Chromosomal evolution and speciation in the Gyratrix hermaphroditus species complex (Platyhelminthes,Kalyptorhynchia): karyometric and morphological analyses of fifteen fresh-water populations from Western Europe

Fifteen fresh-water populations of the Gyratrix hermaphroditus species complex from Western Europe (Italy, Spain, France) were karyometrically analysed. All shared the same chromosome number 2n = 4. Three distinct karyotypes were recognized on the basis of the different values of the centromeric index of chromosome 2, respectively metacentric, intermediate between meta- and submetacentric, and sub-telocentric. The morphology of chromosome 1 (an isobrachial metacentric) and the length ratio of chromosomes were on the contrary constant in all the populations investigated.Differences in the size of male cuticular organs were observed accompanying the karyological differentiation.The fifteen populations are interpreted as representing at least three sibling-species.Chromosomal evolution and phylogenetical relationships with the marine species of the Gyratrix hermaphroditus complex are discussed.     

Karyology of a marine population of Gyratrix hermaphroditus (Turbellaria,Rhabdocoela) and chromosomal evolution in this species complex

Karyology and reproductive biology of a marine population of the species complex Gyratrix hermaphroditus, from Roscoff (Brittany, France), have been investigated. A diploid complement of six chromosomes was determined from spermatogonial mitotic figures. One chromosome pair is metacentric, the second is intermediate between meta- and submetacentric, and the third is subtelocentric.In this population, regular meiosis occurs in both female and male germ lines, and the animals reproduce only by means of amphimictic eggs. Certain specimens of the population showed the elimination of one of the three bivalents during the first meiotic division in spermatogenesis. It seems that such animals produce normal and aneuploid sperm simultaneously; the aneuploid sperm are not capable of fertilization.The Roscoff population differs in its karyotype (2n = 6) from freshwater populations, which are either diploid (2n = 4) or polyploid (3n = 6, 4n = 8). These results suggest that aneuploidy played a role in the differentiation of freshwater populations from an originally marine species complex.     

How many species of Paradoxurus civets are there? New insights from India and Sri Lanka

Using molecular data and morphological features, we investigated the species limits and genetic diversity among populations of the Asian palm civets of the genus Paradoxurus. Our main objectives were to determine the number of species within Paradoxurus hermaphroditus and to test the validity of the newly proposed species within Paradoxurus zeylonensis. Fragments of two mitochondrial (Cytochrome b, Control Region) and one nuclear (intron 7 of the beta fibrinogen) markers were sequenced from 128 individuals of P. hermaphroditus, P. zeylonensis and Paradoxurus jerdoni. DNA sequences were analysed using phylogenetic and haplotype network methods. Our analyses confirmed that P. hermaphroditus comprises three major clades, which should be recognized as separate species: P. hermaphroditus (Indian and Indochinese regions), Paradoxurus musangus (mainland Southeast Asia, Sumatra, Java and other small Indonesian islands) and Paradoxurus philippinensis (Mentawai Islands, Borneo and the Philippines). Furthermore, we have proposed that there are two subspecies within both P. musangus and P. philippinensis, and there might be at least two or three subspecies within P. hermaphroditus. We found a very low genetic diversity and no geographical structure within P. zeylonensis and did not find any support for splitting P. zeylonensis into several species nor subspecies. Finally, we confirmed that P. jerdoni and P. zeylonensis are sister species.     

Characteristics of spermiogenesis and mature spermatozoa in Gyratrix hermaphroditus (Plathelminthes, Rhabdocoela, Kalyptorhynchia)

 Some events of spermiogenesis and the submicroscopic anatomy of male gametes in Gyratrix hermaphroditus are described. Special features occurring during the steps of sperm cell maturation are the development of delicate rootlet-like and spike-shaped structures attached to the basal bodies, an anchor apparatus of cilia staying near the tip of the outgrowing spermatids, the differentiation of a single mitochondrial rod, and the processes of compartmentalizing the chromatin resulting in the formation of extranuclear lamellar stacks. Mature spermatozoa are especially characterized by two incorporated axonemes, a string of large mushroom-shaped dense bodies in the anterior section of the cell, and two lateral rows of nuclear lamellar stacks in the median cell segment. Based on the present findings, the process of spermiogenesis and the organization of spermatozoa in the ground pattern of the monophylum Kalyptorhynchia are reconstructed. Apparently, the sister group of the Kalyptorhynchia can be found within the other free-living and symbiotic Rhabdocoela; the hypothesis of a sister group relationship with the Neodermata is not corroborated. Probably, spermatozoal characteristics can contribute to a discrimination between distinct populations of G. hermaphroditus which is proved to be a complex of several sibling species. Accepted: 25 June 1998     

Serotonergic and SCP<Subscript>b</Subscript>-like innervation of the atrial complex in <Emphasis Type="Italic">Gyratrix hermaphroditus</Emphasis> (Platyhelminthes,Kalyptorhynchia) revealed with CLSM

The platyhelminth reproductive system is a complex series of canals, glands, and sclerotic components that figure prominently in our understanding of reproductive physiology, taxonomy, and evolution of the Platyhelminthes. Yet, there is limited information on its innervation, especially for free-living species of Rhabdocoela, the most speciose and ecologically diverse assemblage. Here, innervation of the reproductive system in the common marine kalyptorhynch, Gyratrix hermaphroditus, is studied using confocal laser scanning microscopy (CLSM), fluorescent phalloidin, and antibodies to serotonin (5HT) and small cardioactive peptide b (SCP_b). Results show that portions of the female atrial system (uterus, female gonopore) are innervated by peptidergic (SCP_b) neurons that may function to control muscles involved in egg movement. In contrast, portions of the male atrial system (male atrium, male gonopore) are innervated by both peptidergic and serotonergic neurons. These neurons form a complex series of hoops around the musculature of the male atrium that houses prostate stylet type II. It is hypothesized that 5HT is the primary myoexcitatory neurotransmitter, and that it acts either synergistically with SCP_b to trigger muscle contractions of the male atrium and protract the copulatory stylet, or that SCP_b plays an inhibitory role during contraction of the male atrium. A comparison of the distribution of peptidergic and serotonergic neurons in G. hermaphroditus with other free-living species and parasitic rhabdocoels (Neodermata) reveals similarities in innervation of comparable (though probably not homologous) reproductive organs. These results suggest that the atrial systems of phylogenetically diverse species may share a common neuronal physiology despite their structural differences and potential independent evolutionary origins within the Platyhelminthes.     

A new freshwater folliculinid (Ciliophora) from the karstic region of Croatia

Members of the genus Lagotia (Folliculinidae, Ciliophora) were known only from marine habitats where they are widely distributed. In running waters on travertine barriers in the karstic region of Croatia we found a freshwater species, Lagotia dinaridica n. sp., which exhibits several characteristics that set it apart from its congeners.     

Vegetative propagation by fragmentation of Gelidium sclerophyllum (Gelidiales,Rhodophyta)

Results of the first phase of production of "seed" for a project of marine culture of species of Gelidiales are presented. Vegetative fragments of different sizes and from different parts of the thallus of Gelidium sclerophyllum from the Mexican tropical Pacific coast were cultured under nine treatments with different N and P concentrations. In treatments without N all fragments died; good vegetative growth was present in all remaining treatments. Growth in length and branch production were favored in medial fragments, and rhizoid production was favored in apices. Growth in length and biomass increase were stimulated by high N concentrations (2–3 mM), and branch and rhizoid production were stimulated by high P concentrations (100–150 µM).     

Development and characterization of ten novel microsatellite markers from olive ridley sea turtle (Lepidochelys olivacea)

Olive ridleys, one of the widely distributed marine turtle species has undergone declines in recent years due to multiple anthropogenic factors warranting conservation efforts for which assessment of genetic variability in existing populations become critical. Here we describe development of ten new microsatellite markers from a short sequence repeat-enriched partial genomic DNA library, which are found to be highly informative for genetic studies. Eight of these markers when tested on 83 olive ridley turtles revealed high allelic diversity (4–27 alleles per marker), and high observed and expected heterozygosity estimates that ranged from 0.29 to 0.82 and 0.62 to 0.94, respectively. Two microsatellites were monomorphic in the tested olive ridley samples, but were found to be informative/polymorphic when tested on related marine turtle species. More importantly, nine of the new markers showed robust cross-species amplifications in three related species Dermochelys coriacea, Chelonia mydas and Eretmochelys imbricata. Thus, this study describes ten new microsatellite markers and also demonstrates their potential as efficient genetic markers in studies related to parentage analysis, population structure, phylogeography and species relationships of olive ridleys and other marine turtle species.     

Two new species of the genus Rhynchonema (Nematoda,Xyalidae) from Amakusa,south Japan

During the ecological studies on marine nematodes in the intertidal zone and an adjacent seagrass Zostera marina Linne bed in Amakusa, south Japan, two new species of the genus Rhynchonema Cobb, 1920 were found. They are described as R. kikuchii and R. amakusanum. Up to now, Rhynchonema species were unknown from Japan.     

The genetic status and conservation management of two cultivated bulb species extinct in the wild: Tecophilaea cyanocrocus (Chile) and Tulipa sprengeri (Turkey)

Tecophilaea cyanocrocus and Tulipasprengeri are both extinct in the wild as aresult of overharvesting by commercialcollectors. Stocks of both species survive incultivation in amateur collections, commercialnurseries and botanic garden collections.Whilst both species share a similarconservation history, the distribution ofgenetic diversity within cultivated materialwas unknown. To support the long-termconservation of both species the geneticdiversity of the surviving stocks was assessedusing amplified fragment length polymorphisms(AFLPs). This study revealed different geneticstructures for the two bulb species. The Kewcollections of T. cyanocrocus, originallyobtained from a commercial nursery, VanTubergen, are genetically highly uniform andthe addition of samples from other collectionshas dramatically increased the level ofvariation. In contrast, the collections ofT. sprengeri held at Kew since the early20th century include representativegenotypes covering the whole range of geneticvariation found in this species and areprobably the source of all other cultivatedmaterial. The results are discussed in relationto the management of these species to ensuretheir continued survival in cultivation.     

Doxomysis algoaensis, a new mysid species (Crustacea: Mysidacea) from Algoa Bay (South Africa)

Doxomysis algoaensis sp.nov. is described from Algoa Bay, South Africa where it is common in nearshore marine waters just beyond the breaker line. D. algoaensis sp.nov. is morphologically similar to D. australiensis, but can readily be distinguished by the shape and armature of the telson and the length of the exopod of the fourth male pleopod. The apical cleft is one fifth the telson length in the former species and one third the length in D. australiensis. The telson apex on each side of the cleft is also armed with five and four stout spines in the two species, respectively. In D. australiensis, the exopod of the fourth male pleopod is almost three times the length of the endopod; in D. algoaensis sp.nov., the exopod is only slightly longer than the endopod. Other distinctive features of D. algoaensis sp.nov. include the maxillary palp, which is only slightly broader than long and the greater number of spines on the endopod of the uropod.     

Karyotaxonomic analysis of the genusBunium (Umbelliferae)

Chromosome numbers are reported for 23 species of the genusBunium, distributed mainly in Middle Asia and Transcaucausia. The great diversity of basic chromosome numbers is unusual for a genus ofUmbelliferae: x = 11, 10, 9, 8, 7, and 6. This series obviously has to be interpreted as descending dysploidy. Infraspecific aneuploidy was found in three species, but there are no B-chromosomes or polyploidy. The karyotypes of 18 species are described, using two morphometric chromosome parameters, centromere position and relative length (Table 2), and illustrated (Figs. 2–4). Species vary greatly in their karyotype asymmetry (20.4–45.9%). Chromosome numbers and structures appear to be useful in the karyotaxonomical and biogeographical analysis of the genus.     

Fungal Infection of Mantis Shrimp (<Emphasis Type="Italic">Oratosquilla oratoria</Emphasis>) Caused by Two Anamorphic Fungi Found in Japan

Two fungal pathogens of the mantis shrimp (Oratosquilla oratoria) in Yamaguchi and Aichi Prefectures, Japan are described as the new species Plectosporium oratosquillae and Acremonium sp. (a member of the Emericellopsis marine clade). Both fungi infect the gills of the mantis shrimp, which become brown or black due to melanization. The former species is characterized by its slow growth on artificial seawater yeast extract peptone glucose (PYGS) agar, pale yellow to pale orange or grayish yellow colonies, short cylindrical solitary phialides with a wavy tip, and one-celled ellipsoidal conidia. Although lacking the two-celled conidia demonstrated by the type species Plectosporium tabacinum, the taxonomic placement of the new species was confirmed by DNA sequence analysis of the internal transcribed spacer (ITS) region of ribosomal DNA (ITS1, 5.8S rDNA and ITS2). Acremonium sp., the other causal pathogen, differs from P. oratosquillae by its fast growth on PYGS agar, pale orange to salmon-colored colonies, long, slender conidiophores consisting of solitary phialides with tips lacking an undulate outline, and typically cylindrical conidia. Analysis of ITS and β-tubulin gene sequences placed this fungus within the phylogenetically distinct Emericellopsis (anam. Acremonium) marine clade. Various physiological characteristics of both pathogens were also investigated. This is the first report of a fungal infection found on the mantis shrimp in Japan.     

Comparative floral morphometrics in day-flowering, night-flowering and self-pollinated Caryophylloideae (Agrostemma, Dianthus, Saponaria, Silene, and Vaccaria)

Morphological variation of flowers with different pollination modes was studied in 53 species representing five genera (Agrostemma, Dianthus, Saponaria, Silene s.l., Vaccaria) of the subfamily Caryophylloideae. All species were classified a priori as either diurnal, nocturnal, or selfing. Canonical discriminant analysis (CDA) of 13 floral characters revealed significant correlations between floral morphometry and pollination modes. A CDA of taxonomical groups represented with more than three species (Dianthus, and subgroups of Silene s.l.: Lychnis, Silene s.str., and Viscaria), revealed that the discrimination between these four taxa, based on the same floral characters, is also well supported. Main factors for the discrimination of the species with different pollination modes were characters that (a) define nectar accessibility (calyx length, calyx tooth length), (b) are related to the positioning of anthers and stigmatic areas in relation to pollinators and (c) are important for the visual attractiveness of flowers (plate width). The functional distance between the nectar source (anthophore base) and contact zone with pollinators (style tips), given as the sum of anthophore length, ovary length, and style length (hence called AOS-complex) is better correlated with the calyx length than the single characters. Further, the total AOS-complex length differs significantly between pollination modes suggesting that these characters form a functionally linked complex that is related to the pollen placement on, and stigma contact with, the pollinator's body. However, the contribution of anthophore and style length to the total AOS-complex differed significantly between Silene s.l. and other taxa indicating that the taxonomic groups follow different evolutionary ways for the construction of the functionally linked AOS-complex.     

Pollination of sevenPlectranthus spp. (Lamiaceae) in southern Natal, South Africa

The genusPlectranthus (Lamiaceae) shows remarkable radiation on the sandstones of southern Natal and northern Transkei in South Africa, where six endemic species occur. Two of these endemic species,P. hilliardiae andP. oribiensis, are included in this study, as well asP. reflexus, for which only limited data are available. The other species that were studied areP. ambiguus, P. ciliatus, P. ecklonii, P. madagascariensis andP. zuluensis. Four of these taxa,P. ambiguus, P. hilliardiae, P. reflexus andP. saccatus var.longitubus, have uniquely long corolla-tubes (20–30mm) and this is related to pollination by nemestrinid flies of the genusStenobasipteron that have proboscides of similar length. Other nemestrinid species of the genusProsoeca have shorter proboscides and pollinate two species ofPlectranthus with shorter corolla-tube lengths (6–15mm). Acrocerid flies, tabanid flies and anthophorid bees are also important visitors to these species. This study on the pollination of seven species of varying corolla-tube lengths shows a correlation between floral tube length and proboscis length of insect visitors, many of which are recorded for the first time as pollinators ofPlectranthus.     

Restriction fragment length polymorphisms of the phytohemagglutinin genes inPhaseolus andVigna (Leguminosae)

Restriction fragment length polymorphisms at the phytohemagglutinin (PHA) locus were determined among 21 genotypes ofPhaseolus vulgaris, P. coccineus, P. acutifolius, P. lunatus, and threeVigna species, using five restriction enzymes and one double digestion, in order to provide molecular evidence for their genetic relatedness. The dissimilarity between genotypes was estimated from binary RFLP data. The dissimilarity was high among species (from 0.75 to 0.95), and of variable extent among genotypes of the same species (0.33–0.89). InP. vulgaris, two different DNA hybridization patterns were found, giving further evidence for two major gene pools in that species. The restriction patterns ofP. vulgaris var.aborigineus, the putative ancestral form ofP. vulgaris, exhibit clear homology toP. vulgaris genotypes. An undefined landrace from Taiwan could be identified as aP. vulgaris genotype. RFLP-based trees for the phytohemagglutinin genes of the species studied were computed with several distance matrix and parsimony methods.     

Morphometric analysis of the genusHemerocallis L. (Liliaceae) in Korea

To better understand the patterns of variability and distributions ofHemerocallis in Korea, 53 locations were visited and measurements of 19 morphological and phenological characters were taken on plants directly from their natural habitats. For morphometric analysis, 10 plants from each of 34 populations and five herbarium specimens ofH. middendorffii were used and the data from 12 quantitative characters was analyzed using univariate analysis. Except the littoral populations of Cheju, Hong, Taehuksan, and Sohuksan Islands (H. hongdoensis M. Chung & S. Kang), three peninsular KoreanHemerocallis species can be recognized mainly in South Korea:H. hakuunensis Nakai (=H. micrantha Nakai, growing on southern, central, and northwestern Korea);H. thunbergii Baker (=H. coreana Nakai, found on southeastern and central Korea); andH. middendorffii Tr. et Mey. (central and northeastern Korea). Morphological and phenological features contributing to recognition of the three groups were; color of perianth, shape of roots, shape of inflorescence, flowering time, odor, length of inflorescence, width of the lowest bracts, length of perianth tube enclosing a ovary, width of the inner perianth lobes. Natural hybridization seems to be rare in KoreanHemerocallis. It appears that the KoreanHemerocallis species are relatively well characterized by their distribution patterns, phenology, and habitats compared with the JapaneseHemerocallis species.     

Chromosomal evolution of the Hawaiian Telmatogeton (Chironomidae,Diptera)

It is only in the Hawaiian Islands that species of the otherwise marine genus Telmatogeton have evolved into freshwater. An analysis of polytene chromosomes and karyotypes of two marine species and five freshwater species revealed that paracentric inversions and centric fusions were important in chromosomal evolution. The sequence of polytene chromosome bands common to most species, established as the Telmatogeton standard sequence, is found in a population of T. torrenticola from West Maui. Most species and other populations of T. torrenticola may be derived from the standard sequence by paracentric inversions. Similarities with the standard band sequence places T. japonicus (n=7) rather than T. pacificus (n=4) in the proposed phylogeny as the species closest to the marine ancestor of the freshwater species. One of three species (T. fluviatilis from Oahu, T. torrenticola from West Maui, or an undescribed species from East Maui), each with seven pairs of chromosomes is considered to be closest to the original freshwater species. T. torrenticola is a complex species in which there is an accumulation of fixed inversions and centric fusions in stepwise fashion in populations from west to east (West Maui n=7; East Maui n=6; Kohala Mountains n=5 and Mauna Kea n=4 both from the island of Hawaii). The population of T. torrenticola from Molokai has a reduced chromosome number (n=4) and fixed inversions. T. abnormis and T. hirtus, the only species which exhibit differentiated sex chromosomes, may be derived from the standard sequency by paracentric inversions. T. abnormis (n=4) has a simple XY system and T. hirtus (n=3/4) has a complex XY₁Y₂ system. Unique sequences of bands, differences in staining intensity of puffs and bands, and an inversion form the basis for the differentiation of the various Y-chromosomes in these species.     

Description ofGerres chrysops sp. nov. From Thailand and redescription ofGerres setifer (Hamilton, 1822) andG. Decacanthus (Bleeker, 1865) (perciformes: Gerreidae)

Gerres chrysops, a new gerreid species from the Gulf of Thailand, is described on the basis of 29 specimens, 58–83 mm in standard length (SL). A small-sized species (less than 100 mm SL), it is characterized by a silvery-gold sheen on the head and trunk, vivid yellow or yellowish-hyaline fins in life, two supraneural bones (formula 0/0/2/) and dorsal fin rays usually IX, 10. The new species is similar toG. decacanthus (Bleeker, 1865) andG. setifer (Hamilton, 1822), which are redescribed. being similarly small valid gerreid species characterized by two supraneural bones. Together, the three species comprise “theGerres setifer complex.”Gerres chrysops differs from bothG. decacanthus andG. setifer in life and fresh colors, the body being silvery-gold with vivid yellow or yellowish dorsal, caudal, anal and pelvic fins, and yellowish-hyaline pectoral fins (vs. silver body with hyaline fins in the latter two species).Gerres setifer differs fromG. chrysops andG. decacanthus in having the last dorsal fin spine longer than the penultimate spine (vs. almost same length or shorter), usually ten dorsal fin spines and nine soft dorsal rays (vs. usually IX, 10), and 8 or 9 lower series gill rakers (vs. usually 7).Gerres decacanthus differs fromG. chrysops andG. setifer in having a shorter head, lesser body depth at the first anal fin spine base, lesser body width at the pectoral fin base, and shorter second dorsal and third anal fin spines. The new species is currently known only from Angsilla, near Bangsaen, and around Si Chang Island, northeastern Gulf of Thailand.Gerres decacanthus inhabits southern Chinese waters andG. setifer is currently known from the Bay of Bengal to the Andaman Sea.     

Evolutionary history of the Paradoxurus palm civets – a new model for Asian biogeography

Aim Using molecular data and dental features, we investigated the genetic and morphological diversity among species of palm civets in the genus Paradoxurus, with a focus on the common palm civet, Paradoxurus hermaphroditus (Carnivora, Viverridae), in order to address biogeographic scenarios and provide recommendations for a taxonomic revision. Location Asia: Pakistan to the Lesser Sunda Islands. Methods We investigated the genetic diversity within Paradoxurus using two mitochondrial (cytochrome b, control region) and one nuclear (intron 7 of the β‐fibrinogen) markers. We used samples from 85 individuals of P. hermaphroditus (including 20 museum specimens) and one representative of each of the other species in the genus Paradoxurus: Paradoxurus jerdoni and Paradoxurus zeylonensis. DNA sequences were analysed using phylogenetic and haplotype network methods, and divergence dates were estimated for the clades retrieved. Furthermore, we examined dental characters from a large series of specimens and compared the morphological variation with the molecular data. Results Our phylogenetic analyses revealed that P. hermaphroditus is paraphyletic. We identified three major lineages distributed: (1) in the Indian subcontinent, south China, Hainan and in areas above 200 m in Indochina; (2) in Peninsular Malaysia, Java, Sumatra and in areas below 200 m in Indochina; and (3) in Borneo, the Philippines and the Mentawai archipelago. Our morphological observations were congruent with these three molecular lineages. Divergence date estimates inferred a Pliocene origin for Paradoxurus (2.8–5.7 Ma), with the three main clades diversifying from the mid–Early Pliocene to the end of the Pliocene. We suggest that the flooding of the Isthmus of Kra during the Pliocene was a major event shaping the diversification of Paradoxurus palm civets. We also hypothesize that the elevational segregation of the two lineages on the mainland could have resulted from the vegetational changes that were induced by Late Pliocene glacial episodes. Main conclusions The Isthmus of Kra is a major boundary between two major lineages of P. hermaphroditus. There is a need for a taxonomic revision for P. hermaphroditus, and we suggest that this species should be split into at least three species.