Analysis of Allelic Diversity and Genetic Relationships Among Cultivated Mangosteen (<Emphasis Type="Italic">Garcinia mangostana</Emphasis> L.) in Java,Indonesia Using Microsatellite Markers and Morphological Characters

The genetic variation and relationships of the mangosteen (Garcinia mangostana L.) were observed across mangosteen populations in Java, Indonesia using newly identified microsatellite loci and morphological characters. In this study, we developed an improved protocol to isolate microsatellite loci, named Selective Repeats from AFLP Sequence, by using a hybridized membrane. Twenty microsatellite loci were evaluated using 78 individuals from five mangosteen populations, and we successfully amplified four closely related Garcinia species, including G. malaccensis, G. hombroniana, G. celebica, and G. porrecta. Eight loci were monomorphic and the others were polymorphic. Sixty-nine alleles were found, with 3.491 per locus on an average. Genetic diversity (H?) was calculated with an average across loci within population (H?_S) as 0.39, an average loci across many populations (H?_T) as 0.444, and genetic differentiation (F?_ST) as 0.147. Furthermore, based on morphological characters, mangosteen individuals from four populations including Leuwiliang, Wanayasa, Puspahiang, and Kaligesing also had morphologically distinct fruit weight, rind weight, and rind thickness among populations. The study also elucidated the dispersal pattern of mangosteen in Java; the source of the genotype mangosteen in Java population was the Wanayasa population. In addition, we found evidence of tetraploidy in mangosteen. These results have potential applications in future breeding, conservation studies, and genetic assessment of mangosteen and their closely related species.     

Contributions to the cytotaxonomy of the Commelinaceae*

Further material of Gibasis geniculata (Jacq.) Rohw. (syn. Tradescantia geniculata Jacq.) and other Gibasis species collected in the field has been studied. The existence of several species in our earlier experimental material is confirmed. These include G. geniculata itself (2n= 32 or 48 small chromosomes), G. oaxacana D. R. Hunt (2n= 16 small chromosomes) and G. schiedeana (Kunth) D. R. Hunt, which has two chromosome forms, 2n= 10 and 2n= 16, both with large chromosomes. These forms are diploid and tetraploid based on x= 5 and x= 4 respectively and show a Robertsonian relationship with each other. The cytology of tetraploid (2n= 20) G. karwinskyana is confirmed and that of a diploid form (2n=10) described. The recently described G. consobrina D. R. Hunt (2n= 20) is shown to be cytologically comparable with G. karwinskyana, but to differ in significant details. Next, the cytology of G. pulchella (Kunth) Rafin., the type species of Gibasis, is described and also that of the allied G. matudae D. R. Hunt. Both G. pulchella (2n= 10, 15) and G. matudae (2n= 10) show interchange heterozygosity, with complete rings of ten chromosomes at meiosis in some plants of G. pulchella. Preliminary comments are made on the cytology of G. aguensis (Standl. & Steyerm.) Rohw. (2n= 10), another ally of G. pulchella, and on the G. linearis group, where the basic number appears to be x= 6 but two karyotype patterns have been found. A discussion of chromosome architecture in Gibasis in relation to the taxonomy of the genus concludes the paper.     

Systematic Status of the Glycine tomentella and G. tabacina Species Complexes (Fabaceae) Based on ITS Sequences of Nuclear Ribosomal DNA

Glycine was reconstructed based on nucleotide sequences of the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA to examine the systematic status of the G. tomentella and G. tabacina species complexes. Rooted at the subgenus Soja (2 species, 7 accessions), parsimony analysis was conducted for 17 species (31 accessions) of the subgenus Glycine, including 9 and 6 populations of G. tomentella s.l. and G. tabacina s.l., respectively. The nrlTS phytogeny indicated polyphyly of G. tomentella s.l. as well as G. tabacina s.l. In the G. tomentella species complex, larger legumes, narrower leaflets, and deflexed indumentum hairs differentiated G. dolichocarpa from G. tomentella s.s. The tetraploid G. dolichocarpa (2n=80) and aneuploid G. tomentella (2n= 38) represented independent lineages from another clade of the remaining diploid (2n=40) and tetraploid species with a DD genome type. Tetraploid G. tabacina (2n=80) was closely related to G. dolichocarpa instead of the diploid G. tabacina (2n=40) with a BB genome type. The nrlTS phytogeny suggested allopolyploidy of G. dolichocarpa and of the tetraploid G. tabacina, both of which possibly share a common parental species with an AA genome type. Their phylogenetic affinity also indicated biased inheritance of the nrDNA ITS and a possible dominant role of the AA genome. Phylogenetically, G. dolichocarpa and allotetraploid G. tabacina should be recognized as distinct species. Received 12 February 2001/ Accepted in revised form 17 August 2001     

Comparative morphometry as a diagnostic tool applied to species delimitation in the genus Gastridium P. Beauv. (Poaceae)

Based on critical examination of both fresh and dried specimens and a morphometric analysis of herbarium specimens we performed a comparative study of the three closely related Gastridium (Poaceae, Tribe: Poeae, Subtribe: Agrostidinae) species G. ventricosum, G. phleoides and G. scabrum collected in Italy. The study aimed to provide new diagnostic tools to improve the taxonomy of the genus and confirm species delimitation. Variation in floral morphology in the three species was explored using seven quantitative and ten qualitative characters measured on 318 spikelets, both awned and unawned. Statistical methods, including principal components analysis (PCA) and non‐metric multidimensional scaling (NMDS), were used. Numerical analyses showed a general distinctness of spikelets and florets, consistent with the three studied species, and revealed among and within species variation patterns in both spikelet types. Accordingly, the most informative quantitative characters appeared to be the width and length of glumes and the length of awns, useful to distinguish G. phleoides which has the narrowest glumes and longer awns than the other species, while G. scabrum has the widest glumes. The most informative qualitative characters appeared to be the presence/absence of thick hairs or minute tubercles on the upper side of the lemma. The autonomy of the three studied species was confirmed. The taxonomic significance of the results is briefly discussed, and notes on the species distribution are included. Some nomenclatural and taxonomic notes on G. scabrum, of which literature still showed evidence of a general unawareness, are provided.     

Genomic relationships in the genus Glycine (Fabaceae: Phaseoleae): use of a monoclonal antibody to the soybean Bowman-Blrk inhibitor as a genome marker

We investigated the use of a monoclonal antibody (MAb 238) to the soybean Bowman-Birk inhibitor (BBI) to verify and understand the intergenomic relationships among the wild perennial Glycine species. Competitive enzyme linked immunosorbent assay and western blot screening studies revealed that the accessions of B-genome (G. latifolia, G. microphylla, and G. tabacina, 2n = 40) and C-genome (G. curvata and G. cyrtoloba) species did not contain the MAb 238 crossreactive proteins (BBI-nulls). By contrast, all the A-genome (G. argyrea, G. canescens, G. clandestina, and G. latrobeana), E-genome (G. tomentella, 2n = 38), and F-genome (G. falcata) species, G. arenaria (genome unknown), and the polyploid (2n = 78,80) G. tomentella accessions were BBI-positive. The D-genome G. tomentella (2n = 40) and tetraploid G. tabacina (2n = 80) contained both BBI-null and BBI-positive type accessions. Among the recently described species, G. hirticaulis (2n = 40), G. lactovirens, and G. pindanica contained the MAb 238 crossreactive proteins while G. albicans did not. Glycine hirticaulis, G. pindanica, and G. tomentella (2n = 38) displayed highly similar MAb 238 crossreactive isoelectric focusing banding patterns, indicating that they are genomically close to each other. Glycine hirticaulis was found to have both diploid (2n = 40) and tetraploid (2n = 80) cytotypes. We demonstrated that the MAb 238 was specific to the trypsin inhibitor domain of the BBI. The MAb 238 clearly reflected all the previously established relationships in the genus Glycine, validating its use as a genome marker.     

Some generic concepts in Compositae – Gnaphaliinae

The species of Gnaphalium, Helichrysum and related genera in southern Africa have been surveyed and a number of groups of allied species detected in which the ratio of female to hermaphrodite flowers ranges from a majority of female to a majority of hermaphrodite and in some groups there are even homogamous species. Thus, the failure of the old sex-ratio character used to separate Gnaphalium and Helichrysum is confirmed. The validity of these groupings has been tested by studying detailed characters such as the structure of the stereome (whether undivided or fenestrated) and the type of achenial hair, as well as the more usual floral characters. Most of these plants have leaves with flat or revolute margins and the occasional occurrence of involute margins is taxonomically important. Gnaphalium L. (type G. uliginosum L.) has been redefined to include Amphidoxa DC. and Demidium DC. (female flowers epappose) and species with more hermaphrodite flowers then female. The stereome is undivided. Helichrysum remains a massive and highly diverse genus and includes Leontonyx Cass. The stereome is normally fenestrated and the receptacle epaleate. One or two species have paleae, but Rhynea with receptacle paleate and stereome undivided is maintained. A small group of species with undivided stereomes, centred on H. marifolium D.C., has had to be retained in Helichrysum because of the existence of linking species. Undivided stereome and subglobose achenial hairs enjoin the transfer of H. vestitum (L.) Willd. and two allies, as well as the group of species centred on H. paniculatum (Thunb.) Thunb., to Helipterum, despite the lack of plumose pappus. Edmondia Cass. (H. sesamoides (L.) Willd.) with distinctive habit, involute leaves and no close relatives in Helichrysum is reinstated. Pseudognaphalium Kirp., to which only P. oxyphyllum (DC.) Kirp. had previously been assigned, is extended to include, in its typical subgenus, Hypelichrysum Kirp. and a number of American species of Gnaphalium, as well as the African G. undulatum L. and G. oligandrum (DC.) Hilliard & Burtt, and the Asiatic G. hypoleucum DC. and G. chrysocephalum Franch. A new subgenus, Laphangium, is created to accommodate G. luteo-album L. and its allies. Achyrocline (Less.) DC. has been maintained, as is done in South America, and some African species placed under Helichrysum by Moesert 1910) are returned to it. Two new genera are recognized among gnaphalioid plants with undivided stereomes: Troglophyton (based on G. capillaceum Thunb.) with about seven species and Plecostachys (based on G. serpyllifolium Berg, and including G. polifolium Thunb.). Both genera straddle the old numerical boundary between Gnaphalium and Helichrysum. The monotypic Helichrysopsis Kirp. is maintained. Plants of gnaphalioid aspect with fenestrated stereomes necessitate a further new genus: Vellereophyton (type G. dealbatum Thunb., better known as G. candidissimum Lam.) with seven species. Eriosphaera Less, and Lasiopogon Cass, are redefined and enlarged to include species previously placed in Gnaphalium. Metalasia and Lachnospermum are redefined, with the transfer of the only large-headed species remaining in Metalasia to Lachnospermum. Two new genera are created for plants in this affinity previously misplaced in Helichrysum. Atrichantha is based on H. gemmiferum Bol. and one new species is described. Dolichothrix is established for the anomalous H. ericoides (Lam.) Pers. Ifloga is briefly investigated and it is shown that the distinction between sect. Ifloga and sect. Eutrichogyne Bentham must be based on characters of the capitulum, not just on annual versus subshrubby habit. Lasiopogon molluginoides DC. (=Comptonanthus Nordenstam) is transferred to Ifloga and the structure of the capitulum in the genus is discussed. Some earlier work on this group of plants is briefly summarized, the phytogeographical interest of the proposed arrangement is discussed and some general conclusions offered.     

A REAPPRAISAL OF THE SUBGENUS GLYCINE

The genus Glycine Willd. is divided into three subgenera, Glycine Willd., Soja (Moench) F. J. Herm., and Bracteata Verdc. Six species are currently recognized in the subgenus Glycine: G. canescens F. J. Herm., G. clandestina Willd., G. falcata Benth., G. latrobeana (Meissn.) Benth., G. tabacina (Labill.) Benth., and G. tomentella Hayata. Distribution of the subgenus extends from south China to Tasmania and includes several Pacific islands. A collection of these species was examined cytologically and morphologically in an attempt to evaluate existing variability between and within taxa. Chromosome counts confirmed G. canescens, G. clandestina and G. falcata to be diploid with 2n = 40. Both tetraploids (2n = 80) and diploids were found in G. tabacina, the latter restricted to Australia. Glycine tomentella accessions were primarily tetraploid, but several collections from New South Wales, Australia, were found to be aneuploid with 78 chromosomes. One collection was aneuploid at the diploid level with 38 chromosomes. Meiosis appeared normal in the aneuploids with regular bivalent formation. Several accessions previously identified as G. tomentella were diploid. Seed of G. latrobeana was not available for analysis. Numerical techniques in the form of cluster analysis and principal components analysis were applied to morphological data on vegetative and inflorescence characters obtained from each collection. Numerical analysis grouped the accessions essentially according to current species delimitations with some exceptions. Glycine tabacina specimens from Taiwan approached G. clandestina in several characteristics. The diploid G. tomentella specimens formed a separate cluster and appeared morphologically distinct from the remaining taxa.     

Phylogenetic systematics and historical biogeography of the Neotropical electric fish Gymnotus (Teleostei: Gymnotidae)

Phylogenetic interrelationships of the Neotropical electric fish genus Gymnotus are documented from comparative study of phenotypic data. A data matrix was compiled of 113 phenotypic characters for 40 taxa, including 31 recognized Gymnotus species, six allopatric populations of G. carapo, two allopatric populations of G. coropinae, and three gymno‐tiform outgroups. MP analysis yielded 15 trees of equal length, the strict consensus of which is presented as a working hypothesis of Gymnotus interrelationships. Diagnoses are presented for 26 clades, including three species groups; the G. cylindricus group with two species restricted to Middle America, the G. pantherinus group with 12 species in South America, and the G. carapo group with 16 species in South America. The basal division of Gymnotus is between clades endemic to Middle and South America. Both the G. pantherinus and G. carapo groups include trans‐Andean sister‐taxon pairs, suggesting a minimum date for the origins of these groups in the late Middle Miocene (c. 12 Ma.). The geographically widespread species G. carapo is paraphyletic. Analysis of character state evolution shows characters of external morphology are more phylogenetically plastic and provide more phylogenetic information in recent branches than do characters of internal morphology, which themselves provide the more information in deeper branches. Nine regional species assemblages of Gymnotus are recognized, none of which is monophyletic. There are at least two independent origins of Gymnotus species in sediment rich, high conductivity, perennially hypoxic whitewater floodplains (varzea´) derived from an ancestral condition of being restricted to low conductivity non‐floodplain (terra firme) black and clearwater rivers and streams. These phylogenetic, biogeographic and ecological patterns suggest a lengthy and complex history involving numerous instances of speciation, extinction, migration and coexistence in sympatry. Evolution in Gymnotus has been a continent‐wide phenomenon; i.e. Amazonian species richness is not a consequence of strictly Amazonian processes. These patterns are similar to those of other highly diverse groups of Neotropical fishes and do not resemble those of monophyletic, rapidly generated species flocks.     

An integrative approach for species delimitation in the spider genus Grammostola (Theraphosidae,Mygalomorphae)

The mygalomorph genus Grammostola (family Theraphosidae) is endemic to South America. The species Grammostola anthracina is one of the largest spiders in Uruguay and reputed to be the longest lived tarantula in the world. This nominal species has two distinct colour morphs comprising black and reddish‐brown forms with controversial taxonomic status. Here, we present a phylogenetic study based on molecular characters (cytochrome c oxidase subunit I) of haplotypes of G. anthracina and closely related species. Our analysis together with new morphological data and biogeographical information indicates that the two morphs of G. anthracina constitute different species that are not sister to each other. Consequently, a new species, Grammostola quirogai is described, diagnosed and illustrated to encompass the black morph. Phylogenetic relationships and new taxonomic characters for Grammostola species included in this study are discussed.     

Studies on the diversity of the distinct phylogenetic lineage encompassing Glomus claroideum and Glomus etunicatum

Morphological and molecular characters were analysed to investigate diversity within isolates of the Glomus claroideum/Glomus etunicatum species group in the genus Glomus. The inter- and intra-isolate sequence diversity of the large subunit (LSU) rRNA gene D2 region of eight isolates of G. claroideum and G. etunicatum was studied using PCR-single strand conformational polymorphism (SSCP)-sequencing. In addition, two isolates recently obtained from Southern China were included in the analysis to allow for a wider geographic screening. Single spore DNA isolation confirmed the magnitude of gene diversity found in multispore DNA extractions. An apparent overlap of spore morphological characters was found between G. claroideum and G. etunicatum in some isolates. Analysis of the sequence frequencies in all G. etunicatum and G. claroideum isolates (ten) showed that four LSU D2 sequences, representing 32.1% of the clones analysed for multispore extraction (564) were found to be common to both species, and those sequences were the most abundant in four of the ten isolates analysed. The frequency of these sequences ranged between 23.2% and 87.5% of the clones analysed in each isolate. The implications for the use of phenotypic characters to define species in arbuscular mycorrhizal fungi are discussed. The current position of G. claroideum/G.etunicatum in the taxonomy of the Glomeromycota is also discussed.     

A multiple sex-chromosome system in Antarctic ice-fishes

Summary We have studied the chromosomes of 11 of the 15 known species of the notothenioid family Channichthyidae, the specialized whiteblooded Teleosts endemic to the Southern Ocean (ice-fishes). In the female sex, all studied species have the same diploid number of forty eight mostly acrocentric (uniarmed) chromosomes; however there is an interspecific variability in the chromosome morphology, type and quantity of repetitious DNA (usually seen as heterochromatin) localization of silver-stained nucleolar organizers. At least five of the studied species show a multiple sex-chromosome system possibly originated by the translocation of an autosome on an early Y gonosome morphologically similar to the X: the digametic males (2n = 47) show a X₁Y X₂ and the homogametic females (2n = 48) a X₁X₁X₂X₂ gonosomic constitution. This peculiar sex determining mechanism, otherwise rare in Teleosts, can be considered apomorphic in the same way as other morphofunctional characters usually interpreted as adaptive in these fishes.Some of the data presented here were collected during the European Polarstern Study (EPOS) sponsored by the European Science Foundation     

Fascicled Androecia in Dilleniidae and Some Remarks on the Garcinia Androecium

In Dilleniidae, stamen fascicles are interpreted either as phylogenetically secondary structures (derived from a single stamen primordium by dédoublement), or as the most archaic type of androecial organs in angiosperms. In context with an assumed high plasticity in the flowers at the beginning of angiosperm evolution (Endress, 1987 a, b), fascicled androecia can also be regarded as having coexisted with “magnolioid” spiral androecia since early in the evolution. On the basis of this assumption, it is easy to link the Dilleniidae, via their basal group, the Paeoniaceae, to the Magnoliidae. In Paeonia, the stamen clusters continue the spiral arrangement of the perianth members, with “limiting divergence” (Hiepko, 1964). Our investigations of Paeonia officinalis show that the fascicle primordia follow the spiral not only in their position, but also in their temporal sequence. In some Theaceae (Stewartia) and Clusiaceae (species of Garcinia) the divergence angle changes to 2/5, resulting in an epipetalous position of the stamen fascicles. The transition from fascicled (complex) androecia to simple ones (one- or two-whorled) has occurred repeatedly within the Dilleniidae. A great diversity of androecial structures, based on a fascicled androecium, can be found in the palaeotropical genus Garcinia (Clusiaceae), which comprises about 200 species. Some of these forms, including exceptional ones, are presented in this article. The diversity in the androecium in Garcinia can be interpreted phylogenetically as a secondarily increased plasticity, resulting in morphological curiosities.     

Morphological variability of potato cyst nematodes (Globodera spp.) in the Canary Islands

Thirty populations of potato cyst nematode (Globodera spp.) from the Island of Tenerife and two populations from the UK were assessed for several morphometric and non-morphometric characters thought to discriminate between the species G. rostochiensis and G. pallida. Also 200 cysts from each population were analysed by isoelectric focusing of soluble proteins. Correlation analysis, analysis of variance and principal component analysis were used to investigate relationships between the morphometric characters, how the relationships varied between species and between populations, and which characters were most useful for discriminating between species. The two species differed significantly for each of the four morphometric characters: stylet length, fenestra length, anus-fenestra distance and the number of ridges. The stylet length and fenestra length also showed differences between populations of G. rostochiensis while stylet length and number of ridges showed differences between populations of C. pallida. In general, populations of G. pallida showed greater variation than populations of C. rostochiensis. Principal component analysis of the population means indicated that over 73% of the variation in the characters could be explained by the contrast of stylet and fenestra lengths against the anus-fenestra distance and number of ridges. A plot of the first two principal components separated the two species. Stepwise discriminant analysis provided a linear combination of these four variables which discriminated between the species. Stylet length was found to be the most useful characteristic for distinguishing the species whilst anus-fenestra distance was the least useful.     

Deoxyribonucleic acid base composition and taxonomy in the genus Geotrichum Link

The molar percentage of guanine + cytosine (% GC) in the DNA from 25 yeast-like fungi related to the genus Geotrichum has been measured. This criterion together with the biochemical characteristics allow the division of the species into 3 groups: one group resembling G. candidum (% GC: 31.5–42) and containing 8 imperfect and 6 perfect forms. In this group only G. capitatum has a lower % GC (31.5). A second group of 6 species resembling the genus Trichosporon (57–60%) and a third intermediate category of 4 species (42–53.5%) were observed. The GC-values correlate well with biochemical characters, e.g. the presence of urease, the number of organic substrates assimilated, and also with the presence of characters typical of the Ascomycetes or the Basidiomycetes.This work was supported by I.N.S.E.R.M. (Institut National de la Santé et de la Recherche Médicale) C.R.L. n^o 76 1 153 8.     

Encephalon gross morphology of the cichlid Geophagus sveni (Cichlidae: Geophagini): Comparative description and ecological perspectives

The encephalon gross morphology of Geophagus sveni is described, compared between male and female specimens and discussed in relation to evolutionary, ecological and behavioural aspects. The Student's t-test revealed that there are no sexual dimorphism regarding the volume or linear measurements obtained from the main encephalon subdivisions (telencephalon, tectum mesencephali, cerebellum, gustative lobes, hypothalamus and hypophysis) in proportion to encephalon length, which is congruent with the absence of external dimorphic characters and presence of biparental care behaviour. In all specimens examined, the tectum mesencephali is the largest structure of the encephalon, which may be explained by feeding habit and by the importance of the vision center in a social context (i.e., brood guarding and territory defence, which are common behaviours in cichlids). Also, the lobus vagi is more developed than usual for other teleosts, which may be explained by the presence, in G. sveni as well as in other Geophagini species, of a differentiated pharyngeal apparatus, probably an adaptation to winnowing, a specialized feeding habit. The little intraspecific variation in neuroanatomical characters observed herein indicates a possible source of morphological characters to be explored in cichlid phylogeny.     

A revision of the genus Monstrillopsis Sars (Crustacea: Copepoda: Monstrilloida) with description of a new species from Chile

In the course of a comparative analysis of the species included in the copepod genus Monstrillopsis Sars, a new species of the genus was discovered. The new species, Monstrillopsis chilensis n. sp., was found in a plankton sample collected in coastal waters off central Chile in the Southeastern Pacific. The adult female is similar to females of the type species M. dubia Scott and related forms but differs from its congeners by a combination of characters including: (1) a pair of small cephalic processes present between bases of antennules; (2) deep corrugations present on proximo-lateral surface of fifth pedigerous somite; (3) exopodal lobe of fifth leg not distally elongated; and (4) endopodal lobe of fifth legs short. This is the first record of the genus in the Southeastern Pacific and the second record of any monstrilloid in Chile. A comparative analysis of the species assigned to Monstrillopsis allowed a clarification of the generic characters and limits. The most important apomorphic characters in the genus are the presence of four caudal setae and modifications of the male antennule; other potentially useful characters are the pattern of antennule segmentation and the uniform presence of a reduced inner lobe of the fifth leg. The female of Monstrilla reticulata Davis should be retained in Monstrilla, but the purportedly conspecific male is probably a Monstrillopsis. Monstrillopsis ciqroi Suárez-Morales and M. angustipes Isaac should be transferred to Monstrilla. Haemocera filogranarum Malaquin is probably a species of Monstrillopsis. Monstrillopsis zernowi is tentatively retained in this genus as an aberrant form. A key for the identification of the species of this genus is included.     

Polycentropus cianficconiae sp. n. from Sicily and Descriptions of the Females of the Polycentropus Species Found in Sicily (Trichoptera,Polycentropodidae)

The male and female of Polycentropus cianficconiae sp. n. (Peloritani Mountains, Northeast Sicily) are described. This new species is related to P. divergens Mosely, 1930, of which the variability of some characters of male genitalia is examined. Female genitalia of the new species are compared to those of P. divergens, P. malickyi Moretti, 1981 and P. mortoni Mosely, 1930. The characters of female genitalia are useful for evaluating the relatedness and differences among the species. They agree with the findings of the study on male genitalia.     

Patterns of cryptic hybridization revealed using an integrative approach: a case study on genets (Carnivora, Viverridae, Genetta spp.) from the southern African subregion

Recent years have seen the development of molecular‐based methodologies to investigate hybridization and its impact on the evolutionary process. However, morphological characterization of hybrid zones has only scantily been considered, especially in zootaxa. Thus, the level of congruence between molecular and morphological characters when attempting to detect hybrids remains a poorly tackled area. The genets (genus Genetta) provide an ideal case study for further investigation of the respective contribution of morphology and DNA in hybrid zone characterization because (1) their morphology has recently been exhaustively explored and (2) the existence of hybrid zones in southern Africa was proposed in the literature. We assessed levels of hybridization among the southern African genets, and questioned the role of ecological factors on the hybridization patterns detected. We used an integrative approach involving nine discrete morphological characters and a diagnostic discriminant function, geometric morphometrics and sequences of cytochrome b including collection specimens. The combination of independent materials allowed us to accurately reassess the level of hybridization in southern African genets, and revealed cryptic, interspecific gene flows. Morphology unambiguously detected a low number of G. maculata × G. tigrina hybrids and rejected the hypothesis of a large intergradation zone in KwaZulu‐Natal, thus supporting the species status of the two genets. Cytochrome b analyses revealed: (1) cryptic, massive hybridization between G. tigrina and the sympatric G. felina, and (2) a trace of reticulation (one sequence) between G. tigrina and the allopatric G. genetta. The type specimen of G. mossambica Matschie, 1902 is considered to be a morphological hybrid between G. maculata and G. angolensis. Remarkably, the morphological approaches (discrete characters and morphometrics) proved complementary to conclusions derived from cytochrome b sequences. Whilst morphometrics was generally unable to accurately identify all putative hybrids, this approach revealed diagnostic cranial shape differences between recognized species as well as the cryptic G. ‘letabae’ (included in the super‐species G. maculata). Morphometrics also confirmed the diagnostic value and age dependency of discrete characters. Our integrative approach appeared necessary to the detection of cryptic hybridizations and to the comprehensive characterization of hybrid zones. The recurrent detection of hybrids exhibiting tigrina‐like coat patterns may suggest (1) asymmetric hybridization of G. tigrina males to females of other species and (2) positive selection for tigrina‐like phenotype in South African habitats, but these hypotheses will have to be further tested using other sources of evidence. Despite the precise mosaic of hybrid zones identified in southern African genets, the environmental factors that shape patterns of distribution of hybrids remain unclear. Nevertheless, in the light of our range reassessment, it appears that seasonality of precipitation and periods of annual frost may play stringent roles in the distribution of genets. The complementarity of our results based on morphology and molecules is regarded as encouraging for the further development of integrative approaches in order to better understand the complex phenomena that underlie hybridization processes. © 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 86 , 11–33.