves) Sakayama originally identified by LM includes at least two distinct species, characterized by W‐SG and S‐SG types of IMOW, respectively.     

HYBRIDIZATION AND CHLOROPLAST DNA VARIATION IN A PINUS SPECIES COMPLEX FROM ASIA

Heterologous hybridization of chloroplast DNA (cpDNA) involving 30 endonucleaseprobe combinations was used to analyze cpDNA variation in multiple individuals and populations of Pinus tabulaeformis (Carr.), Pinus yunnanensis (Franchèt) and Pinus massoniana (Lamb.). Restriction fragment patterns detected by several combinations distinguished among the three species. The obtained cpDNA markers were subsequently used to examine cpDNA variation of Pinus densata (Masters), a putative tertiary hybrid between P. tabulaeformis and P. yunnanensis. The analysis demonstrated that P. densata populations harbor three different haplotypes. Two of these haplotypes are characteristic of P. tabulaeformis and P. yunnanensis. However, the third haplotype found in P. densata appears to be absent in other extant Asian Pinus species. It is suggested that the observed cpDNA composition of P. densata populations is a result of past hybridization involving P. tabulaeformis, P. yunnanensis, and a third unknown or extinct taxon. Chloroplast DNA polymorphism in P. densata was much greater than that for nuclear allozyme markers in this and the other Pinus species. Population differentiation was also substantial in P. densata and exceeded that for allozyme markers. In contrast, no cpDNA polymorphism was detected in populations of P. tabulaeformis, P. yunnanensis, and P. massoniana. The study suggests that interspecific gene exchange may lead to the creation of stable cpDNA polymorphism in conifer hybrids.     

湖南、江西普通野生稻居群变异的数量分类研究

本文运用数量分类方法,对产于湖南、江西两省的普通野生稻居群和性状进行了聚类分析。结果初步揭示了分布于我国北界的普通野生稻居群的类型和性状间的关系,表明环境因素是决定居群类型的重要原因;东乡县两居群高度游离于所有其它居群,充分说明了其高度的特殊性和它们之间的遗传异质性。     

THE CONTRIBUTION OF MATING SYSTEM VARIATION TO REPRODUCTIVE ISOLATION IN TWO CLOSELY RELATED CENTAURIUM SPECIES (GENTIANACEAE) WITH A GENERALIZED FLOWER MORPHOLOGY

In closely related plant species that display strong similarities in phenology and pollinator communities, differences in breeding system and associated shifts in floral traits may have important effects on the magnitude and direction of heterospecific pollen flow and hybridization. Here, we quantified the strength of several pre‐ and postzygotic barriers acting between the facultatively outcrossing Centaurium erythraea and the predominantly selfing C. littorale via a suite of experiments, and estimated the frequency of hybridization in the field using molecular markers. The reproductive barriers primarily responsible for preventing hybridization were essentially prezygotic and these acted asymmetrically. Due to differences in floral display, pollen production, and pollen transfer rates, heterospecific pollen flow occurred predominantly from C. erythraea to C. littorale. In C. littorale, on the other hand, close anther–stigma positioning and resulting higher capacity for autonomous selfing functioned as an efficient barrier to counterbalance the higher risk for hybrid mating. In both species the action of all reproductive barriers resulted in a small opportunity for hybrid establishment, which was confirmed by the occurrence of only ～1% putative hybrids in the field. Our findings confirm that differences in breeding system affect heterospecific pollen transfer patterns and that autonomous selfing may efficiently prevent hybridization.     

POTENTIAL UTILITY OF MITOCHONDRIAL CYTOCHROME B AND ITS MRNA EDITING IN RESOLVING CLOSELY RELATED DINOFLAGELLATES: A CASE STUDY OF PROROCENTRUM (DINOPHYCEAE)1

Difficulties often occur in separating closely related dinoflagellate species. In this study, the potential utility of mitochondrial cytochrome b (cob) gene sequence and mRNA editing characteristics was assessed using Prorocentrum Ehrenberg as a model. The cob sequences and the patterns of their mRNA editing were analyzed for several Prorocentrum taxa. Results revealed little difference in cob sequence and mRNA editing characteristics between geographic populations of P. minimum (Pavillard) Schiller, while a notable difference was detected between different species (P. minimum and P. micans Ehrenberg). Furthermore, these P. minimum populations consistently formed a tight cluster on phylogenetic trees inferred from cob sequences as well as mRNA editing characteristics, whereas different Prorocentrum species were well separated, with a genetic distance of 0.0042±0.0024 for the former and 0.0141±0.0012 for the latter (P<0.01; two‐tailed t‐test). When the analysis was applied to the case of P. donghaiense Lu et Goebel and CCMP1517 strain of P. dentatum Stein, no differences were detected between these two taxa with respect to cob mRNA editing pattern and only small differences equivalent to those between P. minimum populations were detected in terms of cob sequence. On the cob sequence‐ and editing‐based phylogenetic trees, P. donghaiense and P. dentatum CCMP1517 consistently clustered together at a position sister to P. minimum. The results suggest that cob, combined with its mRNA editing, can potentially be a useful delineator of Prorocentrum species, and that P. donghaiense and P. dentatum CCMP1517 are most likely the same species and both are closely related to P. minimum.     

海南西部近岸浮游植物的周年变化及主要关联因素

报道了2008—2009年4季度海南西部近岸浮游植物群落的周年动态并探讨其主要关联因素。165份样品经鉴定共有浮游植物4门74属155种(含5变型和2变种),周年平均丰度为(6.36±4.75)×103cells/L。硅藻在物种组成和丰度上均占绝对优势,其次为甲藻,蓝藻(束毛藻)在7月增殖。主要优势种为菱形海线藻(Thalassionema nitzschioides)、奇异棍形藻(Bacillaria paradoxa)、具槽帕拉藻(Paralia sulcata)、旋链角毛藻(Chaetoceros curvisetus)、笔尖根管藻(Rhizosolenia styliformis)、束毛藻(Trichodesmium spp.)、海洋原甲藻(Prorocentrum micans)等。物种组成的季节差异较大,10月浮游植物种类贫乏,1月次之,4月、7月最丰富。丰度10月最高,季节差异并不明显。束毛藻在4月、7月呈斑块状群聚分布。浮游植物周年平均丰度并不高(<1.0×104cells/L)。不同季节优势种有明显的交错和变化,菱形海线藻、奇异棍形藻、具槽帕拉藻为全年优势种。浮游植物物种多样性指数和均匀度都表现出较高的值,均匀度与多样性指数的季节变化特点基本一致,群落多样性高的季节物种均匀度也好。物种多样性指数指示调查区水体遭受污染程度低,水质状况优。调查区各季节的浮游植物丰度与温度之间无显著的相关关系,1月丰度与盐度则呈密切负相关关系。10月浮游植物丰度与无机氮(DIN)呈密切的正相关关系。7月浮游植物丰度与活性磷酸盐(PO4-P)呈密切的负相关关系。浮游动物对浮游植物的摄食压力直接影响到后者的丰度变动,并伴随着海区生态系统的相关复杂现象及生物学过程的作用。     

MAJOR DEVELOPMENTAL REGULATORS AND THEIR EXPRESSION IN TWO CLOSELY RELATED SPECIES OF PORPHYRA (RHODOPHYTA)1

Little is known about the genetic and biochemical mechanisms that underlie red algal development, for example, why the group failed to evolve complex parenchyma and tissue differentiation. Here we examined expressed sequence tag (EST) data from two closely related species, Porphyra umbilicalis (L.) J. Agardh and P. purpurea (Roth) C. Agardh, for conserved developmental regulators known from model eukaryotes, and their expression levels in several developmental stages. Genes for most major developmental families were present, including MADS‐box and homeodomain (HD) proteins, SNF2 chromatin‐remodelers, and proteins involved in sRNA biogenesis. Some of these genes displayed altered expression correlating with different life history stages or cell types. Notably, two ESTs encoding HD proteins showed eightfold higher expression in the P. purpurea sporophyte (conchocelis) than in the gametophyte (blade), whereas two MADS domain‐containing paralogs showed significantly different patterns of expression in the conchocelis and blade respectively. These developmental gene families do not appear to have undergone the kinds of dramatic expansions in copy number found in multicellular land plants and animals, which are important for regulating developmental processes in those groups. Analyses of small RNAs did not validate the presence of miRNAs, but homologs of Argonaute were present. In general, it appears that red algae began with a similar molecular toolkit for directing development as did other multicellular eukaryotes, but probably evolved altered roles for many key proteins, as well as novel mechanisms yet to be discovered.     

INTRASPECIFIC VARIATION IN THE DINOFLAGELLATE PERIDINIUM VOLZIP1

Clonal isolates of Peridinium volzii Lemmerman were analyzed morphologically and biochemically. Morphological observations at the light microscope level show the clones to be different varieties and forms of the same species. Biochemical analysis by enzyme electrophoresis and flow cytometric determination of nuclear DNA quantities indicates that these isolates are genetically heterogeneous without any clear correlation existing between morphological variation and biochemical variation. Isozyme analysis, however, indicates that strains from the same location are generally more related to each other than they are to isolates with other geographical origins. In general, our results suggest the presence of genetic redundancy and a multiclonal origin for individuals of the same species present in the same locale.     

LINKAGE OF 5S RIBOSOMAL DNA TO OTHER rDNAS IN THE CHROMOPHYTIC ALGAE AND RELATED TAXA1

Gene organization within the nuclear ribosomal DNA cistron and linkage of the 5S rDNA gene to the cistron were surveyed in 20 taxa of protists representing most of the Chromophyta (stramenopiles) and representatives of the Dinophyceae (alveolata) and Euglenophyceae. The intergenic spacer, which separates adjacent cistrons, was first PCR-amplified from total DNA using primers anchored in the 3’end of the large subunit and the 5’end of the small subunit in the next downstream cistron. Presence of the 5S gene in the cistron was determined by a second round of PCR using primers anchored in the large subunit and the 5S gene. where 5S-linked rDNA was not detected in the cistron, the presence of 5S tandem repeating units were confirmed by the PCR of 5S-5S fragments from the total DNA. Results show that most of the Chromophyta, as well as Opalina, Proteromonas (colorless stramenopiles), Dinophyceae, and Euglenophyceae have a 5S-linked type of rDNA organization. In contrast, only tandem repeats of 5S rDNA were detected in Bacillariophyceae and Synurophyceae. The occurrence of 5S-unlinked rDNA is hypothesized to be the result of secondary transfer from an ancestral, linked 5S type. The 5S-linked type of rDNA organization is apparently common in protists. Given the fact that most of these protists have mitochondria with tubular or discoid cristae, as compared with flattened cristae common in higher plants and animals, we conclude that the 5S-linked type of rDNA diverged at a very early stage in the evolution of eukaryotes.     

Multivariate morphometric analysis of the Potamogeton compressus group (Potamogetonaceae)

The Potamogeton compressus group is a complex of three to five closely related species with a circumpolar distribution in the Northern Hemisphere. Multivariate morphometric analyses (principal component analysis, cluster analysis, canonical and classificatory discriminant analyses) were used to elucidate the patterns of variation within this group and to test the morphological differentiation of the species recognized in the current literature. From the entire distribution range, 156 specimens of the group were included in the numerical methods. Results from morphological comparison are discussed in relation to molecular data, reproductive behaviour and geographical distribution. Morphometric analyses provided evidence that this complex can be clearly divided into three groups, one of which was subdivided mainly on the basis of allopatric occurrence and genetic differentiation. These groups correspond to four species accepted here: P. acutifolius (temperate regions of Europe), P. compressus (boreal and temperate regions of Europe and Asia), P. manchuriensis (northeastern China and Russian Far East) and P. zosteriformis (boreal and temperate regions of North America). Two species, P. acutifolius and P. compressus, are partly sympatric, but clearly differentiated morphologically and genetically, and effectively isolated reproductively. Endemic P. manchuriensis is characterized by a unique combination of characters and an occurrence in a limited geographical area. Allopatric P. zosteriformis is weakly differentiated morphologically from P. compressus, but differs markedly in molecular markers correlated with geographical differentiation. It may represent a cryptic species. In contrast, a recently suggested concept of southern Siberian P. henningii was not supported by our analyses. Plants so named are considered here as slender phenotypes of the widespread and variable P. compressus. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 170 , 112–130.     

LIFE-CYCLE COMPONENTS OF SELECTION IN ERIGERON ANNUUS: II. GENETIC VARIATION

Genetic variation for seedling and adult fitness components was measured under natural conditions to determine the relative importance of the seedling stage for lifetime fitness in Erigeron annuus. Variation in lifetime reproductive success can result from both the persistent effects of genetic variation expressed among seedlings and from variation in adult fitness components. Analysis of covariance was used to separate the stage specific from the cumulative effects of genetic variance expressed earlier in the life cycle. E. annuus produces seeds through apomixis, which allowed measurement of the fitness of replicate genotypes from germination through the entire life cycle. There were significant differences among genotypes for date of emergence, seedling size, survivorship and fecundity, but heritabilities were low, indicating slow response to selection. For all characters, environmental components of variance were one to two orders of magnitude larger than genetic variance components, resulting in broad sense heritabilities less than 0.1. For seedling size and fecundity, all of the genetic variance was in the form of genotype-environment interactions, often with large negative genetic correlations across environments. In contrast, genotypes differed in mean survivorship through one year, but there were no genotype-environment interactions for viability. Genetic differences in viability were primarily expressed as differences in overwinter survivorship. Genotype × environment interactions among sites and blocks were generated early in the life cycle while the genotype × environment interactions in response to competitive environment (open, annual cover, perennial cover) first appeared in adult fecundity. Genetic variation in lifetime fitness was not significant, despite a fourfold difference in mean fitness among genotypes.     

APPARENT SELECTIVE NEUTRALITY OF MITOCHONDRIAL DNA SIZE VARIATION: A TEST IN THE DEEP-SEA SCALLOP PLACOPECTEN MAGELLANICUS

Animal mitochondrial DNA (mtDNA) is believed to have evolved under intense selection for economy of the size of the molecule. Among scallop species mtDNA size may vary by a factor of two and among conspecific individuals by as much as 25%. We have examined the possibility that large mtDNA size differences may be associated with fitness in the deep sea scallop Placopecten magellanicus by comparing shell lengths of individuals with different copy numbers of a large mtDNA repeated sequence. Among juvenile cohorts of same age, shell length is known to be a good index of overall fitness in marine bivalves and it is shown here to be affected by differences in nuclear genotype, expressed as the degree of enzyme heterozygosity. We have observed no correlation between shell length and mtDNA length and interpreted this to mean that variation in the size of animal mtDNA is effectively neutral to the forces of natural selection acting on the individual. This type of mtDNA variation must, therefore, be explained in terms of biases in the molecular mechanisms causing expansion or contraction of the molecule, differential replication rates of mtDNA molecules of different size, and the stochastic assortment of mtDNA size classes among individuals.     

INTERSPECIFIC AND INTRASPECIFIC MORPHOLOGICAL VARIATION IN MANATEES (SIRENIA: TRICHECHUS)

The three living species of Trichechus are clearly defined and well exemplify the degree of variability and taxonomic value of morphological characters in a well-understood mammalian genus. Statistical analysis of the largest sample of manatee skulls yet studied has allowed us to identify small suites of characters that effectively distinguish these species. The two subspecies of T. manatus proposed by Hatt (1934) can likewise be distinguished, and their use as taxonomic categories seems justified. This suggests that the cool winters of the northern Gulf Coast, on the one hand, and the deep water and strong currents of the Straits of Florida, on the other, are effective barriers to gene flow between Florida and Antillean manatees. Alleged taxonomic distinctions within T. senegalensis , however, have no demonstrated basis. No significant sexual dimorphism was detected in skulls of any of the species. Many interspecific differences can be correlated with feeding ecology, but others remain unexplained. Features of tooth crown morphology are among the most constant characters examined, but some osteological characters are equally good. T. manatas and T. senegalensis (which are phenetically the most similar) also seem to share a more recent common ancestor than either does with T. inunguis. However, the three species probably separated from each other at nearly the same time. T. inunguis has since become the most derived species, while T. senegalensis has changed the least.     

A UNIQUE MITOTIC VARIATION IN THE MARINE DINOFLAGELLATE OXYRRHIS MARINA (PYRROPHYTA)1

Mitosis is described in the flagellate Oxyrrhis marina Dujardin and is compared in related genera. Dense plaques develop in the nuclear envelope at prophase and give rise to an intranuclear spindle. Some of the microtubules associate with the chromosomes while others extend across the nucleus. The basal bodies migrate toward the poles early in division and retain a position lateral to the nuclear poles throughout mitosis. Microtubules are not present between the nucleus and the basal bodies. The nucleolus is persistent and elongates throughout anaphase and telophase. Chromosomal separation is accomplished by sliding of non-chromosomal microtubules and by elongation of the nuclear envelope rather than by shortening of the spindle microtubules. The nuclear envelope begins to constrict in the center early in anaphase. Continued constriction of the envelope and elongation of the nucleus leads to the formation of a dumbbell-shaped nucleus by late telophase. Mitosis culminates by the constriction of the nucleus into two daughter nuclei. The taxonomic position of Oxyrrhis marina is discussed in light of these findings.     

SPATIAL AND TEMPORAL VARIATION OF EPIPSAMMIC DIATOMS IN A SPRING-FED BROOK1

Epipsammic diatom communities were sampled and quantified bimonthly from June 1983 to April 1985 at four sites in a small brook in northwest Ohio. The primary objective was to determine the extent of temporal and spatial variability in the epipsammon. The four sites, approximately 250 m apart, differed in current velocity, illumination and the amount of sand-associated detritus. Diatoms were significantly more abundant in the portion of the brook flowing through an unshaded marsh than at three heavily shaded sites located upstream. Fragilaria leptostauron (Ehr.) Hust. and Achnanthes lanceolata var. dubia Grun. dominated the epipsammic assemblage at all sites throughout the entire study period. Only Meridion circulare (Grev.) Ehr. displayed a marked seasonal distribution. Although the same species were generally found at all sites, there were sufficient differences in relative abundance that communities could be discriminated according to site. Using two canonical variate axes, all 48 samples were correctly assigned to their proper sampling site based on community composition. Data from this epipsammic assemblage support the idea developed from the river continuum concept that species comprising riverine benthic assemblages continually persist and rarely become completely absent.