Comparison of genetic variation and differentiation using microsatellite markers among three rare threatened and one widespread toad lily species of Tricyrtis section Flavae (Convallariaceae) in Japan

Genetic diversities were examined using six microsatellite markers amplifiable in three rare and one widespread species of Tricyrtis section Flavae, which are endemic to Japan. Contrary to a general expectation, the three rare species, Tricyrtis flava, Tricyrtis ohsumiensis and Tricyrtis perfoliata, have comparable genetic variation at the species level to that of the widespread Tricyrtis nana. This is probably because T. nana has not sufficiently recovered genetic diversity from the bottleneck at speciation or because recent range contractions have occurred in the three rare species. Genetic diversity at the population level was smaller in the putative selfing species T. nana than in the other three outcrossing species. Compared with a preceding study using allozyme markers, the genetic diversity in microsatellite loci was considerably larger, probably resulting from higher mutation rates at the microsatellite loci. Owing to the high genetic diversity of the microsatellite markers, genetic differentiation among populations could be estimated even in T. nana with little allozyme polymorphism.     

Impatiens macrantha (Balsaminaceae) sp. nov., a new species from limestone areas in Guangxi,China

Impatiens macrantha S. X. Yu&Ying Qin (Balsaminaceae), a new species from Guangxi, China, is described and illustrated. Phylogenetic analysis based on molecular data shows the new species to belong to I. subg. Clavicarpa and to be closely related to I. tubulosa with which it shares succulent stems and racemose inflorescences, but it can be easily distinguished by having a 4 – or 5 – ridged stem, obovate or elliptic leaf blade, and saccate lower sepal. Furthermore, I. macrantha is distinguishable from other Impatiens in China by its obviously larger leaves and flowers. The evidence from morphology and molecular data both support I. macrantha as new to science.     

大花地不容块根生物碱成分的研究

为研究大花地不容块根的生物碱成分,该研究采用酸提碱沉法从大花地不容块根中提取出总生物碱,通过硅胶柱色谱法和制备液相色谱技术对大花地不容块根的总生物碱提取物进行分离纯化,并利用波谱学手段对化合物进行结构鉴定。结果表明:(1)从总生物碱提取物中分离得到11个化合物,分别鉴定为青藤碱(1)、青风藤碱(2)、斯帝酚灵碱(3)、瑞枯灵(4)、异紫堇定(5)、紫堇单酚碱(6)、巴婆碱(7)、sukhodianine(8)、荷包牡丹碱(9)、7-氧代克班宁(10)和巴马汀(11)。(2)体外细胞毒性测试显示,大花地不容总生物碱和其主成分青藤碱对人类肺癌细胞A549的IC₅₀值分别为7.5×10^-4 g·mL^-1和6.59×10^-9 g·mL^-1。化合物2、3、4、7、8、9和10系首次从大花地不容中分离得到。大花地不容块根中含有吗啡烷、原阿朴啡、阿朴啡、苄基四氢异喹啉和原小檗碱5种类型的生物碱。     

Occurrence of the endangered species Nitellopsis obtusa (Charales,Charophyceae) in western Japan and the genetic differences within and among Japanese populations

Nitellopsis obtusa (Charales, Charophyceae) are widely distributed from Europe to Asia; however, this species has been recorded in only the five lakes in central Honshu in Japan. This species was thought to be extinct in Japan, but was rediscovered in limited areas of Lake Kawaguchi in central Honshu. More recently, we discovered more Japanese populations of N. obtusa in Lake Biwa in western Honshu, and it became clear that the species had a broader distribution in Japan than originally believed. In addition, although only male or female thalli have been collected at each lake, both male and female thalli were found from Lake Biwa. This is the first report of a potentially sexual population of N. obtusa in Japan. The DNA sequences of three chloroplast DNA markers, including both coding and non‐coding regions, were identical in all specimens from Lake Kawaguchi and Lake Nojiri (Central Honshu), and differed from those of Lake Biwa and German specimens. Although Japanese and German specimens were genetically similar, Japanese specimens displayed considerable genetic diversity according to locality.     

Genetic structures of allopatric and sympatric populations in Pseudorasbora pumila pumila and Pseudorasbora parva

Pseudorasbora pumila pumila was collected to investigate its genetic relationships from 11 localities in Hokkaido and eastern Honshu of Japan with Pseudorasbora parva from 6 localities in Hokkaido, eastern Honshu, and Polovynka River (a tributary of the Amur River in eastern Russia) in 1996–1998. In Hokkaido and Akita prefectures (Honshu), three P. pumila pumila populations were distributed with P. parva sympatrically. Samples of P. pumila pumila and P. parva were analyzed based on 27 allozyme loci. Genetic differentiation between P. pumila pumila and P. parva (D = 0.522–0.622, average 0.601) was almost equal to or greater than that between other congeneric fish species previously reported. In the three sympatric populations of two species, which had arisen from artificial transplantation of the latter into localities of the former, the individual frequency composition of genetic types (P. pumila pumila, P. parva, F₁ hybrid, and later filial generation hybrid) were significantly different from each other. Differing from the previous studies, F₁ hybrids were fertile and coexisting populations of the two species included the F₁ hybrid and the later filial generation hybrid. These results suggest that genetic population dynamics of sympatric populations of the two species would be influenced not only by hybridization but also by ecological and environmental factors. Alternatively, the existence of populations that possessed both the alleles specific to P. pumila pumila and those specific to P. parva may indicate that introgressive hybridization, instead of species displacement, has occurred between the two species under a certain condition.     

广西罂粟科一新记录属——黄药属

报道了广西罂粟科一新记录属——黄药属(Ichtyoselmis LidénFukuhara)及一新记录种黄药[Ichtyoselmis macrantha(Oliver)Lidén]。     

百合科两个分类群学名的订正

笔者在编研《中国生物物种名录》第1卷第3分册(百合科——五桠果科)的过程中,发现百合科两个物种在《中国植物志》和Flora of China中均使用了错误的名称,需要做出新处理。其中,有斑百合的正确名称应为Lilium concolor var.pulchellum(Fisch.)Baker,而非《中国植物志》使用的晚出等名Lilium concolor var.pulchellum(Fisch.)Regel,黄花油点草的正确名称应为Tricyrtis maculata(D.Don)J.F.Macbr.,而非Flora of China使用的Tricyrtis pilosa Wall.。     

Successional Position of Dry Andean Dwarf Forest Species as a Basis for Restoration Trials

The successional affinity of nine woody species was inferred from the structure, diversity and disturbance history of the vegetation where these occurred. This was done in order to obtain a basis for a restoration experiment, currently in execution, in the dry Andean dwarf forest zone on the edge of the High Plain of Bogotá (Colombia), at 2600–2950 m.a.s.l. We laid out 101 relevees in grassland and shrubland types in different stages of recovery, and in relatively little disturbed endemic Condalia thomasiana dwarf forest. The disturbance history of sites over the last ~60 years was inferred from aerial photograph series (1941–1991). CCA and logistic regression were applied to relate species composition to diversity, environment and disturbance history. All species showed a preference for certain structural groups. Also, a clear relation between species occurrence and vegetation diversity was found. Baccharis macrantha, and Dalea coerulea appeared relatively tolerant to grazing, while the remaining seven species reacted negatively. Soil clay content, base availability and organic carbon content was also an important factor for occurrence of each species. Invasion of grasslands by woody species is pioneered by Baccharis macrantha and followed by Dodonaea viscosa. Dalea coerulea was predominantly found on truncated clayey soils, which will probably not support Condalia dwarf forest. The hypothesized classification of the nine planted species to either pioneers or late-successional was fine-tuned. This exploratory study will be of use in the set-up of future succession-based restoration experiments, and for converting exotics afforestations to natural vegetation.     

Characterization of six microsatellite markers in Trillium camschatcense using a dual‐suppression‐polymerase chain reaction technique

Trillium camschatcense is a herbaceous perennial plant distributed in Hokkaido and northern Honshu, Japan. Geographical variations in the breeding system (partial selfing or obligate outcrossing) are reported in the populations of Hokkaido. We isolated six polymorphic microsatellite loci from this species. The number of allele per locus ranged from four to 12, whereas the expected heterozygosities ranged from 0.69 to 0.83. These markers may allow further investigations to reveal the evolutionary and ecological processes of mating system in T. camschatcense.     

Isolation and characterization of microsatellite loci in the threatened wild toad lily Tricyrtis flava (Liliaceae)

Twelve polymorphic microsatellite loci were isolated and characterized from the threatened toad lily Tricyrtis flava. The number of alleles ranged from three to 17, and the observed and expected heterozygosities ranged from 0.35 to 0.96 and from 0.50 to 0.93, respectively, in a population of T. flava. We also tested cross‐species amplifications of the markers in three other species of Trycyrtis sect. Flavae. Most of 12 markers developed for T. flava were successfully amplified in the species of sect. Flavae and will be useful for population genetic studies of these species.     

Interpopulation variation of a saurian malaria, Plasmodium sasai telford & ball, 1969, in three host species distributed within a range of 24° north latitude

Telford S. R., jr. 1982. Interpopulation variation of a saurian malaria. Plasmodium sasai Telford & Ball, 1969, in three host species distributed within a range of 24° north latitude. International Journal for Parasitology12: 17–22. The saurian parasite P. sasai has been found in four populations of three host species which range from Niigata Prefecture, Honshu to Bangkok, Thailand, a distance exceeding 4600 km and 24° north latitude. Variation of P. sasai within samples from the type population exceeds that found among samples from all three hosts, Takydromus tachydromoides (Honshu), T. smaragdinus (Amami Island, Ryukyus) and T. sexlineatus (Thailand). Plasmodium sasai was associated with Trypanosoma sp., Schellackia sp. and Pirhemocyton in two or more host species in ricefield-margin habitat bordered by shrub-like or heavier vegetation. This association possibly represents persistence of a host-parasite complex which evolved with an ancestral lacertid stock that later diversified into the present Takydromus species.     

A comparative floral and pollination biology ofTricyrtis flava maxim.,T. nana yatabe andT. ohsumiensis

The floral and pollination biology of three closely related species,Tricyrtis flava, T. nana andT. ohsumiensis, were comparatively investigated. The primary pollinator wasBombus diversus in all the three species observed, andAmegilla sp. also acted forT. ohsumiensis. The flowers ofT. flava andT. ohsumiensis bloom for two days and are protandrous. Thus autogramy seems to be prevented in these species when the larger bees forage on them, though geitonogamy may also occur. On the other hand,T. nana appears to be a primarily self-pollinating species. The flowers of this species open only during one day and are homogamy. The stigmata seem to receive much pollen of their own flower by the visit of bumblebees. Moreover, many flowers ofT. nana fruited without any visit of pollinators.T. nana has many features characterizing the autogamous derivatives.     

Geographic distribution and origin of the chloroplast T/C‐type in Quercus mongolica var. crispula in northeastern Japan

In order to investigate the historical expansion of common oaks (Quercus sect. Prinus) in the northeastern part of Japan, the relation between the chloroplast haplotypes (I and II) in Quercus mongolica var. crispula and the chloroplast types (T‐ and C‐type) were examined. Complete linkage between haplotype II and chloroplast C‐type was found. The chloroplasts examined in the oak species collected from Sakhalin and Primorski Krai, Russia, and Harbin, China were all T‐type. This suggests that the T to C mutation had occurred in haplotype II in Japan. Neither of haplotype I nor haplotype II was found outside Japan, suggesting both occurred in Japan independently from an ancestral haplotype VI. Haplotype I, which has been known only in Mount Hayachine within Honshu, is distributed southward to the Kanto district, where refugia might have occurred during glaciations.     

兰科带唇兰属一新异名

经过对标本馆馆藏标本的研究,确认原四川特有的峨眉带唇兰[Tainia emeiensis (K. Y. Lang) Z. H. Tsi]与大花带唇兰(T.macrantha Hook. f.)为同种植物,因此予以归并。     

Development of microsatellite markers for an extremely limited distributed rare diving beetle species,Acilius kishii,and a widely distributed species,Acilius japonicus (Coleoptera: Dytiscidae)

Acilius kishii Nakane, 1963 (Coleoptera: Dytiscidae) is an endangered diving beetle species distributed in only one location, Lake Yashaga‐Ike, Honshu Island, Japan. Acilius japonicus, which is related to A. kishii, is distributed widely in northern Honshu Island and Hokkaido Island in Japan. In this study, we identified 14 microsatellite loci for A. kishii and A. japonicus, including both polymorphic and monomorphic loci, using the next‐generation sequencing method. We observed that 5 and 10 loci showed polymorphisms in 31 and 32 individuals of A. kishii and A. japonicus, respectively. The observed and expected heterozygosities were 0.00–1.00 and 0.00–0.74, respectively. These microsatellite loci could be useful for future conservation genetic studies, including monitoring of genetic diversity and extinction risk of A. kishii.     

Molecular phylogeny of the genus Apatrobus (Coleoptera: Carabidae: Patrobinae) in western Japan

We performed a molecular phylogenetic analysis of the ground beetles Apatrobus (Carabidae), endemic to Japan, using the mitochondrial cytochrome c oxidase subunit I (COI) and the nuclear 28S rRNA (28S) genes. We focused on the species divergence in Kyushu, Shikoku and western Honshu and used 15 of 19 species and three populations with undetermined species in the DNA analysis. The gene trees showed that, of the Apatrobus species studied, A. hayachinensis Nakane from northern Honshu was not included in the monophyletic group of the other Apatrobus species and likely to be of a different genus. Divergence time estimation suggested that Apatrobus species excluding A. hayachinensis diverged 5.2 million years ago and the subsequent divergence of species occurred during the Pliocene and Pleistocene. In each of the main islands, Kyushu, Shikoku and Honshu, two or more distinct lineages occurred and all species had restricted distribution areas, suggesting that ancient dispersal and vicariance among the three main islands resulted in the nested biogeographical pattern of species distribution.     

Genetic diversification of the Kanehira bitterling Acheilognathus rhombeus inferred from mitochondrial DNA,with comments on the phylogenetic relationship with its sister species Acheilognathus barbatulus

The Kanehira bitterling, Acheilognathus rhombeus, is a freshwater fish, discontinuously distributed in western Japan and the Korean Peninsula. Unusually among bitterling it is an autumn-spawning species and shows developmental diapause. Consequently, the characterization of its evolutionary history is significant not only in the context of the fish assemblage of East Asia, but also for understanding life-history evolution. This study aimed to investigate the phylogeography of A. rhombeus and its sister species Acheilognathus barbatulus, distributed in China, using a mitochondrial analysis of the ND1 gene from 311 samples collected from 50 localities in Japan and continental Asia. Phylogenetic analysis revealed that A. barbatulus is included in A. rhombeus and genetically closer to Japanese A. rhombeus than to Korean A. rhombeus. Divergence of Korean A. rhombeus and A. barbatulus from Japanese A. rhombeus was estimated to be from the late Pliocene (3.44 Mya) and the early Pleistocene (1.98 Mya), respectively. Each event closely coincided with the time of the Japan Sea opening. Japanese A. rhombeus comprised seven lineages: three in Honshu and four in Kyushu. One lineage in central Kyushu was genetically closer to the Honshu lineages than to other lineages in northern Kyushu. Divergence of Japanese lineages was estimated to be from the early to middle Pleistocene (0.55–0.93 Mya), during a period of geological and paleoclimatic change, including volcanic activity. Population expansion in the late Pleistocene (<0.10 Ma) was suggested in many of the lineages, which accords with other freshwater fishes. Biogeographically the ancestral A. rhombeus/A. barbatulus was likely to have repeatedly colonized Japan from the continent through land bridges in the late Pliocene and the early Pleistocene. However, the close genetic relationship between Japanese A. rhombeus and A. barbatulus suggests another possibility, with the second colonization occurring in reverse, from Japan to China. The small genetic distance between them indicates that the colonization occurred later than colonization events of other freshwater fishes, including other bitterling species.     

Clonal diversity in the agamospermous polyploids ofTaraxacum hondoense in northern Honshu,Japan

Chromosome numbers and allozyme variations were surveyed in 74 polyploid populations ofTaraxacum hondoense, in northern Honshu, Japan. Most of the populations (94.4%) consisted of triploid (2n=24), indicating the predominance of this ploidy level. Approximately 42.6% were found to contain tetraploid (2n=32), and a few plants were pentaploid (2n=40). Electrophoretic analysis at6 Pgdh-1 revealed twelve phenotypes with four alleles (including one putative null allele). The triploids showed excessive heterozygosity (82.4%) and all of the tetraploids and pentaploids were heterozygote. Phenotype IV was the most frequent and widely distributed in northern Honshu. Forty five percent of the populations were found to contain multiple phenotypes at 6Pgdh-1. A total of 21 clones were distinguished using three polymorphic loci (6Pgdh-1, Got andMdh), and a considerable amount of clonal diversity was detected both within and among polyploid populations ofT. hondoense. Factors causing multiclonality in agamospermous polyploids are discussed.     

Systematic studies on the Pompilidae occurring in Japan: Genus Platydialepis Haupt (Hymenoptera: Pompilidae: Pepsinae)

Generic characters of Platydialepis Haupt, 1941 are presented. A new species of the genus, P. amamiensis sp. nov. from the Amami Islands, the Ryukyus, Japan, is described, and another species of the genus, P. ryoheii Ishikawa, 1957, distributed in Honshu, Shikoku and Kyushu, Japan, is redescribed. Their biological notes are also given. A new combination is proposed: Platydialepis taiwaniana (Tsuneki, 1989) (=Malloscelis taiwaniana Tsuneki).     

Systematic review of the genus <Emphasis Type="Italic">Bothrocara</Emphasis> Bean 1890 (Teleostei: Zoarcidae)

The systematics of the eelpout genus Bothrocara Bean 1890 is reviewed on the basis of 941 specimens. Eight mostly eurybathic, demersal species are recognized, distributed mainly along the continental slopes of the North and South Pacific oceans, with one species entering the South Atlantic. Distributions are: B. brunneum ranges from the Sea of Okhotsk to the Gulf of Panama at depths of 199–1,829 m; B. elongatum ranges from the Gulf of Panama to Chile at depths of 720–1,866 m; B. hollandi ranges from the Sea of Japan to the southeastern Bering Sea at depths of 150–1,980 m; B. molle ranges from the western Bering Sea to the South Atlantic at depths of 106–2,688 m; B. nyx is known only from the eastern Bering Sea at depths of 790–1,508 m; B. pusillum ranges from the northern Bering Sea to British Columbia, Canada, at depths of 55–642 m; B. tanakae is found along the northern coasts of Honshu and Hokkaido islands, Japan, at depths of 274–892 m; B. zestum ranges from the Izu Islands, Japan, and central Honshu, Japan, to the Gulf of Alaska at depths of 199–1,620 m (an unidentifiable specimen from off Taiwan may be B. zestum). The species are distinguished from one another mainly on the basis of head pore patterns, gill raker morphology, coloration and various meristic and morphometric values. A determination key to the species is provided.