Mitochondrial DNA diversity and taxa delineation in the land snails of the Iberus gualtieranus (Pulmonata,Helicidae) complex

Iberus gualtieranus is a species complex of land snails that is endemic to the Iberian Peninsula. The species taxonomy of the group is based merely on the basis of shell morphology, but validity of the existing taxonomy is uncertain. Using mitochondrial DNA (mtDNA) data (cytochrome oxidase I and 16S rRNA sequences) we were able to validate the observed phylogenetic taxa within the I. gualtieranus s.l. complex by means of the analysis of specimens of the different morphospecies, together with the study of topotypes. Strong incongruences were obtained between morphology and molecular data. The Iberus alonensis morphospecies comprised several genetically divergent but morphologically cryptic lineages. Considering (1) the allopatric distribution of the operational taxonomic units (OTUs), (2) the morphological differentiation, (3) the possible occurrence of hybridization among the different lineages, and (4) the strong differentiation of the mtDNA phylogroups, we suggest the main lineages obtained, for the time being, may be treated as evolutionary species. The robust phylogenetic reconstruction obtained allows us to consider I. alonensis s.s., Iberus campesinus, Iberus carthaginiensis, and Iberus gualtieranus s.s. as valid species. Two additional unnominated taxa of the alonensis shell type have also been identified. Further subdivisions are also considered, including Iberus gualtieranus mariae and Iberus gualtieranus ornatissimus. © 2008 The Linnean Society of London, Zoological Journal of the Linnean Society, 2008, 154 , 722–737.     

A COMPARATIVE STUDY OF THE MORPHOLOGY OF THE PULMONATE SNAIL PSEUDOTACHEA LITTURATA (PFEIFFER) AND OTHER SPECIES OF PSEUDOTACHEA, IBERUS AND CEPAEA

A population of Pseudotachea litturata (Pfeiffer, 1851) fromTarifa (Càdiz, Spain) has been studied. The morphologicalresults are compared with those from P. splendida, Iberus gualtierianus,I. alonensis, I. marmoralus, I. guiraoanus and four speciesof the genus Cepaea using, as an exploratory method, the Wagnerparsimony procedure and 18 characters of the shell, genitalsystem and karyotype have been analysed. According to this methodit seems that the taxonomical position of P. litturata in thegenus Pseudotachea is confirmed, and agrees with the phylogeneticalrelationships in this group of species. The genus Cepaea seemsto be well established, although two species groups can be distinguished:C. nemoralis—C. hortensis and C. syluatica—C. vindobonensis.These differ mainly in chromosome number, diverticulum lengthand degree of shell polymorphism. Although the present resultsdo not allow us to clarify the current taxonomical problemswithin the genus Iberus, the species studied seem to belongto a natural group (Received 15 September 1987; accepted 1 January 1988)     

THE GENUS TRICOLIA IN THE EASTERN ATLANTIC AND THE MEDITERRANEAN

The genus Tricolia(Phasianellidae) in the Eastern Atlantic andMediterranean is reviewed on the bases of radular morphology,shell characters and polychromatism. The taxa included within the T. pullus (L.) group are not clearlyseparable and are treated as geographical subspecies:T. pulluspullus (L.), T. pullus picta (da Costa), T. pullus azorica (Dautzenberg),T. pullus canarica Nordsieck. The case of T. tenuis (Michaud)is problematic since it is clearly separable from sympatricT. pullus pullus in most parts of the Mediterranean but transitionalto T. pullus picta, towards the Atlantic. Other taxa are considered as having specific status since theyare consistently separable when found in sympatric populations.These include T. speciosa (Muhlfeldt), T. miniata (Monterosato),T. tingitana n.sp., T. petiti (Craven), T. nordsiecki (Talavera)and T. algoidea (Pallary). The last two species are stated forthe first time as belonging to the Phasianellidae. The genusEpheriella Pallary, based on T. algoidea, is synonymized withTricolia. (Received 6 April 1981;     

Disruptive selection by predation offsets stabilizing selection on shell morphology in the land snail Iberus g. gualtieranus

This work analyses the selection on shell morphology (height and width) in an arid-dwelling land snail with a flattened shell, Iberus g. gualtieranus. The findings show absence of selection, but more detailed analyses, separating the effect of different selective agents, show that there is disruptive selection on shell height caused by predation (by black rats, Rattus rattus). Nevertheless, this disruptive selection was balanced by stabilizing selection caused by other unknown mortality sources with the same strength. The two selective forces acting in opposite directions resulted in an absence of appreciable selection on shell height. This study suggests that it is important to analyse the effect of different sources of selection acting simultaneously on a trait, in order to attain a precise picture of the selective patterns, especially when the whole selection is close to zero, as this does not necessarily imply the absence of selection on the trait.     

Designing ex-situ conservation strategies through the assessment of neutral genetic markers: Application to the endangered Androcymbium gramineum

Recent anthropogenic habitat perturbationsrelated to the unsustainable development of thetwo areas of distribution of Androcymbiumgramineum in Almería (South of Spain) andthe Atlantic coast of Morocco are outweighingthe intrinsic biological survival assets ofthis narrow Ibero-Moroccan endemic. We usedpopulation genetic data on 18 isozyme loci for13 populations to design a comprehensivesampling strategy for ex-situ conservation thatstraddles the results of independenttheoretical developments and the indications ofseveral genetic polymorphism parameters.Regressions based on the probabilities of lossestimate that sampling one population would beinsufficient to represent the variationattributable to the rare alleles found in thepopulations surveyed from Almería orMorocco and suggest that the relationshipbetween the number of populations sampled ineither area should conform to the proportion0.6:0.4, respectively. The estimate of thenumber of populations that need be targeted torepresent 99% of the genetic variationdetected indicates that it would be necessaryto sample eight populations intensively. Fivepopulations from Almería and three fromMorocco were selected through the ranked valuesof the average number of alleles per locus andthe expected heterozygosity on the grounds thatthese parameters provide unambiguousindications of polymorphism that, in the caseof heterozygosity, are less likely to beinfluenced by sampling error. Spatialautocorrelation surveys in the largestpopulation known indicate that seeds should becollected at a minimum separation of 15 m toavoid the sampling of close relatives.     

Genetic Diversity inBrassica oleraceaL. (Cruciferae) and Wild Relatives (2n=18) using Isozymes

Genetic diversity in 36 populations of wild taxa of theBrassicaoleraceaL. group (2n=18) and two cultivated forms was studiedusing isozyme variation at 11 loci for five enzyme systems (IDH,6-PGD, PGM, PGI, MDH). Mean values for the percentage of polymorphicloci and expected heterozygosity were 54% and 0.224, respectively.Statistically significant differences among allele frequencieswere found with the 6-PGD isozyme system. Intrapopulationalgenetic diversity was 67% while interpopulational genetic diversitywas only 33%. The dendrogram obtained, using genetic distancesamong taxa, showed three different groups. With the exceptionofB. incana,they agree to the already accepted relationshipsamong the 14 taxa studied: the West Mediterranean group, withB.oleracea, B. alboglabra, B. bourgeauiandB. incana; another groupof species growing in the central Mediterranean area, whichincludesB. villosa, B. villosasubsp.drepanensis, B. rupestris,B. macrocarpa(the four taxa together withB. incanaare consideredtheB. rupestrisgroup) andB. montana; and finally the Aegeangroup, which includes the three subspecies ofB. cretica.Clearlyseparated wereB. insularisandB. hilarionis, showing the maximumgenetic distance. Separate dendrograms were also obtained forB.oleracea, B. montana, B. creticaandB. rupestrisgroup, and geneticdiversity parameters were estimated. Genetic distances amongB.oleraceapopulations are in the same range as populations oftheB. creticasubspecies. Highest genetic distances were foundamong populations of theB. rupestrisgroup.Copyright 1998 Annalsof Botany Company. Brassica oleraceaL., wild relatives, genetic diversity, genetic resources, isozymes.     

Hydrography and phytoplankton of the Ria de Vigo before and during a red tide of Gymnodinium catenatum Graham

At the end of summer 1986, hydrographic data and phytoplanktonsamples were collected on three occasions in the Ríade Vigo. The phytoplankton composition was studied in relationto the hydrography using principal component analysis and canonicalcorrelation analysis. Upwelling and the response of the planktoncommunity emerge as the main source of variation in the analysesWithin this framework, nutrient regeneration was observed, partlyattributable to ciliates and small flagellates. The second sourceof variation of the phytoplankton was the presence in the latersamples of a Gymnodinium catenatum red tide. Two other phytoplanktonpopulations were clearly differentiated One oceanic, dominatedby Erythropsis sp, Ceratium horridum and Stauroneis membranacea,which practically disappeared when the red Ode was established.The other, dominated by Solenicola setigera, dinoflagellatecysts and elongated diatoms, was a permanent population locatedbelow the populations which responded to upwelling. The penetrationof warm oceanic water into the ría during the secondday of observations altered the distributions of all the planktonicpopulations, isolating them in a downwelling zone. The energyof the oceanic intrusion could have been sufficient to removesediments from the interior of the ría and to stimulatethe formation of the red tide, by confining a water body inthe ría which lost little by mixing. It is likely thatthe resuspension of G.catenatum cysts provided the necessaryinoculum.     

REVISION OF THE GENUS ACAVUS FROM SRI LANKA (GASTROPODA: ACAVIDAE)

The genus Acavus from Sri Lanka includes three species, Acavushaemastoma (Linnaeus), Acavus superbus (L. Pfeiffer) and Acavusphoenix (L. Pfeiffer) with two subspecies, Acavus phoenix phoenix(L. Pfeiffer) and Acavus phoenix castaneus new subspecies.The conchological and anatomical differences between these species,as well as their intraspecific variation are described. Theirdistribution is shown on maps and all locality records are listed.The presence of extensive intraspecific polymorphism, the developmentof geographical subspecies and the presumable hybridizationbetween some species show that the Acavus species are not isolatedrelict species, but that speciation is in progress and thatthe radiation of the Acavus species is probably much youngerthan the radiation of the acavid genera. The three Acavus speciesare not acutely endangered at present. In contrast to manyother land snails, there is no risk of a rapid decline of theAcavus species due to deforestation, because these species alsooccur in synanthropic habitats. (Received 22 December 1998; accepted 2 October 1999)     

THE OPERCULA OF NERITOPSID GASTROPODS AND THEIR PHYLOGENETIC IMPORTANCE

The fossil record of neritopsid opercula and shells shows thatthe shell shape typical for Neritopsidae and Neritidae appearedin the Triassic. The ancestors of Neritimorpha were most probablyforms similar to Naticopsis. The operculum of Recent Neritopsisis composed of two calcitic parts secreted from inside and anaragonitic callus deposited from outside. Similar neritopsidopercula were already present in the Late Triassic. The firstopercula with asymmetrically situated muscle scars, possiblyancestral for neritids, also appeared at that time. (Received 27 May 2004; accepted 8 December 2004)     

ELECTROPHORETIC AND MORPHOMETRIC CHARACTERS IN POPULATION DIFERENTIATION OF THE PEARL OYSTER, PINCTADA RADIATA (LEACH), FROM AROUND BAHRAIN

Variation at the leucine aminopeptidase (Lap), glucose phosphateisomerase (Gpi) and tetrazolium oxidase (To) loci was investigatedin samples of three populations, Al-Mayana (MAY), Shigita (SH)and Mina Salman (MS), of Pinctada radiata from pearl oysterbeds around Bahrain. The To locus was monomor-phic. SignificantLap and Gpi heterozygote deficiencies were evident and it issuggested that these were generated by selection. The MS population,to the East of Bahrain, differed significantly in Gpi allelefrequencies from both Northern populations (MAY, SH) and Nei'sgenetic identity indicates a close relationship between theNorthern populations. Measurements of shell morphometrics were used both as ratiosof one dimension to another, and as regressions of one dimensionon another to examine relatedness between populations. Boththese mor-phometric approaches gave different results from eachother and also differed from the electrophoretic data. It isconcluded that estimates of relatedness in pearl oysters basedon electrophoretic data will be more reliable than those basedon shell shape. (Received 20 November 1990; accepted 12 April 1991)     

Genetic Diversity inBrassica oleraceaL. (Cruciferae) and Wild Relatives (2n=18) using RAPD Markers

This paper presents the evaluation of genetic diversity in 29populations of wild taxa of theBrassica oleraceaL. group (2n=18)and two cultivars, using RAPDs as molecular markers. In a previouspaper (Lázaro and Aguinagalde,Annals of Botany82: 000–000,1998), 11 isozymes were used for the same purpose. Results obtainedwith the two molecular markers (isozymes and RAPDs) are compared.DNA from ten individuals per population was analysed using sixdifferent primers; the 151 detected bands were polymorphic,11 were common to all species, six to all taxa, only one toevery population; and no bands were shared by every individual.The dendrogram obtained using genetic distances clustersB. oleraceapopulationswithB. bourgeaui, B. alboglabra, B. montanaandB. incana. B.insularis, B. macrocarpa, B. villosaandB. rupestrispopulationsform another cluster. Populations ofB. creticaandB. hilarionisformthe third cluster. Genetic diversity inB. oleraceapopulations,theB. rupestriscomplex andB. creticasubspecies was estimatedusing the AMOVA programme; the latter was the most diverse.Copyright1998 Annals of Botany Company. Brassica oleraceaL., wild relatives, genetic diversity, genetic resources, RAPD markers, AMOVA.     

TAXONOMIC REVIEW OF THE TREE SNAIL AMPHIDROMUS ALBERS, 1850 (PULMONATA: CAMAENIDAE) IN THAILAND AND ADJACENT AREAS: SUBGENUS AMPHIDROMUS

Extensive series of Amphidromus Albers, 1850 were collectedfrom low-altitude areas (30–500 m above sea level)in Thailand, Laos, Malaysia, Singapore and Indonesia between1993 and 2004. Data on shell colour, genital and spermatophoremorphology, and radular structure are presented. Five speciesof Amphidromus (Amphidromus) are reviewed, of which four occurin Thailand. Intraspecific variation between populations ofAmphidromus (Amphidromus) atricallosus (Gould, 1843) is considered;three subspecies are accepted and a new subspecies described.Subspecies of A. (A.) inversus (Müller, 1774) and A. (A.)schomburgki (Pfeiffer, 1861) are reconsidered, and a new subspeciesof the latter is described. A dichotomous key to the speciesand subspecies of Amphidromus (Amphidromus) in Thailand is provided. (Received 7 May 2004; accepted 31 March 2005)     

Variation of Nuclear DNA Content in the Biflorus Species of Lonchocarpus (Leguminosae)

This represents the first study of nuclear DNA content in alarge sample (135 spp.) from a tropical arboreal genus, in whicha large proportion of the species were examined (42 spp., 31.1%).Somatic chromosome numbers and 4C-DNA values for 51 taxa ofLonchocarpus are reported. All taxa were diploid with 2 n =22,but their DNA content ranged from 1.92 to 2.86 pg 4C nucleus,corresponding to a 48.95% variation in genome size. In the 74collections studied, no correlation was observed between DNAcontent and habitat altitude. Variation in nuclear DNA contentwas analysed at the level of genus, subgenus, section and subsection.Variation in genome size was also studied within some species,either among widely separated populations or among differentintraspecific taxa. Very little variation in genome size wasdetected between populations, subspecies, and varieties of thesame species. The taxonomic implications of variation in nuclearDNA content are discussed.Copyright 2000 Annals of Botany Company Lonchocarpus (Leguminosae), DNA content, chromosome number.     

HETEROSTOPHIC SHELLS AND PELAGIC DEVOLOPMENT IN TROCHOIDEANS: IMPLICATIONS FOR CLASSIFICATION, PHYLOGENY AND PALAEOECOLOGY

Four species of archeogastropods, presumed members of threesubfamilies of the trochidae, exhibit significant differencesin developmental modes and shell coiling. All four species havelecithotrophic development which is reflected in their inflatedpausispiral protoconchis; however, Margarites marginatus andLirularia succincta have benthic development in gelatinous masses,while Margarites pupillus and Calliostoma ligatum have pelagicembryos and swimming larvae with a potential for dispersal overa period of a week or longer. These modes cannot be deducedfrom the size of the egg, the size or shape of the protoconch,or the size or relative prominence of female pallial reproductivestructures. The protoconch of C. ligatum is orthostrophicallycoiled, but the protoconchs of the other three species are hyperstrophicallycoiled although their teleconchs are orthostrophic. These threetrochoidean species thus share with architectonicoideans, pyramidelloideans,opisthobranchs, and pulmonates the distinctive shell characterof heterostrophy, previously unreported for archaeogastropods. These observations, considered together with others reportedin the literature, show: (1) that developmental mode (pelagicor benthic) cannot be inferred from protoconch appearance ortaxonomy in major trochoidean ganera; (2) that significant dispersalpotential is present in the histories of some trochoidean archeogastropoids;and (3) that character sets (pallial reproductive structures,pairing during spawning, heterostrophic shell coiling) thoughtnot to occur below the mesogastropod level are found in theTrochoidea. These conclusions bring into question the usefulnessof these characters in defining higher gastropod taxa and raiseadditional questions concerning the ancestry of the higher gastropods. (Received 16 February 1989; accepted 25 June 1989)     

Random Amplified Polymorphic DNA Variation among and within Selected Ixora (Rubiaceae) Populations and Mutants

Random amplified polymorphic DNA (RAPD) analysis was used tostudy variation among and within selectedIxora (Rubiaceae) populationsand mutants. Six populations of I. congesta yielded identicalbanding patterns suggesting genetic uniformity of this species.However, six populations of I. coccinea varieties (three red-flowered,two yellow-flowered and one red-flowered wild-type) exhibitedinfraspecific differences in RAPD profiles. Small and largeleaves of an atavistic mutant cultivar of I. coccinea were alsosubjected to RAPD analysis. An extra band was amplified in thelarge leaves that was absent in small leaves, suggesting thatthe phenotypic alteration in this taxon is due to genetic mutationrather than epigenetic changes. Similarly, an extra band wasdetected in the white sectors of I. Variegated compared to thegreen sectors, suggesting that the shoot apical meristems ofthis cultivar exist as a genetic chimera. DNA gel blot hybridizationwas performed to confirm the specificities of selected bands.Our study indicates that differences among individuals of variouspopulations and mutants may be detected using RAPD markers.Copyright 1999 Annals of Botany Company Ixora L., variegated variety, RAPD fingerprinting, DNA gel blot, intraspecific genetic similarity, atavistic mutant.     

赤眼蜂分子鉴定技术研究

通过对6种常见赤眼蜂,即松毛虫赤眼蜂Trichogramma dendrolimi Matsumura、玉米螟赤眼蜂T. ostriniae Pang et Chen、螟黄赤眼蜂T. chilonis Ishii、广赤眼蜂T. evanescens Westwood、甘蓝夜蛾赤眼蜂T. brassicae Bezdenko及食胚赤眼蜂T. embryophagum (Hartig)之核糖体核糖核酸基因第二内部转录区（rDNA-ITS2）的克隆测序,调用GenBank中同源序列,对不同蜂种的rDNA-ITS2序列进行了多重排比和聚类,探讨了rDNA-ITS2用于赤眼蜂属不同种系统进化关系分析及赤眼蜂分子鉴定的可行性。为了考察rDNA-ITS2在赤眼蜂种下水平鉴定上的可能性,作者收集了我国常见的松毛虫赤眼蜂6个地理种群（黑龙江亚布力、吉林长春、吉林仁和、陕西长安、江苏徐州、广东广州）,采用相同方法测定了它们的rDNA-ITS2序列。序列分析结果表明,赤眼蜂种下阶元ITS2序列非常保守,而种间存在明显的遗传差异。通过外群比较发现,rDNA-ITS2只适合于赤眼蜂种一级的分子鉴定。     

Molecular and morphological divergence in a stygobiont gastropod lineage (Truncatelloidea,Moitessieriidae, Paladilhiopsis) within an isolated karstic area in the Mecsek Mountains (Hungary)

Truncatelloid gastropods are one of the most species‐rich subterranean invertebrate groups. Their current taxonomy is based on morphological characters. However, this is not a comprehensive approach and does not take into account the degree of phylogenetic divergence between stygobiont populations inhabiting hydrologically isolated but geographically close caves. We studied two Paladilhiopsis populations of a small and isolated karstic area (Mecsek Mountains, Hungary) with two hydrologically separated cave systems, investigating morphological (shell morphometrics) and genetic (COI, 16S) divergence together. The populations differed both morphologically and genetically: we found strong divergence in the relative width of the shell (best described by the variable “shell angle”) and a 6.4% divergence in COI. This provides strong support for the presence of two distinct taxa; however, it is still doubtful whether they differ at the species or the subspecies level. In one of the caves, we found representatives of both haplotypes (and phenotypes), which can be explained by secondary contact after an allopatric divergence.     

ALTITUDINAL VARIATION AND ADAPTIVE ZONES IN ARIANTA ARBUSTORUM: A NEW LOOK AT A WIDESPREAD SPECIES

Arianta arbustorum is a European land snail, occurring in bothlowland and mountainous regions. Its shell is usually globular,which makes the species aberrant among the Campylaeinae in shellshape, habitat, and wide, not exclusively montane range. Withincreasing altitudes, the shells usually decrease in size andbecome more conical. However, in some mountain stocks thereare populations with depressed shells, provided with an openumbilicus. This form (subspecies) is only known from formerPleistocene refugia. It is hypothesized that in the Pliocene, the primary montaneA. arbustorum, with a depressed shell, entered an additional,adaptive zone, while conquering damp lowland habitats. Thisresulted in an enormous range extension. In the new habitata different (globular) shell shape evolved and the species becamepolytypic. The subsequent glaciations wiped out most of theoriginal montane A. arbustorum with depressed shells. Wherethis had occurred, post-Pleistocene re-colonizing conspecificsnails with globular shells, found vacant territory and coulddevelop the dines mentioned before. The invading populationsdid not regain the original, montane, depressed shell shape.This shape was only retained in a few refugia, where the populationssurvived amidst the glaciers. (Received 9 February 1990; accepted 31 May 1990)     

Subspecies recognition in the house mouse: a study of two populations from the border of a hybrid zone

Mate choice is the outcome of sexual preference for partnerscarrying specific signals. Thus, mating among conspecifics(homogamy) depends on the occurrence of species recognitionsystems. We asked what happens if populations diverge, andwe investigated female sexual preference between two subspeciesof the house mouse in populations from the borders of a hybridzone (Jutland, Denmark). We used choice tests to analyze theoccurrence of recognition signals and to locate these signalsin soiled bedding and urine. Our results show that populationsof the two subspecies can be discriminated on the basis ofurinary signals, suggesting that the latter have diverged. Additionally, these signals seem to have similar features inpopulations of different geographical origins, suggesting thatsubspecific differentiation occurs. This is the first demonstrationthat subspecific recognition through urinary signals occursin the house mouse. However, while Mus musculus domesticusdoes not display a preference, we show that Mus musculus musculusfemales tend to mate with males of the same subspecies. We discuss the different factors that could explain these discrepanciesbetween females of the two taxa: differences in signal perception,evolution at a different pace, or evolution under differentselective pressures in their area of contact. Further, we proposethat the divergence in male signal was at least partly initiatedin allopatry and discuss different evolutionary scenario thatmay explain the patterns observed in Denmark and their relevance to isolation between the two taxa.     

COMPARATIVE ANALYSIS OF KARYOTYPES AND NUCLEOLAR ORGANIZER REGIONS IN DIFFERENT POPULATIONS OF MYTILUS TROSSULUS, MYTILUS EDULIS AND MYTILUS GALLOPROVINCIALIS

The cylotaxonomical characteristics of Mytilus tros-sulus, M.edulis and M. galloprovincialis were studied using karyometricanalysis and silver staining. The karyotype of M. trossulus,reported here for the first time, consists of six metacentricand eight submetacentric-subtelocentric chromosome pairs. Theoccurrence of five metacentric chromosome pairs in M. galloprovincialisdifferentiates this taxon from M. trossulus and M. edulis whichboth have six meta-centrics. The number of submetacentric tosubtelo-centric chromosome pairs was variable between populationsand between taxa. We suggest that this variability has arisenfrom differential chromosome condensation rather than from structuralchange. Intercomparison of chromosome relative length showedstrong similarity among the three taxa. Chromosomal nucleolarorganizer regions (NORs) were identically located on two submetacentric-subtelocentricchromosome pairs in all three taxa. However, M. trossulus showeda characteristic difference in the occurrence of NORs on onemetacentric pair, which was absent in M. galloprovincialis andM. edulis. Variability in the number of NORs per cell was observedin all populations studied. We conclude that the three taxaof Mytilus studied here cannot be differentiated by asinglekaryological character, but that a combination of karyologicalcharacters is virtually diagnostic ^* To whom correspondence should be sent (Received 24 January 1994; accepted 9 March 1994)