The role of triploids in the origin and evolution of polyploids of <Emphasis Type="Italic">Turnera sidoides</Emphasis> complex (Passifloraceae,Turneroideae)

Triploids can play an important role in polyploid evolution. However, their frequent sterility is an obstacle for the origin and establishment of neotetraploids. Here we analyzed the microsporogenesis of triploids (x?=?7) and the crossability among cytotypes of Turnera sidoides, aiming to test the impact of triploids on the origin and demographic establishment of tetraploids in natural populations. Triploids of T. sidoides exhibit irregular meiotic behavior. The high frequency of monovalents and of trivalents with non-convergent orientations results in unbalanced and/or non-viable male gametes. In spite of abnormalities in chromosome pairing and unbalanced chromosome segregation, triploids are not completely sterile and yielded up to 67% of viable pollen. Triploids that originated by the fusion of 2n?×?n gametes of the same taxon showed more regular meiotic behavior and higher fertility than triploids from the contact zone of diploids and tetraploids or triploids of hybrid origin. The reproductive isolation of T. sidoides cytotypes of different ploidy level is not strict and the ‘triploid block’ may be overcome occasionally. Triploids of T. sidoides produce diploid and triploid progeny suggesting that new generations of polyploids could originate from crosses between triploids or from backcrosses with diploids. The capability of T. sidoides to multiply asexually by rhizomes, would enhance the likelihood that a low frequency of neopolyploids can be originated and maintained in natural populations of T. sidoides.     

A multivariate morphometric delimitation of species boundaries in the South American genus <Emphasis Type="Italic">Nicoraepoa</Emphasis> (Poaceae: Pooideae: Poeae)

A multivariate morphometric study of the genus Nicoraepoa was carried out to examine patterns of morphological variation among the species and to identify additional characters to allow a clearer delimitation among species. Nicoraepoa is a taxonomically complex genus comprising seven species endemic to South America. Although most of the species are well delimited, two of them, N. andina and N. chonotica, are morphologically similar and very difficult to separate. The morphological variation among species, with overlapping morphological characters, pseudovivipary and possible hybridization make species boundaries unclear. In the present paper, sixty-seven characters were scored for a total of 216 specimens and analyzed using ordination, cluster and discriminant analyses. Based on multivariate results, we recognized species groups and morphological diagnostic characters that allow clearer species delimitation. As a result, N. chonotica has been transferred to N. andina as a subspecies that inhabit southern Patagonia. Other taxa could be recognized and delimited by diagnostic characters: N. erinacea, N. pugionifolia, N. stepparia, N. subenervis subsp. spegazziniana and N. subenervis subsp. subenervis. Moreover, we discuss the possibility that a new species from Bolivia, based on only one specimen, closely related to N. subenervis should be ascribed in Nicoraepoa.     

Morphological characteristics and intraspecific structure of <Emphasis Type="Italic">Triplophysa orientalis</Emphasis> (Balitoridae: Nemacheilinae)

The morphology and intraspecific structure of Triplophysa orientalis inhabiting the inland basins of Tsaidam (northeastern edge of the Tibetan Plateau) and the basin of the upper reach of the Huang He River in China were studied. Significant morphological differences between the populations from the inland basins and the Huang He River were revealed; the latter were distinguished as a separate subspecies T. o. hwanghoensis subsp. n. Intermediate character of some traits of the new subspecies between the nominative subspecies and T. hutjertjuensis was found; as a result, the status of the latter was reduced to the subspecies level. The synonymy of T. o. elongata with the nominative subspecies was confirmed. The differences between T. orientalis and T. obscura—morphologically poorly distinguishable species, sympatric in the basin of the upper reach of the Huang He River—were defined more exactly.     

Hybridization rate and genotypic diversity of apomictic hybrids between native (<Emphasis Type="Italic">Taraxacum japonicum</Emphasis>) and introduced (<Emphasis Type="Italic">T. officinale</Emphasis>) dandelions in western Japan

Hybridization between the introduced and native plants may enhance invasiveness, especially in asexually reproducing species. Hybrid apomictic dandelions between native (Taraxacum platycarpum and T. japonicum) and exotic (T. officinale) species are distributed widely throughout Japan. To estimate the origin(s) and dispersal of the hybrids, we investigated the hybridization rate and genotypic diversity in mixed populations of T. japonicum, T. officinale and their hybrids at two green parks in western Japan. Among the plants identified as exotics from flower morphology, 86–96% were hybrids by genetic analysis. Genetic data with simple sequence repeat markers revealed a high clonal diversity of the hybrid both within and between populations, indicating multiple origins. A hybrid seed was found from among the 1891 seeds collected from T. japonicum in the parks, indicating ongoing hybridization in the field. T. officinale and hybrids were genetically differentiated between the two parks independent of the ploidy level; the allele frequency of T. officinale and tri- and tetraploid hybrids were similar within each park but different between the two parks. This suggests that the origins of hybrids were similar within the park but different between the parks. Overall, our results suggest that hybridization, including backcross, is an ongoing process, and that genetically diverse hybrids with various origins have been spreading in western Japan, probably because hybridization enhanced invasiveness at native habitat.     

Morphological and molecular investigations of <Emphasis Type="Italic">Gagea</Emphasis> (Liliaceae) in southeastern Kazakhstan with special reference to putative altitudinal hybrid zones

The most important center of speciation in the genus Gagea is thought to be in Central Asia. Here, we focus on species diversity in southeastern Kazakhstan (around Almaty, Ili-Alatau range of the Western Tian-Shan mountains). We studied an elevational transect, reaching from lowland steppes to the alpine zone (500–2750 m a. s. l.), and carried out detailed morphological and molecular investigations for populations of Gagea spp. Nine species were identified in different altitudinal zones; one of these (Gagea almaatensis) is described as new to science. We could detect two altitudinal contact zones between closely related species: G. filiformis and G. granulosa (sect. Minimae), and G. almaatensis and G. kuraminica (sect. Gagea). Morphological and molecular investigations (ITS data and cpDNA networks) indicate ongoing hybridization of co-occurring G. filiformis into G. granulosa and putative bidirectional hybridization events between G. almaatensis and G. kuraminica.     

Anatomical and micromorphological studies on <Emphasis Type="Italic">Teucrium</Emphasis> sect. <Emphasis Type="Italic">Isotriodon</Emphasis> (Lamiaceae) in Turkey with a taxonomic note

In this study, the anatomical features of the leaf and stem, besides the nutlet characteristics of some Teucrium sect. Isotriodon (Lamiaceae) taxa in Turkey, T. montbretii Betham subsp. montbretii, T. montbretii subsp. pamphylicum P. H. Davis, T. odontites Boiss. &; Bal., T. cavernarum P. H. Davis, T. antitauricum T. Ekim, along with an isolated population of T. montbretii (T. montbretii subsp.) were investigated. The anatomical studies revealed that the taxa share generally similar anatomical characters, such as thicker upper leaf cuticles and larger upper leaf epidermal cells compared to lower ones and diacytic to anomocytic stomata on the leaves. However, the portion of the mesophyll occupied by palisade parenchyma and the occurrence of mucilage cells in leaf epidermis shows difference among the taxa. Furthermore, the studied taxa have general stem characteristics of the Lamiaceae family, except for having poorly developed collenchyma at the corners. With the amphistomatic leaves and developed sclerenchymatic tissue in the leaf median vein, T. cavernarum is seperated from the other taxa. Trichome types on the vegetative organs and nutlet shape and sculpturing are generally the same or similar in the studied taxa, but trichomes on the nutlets are different among them. Based on nutlet characteristics and some morphological ones, it was revealed that the isolated population of T. montbretii represent a new subspecies, T. monbretii subsp. yildirimlii M.Dinç &; S.Do?u subsp. nov.     

An overlooked hybrid between the two diploid <Emphasis Type="Italic">Chenopodium</Emphasis> species in Central Europe determined by microsatellite and morphological analysis

The presence and extent of hybridization within the Chenopodium album aggregate (Amaranthaceae) is still unclear. Although many hybrid combinations have been described, their existence in the field has never been systematically studied and verified. The main aim of this study was to ascertain the extent of interspecific hybridization between the diploid species C. ficifolium and C. suecicum using highly variable nuclear microsatellite markers. Due to the absence of such kind of molecular markers for the whole C. album group, we divided the analysis into two steps: (1) Eleven microsatellite loci designed for the closely related species C. quinoa were cross-amplified in five Eurasian species of the C. album diploid–polyploid complex, i.e. C. album s.s. (6x), C. striatiforme (4x), C. strictum (4x), C. ficifolium (2x) and C. suecicum (2x); (2) For the detection of interspecific hybridization between C. ficifolium and C. suecicum, we sampled 480 individuals from five localities in Central Europe. We also investigated morphological differences between the parental taxa and their hybrid and devised a key for their determination. Analysis of variation in microsatellite loci using Bayesian methods, PCoA and Neighbour-joining tree identified 32 F1 hybrids. These F1 hybrids, described here as C. paradoxum Mandák, formed a cluster between well-differentiated parental species, combining the morphological characters of both their parents. Moreover, genetic analyses also recognized several F2 or backcross hybrids, whose delimitation, mainly from C. suecicum and F1 hybrids, based on morphological characters, is problematic.     

The <Emphasis Type="Italic">Bactrocera dorsalis</Emphasis> species complex: comparative cytogenetic analysis in support of Sterile Insect Technique applications

Background

The Bactrocera dorsalis species complex currently harbors approximately 90 different members. The species complex has undergone many revisions in the past decades, and there is still an ongoing debate about the species limits. The availability of a variety of tools and approaches, such as molecular-genomic and cytogenetic analyses, are expected to shed light on the rather complicated issues of species complexes and incipient speciation. The clarification of genetic relationships among the different members of this complex is a prerequisite for the rational application of sterile insect technique (SIT) approaches for population control.

Results

Colonies established in the Insect Pest Control Laboratory (IPCL) (Seibersdorf, Vienna), representing five of the main economic important members of the Bactrocera dorsalis complex were cytologically characterized. The taxa under study were B. dorsalis s.s., B. philippinensis, B. papayae, B. invadens and B. carambolae. Mitotic and polytene chromosome analyses did not reveal any chromosomal characteristics that could be used to distinguish between the investigated members of the B. dorsalis complex. Therefore, their polytene chromosomes can be regarded as homosequential with the reference maps of B. dorsalis s.s.. In situ hybridization of six genes further supported the proposed homosequentiallity of the chromosomes of these specific members of the complex.

Conclusions

The present analysis supports that the polytene chromosomes of the five taxa under study are homosequential. Therefore, the use of the available polytene chromosome maps for B. dorsalis s.s. as reference maps for all these five biological entities is proposed. Present data provide important insight in the genetic relationships among the different members of the B. dorsalis complex, and, along with other studies in the field, can facilitate SIT applications targeting this complex. Moreover, the availability of 'universal' reference polytene chromosome maps for members of the complex, along with the documented application of in situ hybridization, can facilitate ongoing and future genome projects in this complex.

     

Experimental hybridization and an evaluation of the fertility of some forms of the house mouse supraspecies complex <Emphasis Type="Italic">Mus musculus</Emphasis> (Rodentia,Muridae)

The degree of development of the mechanisms of postcopulatory isolation was evaluated on the basis of experimental hybridization of representatives of three subspecies of M. musculus (M. m. musculus, M. m. wagneri, and M. m. gansuensis) and remote populations of the subspecies M. m. musculus. Experimental crosses between the different subspecies and populations indicated the presence of initial stages of postcopulatory reproductive isolation between some forms of house mice. In a number of crosses conducted between different populations and subspecies of M. musculus, asymmetry was observed. In one variant of mating, M. m. musculus (male) × M. m. wagneri (female), a reduced intensity of breeding and nonviability of pups were observed. A decrease in the intensity of reproduction was found in all variants of crosses that used male M. m. musculus from the city of Ishim. These data are assumed to confirm the previous assumption about the hybrid origin of mice inhabiting that city. The results confirm a significant level of divergence of the subspecies M. m. musculus and M. m. wagneri. Thus, initial stages both of post- and precopulatory isolation mechanisms between M. m. wagneri and M. m. musculus were shown.     

A new subspecies of <Emphasis Type="Italic">Centaurea cassia</Emphasis> (Asteraceae) from Turkey

A new subspecies in sect. Jacea (Mill.) DC., Centaurea cassia Boiss. subsp. dumanii M. Dinç, A. Duran &; B. Bilgili subsp. nov., collected by the authors from South Anatolia, is described and illustrated. The new subspecies is restricted to Abies cilicica (Ant. &; Kotschy) Carr. subsp. cilicica forest above Göller Yaylas? (C6 Adana-Kozan). Diagnostic morphological characters from C. cassia subsp. cassia are discussed. The ecology, biogeography and conservation status of the new taxon are also presented.     

Differentiation of the pollen size in five representatives of <Emphasis Type="Italic">Taraxacum</Emphasis> sect. <Emphasis Type="Italic">Palustria</Emphasis>

Measurements of the pollen size in 5 species of Taraxacum sect. Palustria at three levels of ploidy: 2n = 3x = 24 (T. paucilobum), 2n = 4x = 32 (T. vindobonense, T. trilobifolium), 2n = 5x = 40 (T. mendax) and one taxon of unknown number of chromosomes 2n = ? (T. portentosum) are presented in this paper. Obtained results indicate a lack of distinct positive correlation between the pollen size and ploidy in the studied group of plants. Distinct relationship was, however, found between ploidy and the range of pollen size and shape variability. Most variable were the pollen grains of triploid T. paucilobum and the least — those in pentaploid T. mendax. Ranges of pollen variability in tetraploid T. trilobifolium and T. vindobonense and in T. portentosum of unknown number of chromosomes showed intermediate values.     

Investigating species boundaries using DNA and morphology in the mite <Emphasis Type="Italic">Tyrophagus curvipenis</Emphasis> (Acari: Acaridae), an emerging invasive pest,with a molecular phylogeny of the genus <Emphasis Type="Italic">Tyrophagus</Emphasis>

Mites of the genus Tyrophagus (Acari: Acaridae) are among the most widespread and common mites, inhabiting diverse natural and anthropogenic habitats. Some species are pests of agricultural products and stored food and/or live in house dust, causing allergies to humans. We sequenced 1.2 kb of the mitochondrial COI gene for 38 individuals belonging to seven species of Tyrophagus, including T. curvipenis, T. putrescentiae, T. fanetzhangorum, T. longior, T. perniciosus, and T. cf. similis. Molecular phylogenetic analyses (1) recovered two major clades corresponding to the presence or absence of eyespots, and (2) separated all included morphological species. Tyrophagus curvipenis and T. putrescentiae had the lowest between-species genetic distances (range, mean?±?SD): 14.20–16.30, 15.17?±?0.40 (K2P). The highest within-species variation was found in T. putrescentiae 0.00–4.33, 1.78?±?1.44 (K2P). In this species, we recovered two distinct groups; however, no geographical or ecological dissimilarities were observed between them. Based on our analyses, we document important morphological differences between T. curvipenis and T. putrescentiae. For the first time, we record the occurrence of T. curvipenis in the New World and suggest that it may be an emerging pest as it is currently spreading in agricultural produce.     

Geographical Variability of Morphological and Acoustic Signs of the North Populations of the Long Tailed Tit <Emphasis Type="Italic">Aegithalos caudatus</Emphasis> (Passeriformes,Aegithalidae)

The geographic variability of the morphological and bioacoustic characteristics of the long-tailed bushtit, or long-tailed tit (Aegithalos caudatus), was studied, and the validity of the selection of the controversial subspecies Ae c. japonicus Prazak 1897 was evaluated., Ae. c. brachyurus Portenko 1954 and Ae. c. sibiricus (Seebohm 1890), distributed from Scandinavia and Eastern Europe to the Pacific coast. On the basis of a study of nine metric characters and three signs of coloration, we showed that in the space of the range of white-head long-tailed tits from Eastern Europe and Scandinavia to Primorye and Sakhalin, the majority of morphological characters slightly fluctuate or change in a clinal mode; i.e., they change gradually. The vocalization of the nominative subspecies of long-tailed tits is analyzed for the first time. When comparing samples from neighboring regions, no significant differences were revealed, either morphologically and acoustically, so the names Ae. c. sibiricus and Ae. c. brachyurus should be synonymous with the nominative subspecies Ae. c. caudatus. Only the populations of long-tailed tits of the southern Kuril Islands and Hokkaido Islands can be isolated as an independent subspecies Ae. c. japonicus.     

Development of SCoT-Based SCAR Marker for Rapid Authentication of <Emphasis Type="Italic">Taxus Media</Emphasis>

Taxus media is an important species in the family Taxaceae with high medicinal and commercial value. Overexploitation and illegal trade have led T. media to a severe threat of extinction. In addition, T. media and other Taxus species have similar morphological traits and are easily misidentified, particularly during the seedling stage. The purpose of this study is to develop a species-specific marker for T. media. Through a screening of 36 start codon targeted (SCoT) polymorphism primers, among 15 individuals of 4 Taxus species (T. media, T. chinensis, T. cuspidate and T. fuana), a clear species-specific DNA fragment (amplified by primer SCoT3) for T. media was identified. After isolation and sequencing, a DNA sequence with 530 bp was obtained. Based on this DNA fragment, a primer pair for the sequence-characterized amplified region marker was designed and named MHSF/MHSR. PCR analysis with primer pair MHSF/MHSR revealed a clear amplified band for all individuals of T. media but not for T. chinensis, T. cuspidate and T. fuana. Therefore, this marker can be used as a quick, efficient and reliable tool to identify T. media among other related Taxus species. The results of this study will lay an important foundation for the protection and management of T. media as a natural resource.     

Patterns of subspecies genetic diversity among oilseed <Emphasis Type="Italic">Brassica rapa</Emphasis> as revealed by agro-morphological traits and SSR markers

Brassica rapa (2n = 20, AA genome) is an important oil yielding species of the family Brassicaceae and characterized by wide range of genetic and morphological subtypes suitable for cultivation under diverse agro-climatic regions of India. In this study, genetic diversity among three subspecies of B. rapa including yellow sarson, toria and outlier brown sarson was estimated using various agro-morphological traits and simple sequence repeat (SSR) markers. Maximum variability was recorded for siliqua angle (Coefficient of variation = 30.9%), followed by seeds/siliqua (CV = 18.8%), leaf length (CV = 10%) and plant height (CV = 16.8%). Principal component analysis explained more than 50% of the total observed morphological variability for first two components. Of the 107 SSR markers tested, 80 generated reproducible, clear and distinct amplicons of which, 65 (81.25%) were found polymorphic. The number of alleles at each locus ranged from 2 to 7, with an average of 3.03 alleles per marker. A total of 197 alleles were detected at 65 SSR loci with average PIC value of 0.457 and a mean resolving power of 3.04. Neighbor-Joining cluster analysis based on morphological traits and SSR markers separately classified all the 28 genotypes into five major groups. The population structure analysis resulted into three sub-populations with certain extent of admixture among the earlier established taxonomic sub-groups. Twenty-three unique alleles were detected in thirteen B. rapa varieties. The clustering analysis and principal coordinate analysis outlined the genetic relationships among different varieties belonging to the three subspecies of B. rapa. Genetically diverse genotypes as illustrated by score plots and from the clustering patterns brought out the wide range of diversity present among B. rapa genotypes and the underlying options available for selecting parental genotypes for hybridization and developing high yielding cultivars suitable for Indian conditions.     

New species of the tenebrionid-beetle genera <Emphasis Type="Italic">Tagonoides</Emphasis> Fairm., <Emphasis Type="Italic">Gnaptorina</Emphasis> Rtt. and <Emphasis Type="Italic">Agnaptoria</Emphasis> Rtt. (Coleoptera,Tenebrionidae) from China

Six new species Tagonoides belousovi sp. n., T. yunnana sp. n., Gnaptorina australis sp. n., Agnaptoria elongata sp. n. (Yunnan), A. lecta sp. n., and A. ruida sp. n. (Sichuan), and a new subspecies Gnaptorina potanini minxiana subsp. n. are described from China. Morphological adaptations of tenebrionids of the tribe Blaptini associated with phytophagous habit are discussed.     

Unresolved native range taxonomy complicates inferences in invasion ecology: <Emphasis Type="Italic">Acacia dealbata</Emphasis> Link as an example

Elaborate and expensive endeavours are underway worldwide to understand and manage biological invasions. However, the success of such efforts can be jeopardised due to taxonomic uncertainty. We highlight how unresolved native range taxonomy can complicate inferences in invasion ecology using the invasive tree Acacia dealbata in South Africa as an example. Acacia dealbata is thought to comprise two subspecies based on morphological characteristics and environmental requirements within its native range in Australia: ssp. dealbata and spp. subalpina. Biological control is the most promising option for managing invasive A. dealbata populations in South Africa, but it remains unknown which genetic/taxonomic entities are present in the country. Resolving this question is crucial for selecting appropriate biological control agents and for identifying areas with the highest invasion risk. We used species distribution models (SDMs) and phylogeographic approaches to address this issue. The ability of subspecies-specific and overall species SDMs to predict occurrences in South Africa was also explored. Furthermore, as non-overlapping bioclimatic niches between the two taxonomic entities may translate into evolutionary distinctiveness, we also tested genetic distances between the entities using DNA sequencing data and network analysis. Both approaches were unable to differentiate the two putative subspecies of A. dealbata. However, the SDM approach revealed a potential niche shift in the non-native range, and DNA sequencing results suggested repeated introductions of different native provenances into South Africa. Our findings provide important information for ongoing biological control attempts and highlight the importance of resolving taxonomic uncertainties in invasion ecology.     

Chemical and molecular identification of the invasive termite <Emphasis Type="Italic">Zootermopsis nevadensis</Emphasis> (Isoptera: Archotermopsidae) in Japan

Numerous termite species have been introduced outside their native ranges by human transport, and some have become invasive. The dampwood termite Zootermopsis nevadensis (Hagen), which is native to western North America, has been introduced to and become established in Kawanishi City, Hyogo Prefecture, Japan. Zootermopsis nevadensis is subdivided into two subspecies based on cuticular hydrocarbon (CHC) phenotypes: Z. nevadensis nevadensis and Z. nevadensis nuttingi (Haverty and Thorne). Here, we identified Z. nevadensis in Japan as hybrids between the two subspecies. Chemical analysis showed the presence of 7,15-dimethylhenicosane and 5,17-dimethylhenicosane in the CHCs of Z. nevadensis in Japan, corresponding to the CHC phenotype of Z. n. nevadensis. Conversely, all mitochondrial cytochrome c oxidase subunit I sequences of Z. nevadensis in Japan were identical to sequences from Z. n. nuttingi and hybrids between the two subspecies from a native hybrid zone in California, USA. In addition, phylogenetic analysis showed that Z. nevadensis in Japan formed a clade with Z. n. nuttingi and hybrids between the two subspecies. Our results show discordance between the chemical and genetic features of Z. nevadensis in Japan, indicating that individuals of Z. nevadensis in Japan are hybrids between the two subspecies.     

Redescription of <Emphasis Type="Italic">Cottus sibiricus</Emphasis> and an estimation of differences between this species and <Emphasis Type="Italic">Cottus dzungaricus</Emphasis> (Cottidae)

The type specimens of Cottus sibiricus and C. dzungaricus as well as individuals from the Olkha River (Yenisei River basin) are studied. The diagnostic features of C. sibiricus are established. Comparative morphological and molecular analysis of divergence between C. sibiricus and C. dzungaricus demonstrates that the differences lie within the limits of one polymorphous species. The rank of Dzungarian sculpin is lowered to the subspecies, C. sibiricus dzungaricus.     

Speciation of Eversmann and Mongolian hamsters (<Emphasis Type="Italic">Allocricetulus</Emphasis>, Cricetinae): Experimental hybridization

Contrary to the pre-existing concepts based on morphological and karyological data on the impossibility of hybridization between hamster species of the genus Allocricetulus (A. curtatus and A. eversmanni), F1 hybrids in both combinations, and backcrosses, were obtained for the first time. Some restrictions of hybridization between females of A. eversmanni and males of A. curtatus were detected. Analysis of the synaptonemal complex in the hybrids produced by crossing a female of A. curtatus with a male of A. eversmanni demonstarted that a significant portion of the cells was subject to arrest and selection at the stage of meiosis prophase I. Nevertheless, some spermatocytes still generate viable spermatozoa. Thus, the possibility of hybridization in the laboratory between two Allocricetulus species with different numbers of chromosomes may testify to a relatively low divergence between them.