Oospore Wall Ornamentation in the Genus Tolypella (Charales,Charophyceae)

The ultrastructural features of oospore wall ornamentation in the genus Tolypella were examined using scanning electron microscopy (SEM). The taxonomic relationships among several species were discussed on the basis of oospore ultrastructure and measurements. In the case of T. glomerata and T. nidifica, our results support the status of separate species. Close relationships and transitional forms may exist between T. nidifica and T. normaniana, and not only in oospore wall ornamentation. Oospores of T. hispanica exhibited the same distinct type of reticulate oospore wall as previously reported, but our results do not support the recognition of T. hispanica as a separate species. Ultrastructure of the oospore walls of T. prolifera and T. intricata was almost identical, suggesting that these species are closely related. We therefore reject previous suggestions that morphological characteristics of oospores as observed in SEM are sufficient for identification of individual species. Although significant differences were found among oospores in individual species of Tolypella, large variation among populations, and among individuals belonging to the same population, caused substantial overlap among species.     

Gradualistic characean lineages in the Upper Cretaceous–Palaeocene of southern Europe

Two charophyte lineages from the Upper Cretaceous–Palaeocene of southern Europe are described here, constituting a useful biostratigraphic tool for correlating non-marine stratigraphic sequences. The Peckichara pectinata lineage, ranging from the middle Campanian to the middle–upper Maastrichtian, consists of four successive gyrogonite morphotypes that displayed progressively more complex gyrogonite ornamentation and increasingly smaller gyrogonites with time. The Peckichara sertulata lineage, ranging from the middle Campanian to the Danian, consists of three morphotypes that followed a more common evolutionary trend, developing progressively larger gyrogonites without any changes in ornamentation. Intermediate forms between the successive morphotypes suggest that they corresponded to anagenetic varieties of the same evolutionary species rather than to separate species. The different trends observed appear to be palaeoenvironmentally controlled. The grades of the P. pectinata lineage first occurred in permanent lakes that later shifted to fluvial influenced floodplain ponds, this shift accounting for the reduction in gyrogonite size. By contrast, the P. sertulata lineage mainly occurred in permanent lakes, where stable palaeoenvironmental conditions enabled a steady rise in gyrogonite size to increase the space available for the zygote.     

Reproductive trade-off of the copepod Acartia tonsa in a hypersaline estuary of the Southwestern Atlantic. Temporal variations in the morphology of eggs

The purpose of the present work was to study the seasonal variations in egg production, morphology and hatching success in the cryptic species Acartia tonsa, taking into account variations in female size, population abundance and environmental factors in a turbid and hypersaline estuary. Sampling was performed during the austral warm (18–23°C and 32–36 salinity) and cold seasons (5–7°C; 32–38) in Bahía Blanca Estuary (BBE), Argentina, during 2007 and 2009. Field-collected females were incubated in the laboratory simulating in situ environmental conditions, and specimens from fixed samples were measured using optical and scanning electronic microscopy. Acartia tonsa’s marked seasonality in its reproductive traits was found to ensure its permanence in the water column all over the year. During the warm season, small-sized females were observed to invest their energy in the production of subitaneous eggs with high hatching success and smooth appearance (12.95?±?2.38 eggs f^?1 day^?1 and specific egg production rate (SEP) of 16.57%C f^?1 day^?1). During the cold season, females invested C in body mass as well as in the production of resting eggs of three different morphotypes (6.56?±?3.2 eggs f^?1 day^?1 and SEP of 7.37%C f^?1 day^?1). Although these morphotypes were found to show differences in surface ornamentation, they exhibited the same delayed hatching behaviour. The eggs with shorter spines were found to integrate the resting egg bank in BBE. Our findings confirming a delayed egg hatching behaviour and a great tolerance to low temperatures and high salinities in the A. tonsa population in BBE suggest that this possible strain is a valuable phenotype for aquaculture.     

Phenotypic variability in the Caribbean Orange Icing sponge Mycale laevis (Demospongiae: Poecilosclerida)

Sponge species may present several morphotypes, but sponges that are morphologically similar can be separate species. We investigated morphological variation in Mycale laevis, a common Caribbean reef sponge. Four morphotypes of M. laevis have been observed (1) orange, semi-cryptic, (2) orange, massive, (3) white, semi-cryptic, and (4) white, massive. Samples of M. laevis were collected from Key Largo, Florida, the Bahamas Islands, and Bocas del Toro, Panama. Fragments of the 18S and 28S rRNA ribosomal genes were sequenced and subjected to phylogentic analyses together with sequences obtained for 11 other Mycale species and additional sequences retrieved from GenBank. Phylogenetic analyses confirmed that the genus Mycale is monophyletic within the Order Poecilosclerida, although the subgenus Aegogropila is polyphyletic and the subgenus Mycale is paraphyletic. All 4 morphotypes formed a monophyletic group within Mycale, and no genetic differences were observed among them. Spicule lengths did not differ among the 4 morphotypes, but the dominant megasclere in samples collected from Florida and the Bahamas was the strongyle, while those from Panama had subtylostyles. Our data suggest that the 4 morphotypes constitute a single species, but further studies would be necessary to determine whether skeletal variability is due to phentotypic or genotypic plasticity.     

Phylogenetic and populational study of the Tuber indicum complex

When examined using SEM, Chinese samples of Tuber indicum and T. sinense displayed the same ascospore ornamentation as that of T. pseudohimalayense, T. indicum, collected in India by Duthie in 1899, and samples renamed T. himalayense in 1988. The different authors who named the four taxa (T. indicum, T. himalayense, T. sinense, T. pseudohimalyense) described differences in the surface of the peridium which could be considered as usual variations within a single species. We consider T. indicum, T. himalayense, T. sinense and T. pseudohimalayense as one species, T. indicum. Within this T. indicum complex, according to ITS and β-tubulin sequences, there are two groups in China, which could be considered as geographical ecotypes. This study is the first to identify a genetic and phylogeographical structure within the Chinese Tuber species.     

Two new species ofIsoetes (Isoetaceae) from Jeju Island, South Korea

We name and describe two new species,Isoetes jejuensis and I.hallasanensis, from Jeju Island, South Korea. These taxa are compared with related species in East Asia, including I.coreana andI. sinensis. The rugulate megaspore ornamentation ofI. jejuensis differs from those otherIsoetes species. Furthermore,I. hallasanensis varies fromI. asiatica in its microspore ornamentation and chromosome number, although both species have an echinate megaspore ornamentation. These new species are tetraploid (2n = 44), whereasI. coreana is hexaploid (2n = 66).     

Contribution of external morphology in solving a species complex: The case of Prorocentrum micans, Prorocentrum gracile and Prorocentrum sigmoides (Dinoflagellata) from the Mexican Pacific Coast

Prorocentrum micans Ehrenberg, Prorocentrum gracile Schütt and Prorocentrum sigmoides Böhm are planktonic dinoflagellates present along the Mexican Pacific coast. These three species can be treated as a species complex because of similar shape and size. The taxonomy of the group has been based on cell form and size, the presence or absence of apical spines and the ornamentation of the main valves. A morphometric study of samples from 11 locations from the Mexican Pacific coast and from laboratory cultures is presented. The presence of a mucron (small tooth) in the antapical region of P. gracile and P. sigmoides morphotypes was recorded for the first time. Morphotypes were defined using as the main criteria the size of pores in relation to the size of the depressions on the cell’s surface, the presence of a mucron in the posterior end of the cell and the shape of the valve margins. Results show a clear distinction between the morphotype P. micans and the P. gracile–P. sigmoides. P. sigmoides is suggested to be a junior synonym of P. gracile as no convincing characters could distinguish these two morphotypes.     

Effects of fine-grain habitat complexity on egg parasitism by three species of Trichogramma

Effects of microhabitat complexity on host foraging by three species of Trichogramma (T. deion, T. ostriniae, and T. pretiosum) were evaluated under simulated stored product conditions. All three species have been considered as potential biological control agents for the Indianmeal moth, Plodia interpunctella, in retail stores and warehouses. Trials were conducted with single naïve female parasitoids in 10-cm Petri dishes that were either empty, contained flour, or contained millet. Empty 15-cm Petri dishes, which served as a surface area control, comprised a fourth treatment. Females were allowed to forage for sentinel egg disks for 2 h, after which percentages of parasitism and egg mortality were computed. In addition, behavioral observations were made on a subset of the trials. Trichogramma deion parasitized more eggs than the other two species in the empty dishes and in the dish containing flour. For T. deion, rates of parasitism and egg mortality were significantly greater in both the small and large empty dishes than in the small dishes containing flour or millet. Parasitism was consistently low for all three species in the grain-filled dishes. Among species, T. ostriniae spent the most time walking, while T. pretiosum was the most sedentary. In addition, T. pretiosum spent significantly more time on the first egg visited compared with subsequent eggs. T. deion may be the best-suited for use as a biological control agent for P. interpunctella. However, the potential negative effects of fine-grain habitat complexity should be taken into account when developing a release protocol for Trichogramma spp.     

The ankylosaurid dinosaurs of the Upper Cretaceous Baruungoyot and Nemegt formations of Mongolia

The discovery of a new ankylosaurid skull with some unusual features from the Baruungoyot Formation of Mongolia prompted a systematic review of ankylosaurid specimens from the Baruungoyot and Nemegt formations. Dyoplosaurus giganteus was found to possess no diagnostic features and is regarded as a nomen dubium. The holotype of Tarchia kielanae (previously synonymized with Tarchia gigantea) has one autapomorphy, an accessory postorbital ossification with surrounding furrow, and Tar. kielanae is here considered a valid species, making the combination Tar. gigantea unnecessary. An accessory postorbital ossification is also found in the holotype of Minotaurasaurus ramachandrani, and this species is here considered a junior synonym of Tar. kielanae. The newly described skull from the Baruungoyot Formation forms the holotype of a new genus and species, Z araapelta nomadis gen. et sp. nov. , diagnosed by unusual bilayered ornamentation on the squamosal horn and extensive postocular ornamentation. Two distinct tail club handle morphotypes are present in the Nemegt Formation and probably represent two different species. However, it is impossible to assign either tail club morphotype to the single valid species from the formation, Saichania chulsanensis, because of a lack of overlapping material. A revised phylogenetic analysis including newly identified characters found Zaraapelta nomadis to be most closely related to Tar. kielanae. © 2014 The Linnean Society of London     

Ultrastructure of the Secondary Egg Envelope of Cyprinodontidae of the Genus Epiplatys Gill, 1862 (Pisces,Teleostei)

Abstract The structure of the secondary egg envelope of seven species of Cyprinodontidae belonging to the genus Epiplatys was observed by transmission and scanning electron microscopy. The transverse structure of the secondary envelope examined by transmission electron microscopy is a constant feature in all the species studied and represents a good criterion for identification. In contrast, the surface structure of the secondary envelope when observed by scanning electron microscopy exhibits a great diversity of form. It is possible to distinguish each of the seven different species by the pattern of ornamentation, the width of the polygons and the size of the granules comprising them on the secondary egg envelope.     

Lack of association between winter coat colour and genetic population structure in the Japanese hare,Lepus brachyurus (Lagomorpha: Leporidae)

Seasonal changes in fur colour in some mammalian species have long attracted the attention of biologists, especially in species showing population variation in these seasonal changes. Genetic differences among populations that show differences in seasonal changes in coat colour have been poorly studied. Because the Japanese hare (Lepus brachyurus) has two allopatric morphotypes that show remarkably different coat colours in winter, we examined the population genetic structure of the species using partial sequences of the SRY gene and six autosomal genes: three coat colour‐related genes (ASIP, TYR, and MC1R) and three putatively neutral genes (TSHB, APOB, and SPTBN1). The phylogenetic tree of SRY sequences exhibited two distinct lineages that diverged approsimately 1 Mya. Although the two lineages exhibited a clear allopatric distribution, it was not consistent with the distribution of morphotypes. In addition, six nuclear gene sequences failed to reveal genetic differences between morphotypes. Population network trees for 11 expedient populations divided the populations into four groups. Genetic structure analysis revealed an admixture of four genetic clusters in L. brachyurus, two of which showed large genetic differences. Our results suggest ancient vicariance in L. brachyurus, and we detected no genetic differences between the two morphotypes. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 111 , 761–776.     

Fungal communities in mycorrhizal roots of conifer seedlings in forest nurseries under different cultivation systems, assessed by morphotyping, direct sequencing and mycelial isolation

Fungi colonising root tips of Pinus sylvestris and Picea abies grown under four different seedling cultivation systems were assessed by morphotyping, direct sequencing and isolation methods. Roots were morphotyped using two approaches: (1) 10% of the whole root system from 30 seedlings of each species and (2) 20 randomly selected tips per plant from 300 seedlings of each species. The first approach yielded 15 morphotypes, the second yielded 27, including 18 new morphotypes. The overall community consisted of 33 morphotypes. The level of mycorrhizal colonisation of roots determined by each approach was about 50%. The cultivation system had a marked effect on the level of mycorrhizal colonisation. In pine, the highest level of colonisation (48%) was observed in bare-root systems, while in spruce, colonisation was highest in polyethylene rolls (71%). Direct internal transcribed spacer ribosomal DNA sequencing and isolation detected a total of 93 fungal taxa, including 27 mycorrhizal. A total of 71 (76.3%) fungi were identified at least to a genus level. The overlap between the two methods was low. Only 13 (13.9%) of taxa were both sequenced and isolated, 47 (50.5%) were detected exclusively by sequencing and 33 (35.5%) exclusively by isolation. All isolated mycorrhizal fungi were also detected by direct sequencing. Characteristic mycorrhizas were Phialophora finlandia, Amphinema byssoides, Rhizopogon rubescens, Suillus luteus and Thelephora terrestris. There was a moderate similarity in mycorrhizal communities between pine and spruce and among different cultivation systems.Electronic supplementary material Supplementary material is available for this article at and accessible for authorised users     

Pollen diversity in the genus Ptilostemon (Asteraceae,Cardueae) from Italy and its taxonomic and palynoecological implications

The results of a study on pollen morphology of the species of the genus Ptilostemon growing in Italy: Ptilostemon niveus, P. greuteri, P. gnaphaloides, P. casabonae, P. strictus, P. stellatus by light and scanning electron microscopy were presented and discussed including in relation to their taxonomic position. The exine shows two different ornamentation patterns: echinae and scabrae. This last ornamentation pattern together with other features was previously described in the genus Ptilostemon. Of 6 species analysed, P. stellatus is the only one that shows a scabrate ornamentation of exine. Two pollen types were recognized through the exine ornamentation, length of ectocolpus and polar outline: P. stellatus type and P. niveus type. A multivariate analysis (principal component analysis and linear discriminant analysis) was carried out with the aim of examining potential morphological characters which could be used to identify taxa. The data suggest that several characteristics can be used to delimit the species in particular the exine ornamentation. The results of our studies support the actually subgeneric classification, and several features of the pollen grains analysed seem to have a palynoecological role and an important taxonomic significance. A dichotomous key based on palynological data is also given.     

Using palynomorphological characteristics for the identification of species of Alsinoideae (Caryophyllaceae): a systematic approach

Palynological characters of 18 species belonging to seven genera of the traditional subfamily Alsinoideae including Arenaria, Cerastium, Eremogone, Lepyrodiclis, Minuartia, Sabulina and Stellaria were studied in detail using light and scanning electron microscopy. Pollen grains of subfamily Alsinoideae are subspheroidal or prolate, pantoporate and 20.04 to 51.4 µm in size, prominent and sunken apertures uniformly distributed on the pollen surface. Two types of pollen grain ornamentation were observed, i.e. microechinate-punctate or microechinate-perforate. Echini are present on the surface of the pollen of all investigated species with medium, dense, or sparse echinodensity. These species exhibit variation in polar view, equatorial diameter, number of apertures, exine thickness, diameter of pore, appendages per pore, pore ornamentation, echini arrangement, echinidensity and shape of pollen. Based on qualitative characters, a dichotomous key has been developed for quick and easy identification. The present investigation contributes to the systematic approach using palynological characteristics and correct identification of species for members of subfamily Alsinoideae (Caryophyllaceae).     

Morphological and molecular diversity and abundance of tomentelloid ectomycorrhizae in broad-leaved forests of the Hungarian Plain

The most common representatives of tomentelloid ectomycorrhizae (EM) collected in broad-leaved forests (Populus and Quercus) of the Hungarian Plain during a 4-year project are demonstrated. Eighteen specimens of nine tomentelloid EM morphotypes were investigated. Five of these, introduced here for the first time, were characterized by microscopical–morphological and anatomical methods. Molecular identification was carried out using sequence analysis of the nrDNA ITS region. Altogether, 54 ITS sequences (6 previously published and 12 new sequences from our mycorrhizae and 36 from GenBank derived from fruitbodies) were compared by phylogenetic analyses using neighbor-joining, maximum parsimony and maximum likelihood methods. Identification to species level was successful in the case of six EM morphotypes (Tomentella galzinii, T. subtestacea, T. sublilacina, T. pilosa, T. ferruginea and T. stuposa), and the possible taxonomic position of the remaining three was approached. These results are supported by morphology, as compared with literature data. Relative abundance of the EM morphotypes within the soil samples was estimated. Our results confirm that tomentelloid EM are constant, diverse and abundant members of the EM communities in temperate-continental broad-leaved forests.     

The relationship between egg load and fecundity among Trichogramma parasitoids

1. The relationships between parasitoid egg load, size, and age (3–72 h) for Trichogramma minutum, T. platneri, and T. pretiosum, reared from two factitious hosts, Ephestia kuehniella and Sitotroga cerealella, were evaluated to test the hypothesis that 24‐h egg load can be used to estimate the fecundity of Trichogramma parasitoids. 2. Egg load increased in relation to female age over the first 3 days of adult life for all three Trichogramma species to a mean egg storage capacity of 46.7 eggs for T. minutum, 41.1 for T. pretiosum, and 35.7 for T. platneri. At 24 h of age, T. minutum had matured enough eggs to fill 67% of its storage capacity, in comparison with 74% for T. pretiosum and 91% for T. platneri. There was a positive relationship between egg load and parasitoid size for all ages of the three Trichogramma species reared from both hosts (with the exception of T. platneri at 3 h post emergence), accounting for 14–69% of the variance in egg load. 3. The potential fecundity, realised (3 day cumulative) fecundity, and oviposition rate (potential fecundity/longevity) of T. platneri were all related linearly to size‐dependent variation in 24‐h egg load, but only the realised fecundity of T. pretiosum, and none of the reproductive characteristics of T. minutum. It is suggested that 24‐h egg load may not be an accurate measure of egg storage capacity in parasitoids and should be used cautiously to represent fecundity. 4. The potential fecundity of seven Trichogramma species reared from E. kuehniella varied from 55 to 150, but neither potential fecundity nor oviposition rate was related significantly to egg load (represented by eggs laid during first 24 h). Selection to avoid egg depletion in the attack of gregarious hosts appears most likely to account for the variation in potential fecundity among Trichogramma species.     

Mycorrhizal associations in woody plant species at the Mt. Usu volcano,Japan

We investigated the association between ectomycorrhizal (ECM) and arbuscular mycorrhizal (AM) fungi and pioneer woody plant species in areas devastated by the eruption of Mt. Usu, Japan, in 2000. We observed eight woody plant species at the research site, most of which were associated with ECM and/or AM fungi. In particular, dominant woody plant species Populus maximowiczii, Salix hultenii var. angustifolia and Salix sachalinensis were consistently associated with ECM fungi and erratically associated with AM fungi. We found one to six morphotypes in the roots of each ECM host and, on average, two in the roots of each seedling, indicating low ECM fungal diversity. ECM colonization ranged from 17 to 42% of root tips. Using morphotyping and molecular analyses, 15 ECM fungi were identified. ECM fungi differed greatly between hosts. However, Laccaria amethystea, Hebeloma mesophaeum, Thelephora terrestris and other Thelephoraceae had high relative colonization, constituting the majority of the ECM colonization in the roots of each plant species. These ECM fungi may be important for the establishment of pioneer woody plant species and further revegetation at Mt. Usu volcano.     

Comparative egg morphology of six Meccus species and Triatoma recurva (Stål, 1868) Hemiptera: Reduviidae

We investigated the morphology and morphometry of eggs from the colonies of the Entomology Laboratory at ENCB‐IPN belonging to six species of the genus Meccus and Triatoma recurva, using both light and scanning electron microscopy. Egg ornamentation is an important parameter to consider in the differentiation of species. Samples were observed under a scanning electron microscope. Measurements of fifty eggs per species included length, width, and opercular diameter, which were used for the morphometric analysis. The results showed that the seven species display a polygonal ornamentation dominated by hexagons; the operculum shows also a polygonal ornamental characteristic in each species. Morphometry provided valuable information for discriminating between closely related species of the genus Meccus and Triatoma recurva, a species akin to this group, thereby facilitating the complete discrimination of these species.     

Embryonic development of the false limpet Siphonaria lateralis from Atlantic Patagonia

Siphonariids are pulmonate gastropods inhabiting rocky intertidal habitats, and many studies have focused on these false limpets around the world. In the southern South Atlantic, studies on reproduction and development in species of Siphonaria are scarce. We studied the embryonic development and egg masses of Siphonaria lateralis at its northernmost distribution in Atlantic Patagonia. In S. lateralis, as in most species of Siphonaria, individuals spawn benthic egg masses that strongly attach to intertidal rocky substrata. A single spherical egg that measures ~120 µm develops inside the egg capsule of S. lateralis. Considering the relatively small egg size, and reports from previous studies, the developmental modality of S. lateralis might be expected to include a planktotrophic larval phase. However, we found that hatchlings emerged as 1‐mm crawling juveniles, probably owing to the presence of intracapsular fluid, which may provide the energetic requirements for direct development. The embryonic size changed little from the egg to veliger stages, and then increased rapidly until the hatchling stage. We compared development in S. lateralis with development in the sympatric Siphonaria lessonii, in which egg size was reported to be ~80 µm and hatching occurs as planktotrophic veliger larvae. In these two species, spawn and early intracapsular developmental modes are remarkably different; these differences represent contrasting ways to survive in the harsh and physically stressful intertidal Patagonian coasts.