Relationships among Cedrus libani, C. brevifolia and C. atlantica as revealed by the morphological and anatomical needle characters

The main aim of the present study was testing the value of the morphological and anatomical characteristics of the needles in distinguishing Cedrus atlantica, C. libani and C. brevifolia. Nine populations were sampled in their natural habit and 25 characters were used to describe the variation of the brachyblast needles and to analyze the differences between species. The results indicated that morphological and anatomical needle characters provide valuable tools in discrimination of the taxa. The scored differences were statistically significant, as revealed in the Tukey’s t test, discrimination analysis and hierarchical analysis of variation. The results support treating C. libani, C. atlantica and C. brevifolia as independent species.     

Análisis morfométrico de las especies de Echinocereus sección Triglochidiati (Cactaceae) en México

Echinocereus section Triglochidiati has been accepted in the most recent taxonomic treatments of the genus. However, the number of included species has varied from one to 14. This study aims to define the species number of section Triglochidiati in Mexico through multivariate analysis, and to identify quantitative characters that can be used to recognize them. We used canonical discriminant analysis and classification discriminant analysis for 11 putative taxa native to Mexico and 20 morphometric characters. The number of stem ribs, number of central spines, stem diameter, length of radial spines (position 09), length of flower, receptacular tube length / perianth length ratio, and the receptacular tube trichome length permitted the differentiation of six taxa: E. acifer, E. arizonicus, E. coccineus, E. koehresianus, E. polyacanthus and E. scheeri. A key to the species is included.     

Molecular and morphological characterization of two aphid genera,Acyrthosiphon and Aulacorthum (Hemiptera: Aphididae)

The taxonomic histories of two genera, Acyrthosiphon and Aulacorthum, have been controversial because of the ambiguous morphology used as criteria to distinguish these genera. In this study, molecular and morphological characters were analyzed to determine whether the current classification of these genera is reasonable. Two phylogenetic trees based on five genes (COI, COII, srRNA, lrRNA, and EF1a) supported the distinctiveness of Acyrthosiphon and Aulacorthum, and a canonical discriminant analysis using 19 morphological characters showed that the two genera were separated by five morphological characters (HTB, C, Ant.III, HFM, and SIPH). Based on these results, we confirm that the current classification of Acyrthosiphon and Aulacorthum is supported by molecular and morphological characters.     

Reconstruction of the former charophyte community out of the fructifications identified in Albufera de València lagoon sediments

Several decades ago, due to eutrophication, Albufera de València lagoon lost the charophyte meadows that covered the bottom in the past but had never been studied in detail. An area close to the lagoon will be restored now to improve water quality and to recover submerged vegetation. To have information on the background of the charophytes from this area, four 80-87 cm long sediment cores were extracted from the lagoon. The subfossil charophyte fructifications (oospores and gyrogonites) were isolated and identified in two of the four cores. Moreover, sediment core fractions from the third and fourth cores were allowed to germinate under several conditions. The study of the lagoon sediments has revealed that at least 11 morphologies of charophyte fructifications existed in the past. Ten of these were determined at species level: Chara vulgaris, Chara hispida, Chara aspera, Chara baltica, Chara tomentosa, Chara globularis, Lamprothamnium papulosum, Nitella hyalina, Tolypella glomerata and Tolypella hispanica. However, one form of Chara gyrogonite did not fit in any known species. L. papulosum oospores and gyrogonites were the most abundant and concentrated in the deepest sediment layers when the lagoon held brackish water. Large changes in salinity due to freshwater input drove charophyte species substitution (disappearance of L. papulosum). During the oligohaline period C. vulgaris sexual propagules were the most abundant. The last charophyte community before its disappearance due to eutrophication was composed mainly of C. hispida, C. tomentosa and N. hyalina (the two latter with small numbers of oospores in the sediment). To characterize the morphology of the unidentified gyrogonite species, 100 specimens were studied. This type could be a local variety of a more common species, probably C. vulgaris, grown under particular ecological conditions. Old collected oospores (more than 45 years old) germinated and could be identified as C. vulgaris and C. hispida.     

Stepwise and Canonical Discriminant Analysis of Longidorus Species (Nematoda: Longidoridae) from Arkansas

During a 1998-to-2001 survey from Arkansas, nine distinct species of Longidorus were found including five new species. Morphometrics of these nine species were used in a stepwise and canonical discrimination to select a subset of characteristics that best identified each species. Student''s t test was applied to compare Longidorus breviannulatus Norton &Hoffman, 1975; L. crassus Thorne, 1974; L. diadecturus Eveleigh &Allen, 1982; L. fragilis Thorne, 1974; L. biformis Ye &Robbins, 2004; L. glycines Ye &Robbins, 2004; L. grandis Ye &Robbins, 2003; L. paralongicaudatus Ye &Robbins, 2003; and L. paravineacola Ye &Robbins, 2003 to examine interspecies variation and test for the most useful morphometric characters in species discrimination. Most of the morphometric characters were useful to differentiate species, but species identification could not be based on a single character because the morphometric character ranges often overlap. Stepwise discriminant analysis indicated that the guide ring position, head width, tail length, body length, odontostyle length, and anal body width were the most important variables. These were used to generate canonical variables in discriminating the species. The first three canonical variables accounted for 95% of the total variance. The scatterplots by the first three canonical variables grouped and separated the Longidorus species from Arkansas. Stepwise and canonical discriminant analyses were useful for examining the groupings and morphometric relationships of the nine Longidorus species.     

DNA barcoding the genus Chara: molecular evidence recovers fewer taxa than the classical morphological approach

Charophytes (Charales) are benthic algae with a complex morphology. They are vulnerable to ecosystem changes, such as eutrophication, and are red‐listed in many countries. Accurate identification of Chara species is critical for understanding their diversity and for documenting changes in species distribution. Species delineation is, however, complicated, because of high phenotypic plasticity. We used barcodes of the ITS2, matK and rbcL regions to test if the distribution of barcode haplotypes among individuals is consistent with species boundaries as they are currently understood. The study included freshly collected and herbarium material of 91 specimens from 10 European countries, Canada and Argentina. Results showed that herbarium specimens are useful as a source of material for genetic analyses for aquatic plants like Chara. rbcL and matK had highest sequence recoverability, but rbcL had a somewhat lower discriminatory power than ITS2 and matK. The tree resulting from the concatenated data matrix grouped the samples into six main groups contrary to a traditional morphological approach that consisted of 14 different taxa. A large unresolved group consisted of C. intermedia, C. hispida, C. horrida, C. baltica, C. polyacantha, C. rudis, C. aculeolata, and C. corfuensis. A second unresolved group consisted of C. virgata and C. strigosa. The taxa within each of the unresolved groups shared identical barcode sequences on the 977 positions of the concatenated data matrix. The morphological differences of taxa within both unresolved groups include the number and length of spine cells, stipulodes, and bract cells. We suggest that these morphological traits have less taxonomic relevance than hitherto assumed.     

Combined Genetic and Telemetry Data Reveal High Rates of Gene Flow,Migration, and Long-Distance Dispersal Potential in Arctic Ringed Seals (Pusa hispida)

Ringed seals (Pusa hispida) are broadly distributed in seasonally ice covered seas, and their survival and reproductive success is intricately linked to sea ice and snow. Climatic warming is diminishing Arctic snow and sea ice and threatens to endanger ringed seals in the foreseeable future. We investigated the population structure and connectedness within and among three subspecies: Arctic (P. hispida hispida), Baltic (P. hispida botnica), and Lake Saimaa (P. hispida saimensis) ringed seals to assess their capacity to respond to rapid environmental changes. We consider (a) the geographical scale of migration, (b) use of sea ice, and (c) the amount of gene flow between subspecies. Seasonal movements and use of sea ice were determined for 27 seals tracked via satellite telemetry. Additionally, population genetic analyses were conducted using 354 seals representative of each subspecies and 11 breeding sites. Genetic analyses included sequences from two mitochondrial regions and genotypes of 9 microsatellite loci. We found that ringed seals disperse on a pan-Arctic scale and both males and females may migrate long distances during the summer months when sea ice extent is minimal. Gene flow among Arctic breeding sites and between the Arctic and the Baltic Sea subspecies was high; these two subspecies are interconnected as are breeding sites within the Arctic subspecies.     

Multivariate morphometric analyses of the predatory ciliate genus Semispathidium (Ciliophora: Litostomatea), with description of S. longiarmatum nov. spec.

We studied morphometrical variation, species boundaries, and importance of morphometric features for a reliable separation of five African Semispathidium taxa. Altogether, 20 features traditionally used in alpha-taxonomy of the predatory genus Semispathidium were measured or scored on 85 protargol-impregnated interphase specimens, and were analyzed using hierarchical clustering as well as principal component and canonical discriminant analyses. This multivariate approach confirmed that a population found in Botswanan floodplain soil represents a distinct taxon. The new species is described here as S. longiarmatum, using live observation, protargol impregnation, and scanning electron microscopy. Semispathidium longiarmatum strongly resembles S. armatum and S. breviarmatum but it is clearly distinguished from these species by the extrusome pattern. The reliability of S. longiarmatum is also strengthened, according to the canonical discriminant analysis, by several quantitative features, viz., the number of ciliary rows, the length:width ratio of the macronucleus, and the number of dikinetids in brush row 1. Moreover, the present study documents the distinctness of all African Semispathidium species which can be separated by a combination of both qualitative and quantitative (morphometric) features. Consequently, Semispathidium species do not form a continuous complex but fairly discrete clusters in the phenotypic space.     

Species differentiation in the genus Chara (Charophyceae): considerable phenotypic plasticity occurs within homogenous genetic groups

Charophytes are benthic algae with a complex morphology and high phenotypic plasticity. This has led to ambiguities in species delineation. However, until now genetic studies on Chara have been based on samples collected from a restricted geographic range or only included a restricted number of taxa. This may have hindered a general interpretation of the results. We applied barcoding of matK, a rapidly evolving coding section of the plastid genome, in 324 Chara samples collected from 19 countries, in order to test whether the distribution of barcode haplotypes among individuals was consistent with species boundaries as they are currently understood. The phylogenetic tree grouped the 324 Chara individuals, which according to commonly used identification keys represented 29 species, into 12 well-defined groups (i.e. monophyletic morphospecies or groups of morphospecies). Considerable morphological variation occurred within genetically homogeneous groups. This included traits which are commonly used for Chara species determination, such as the length and number of spine cells, the length of stipulodes and bract cells, cortication (tylacanthous, isostichous, aulacanthous and absent cortication), as well as sex differentiation. However, there were also substantial genetic differences among morphologically similar species (e.g. C. virgata – C. globularis – C. connivens). No morphological trait consistently reflected genetic differences. This indicates that morphological traits for specific taxa may serve as diagnostic tools for species delimitation, but that they are not generally suitable for inferring genetic differentiation or phylogenetic relationships. We propose that (i) C. virgata and C. strigosa, (ii) C. liljebladii, C. horrida and C. baltica, and (iii) C. hispida, C. rudis and C. polyacantha are conspecific. Our data also indicate that C. gymnophylla should be divided into tylacanthous forms (which are closely related to C. contraria) and aulacanthous forms (which are related to C. vulgaris).     

A multivariate morphological analysis of the Cardamine concatenata alliance (Brassicaceae)

Principal coordinate analysis using Gower’s similarity coefficient and canonical discriminant analysis were performed using morphological data to assess species distinctness in theCardamine concatenata alliance, comprising the eastern North American dentarias. The results of an overall principal coordinate analysis of the alliance using quantitative and qualitative characters revealed distinct and separate clusters that correspond to five previously recognized species (C. angustata, C. concatenata, C. diphylla, C. dissecta, C. maxima). One additional taxon,C. incisa, was found to group closely with theC. diphylla cluster. In a second principal coordinate analysis including only individuals fromC. diphylla andC. incisa, both are supported as distinct morphological entities. Canonical discriminant analysis of only above-ground quantitative characters was less effective in distinguishing previously recognized taxa, in keeping with the emphasis on qualitative characters in previous taxonomic treatments of the group.Cardamine incisa (Eames) Schumann (pro hybr.) is raised to the rank of species. A key to the species of theCardamine concatenata alliance is provided.     

Speciation of Campylobacter coli, C. jejuni, C. helveticus, C. lari, C. sputorum, and C. upsaliensis by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

Multiple strains of Campylobacter coli, C. jejuni, C. helveticus, C. lari, C. sputorum, and C. upsaliensis isolated from animal, clinical, or food samples have been analyzed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Whole bacterial cells were harvested from colonies or confluent growth on agar and transferred directly into solvent and then to a spot of dried 3-methoxy-4-hydroxycinnamic acid (matrix). Multiple ions in the 5,000- to 15,000-Da mass range were evident in spectra for each strain; one or two ions in the 9,500- to 11,000-Da range were consistently high intensity. “Species-identifying” biomarker ions (SIBIs) were evident from analyses of multiple reference strains for each of the six species, including the genome strains C. jejuni NCTC 11168 and C. jejuni RM1221. Strains grown on nine different combinations of media and atmospheres yielded SIBI masses within ±5 Da with external instrument calibration. The highest-intensity C. jejuni SIBIs were cytosolic proteins, including GroES, HU/HCj, and RplL. Multiple intraspecies SIBIs, corresponding probably to nonsynonymous nucleotide polymorphisms, also provided some intraspecies strain differentiation. MALDI-TOF MS analysis of 75 additional Campylobacter strains isolated from humans, poultry, swine, dogs, and cats revealed (i) associations of SIBI type with source, (ii) strains previously speciated incorrectly, and (iii) “strains” composed of more than one species. MALDI-TOF MS provides an accurate, sensitive, and rapid method for identification of multiple Campylobacter species relevant to public health and food safety.     

Bacterial Community Associated with Healthy and Diseased Reef Coral Mussismilia hispida from Eastern Brazil

In order to characterize the bacterial community diversity associated to mucus of the coral Mussismilia hispida, four 16S rDNA libraries were constructed and 400 clones from each library were analyzed from two healthy colonies, one diseased colony and the surrounding water. Nine bacterial phyla were identified in healthy M. hispida, with a dominance of Proteobacteria, Actinobacteria, Acidobacteria, Lentisphaerae, and Nitrospira. The most commonly found species were related to the genera Azospirillum, Hirschia, Fabibacter, Blastochloris, Stella, Vibrio, Flavobacterium, Ochrobactrum, Terasakiella, Alkalibacter, Staphylococcus, Azospirillum, Propionibacterium, Arcobacter, and Paenibacillus. In contrast, diseased M. hispida had a predominance of one single species of Bacteroidetes, corresponding to more than 70% of the sequences. Rarefaction curves using evolutionary distance of 1% showed a greater decrease in bacterial diversity in the diseased M. hispida, with a reduction of almost 85% in OTUs in comparison to healthy colonies. ∫-Libshuff analyses show that significant p values obtained were <0.0001, demonstrating that the four libraries are significantly different. Furthermore, the sympatric corals M. hispida and Mussismilia braziliensis appear to have different bacterial community compositions according to Principal Component Analysis and Lineage-specific Analysis. Moreover, lineages that contribute to those differences were identified as α-Proteobacteria, Bacteroidetes, and Firmicutes. The results obtained in this study suggest host–microbe co-evolution in Mussismilia, and it was the first study on the diversity of the microbiota of the endemic and endangered of extinction Brazilian coral M. hispida from Abrolhos bank.     

Multivariate analysis of morphological variation among closely related species Bromus japonicus, B. squarrosus and B. arvensis (Poaceae) in comparison with isozyme evidences

Morphological variation and taxonomic continuum of 110 specimens of Bromus arvensis, B. japonicus and B. squarrosus were analysed to assess the morphological variability of these close taxa. The importance of every morphological character was estimated by the GLM procedure. Stepwise discriminant analysis was used in order to find a set of morphological characters that maximises differences among the species. According to this, the most important characters appeared to be the length of anthers, width of lemma and lemma margin angle shape. The most important characters for the determination of Bromus arvensis was the length of anthers and for B. squarrosus width of lemma. According to the UPGMA cluster analysis, canonical discriminant, classificatory discriminant and principal component analyses, all specimens were separated into three moderately distinct groups which corresponded to the three traditional species. The study showed that qualitative characters were the best for the delimitation of the tax a by statistical analyses and demonstrated the incongruity between the pattern of morphological and genetic (allozyme) variation among the three Bromus species.     

Interspecific variation of plant traits associated with resistance to herbivory among four species of Ficus (moraceae)

• Background and aims To understand the defensive characteristics of interspecies varieties and their responses to herbivory damage, four species of Ficus plants (Ficus altissima, F. auriculata, F. racemosa and F. hispida) were studied. They were similar in life form, but differed in successional stages. Of these, Ficus altissima is a late successional species, F. hispida is a typical pioneer and F. auriculata and F. racemosa are intermediate successional species. We addressed the following questions: (1) What is the difference in plant traits among the four species and are these traits associated with differences in herbivory damage levels? (2) What is the difference in the damage-induced changes among the four species?• Methods Herbivory damage was measured in the field on randomly planted seedlings of the four species of the same age. Defences to herbivory were also tested by feeding leaves of the four species to larvae of Asota caricae in the laboratory. A total of 14 characters such as water content, thickness, toughness, pubescence density on both sides, leaf expansion time, lifetime and the contents of total carbon (C), nitrogen (N), phosphorous (P), potassium (K), magnesium (Mg) and calcium (Ca) were measured. Leaf calcium oxalate crystal (COC) density, total Ca and N content, leaf toughness and height were measured to investigate induced responses to artificial herbivory among the four species.• Key results and conclusions Herbivory damage in the four studied species varied greatly. The pioneer species, F. hispida, suffered the most severe herbivory damage, while the late successional species, F. altissima, showed the least damage. A combination of several characteristics such as high in content of N, Ca and P and low in leaf toughness, lifetime and C : N ratio were associated with increased herbivore damage. The late successional species, F. altissima, might also incorporate induced defence strategies by means of an increase in leaf COC and toughness.Key words: Calcium oxalate crystals, defensive characteristics, Ficus; herbivory, induced defence     

A phenetic study of Cassia sensu lato (Leguminosae-Caesalpinioideae: Cassieae: Cassiinae) in Thailand

Cassia L. sensu lato, a large heterogeneous genus of flowering plants, occurs naturally in the tropics around the world. Recent works based on floral morphology have supported a division of this genus into three genera, namely Cassia L. s. str., Chamaecrista Moench and Senna Mill. In order to investigate this new classification, 508 specimens of 18 taxa of the genus Cassia s.l. grown in Thailand were analyzed using cluster analysis and canonical discriminant analysis. The total 32 vegetative and reproductive morphological characters were employed in these analyses. In cluster analysis, Cassia s.l. can be separated into four groups, respectively viz. Chamaecrista, Senna alata, Senna and Cassia s /str. The four-cluster grouping is discussed. From a canonical discriminant analysis using the four-cluster grouping as a priori groups, it can be concluded that Cassia s. str., Senna, and Chamaecrista are indeed distinct taxa. The three most important characters that separate the three genera are filament length, fruit length, and ovary stalk length. These quantitative characters, together with some qualitative characters, were useful in constructing an identification key to these genera. Among the three genera, it was also found that Senna is rather a heterogeneous taxon. The difference between the studied species was discussed.     

Freshwater sponges (Porifera: Spongillidae) from Panamá

Aquatic habitats in the Republic of Panamá were examined for freshwater sponges during 21 Feb. to 6 March 1986. Eunapius carteri, Trochospongilla leidyi, and Trochospongilla horrida were found in the northern canal area. This is the first report of E. carteri and T. horrida from Central America, and the first record of E. carteri from the Western Hemisphere. Taxonomic characters used to distinguish T. leidyi and T. horrida from similar South American species are discussed.     

Iridoids and alkaloids from Castilleja host plants for platyptilia pica. Rhexifoline content of P. pica

Iridoid and alkaloid analyses were conducted on Castilleja sulphurea, C. occidentalis, C. rhexifolia and C. hispida (Scrophulariaceae). Pyrrolizidine alkaloids were found in C. rhexifolia and some C. sulphurea populations, but other C. sulphurea populations had quinolizidine alkaloids or none at all. No C. occidentalis populations contained alkaloids. C. hispida was found to contain lamprolobine and the quinolizidine alkaloid anagyrine. All taxa contained the pyridinemonoterpene rhexifoline. The iridoid content of the Castilleja species were all qualitatively similar. Major iridoids were aucubin, catalpol, penstemonoside and shanzhiside methyl ester, with traces of 8-epiloganin and gardoside methyl ester. Larvae of Platyptilia pica (Pterophoridae) hosted by Castilleja were found to excrete and not sequester iridoids. The adult moths contained rhexifoline alkaloid, but at a low concentration level. Systematic implications of the results for Castilleja are discussed.     

Long-term changes in macrophyte vegetation after reduction of fish stock in a shallow lake

Starting in the middle of the 1970s, submerged macrophytes began to disappear from shallow Lake Warniak due to feeding pressure by grass carp (Ctenopharyngodon idella). In the middle of the 1980s, the lake was stocked with seston-feeding silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis). By 1993, the biomass of silverhead carp and bighead carp had declined. This allowed charophytes to recolonize the bottom of the lake. The main charophyte species at this time were Chara globularis and Chara rudis. Since then, five other stonewort species have been found: Chara contraria, C. filiformis, C. tomentosa, C. aspera and Nitellopsis obtusa. Seventeen species of aquatic angiosperms have also been found. There were distinct changes in the relative abundance and spatial distribution of particular species. C. rudis developed most intensely in the shallow parts of the lake near the southern and western shores. C. globularis gradually took over the deeper central part of the lake. In 2001, C. rudis began to retreat again. The relative abundance and spatial distribution of charophytes was correlated to water clarity (r = 0.87, p < 0.05), total phosphorus level (r = −0.78; p < 0.05), and chlorophyll a content (r = −0.79; p < 0.05).