Morphological and molecular variation in Mitchella undulata,with special reference to the systematic treatment of the dwarf form from Yakushima

Morphological and molecular variation in Mitchella undulata Siebold et Zucc. was examined to evaluate the genetic basis for recognizing the dwarf variety, M. undulata var. minor Masamune. Considerable variation in leaf size in M. undulata, but no obvious morphological discontinuities, were found between the normal and dwarf varieties. Instead, a weak cline running from the Pacific Ocean to the Sea of Japan was found. Anatomical observations of leaf blades revealed that the large variation in leaf size can be attributed to variation in the number of leaf cells and not to differences in cell size. A molecular analysis based on sequences of rDNA internal transcribed spacer regions indicated that there were two major genotypes in M. undulata with minor variation in haplotypes resulting from additional substitutions or putative recombination. The dwarf form from Yakushima was neither genetically uniform nor apparently differentiated from other populations. From these results, we conclude that the dwarf form of M. undulata should be treated at the rank of forma.     

Spiranthes sinensis var.amoena in Japan contains two seasonally differentiated groups

Specimens ofSpiranthes sinensis var.amoena in four Harbaria in Japan and living materials from four localities were examined for the flowering time. As a result, two distinct types, flowering in spring and autumn respectively, are recognized.     

Morphological variation in leaf shape in <Emphasis Type="Italic">Ainsliaea apiculata</Emphasis> with special reference to the endemic characters of populations on Yakushima Island,Japan

We analyzed leaf shape variations in Ainsliaea apiculata Sch. Bip. to evaluate the uniqueness of morphological characters in populations on Yakushima Island, Kagoshima Prefecture, Japan. Leaf size and shape from populations on Yakushima Island (n = 300) were compared with those from populations in other areas of Japan (n = 300). A considerable amount of variation occurred in leaf size in A. apiculata populations both on Yakushima Island and elsewhere, but clear discontinuities in leaf size were not detected. Some variants previously thought to be endemic to Yakushima Island, i.e., A. apiculata var. acerifolia and A. apiculata var. rotundifolia, were also found in other locations in Japan. Moreover, these leaf types were found to be continuous with the typical leaf shape of A. apiculata var. apiculata via various intermediate types, suggesting the need for future revision of these taxa. Based on these results, we reevaluated the uniqueness of the Yakushima populations of A. apiculata in terms of leaf variation. The uniqueness of the Yakushima populations was defined by a more diverse leaf shape than found in populations from other areas.     

Cheek-pouch dispersal of seeds by Japanese monkeys (Macaca fuscata yakui) on Yakushima Island, Japan

Seed dispersal by Japanese monkeys (Macaca fuscata yakui) via cheek-pouch was studied in a warm temperate evergreen forest on Yakushima Island. Plant list was compiled based on a study during 1986–1995, of which troops of monkeys have been habituated without artificial feeding. We followed the well-habituated monkeys in 1993 and 1994 to observe the feeding behavior and their treatments of fruits and seeds, and collected seeds dispersed by monkeys to record the distance carried from the mother trees. We checked the difference of germination ratio between seeds dispersed via cheek-pouch and seeds taken from mother trees by sowing experiments. Seeds and acorns of 22 species were observed to be dispersed via cheek-pouch of monkeys. Among them, three species with acorns were never dispersed via feces, and 15 species with drupes were seldom dispersed via feces. Plant species of which seeds are dispersed only via cheek-pouch had larger seeds than those of dispersed both via cheek-pouch and via feces, and typically had only one or two seeds in a fruit. As for one of cheek-pouch dispersal species,Persea thunbergii, the mean distance when seeds were carried from the mother trees via cheek-pouch was 19.7 m, and the maximum distance was as long as 105 m although more than 80% of seeds were dispersed within 30 m from mother trees. And 82% of seeds dispersed via cheek-pouch germinated. The easy separation of seeds from other parts of the fruit seems to facilitate cheek-pouch dispersal more than dispersal via feces. Cheek-pouch dispersal by monkeys has possibly enhanced the natural selection for larger seeds which bring forth larger seedlings with high shade-tolerance. In conclusion, cheek-pouch dispersal by monkeys is quite an important mode for trees in the mature stand in a warm temperate evergreen forest on Yakushima Island.     

Mycorrhizal differences between genuine roots and tuberous roots of adult plants ofSpiranthes sinensis var.amoena (Orchidaceae)

Genuine roots ofSpiranthes sinensis var.amoena were infected with the mycorrhizal fungusRhizoctonia repens immediately after root formation in autumn. Infection by the mycorrhizal fungus extended, reaching a maximum the following early summer. The amount of living mycorrhizal fungus in the genuine roots dramatically declined in the flowering season, and then the roots decomposed. Tuberous roots were formed in spring. Mycorrhizas were limited to local infections and did not spread along the roots. The infection level of living mycorrhizal fungus in the tuberous roots was less than in the genuine roots throughout the year. The amount of dead fungal coils in the tuberous roots increased as the tuberous roots aged. The mycorrhizal characteritics of tuberous roots ofS. sinensis var.amoena were totally different from those of genuine roots although the tuberous roots morphologically resembled the genuine roots. Contribution No. 96, Laboratories of Plant Pathology and Mycology, Institute of Agriculture and Forestry, University of Tsukuba.     

Leaf-shape variation of Paederia foetida in Japan: reexamination of the small, narrow leaf form from Miyajima Island

Variations in Paederia foetida L. leaf shape were examined to evaluate the taxonomic validity of the small, narrow leaf form of P. foetida f. microphylla Honda from Miyajima Island, Honshu, Japan. There is considerable variation in P. foetida individuals in terms of leaf size and leaf index (leaf length:leaf width ratio). On Miyajima Island, some individuals have narrow leaves with a high leaf index value, a phenotype represented by the type specimen of P. foetida f. microphylla, and some do not. Given that the leaf size of individuals from Miyajima Island is smaller than that of individuals from other localities in Japan, and that the small leaf phenotype is stable even under cultivation, P. foetida f. microphylla is classified as the form having the smallest leaf size. Anatomical examination of leaf blades revealed that the large variation in leaf size was attributable to variation in the number of leaf cells but not to differences in cell size or cell shape. Based on these results, we discuss the endemism of P. foetida f. microphylla.     

Molecular phylogeny of Cypripedium (Orchidaceae: Cypripedioideae) inferred from multiple nuclear and chloroplast regions

A molecular analysis was performed on 56 taxa in the orchid genus Cypripedium using nrDNA ITS and five chloroplast regions (trnH-psbA, atpI-atpH, trnS-trnfM, trnL-F spacer, and the trnL intron). The genus Cypripedium was confirmed as monophyletic. Our data provided strong support for monophyletic grouping of eight infrageneric sections (Subtropica, Obtusipetala, Trigonopedia, Sinopedilum, Bifolia, Flabelinervia, Arietinum, and Cypripedium) defined in earlier taxonomic treatments, and paraphyletic grouping of two sections (Irapeana and Retinervi). Within the genus Cypripedium, the first divergent lineage consisted of two Mesomaerican species, and subsequently the Cypripedium debile lineage from eastern Asia was split. Our study did not support the notion that two Asian species (Cypripedium subtropicum and Cypripedium singchii) were closely related to either Mesoamerican Cypripedium irapeanum or North American Cypripedium californicum, as indicated by previous interpretations based on morphological evidences. In addition, one pair of vicariant species, Cypripedium plectrochilum (eastern Asia) and Cypripedium arietinum (North America), unique to section Arietinum, was confirmed. Furthermore, within the monophyletic section Cypripedium two previously recognized subsections, Cypripedium and Macrantha, were shown to be paraphyletic. Our results suggested that this section split into two groups based on distribution (North America vs. Eurasia) instead of such previously used, morphological traits as flower color, and the shape of the lips (labellum) and lateral petals.     

Molecular phylogenetics of subtribe Aeridinae (Orchidaceae): insights from plastid matK and nuclear ribosomal ITS sequences

We conducted phylogenetic analyses using two DNA sequence data sets derived from matK, the maturase-coding gene located in an intron of the plastid gene trnK, and the internal transcribed spacer region of 18S–26S nuclear ribosomal DNA to examine relationships in subtribe Aeridinae (Orchidaceae). Specifically, we investigated (1) phylogenetic relationships among genera in the subtribe, (2) the congruence between previous classifications of the subtribe and the phylogenetic relationships inferred from the molecular data, and (3) evolutionary trends of taxonomically important characters of the subtribe, such as pollinia, a spurred lip, and a column foot. In all, 75 species representing 62 genera in subtribe Aeridinae were examined. Our analyses provided the following insights: (1) monophyly of subtribe Aeridinae was tentatively supported in which 14 subclades reflecting phylogenetic relationships can be recognized, (2) results are inconsistent with previous classifications of the subtribe, and (3) repeated evolution of previously emphasized characters such as pollinia number and apertures, length of spur, and column foot was confirmed. It was found that the inconsistencies are mainly caused by homoplasy of these characters. At the genus level, Phalaenopsis, Cleisostoma, and Sarcochilus are shown to be non-monophyletic.     

Evaluation of morphological and molecular variation in Plantago asiatica var. densiuscula, with special reference to the systematic treatment of Plantago asiatica var. yakusimensis

Morphological and molecular variations in Plantago asiatica L. var. densiuscula Pilg. were analyzed to evaluate the genetic basis for recognizing the dwarf variety P. asiatica var. yakusimensis (Masam.) Ohwi. Considerable variation in the leaf size of P. asiatica var. densiuscula was observed, and no morphological discontinuities were found between the dwarf types of P. asiatica var. densiuscula and P. asiatica var. yakusimensis. Morphological analysis of plants grown under standardized conditions revealed that both environmental plasticity and genetic differentiation contributed to the dwarfisms. Molecular phylogenetic analysis of rDNA internal transcribed spacer (ITS) regions and the SUC1 locus encoding a sucrose transporter revealed that P. asiatica var. yakusimensis was genetically unique although the differentiation level was low. From the above results, we concluded that P. asiatica var. yakusimensis should be reduced to a form of P. asiatica var. densiuscula. Furthermore, the geographic distribution of the SUC1 genotype suggested multiple origins of dwarves, and possible hypotheses for the origins of dwarves are discussed.     

Molecular phylogeny, diversification and character evolution in Lupinus (Fabaceae) with special attention to Mediterranean and African lupines

Phylogenetic relationships within the complex genus Lupinus are estimated from internal transcribed spacer (ITS) sequences of the nuclear ribosomal DNA repeat. The molecular data supports Lupinus as a distinct monophyletic group within the tribe Genisteae. Different geographical lineages are revealed within Lupinus, which are each restricted to either the Old or the New World. In the New World, the ITS data support an eastern-western geographic disjunction of the lupines and the recognition of some well-supported clades. In the Old World, almost all the previously recognized taxa are taxonomically well differentiated. The homogeneous African rough-seeded lupines, Scabrispermae, are strongly supported as a monophyletic group, which is distinct from the diverse and heterogeneous circum-Mediterranean smooth-seeded ones. The latter appear to have evolved as two lineages, in which are revealed some intersectional relationships. Also ITS data allow the assessment of the phylogenetic position of the newly discovered species, L. anatolicus (in the Old World) and L. jaimehintoniana (the Mexican tree lupin). The ITS phylogeny suggests a rapid initial radiation of the lupines subsequent to their divergence from a common ancestor. Moreover, the results indicate that the annual and perennial habits have evolved many times in Lupinus and suggest a role for generation time in affecting the evolutionary history of lupines. Data on adaptive processes and character evolution are re-examined and discussed in the light of the ITS phylogeny.These studies were supported by the research unit UMR-CNRS 6553 Ecobio – University of Rennes (France), and by the NSERC of Canada (grant to R.J. Bayer, at the University of Alberta). They are greatfully acknowledged. All the persons that have contributed in different ways to this work on Lupinus, summarized in this presentation, are greatly thanked. Particularly, we would like to mention Malika Aïnouche, Roland Greinwald, André Huon, W.K. Swiecicki, Billie L. Turner and Ludger Witte for their contributions.     

The antennae of neopseustid moths: Morphology and phylogenetic implications, with special reference to the sensilla (Insecta, Lepidoptera, Neopseustidae)

The antennae of Lepidoptera Neopseustidae were examined with the scanning electron microscope. The studied species, Nematocentropus cfr. omeiensis, Neopseustis meyricki, Synempora andesae, Apoplania valdiviana and Apoplania penai possess nine types of antennal flagellum sensilla: multiporous large sensilla basiconica, multiporous thin sensilla basiconica, multiporous small sensilla basiconica, multiporous sensilla trichodea, multiporous sensilla coeloconica; uniporous sensilla chaetica; aporous sensilla chaetica, aporous stylus-shaped sensilla chaetica, aporous sensilla styloconica.The multiporous sensillum type here termed “multiporous large sensillum basiconicum” is unknown from other Lepidoptera and probably constitutes an autapomorphy of the family Neopseustidae. This sensillum type is remarkable by having a single base in female Apoplania and Synempora while in male Apoplania it has a bifid or trifid base, and in male Synempora it is composed of two or three incompletely separated hairs. This may be the first recorded example of a sexually dimorphic lepidopteran sensillum type. The stylus-shaped sensillum chaeticum is a primitive type which occurs only in some lower Lepidoptera.     

Population density and group composition of Japanese sika deer (<Emphasis Type="Italic">Cervus nippon yakushimae</Emphasis>) in an evergreen broad-leaved forest in Yakushima,southern Japan

The population density of Japanese sika deer (Cervus nippon yakushimae Kuroda and Okada) in an evergreen broad-leaved forest in Yakushima, southern Japan, was surveyed over 4years from 1998 to 2001. Two approximately 50ha study sites, Hanyama and Kawahara, were established with a total of 4km of census trails at each site. The estimated densities of sika deer at the two sites were 43–70 deerkm^–2 at Hanyama and 63–78 deerkm^–2 at Kawahara, although these values might be underestimates. The adult sex ratio (number of adult males:number of adult females) ranged from 0.6 to 1.0 at Hanyama, and from 0.4 to 0.9 at Kawahara. Mean group size was 1.9 deer (male group, 1.5 deer; female group, 1.6 deer; mixed group, 3.6 deer). The population density of sika deer was relatively high compared to other sites in Japan, with the exception of very small (<10km²) islands. Possible explanations for this naturally high density of sika deer in an evergreen broad-leaved forest in Yakushima are discussed.     

Ypsilonidium bananisporum sp. nov. (Ceratobasidiales) from Iriomote Island, Japan

Ypsilonidium bananisporum sp. nov. belonging to Ceratobasidiales is described and illustrated. This fungus has all the characteristics of the genusYpsilonidium including reticulate-hypochnoid basidiomes, broad hyphae branching at right angles, broadly clavate basidia with two sterigmata, and basidiospores germinating by repetition. It differs from all hitherto known species in the genus by producing suballantoid to banana-shaped basidiospores, measuring 19.5–22×5.5–6 μm. Contribution No. 314 of the Tottori Mycological Institute.     

Taxonomical study ofAster leiophyllus complex (Compositae) in Kanto district,Japan, with special reference to ploidy level

TheAster leiophyllus complex is complicated taxonomically and includes many related taxa representing polyploid series. It has been reported that this group is diversified throughout the Kanto district, central Japan, and there are three taxa endemic to the Kanto district. However, as neither morphological variation range nor distribution pattern is sufficiently investigated, taxonomic confusion sometimes occurs. In this study, morphological variation was examined in relation to the ploidy level, and taxonomic consideration was given. As a result, four morphologically recognized taxa occur in the Kanto district;A. leiophyllus (2x, 6x),A. leiophyllus var.sawadanus (2x, 3x, 4x),A. semiamplexicaulis (2x), andA. sugimotoi (4x, 5x). The distribution ranges ofA. leiophyllus var.sawadanus, A. semiamplexicaulis andA. sugimotoi are small, whileA. leiophyllus is distributed over most of the Kanto district. Morphological and cytological observations indicate that one of the reasons of the taxonomic confusion is the presence of presumed hybridization betweenA. leiophyllus and other taxa.     

Taxonomic status of Monotropastrum humile, with special reference to M. humile var. glaberrimum (Ericaceae, Monotropoideae)

Taxonomic treatment of the achlorophyllous monotropoid plant Monotropastrum humile is still unclear and confusing because of the lack of detailed morphological analyses and molecular phylogeny. In particular, the taxonomic status of a glabrous variety, M. humile var. glaberrimum, is under debate. Our detailed examination of the morphological characteristics of living plants revealed that M. humile var. glaberrimum can be easily distinguished from the putative conspecific taxon M. humile var. humile by characteristics not previously recognized, namely the shape and color of the floral disc. Most morphological features characterizing Cheilotheca were also found in M. humile var. glaberrimum. Moreover, there was considerable nucleotide differentiation in the internal transcribed spacer (ITS)2 sequences of M. humile var. humile and var. glaberrimum. Molecular analysis of the phylogenetic relationship of M. humile var. humile, var. glaberrimum, and other monotropoids using ITS2 sequences showed that two varieties of M. humile formed a monophyletic clade with a member of a different genus, Monotropa L., but obvious phylogenetic relationships among these three taxa were not obtained. Thus we conclude that Monotropastrum humile var. glaberrimum should be treated as a distinct species. However, the generic affiliation of M. humile var. glaberrimum could not be determined because of its intermediate character state combination and the insufficient characterization of related species. We strongly suggest that Monotropastrum as a whole needs re-evaluation.     

Addition and reexamination of Japanese species belonging to the genus Cercospora and allied genera. IX. Newly recorded species from Japan (4)

Pseudocercospora cyatheae C. Nakash. & S. Inaba on Cyathea sp. as a new species is described. Three species belonging to the genus Cercospora and allied genera are newly added to the mycoflora of Japan. They are Cercospora armoraciae on Armoracia rusticana, Passalora passaloroides on Amorpha fruticosa, and Pseudocercospora nogalesii on Cytisus scoparius.     

浙江西闪岛三突多足摇蚊亚属一新种（双翅目：摇蚊科）

记述采自浙江省西闪岛的三突多足摇蚊亚属1新种：杯状三突多足摇蚊Polypedilum (Tripodura) cypellum sp. nov.。该新种与哈氏三突多足摇蚊P. (T.) harteni Andersen & Mendes, 2010相似,区别于本亚属其它已知种的主要特征是具高脚杯状的上附器。文中提供了雄成虫的翅、前足胫节端部、生殖节、上附器等特征图。     

Phalaenopsis bellina (Rchb.f.) Christenson,a segregate from P. violacea Witte (Orchidaceae: Aeridinae)

A reevaluation of floral and vegetative morphology together with data from analysis of floral fragrance and flavonoid co-pigment chemistry suggest that specific status is warranted for the two kinds ofPhalaenopsis violacea Witte known informally as the “Bornean type” and the “Malayan type.”Phalaenopsis violacea var.bellina Rchb.f. is elevated to specific rank, providing a name for the “Bornean type” plants.     

江西虾脊兰属一新变型——异钩距虾脊兰

描述了产自江西的兰科(Orchidaceae)虾脊兰属(Calanthe R. Br.)植物一新变型——异钩距虾脊兰 (Calanthe graciliflora f. jiangxiensis B. Li, L. J. Kong et B. Y. Yang),并绘制了线条图。该新变型与原变型钩距虾脊兰(C. graciliflora Hayata)的主要区别为距末端分裂为2个不等的短尖。     

Cardicola opisthorchis n. sp. (Trematoda: Aporocotylidae) from the Pacific bluefin tuna, Thunnus orientalis (Temminck & Schlegel, 1844), cultured in Japan

A new aporocotylid blood fluke is described, based on specimens from the ventricle of the Pacific bluefin tuna, Thunnus orientalis (Temminck et Schlegel), cultured in Wakayama and Nagasaki Prefectures, Japan. The new species is morphologically similar to the members of the genus Cardicola Short, 1953, but shows distinct differences in the body form, location of the testis and the orientation of the ootype. The body of the new species is long and slender, whereas other Cardicola species are small and generally lanceolate. The testis is mostly located posterior to the caeca and anterior to the ovary, occupying 31–45% of body length, in contrast to the known Cardicola species, whose testis is typically intercaecal. The ootype is oriented anteriorly, while in most congeners, it is directed posteriorly or horizontally. Phylogenetic analyses of this aporocotylid, together with Cardicola orientalis Ogawa, Tanaka, Sugihara et Takami, 2010 from the same host, were conducted based on DNA sequences of the ITS2 rDNA and the 28S region of ribosomal RNA. The analyses revealed that the new blood fluke belongs to the genus Cardicola despite the marked morphological differences. Thus, this aporocotylid is named Cardicola opisthorchis n. sp. and the generic diagnosis is emended in this paper. In addition, 100% identity among the ITS2 sequences from the present species, Cardicola sp. from T. orientalis in Mexico and Cardicola sp. from the northern bluefin tuna, Thunnus thynnus (Linnaeus) in Spain suggests that C. opisthorchis n. sp. has a broad geographical distribution and that it infects both the Pacific and northern bluefin tuna.