Molecular evidence for the taxonomic identity of Korean Adonis (Ranunculaceae)

 Although Korean Adonis has been traditionally recognized as Adonis amurensis Regel and Radde with various infraspecific taxa described, its taxonomic identity is still in dispute. We investigated the genetic variation in 60 individuals from 12 populations in Korea to elucidate the taxonomic identity of the Korean Adonis complex. Random amplified polymorphic DNA (RAPD) analysis revealed that Korean Adonis comprises three species: A. amurensis, A. pseudoamurensis, and A. multiflora. Adonis amurensis is distributed in northern central inland regions of Korea and A. pseudoamurensis is found in southern parts of the Korean peninsula. Adonis multiflora grows only on Cheju Island, which is the southernmost part of Korea. Phylogenetic analysis of nuclear ribosomal internal transcribed spacer (ITS) sequences partially supported the presence of three Adonis taxa in Korea as detected by RAPD analysis. The Adonis population on Jangbong Island located in the West Sea, which was referred to as A. pseudoamurensis on the basis of morphological examination, was separated from the other populations of A. pseudoamurensis. Otherwise, the molecular evidence is well congruent with the recent morphological study that proposes that Korean Adonis consists of these three species. Received: November 6, 2001 / Accepted: February 26, 2002     

Genetic relationships among Korean brown frog species (Anura,Ranidae), with special reference to evolutionary divergences between two allied species Rana dybowskii and R. huanrenensis

Allozyme analysis for 41 populations of brown frog species, Rana dybowskii, R. huanrenensis, and R. amurensis from Korea and three reference species (Chinese R. chensinensis and Japanese R. dybowskii and R. tsushimensis), were performed to clarify taxonomic status of Korean brown frogs. The level of average genetic differentiation (Nei's D) among local populations of each species in Korea was very low (D<0.01 2) and Korean and Japanese R. dybowskii also showed conspecific level of differentiation (D=0.070). Whereas, much larger, discrete genetic differences were detected in the interspecific comparisons (D>0.370). In the genetic relationships among five species examined, the 24 chromosome brown frogs (R. dybowskii, R. huanrenensis, and R. chensinensis) did not form a monophyletic group. Rana dybowskii with the chromosome number of 2n=24 was grouped together with R. amurensis with the chromosome number of 2n=26. The hypothesis of reversal change from 24 to 26 in Korean R. amurensis seems to better explain the phylogenetic relationships of east Asian brown frogs than the assumption of parallel reduction in chromosome number from 2n=26 to 24 in R. dybowskii and in the common ancestor of R. huanrenensis and R. chensinensis. The genetic, morphological, and reproductive divergences between Korean R. dybowskii and R. huanrenensis were compared.     

Phylogenetic and conservation status of the endangered terrestrial orchid Nervilia nipponica (Orchidaceae) in Korea

The putative Japanese endemic Nervilia nipponica was discovered recently on Jeju Island in Korea. However, Japanese populations of N. nipponica have since been shown to comprise two polyphyletic taxa. To confirm the taxonomic status of the Korean Nervilia populations, we compared ITS1, matK and trnL intron sequence data with that from the two cryptic Japanese taxa. Our results confirmed all Korean populations as belonging to the true N. nipponica s.s, which must shed its endemic status. Korean accessions showed no distinct genotypic patterning, with samples from all populations and representing two distinct colour forms forming a single monophyletic clade. The extremely low level of genetic diversity detected within N. nipponica on Jeju Island underscores the urgent need for measures to secure the species from local extirpation. We conclude that N. nipponica is a priority for conservation and recommend that systematic controls and sustained efforts are required to ensure its protection.     

毛榛复合体（桦木科）多变量形态学研究及分类学处理

Corylus mandshurica Maxim. & Rupr. 和 C. sieboldiana Blume 在中国和俄罗斯一直被处理为独立的种。本文对来自不同标本室的这两个种的腊叶标本及野外采集的大量叶片进行了形态学分析,以确定形态分化是否足够成为建立种的依据。对采自中国、韩国和日本覆盖两个种分布区的153份标本的18个形态学性状进行了统计,以构建数据矩阵用于主成分分析。结果表明,包含C. sieboldiana 复合体的数据单位（entities）在所有的形态学性状上均呈现出广泛的重叠区域。叶片和果实性状分析结果表明类群间缺乏分异性。基于形态学性状不足以把 C. mandshurica 从C. sieboldiana 中分离出去,并给予种的等级,也就是说这两个类群在形态上没有完全分化,而且它们的分布区也是相连的。 因此,应该把C. mandshurica作为C. sieboldiana 的种下分类群处理。C. sieboldiana在韩国南部和日本的具短总苞的植物常常被处理为独立的种或变种,但实际上应该作为C. sieboldiana 内的变型处理,因为总苞性状高度变异,即使在同一个体上也是如此。     

A multivariate morphometric study on Corylus sieboldiana

Previous taxonomic treatments in Russia and China have considered Corylus mandshurica and C. sieboldiana to be distinct as independent species. A morphometric analysis was conducted to determine if the morphological differentiation from these taxa warrants specific taxonomic recognition with a large sample of field-collected leaves as well as specimens from several herbaria of northeastern Asia. One hundred and fifty two individuals representing Chinese, Korean, and Japanese geographic ranges of the species were scored for 18 morphological characters and the data matrix was used for principal components analysis. The entities that comprise C. sieboldiana complex exhibit widely overlapping ranges in all morphological attributes. The leaf and fruit data may reflect a lack of divergence between taxa. Therefore, we do not regard these to be sufficient for taxonomic splitting of C. mandshurica from C. sieboldiana to warrant the designation of the rank of a species based on morphology. The two taxa were not morphologically well differentiated and their ranges of distribution come together. Therefore, C. mandshurica should be recognized as an infraspecific taxon of C. sieboldiana. C. sieboldiana with short involucres in southern Korea and Japan is often treated as an independent species （C. hallaisanensis） or a variety (C. sieboldiana var. brevirostris), but should be only recognized as a form of C. sieboldiana because this involucral character is highly variable even within the same individual.     

A palynotaxonomic study of Korean<Emphasis Type="Italic">Adonis</Emphasis> (Ranunculaceae)

The pollen morphology of KoreanAdonis was examined to clarify its specific recognition. Pollen grains were divided into two major types, according to the size and number of their echinae; those with many small echinae were further subdivided into two subtypes: 2 to 3 aligned foveolae vs. unaligned foveolae. The three types of pollen morphology determined here agree well with recent taxonomic treatments of KoreanAdonis. Those analyses, based on morphological and molecular properties, previously recognizedAdonis amurensis, Adonis multiflora, andAdonis pseudoa-murensis.     

Molecules and morphology in concert: tests of some hypotheses in arctic Potentilla (Rosaceae)

We developed a combined molecular and morphological approach to unravel complex variation at low taxonomic levels, exemplified by some arctic members of Potentilla. Twenty-one populations from Svalbard were analyzed for random amplified polymorphic DNAs (RAPDs) and 64 morphological characters to test the hypotheses that (1) the P. nivea complex (section Niveae) consists of three taxa (P. chamissonis, P. insularis, and P. nivea), (2) three "eco-morphotypes" in P. pulchella (section Multifidae) should be considered different taxa, and (3) P. insularis originated as an intersectional hybrid (Niveae × Multifidae). Twenty-two RAPD multilocus phenotypes were observed in the 136 plants analyzed based on 35 markers. Three fairly distinct groups of RAPD phenotypes were identified in the P. nivea complex based on multivariate analyses and an analysis of molecular variance (AMOVA; 77.6% among-group variation). The variation within the P. nivea complex was more or less continuous in multivariate analyses of the morphological data. We identified, however, several individual morphological characters that separated unambiguously among the three groups of RAPD phenotypes, revealing that these groups correspond to the previously hypothesized taxa. Many identical RAPD multilocus phenotypes were observed in the "eco-morphotypes" of P. pulchella, suggesting that its conspicuous morphological variation is caused by plasticity or by genetic variation at a small number of loci. The hypothesis of the hybrid origin of P. insularis was not supported by the RAPD data. Overall, very little RAPD variation was observed within populations of the four taxa (2.1-16.7% in AMOVA analyses; average genotypic diversity, D, was 0.10-0.30). We conclude that detailed, concerted analysis of molecules and morphology is a powerful tool in low-level taxonomy.     

Molecular Phylogeny among Local Populations of Formica japonica (Hym., Formicidae) in Korea

ABSTRACT Not only were used Random amplified polymorphismic DNA(RAPD) markers as input for an analysis of molecular variance (AMOVA) and cluster analysis to describe the population genetic structure, but also morphometric analysis of Formica japonica in Korea in order to investigate the relationships between local populations according to altitude. The ten populations of F. japonica in Korea did not showed any relationships according to altitude in morphological analysis, while they showed distinct relationship in RAPD analysis. It was noticed that they were clearly separated as four groups according to altitude. The first was Jirisan-Gachilbong- Gyebangsan population, the second Iksansi-Odongdo, the third Deukyusan-Hallasan, the fourth Siheungsi- Seonamsa-Jinjusi. The first group was high mountainous, the second flat and low altitude, the third high mountainous, and the fourth flat and low altitude. They occur from low altitude to comparative high altitude 1,400 m in Korea. Actually, it was found that the relationships of 10 local populations of Korean Formica japonica were not related to morphological evolution, but related to molecular evolutionary data evolved from RAPD analysis.     

Genetic differentiation of piscivorous chub (genus Opsariichthys) in Japan,Korea and Russia

A comparison of allozyme variation, restriction fragment length polymorphisms in the mitochondrial DNA and partial sequences of the ND II gene (496 bp) was made among two lacustrine populations of the piscivorous chub (Opsariichthys uncirostris uncirostris) in Japan, four fluvial populations in Korea, one lacustrine population in Russia and one specimen from the Amur River (O. u. amurensis). All analyses indicate that the Japanese populations of piscivorous chub are separable from the Asian mainland populations of Korea and Russia. The latter populations were further divided into the Korean and Russian fish. Although opinions are divided on the phylogenetic position of the population in Lake Mikata, Japan, which shows unique morphological traits intermediate between those of the population in Lake Biwa and the mainland populations, the current analysis indicates a closer relationship to the population in Lake Biwa.     

Morphometric and Isozyme Analysis of the Genus Hosta (Liliaceae) in Korea

Abstract The hostas native to Korea have never been carefully reviewed taxonomically and their patterns of variability are not well understood. For morphological and electrophoretic studies, samples of rootstocks were taken from 45 Korean Hosta populations and from two populations of H. tsushimensis N. Fujita on Tsushima Island, Japan. For morphometric analysis, ten plants from each of twenty populations were grown and observed under greenhouse conditions. Fifty-one floral and vegetative characters were examined on each plant and the data were analyzed using principal components and cluster analysis. Six Korean species can be recognized: H. yingeri S. B. Jones (Taehuksan, Sohuksan, and Hong islands); H. capitata (Koidz.) Nakai (southern Korea); H. clausa Nakai (central and northern Korea); H. minor (Baker) Nakai (southeastern Korea including Wan and Kojae islands); H. venusta F. Maekawa (Cheju Island); and H. jonesii M. Chung (southern islands). Morphological features contributing to recognition of the natural groups were: ridges on the scape; the width of inner and outer floral lobes; bracts pappillous at apex; length and color of the bracts; length of inflorescences; the withering condition of the bracts; scabrous nerves on the lower leaf surface; and size of the leaves. Isozyme analyses indicate a recent origin of H. venusta from H. minor. Although hostas are easily hybridized, natural hybridization seems to be rare in Korea. The morphological and isozyme studies identified no hybrids and the species are well characterized by their distribution patterns, phenology, and habitats.     

Genetic diversity in populations of Gentoo penguins (Pygoscelis papua)

RAPD analysis was used to examine the extent of genetic polymorphism in two populations of Gentoo penguin (Pygoscelis papua) from Antarctic Islands (Petermann and Livingston). The chosen two of three 10 mer oligonucleotide primers accordingly to preliminary results showed different levels of polymorphism in Gentoo penguins at Petermann Island (from 23.53 to 42.86%) and Livingston Island (from 52.94 to 57.14%). Nei's similarity coefficients were in range from 0.5606 (when Gentoo genome profiles were compared with RAPD profiles of two related penguin species: Pygoscelis adeliae (Adelie) and Pygoscelis antarctica (Chinstrep)) to 0.9281 among observed Gentoo penguin populations. Nei's distances values ranged from 0.0746 to 0.5787 among the populations and species. The obtained results will be used for further estimation of genetic diversity of Gentoo penguins and determination of their taxonomic status.     

Molecular and morphological variation of rare endemic oncocyclus irises (Iridaceae) of Lebanon

Oncocyclus irises endemic to Lebanon form a complex of three closely related taxa replacing each other over short geographical distances in a linear habitat. In order to characterize the appropriate taxonomic levels and to assess their conservation status, we investigated patterns of phenotypic variability and the partitioning of genetic variation within and among populations using random amplified polymorphic DNA (RAPD) markers. Multivariate analysis (principal components analysis and multiple correspondence analysis), based on 16 quantitative and six qualitative characters, revealed no separation between populations or taxa. Moreover, no morphological character could be used to define clear boundaries between populations/taxa. The genetic characterization revealed high levels of polymorphism and diversity (H_s). Principal components analysis showed population delimitations, but no groupings reflecting the currently defined taxa could be identified. Both morphological and genetic data showed that Lebanese oncocyclus irises could not be fitted into clear taxonomic boundaries. Consequences for conservation are discussed. © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 159 , 123–135.     

Taxonomic relationships in East AsianVicia species with unijugate leaves based on random amplified polymorphic DNA markers

Random amplified polymorphic DNA(RAPD) markers were investigated to clarify the taxonomic positions ofVicia linearifolia andV. bifolia, and to assess the genomic diversity among the 9 populations ofV. unijuga, each of which represents a geographical variation or infraspecific taxa in southern Korea. These species are characterized by unijugate leaves in East Asia and have been controversial as to infra-or interspecific classification. The polymorphic markers among the populations examined were observed for fifteen decamer primers. The degree of band sharing was used to calculate genetic similarity between populations, and a phenogram using UPGMA cluster analysis was generated based on the Dice similarity coefficient. The taxa studied were divided into two main groups and the populations ofV. unijuga were all grouped together in the phenogram. The genetic similarities ofV. unijuga were very high among the populations and did not show distinctions between the infraspecific taxa, although the populations of Mt. Odae and adjacent areas in eastern Korea were different from others of the species.V. linearifolia fell within the range of the genomic variation among the populations ofV. unijuga, whileV. bifolia was grouped withV. venosa var.cuspidata having multijugate leaves rather thanV. unijuga. The result from studying RAPD markers suggested thatV. linearifolia should be integrated intoV. unijuga and that species with unijugate leaves ofV. bifolia andV. unijuga are polyphyletic.     

Morphological and isozyme divergence in Korean Hepatica sensu stricto (Ranunculaceae)

 Morphological and electrophoretic data were studied to examine species delimitation, patterns of morphological and genetic variation in three Korean Hepatica including two endemics, H.␣maxima and H. insularis. Based on a phenogram using 15 morphological characters, taxa were distinct; it was consistent with the phenogram based on genetic distance. In the enzyme electrophoresis study, the genetic identities suggested that three taxa were genetically divergent enough to be recognized as different species, falling within the range expected␣for congeners. The genetic identity between H.␣asiatica and H. insularis was higher than the values between these two taxa and H.␣maxima, a restricted endemic of Ulleung Island. The least genetic variation was found in H. maxima and the greatest in widespread H. asiatica. These data are consistent with theoretical expectations that small populations are more likely to be genetically depauperate. Received November 13, 2001; accepted May 10, 2002 Published online: December 11, 2002     

Evaluating Genetic Diversity for the Conservation of the Threatened Galapagos Endemic Calandrinia galapagosa (Portulacaceae)

Calandrinia galapagosa (Portulacaceae) is a threatened plant endemic to San Cristobal Island, Galapagos. The species is threatened by goat herbivory: less than 1300 individuals remain in nine populations, three of which have been fenced for protection. It is not known whether these three populations are genetically representative of the species at large or whether the three distinct color morphs exhibited by the species represent different taxonomic units. We measured the frequency of 113 polymorphic amplified fragment length polymorphism markers in 189 specimens representing three different phenotypes from all nine populations to relate the genetic structure of the species to both its geological history and morphological variation. The analysis revealed high diversity within populations and low, but significant, differentiation between populations. In addition, no genetic diversification was observed across color morphs. We conclude that (1) regular gene flow has historically occurred between populations, and (2) individual color morphs are not separate taxa. We identify another population that should be fenced to ensure maximal protection of genetic variation. Our research serves as a case study of using population molecular genetics for optimizing protection strategies for rare plants in tropical archipelagoes affected by introduced herbivores.     

Essential oil composition and preliminary molecular study of four Hungarian Thymus species

Chemical and genetic differences of twenty taxa belonging to four Thymus species were studied in order to determine whether molecular characters and essential oil components could be used as taxonomic markers and to examine the correlation between them. Plant samples, representing different taxa and geographic regions, were collected from experimentally grown populations. Essential oil samples were analysed by GC/MS and cluster analysis of volatile composition resulted in segregation of thymol chemotypes from sesquiterpenic ones. Thymol was characteristic for all the populations of Thymus glabrescens and T. pannonicus as well as for certain taxa belonging to T. praecox and T. pulegioides. Sesquiterpenes occurred in only two taxa of T. glabrescens, in each sample of T. praecox and in three taxa of T. pulegioides. Plant samples were analysed by random amplified polymorphic DNA (RAPD). The obtained dendrogram revealed high gene diversity. The 13 primers resulted 114 polymorphic RAPD bands, and the average percentage of polymorphism was 80.8%. The RAPD dendogram showed separation neither at interspecific nor at interpopulational levels. Therefore, further specific molecular studies involving more taxa are suggested. Partial correlation have been found between molecular and chemical assessments.     

Distinct specific status of the Korean brown frog, Rana amurensis coreana (Amphibia: Ranidae)

We used morphological and genetic data to assess the taxonomic status of Rana amurensis coreana and R. a. amurensis. Morphological comparisons revealed these two subspecies to be different from each other in size of body, nature of tubercles on dorsal stripe, degree of development in toe webbing, and condition of lateral spots of trunk. They were also different in sequence of mitochondrial 16S rRNA gene, with genetic distance as large as those observed among different species of brown frogs. Therefore, Korean populations previously considered a subspecies of R. amurensis should be regarded as a distinct species (=R. coreana).     

The rDNA Internal Transcribed Spacer Region as a Taxonomic Marker for Nematodes

The ITS region from a wide taxonomic range of nematodes, including secernentean and adenophorean taxa, and free-living, entomopathogenic, and plant-parasitic species, was evaluated as a taxonomic marker. Size of the amplified product aided in the initial determination of group membership, and also suggested groups that may require taxonomic reevaluation. Congeneric species often displayed identically sized ITS regions, but genera such as Pratylenchus and Tylenchorhynchus had species with large differences in size. ITS heterogeneity in individuals and populations was identified in several nematode taxa. PCR-RFLP of ITS1 is advocated as a method of taxonomic analysis in genera such as Helicotylenchus that contain numerous species with few diagnostic morphological characteristics.     

侧金盏花属植物成分及药理研究进展

以侧金盏花为代表的侧金盏花属植物在中国分布广泛,用于治疗心衰和心律不齐时具有作用快、蓄积小和毒性小等优点。本文综合了该属植物的生药、活性成分、药理作用、制剂及临床应用方面的研究进展。