Reinstatement of Myelophycus caespitosus (Scytosiphonaceae,Phaeophyceae) from Japan

Reinstatement of Myelophycus caespitosus Kjellman (Ectocarpales s.l., Phaeophyceae) described from western Japan is proposed based on the comparisons of DNA sequences of Japanese Myelophycus specimens, the type specimen of Chordaria simplex and lectotypified specimen of Myelophycus caespitosus. In the genetic analyses using mitochondrial cox1 and cox3, chloroplast atpB, psbA and rbcL DNA sequences, the specimens morphologically referable to M. simplex formed two distinct clades (clade‐1 and clade‐2) supported by high statistical values. Clade‐1 was distributed on the western coast of Japan (Honshu, Shikoku and Kyushu) from the Kii Peninsula to Tsushima and the Pacific coast of Aomori, northeastern Honshu, and clade‐2 on the Pacific coast of central Honshu from Sanriku to the Kii Peninsula. Based on the cox3 DNA sequences and the location of the type locality at Shimoda, Izu Peninsula, clade‐2 was concluded to correspond to true M. simplex. There were no recognizable differences in the representative morphological features (height and diameter of sporophytes and gametophytes, number of cells comprising cortical, subcortical and medullary layers, thickness of plurilocular gametangia, or length and diameter of unilocular zoidangia) between the specimens included in the two taxa. Analyses of the specimens of the two taxa, including sympatric populations on the Kii Peninsula, using a nuclear genetic marker ocm3 did not suggest any genetic exchanges between the two taxa. On the other hand, cox3 gene sequence of the voucher specimen of Myelophycus caespitosus collected from Goto, Kyushu, western Japan housed in the UPS herbarium was included in clade‐1. In conclusion, we propose the reinstatement of M. caespitosus and to lectotypify the specimen in UPS. Korean Myelophycus specimens reported from Cheju Island and Wando were considered to belong to M. caespitosus based on the reported rbcL sequences.     

Mitochondrial phylogeography of the Japanese pond turtle,Mauremys japonica (Testudines,Geoemydidae)

The phylogeography of the Japanese pond turtle, Mauremys japonica (Temminck and Schlegel, 1835), which is an endemic species in Japan, was studied by analyzing the variation in two mitochondrial DNA sequences, the cytochrome b gene and the control region. These analyses suggest that M. japonica comprises two major groups. The first one was found in the more eastern region, eastern Honshu Island and Shikoku Island, while the second was found in a western region, Kyushu Island and the Chugoku District (the westernmost part of Honshu Island). The boundary between the two groups is located in the Chugoku District. The nucleotide and haplotype diversities were very low, and these low diversities seem to have been caused by a bottleneck in the last glacial age. These results suggest that this species survived the last glacial period in two refugia, one in the central part of Honshu Island and the other one in Kyushu Island. Subsequently, population expansion took place in the postglacial period, and the groups from the two refugia extended their distribution ranges to the present boundary in the Chugoku District which represents a secondary contact zone.     

Geographic allozyme variation in the Japanese clouded salamander, Hynobius nebulosus (Amphibia: Urodela)

Using starch gel electrophoresis, geographical enzyme variation was surveyed in 46 populations of the Japanese salamander Hynobius nebulosus , which occurs widely in western Japan. This species exhibits substantial local genetic differentiation and is more diverse in inland regions than on small islands. In several different analyses, two groups of populations, one from Kyushu Island to the westernmost part of Chugoku district, Honshu (western group), and another from Shikoku through Kinki to Chubu district, Honshu (eastern group), were consistently recognized. Most of the remaining populations were placed in two less clearly-defined groups: the montane group from the Chugoku Mountains and the Chugoku group from the coastal regions of Chugoku district. The divergence patterns of H. nebulosus are thought to be related to the geological history of its range. The populations near the boundary of the western group were found to share several alleles that were predominant on both sides of the boundary, indicating past secondary contact. Differentiation in the montane group did not follow the isolation by distance model, probably because several populations included were genetically mixed with lowland populations of the Chugoku group.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 89 , 311–330.     

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.     

Variation in flash speed of Japanese firefly,Luciola cruciata (Coleoptera: Lampyridae), identifies distinct southern “quick‐flash” population on Goto Islands,Japan

Luciola cruciata, a bioluminescent firefly endemic to Japan, communicates with species‐specific flashing patterns. Previous studies reported that flying males possess three distinct flashing frequencies: slow (4 s), intermediate (3 s) and fast (2 s). Our study focused on geographical variations in flashing patterns in the northwestern part of the Kyushu region of southern Japan (including its isolated islands) and their relationship to the mitochondrial COII region of firefly DNA. We found that “quick‐flash (1 s)” fireflies occur in the Goto Islands, at the western tip of Kyushu, and that, although they possess a unique haplotype, they do not differ much genetically from “fast‐flash” fireflies inhabiting mainland Kyushu. In contrast, although fireflies inhabiting the Tsushima and Iki Islands also possess a unique haplotype, their flashing patterns were the same (fast‐flash) as fireflies inhabiting mainland Kyushu. We determined that flash patterns of L. cruciata in Japan should be classified according to four flashing patterns: slow, intermediate, fast and quick.     

Occurrence of two types of Hynobius naevius in Northern Kyushu, Japan (Amphibia: Urodela)

A survey to examine genetic variation among Hynobius naevius from four localities of Fukuoka Pref., northern Kyushu, Japan, resulted in the detection of two, sympatric, genetic types (A and B) that are clearly different in the allelic frequencies of four loci (ACOH-A, ACOH-B, ADH-A, and SOD-A) in each locality. Morphological investigations between the two genetic types also proved that they are clearly discriminated; the type A is about 75 mm in SVL, lacks mottling pattern on bluish purple dorsum, and possesses relatively short vomerine teeth series, while the type B is about 60 mm in SVL, and has light mottling on reddish purple ground color. These results strongly suggest that reproductive isolation occurs between these two types, and that they could be regarded as separate species. Populations from Toyota-cho, western Honshu, and Yabe-machi, central Kyushu, both close to Fukuoka Pref., were very similar to the types A and B, respectively. From these results, we consider that two evolutionary lineages that first evolved allopatrically in western Honshu and southern Kyushu secondarily contacted and became sympatric in the region of northern Kyushu.     

Phylogenetic relationships of Hynobius naevius (Amphibia: Caudata) as revealed by mitochondrial 12S and 16S rRNA genes

Using mitochondrial 12S and 16S rRNA sequences, we investigated phylogenetic relationships among populations of the endemic Japanese salamander Hynobius naevius. Monophyly of this species was recovered only in the maximum parsimony tree and was unresolved in maximum likelihood and Bayesian trees. Instead the following four haplotype clades consistently emerged clearly: Clade 1 from northwestern Kyushu, Clade 2 from Chugoku and northeastern Kyushu, Clade 3 from western Shikoku and Kyushu, and Clade 4 from Chubu-Kinki and central-eastern Shikoku. Of these, Clades 1 and 2, and Clades 3 and 4, respectively, correspond to Groups A and B previously recognized from the analyses of allozyme data in this species, but monophyly of these groups was not strongly supported. Unlike the previous results, the western and eastern samples from Shikoku did not form a clade, and were grouped with Kyushu-B in Clade 3 and Chubu-Kinki in Clade 4, respectively. The reason for this conflict between mtDNA and allozyme results is unknown, but might be related to retention of ancestral mtDNA polymorphism in Shikoku populations. Nearly simultaneous divergence of as many as four lineages in wide-ranging H. naevius is inferred for the late Miocene-Pliocene history of this taxon.     

Development of microsatellite markers for Pasania edulis (Makino) Makino, one of the dominant species of lucidphyllous forests in southern Kyushu, Japan

Pasania edulis (Makino) Makino is a Fagaceous tree species endemic to Japan and one of the dominant species in lucidphyllous forests in southern Kyushu, Japan. Recently, P. edulis was attacked by ambrosia beetles, Platypus quereivorus (Murayama) and mass mortality occurred in several areas in southern Kyushu. We isolated and characterized ten microsatellite loci in this species to reveal the genetic structure and gene flow. The number of alleles ranged from 3 to 11 and expected heterozygosities from 0.3761 to 0.8346. The markers described here will be useful for investigating the genetic diversity, genetic structure and gene flow, and planning for conservation of viable population of P. edulis.     

Life history and host specificity of the Japanese flea beetles Trachyaphthona sordida and T. nigrita (Coleoptera: Chrysomelidae), potential biological control agents against skunk vine, Paederia foetida (Rubiaceae), in the southeastern parts of the United States and Hawaii

Skunk vine, Paederia foetida (Rubiaceae), is native to Asia and has been recognized as an invasive weedy vine of natural areas in Florida and Hawaii. Two insects, Trachyaphthona sordida and Trachyaphthona nigrita (Coleoptera: Chrysomelidae) from Japan are being considered as potential biological control agents against skunk vine. To gather fundamental information on their biology, we carried out field surveys and laboratory experiments in Kyushu, southern Japan, between 2003 and 2006. We found that T. sordida is commonly distributed in Kyushu and T. nigrita is restricted to the southern parts of Kagoshima Prefecture on the southern part of Kyushu. These species are fundamentally univoltine and adults appear in late April to early July. Trachyaphthona sordida overwinters as mature larvae and T. nigrita as mature larvae or rarely as adults. Larvae of both species feed on fine roots of P. foetida in the field and Serissa foetida (Rubiaceae) under rearing conditions, and they appear to have tribe‐level host specificity in their host range. On the basis of these results, we suggest that both species are suitable as biological control agents.     

Seed populations of a harmful unarmored dinoflagellate Cochlodinium polykrikoides Margalef in the East China Sea

An unarmored dinoflagellate Cochlodinium polykrikoides has formed red tides responsible for fish mass mortalities especially in coastal areas of western Japan and southern Korea almost every summer to autumn. In laboratory culture, the optimum temperature for growth of the species is ca. 27 °C. Since the species cannot survive in water of temperatures of less than 10 °C, it was considered to over-winter in some certain regions as a motile form or resting cyst, and expand its distribution after the temperature increases to a level tolerable for growth. To determine the over-wintering regions and migration pattern of C. polykrikoides, occurrences of the motile cells were surveyed in the coastal and offshore areas of western Kyushu, Japan and south coast of the Korean Peninsula from April 2006 to August 2008. Cells of C. polykrikoides were found at 14 sites during the investigated period. Motile cells occurred throughout the year in Usuka Bay, Hirado of West Japan. From offshore regions of the Goto Islands and off Shin-Nagasaki Fishing Port, motile cells of C. polykrikoides were first detected from late May, and continuously occurred until February in Nama Bay of the Kami-Goto Islands. This first appearance was before red tides of C. polykrikoides reported at coastal areas in western Kyushu. In Korea, this species was first observed in May and disappeared after October in 2007. These occurrence patterns imply that Usuka Bay in Hirado is one of the over-wintering regions in western Kyushu, and also this species is possibly transported into the northern part of the East China Sea by the Tsushima Warm Current every year.     

Genetic relationships among the Japanese and Korean striated spined loach complex (Cobitidae: Cobitis) and their phylogenetic positions

The striata complex, a group of spined loaches included in the genus Cobitis and characterized by a striped coloration pattern on the lateral midline, is distributed in rivers in northeastern Asia to western Japan. The complex comprises 2 continental species (Cobitis tetralineata and Cobitis lutheri) and 3 Japanese races of species rank (large race, middle race, and small race), the small race further comprising 6 local forms of subspecific rank (Tokai form, Biwa form, Yodo form, Sanyo form, San-in form, and Kyushu form). Previous karyological studies have revealed that the large race is an allotetraploid, the others being diploid. In this study, mitochondrial (mt) DNA analyses were conducted for 30 diploid populations of the Cobitis striata complex from Japan and Korea to examine: (1) their phylogenetic relationships and the position of the complex among the major lineages of Cobitis; and (2) the genetic relationships among the Japanese and Korean populations. The results, based on cytochrome b sequences (724 base pairs) analyzed with those of the main lineage of European and Japanese Cobitis, indicated that the striata complex should be considered as a monophyletic group, which evolved in northeastern Asia. Initially considered as a subspecies of Cobitis taenia, widely distributed from Europe to Asia, the striata complex does not have a sister-relationship with the former. Although the Korean species C. tetralineata was formerly believed to be closely related to the middle race in Japan, and a second continental species, C. lutheri, closely related to the Kyushu or San-in forms of the small race in Japan, the trees resulting from the present study revealed that the two Korean species were clustered with each other and separated from all Japanese races.     

Cladosiphon umezakii sp. nov. (Ectocarpales, Phaeophyceae) from Japan

The new species Cladosiphon umezakii Ajisaka (Ectocarpales, Phaeophyceae) is described from Japan based on morphology and DNA sequences. The species resembles Cladosiphon okamuranus Tokida in its gross morphology; somewhat slimy, cylindrical, multiaxial and sympodial erect thallus, arising from a small disc‐shaped holdfast, and branching once to twice. However, C. umezakii has considerably longer assimilatory filaments (up to 840 μm long, composed of up to 90 cells) than any known taxa of the genus. The species is a winter to spring annual, growing on lower intertidal to subtidal rocks of more or less exposed sites on the north‐eastern coast of Kyushu and on both the Pacific and the Sea of Japan coasts of Honshu. Specimens from the Sea of Japan coast had both unilocular and plurilocular zoidangia, whereas those from Kyushu and from the Pacific had only unilocular zoidangia. Unilocular zoidangia were formed on the basal part of assimilatory filaments, and plurilocular ones were transformed from the distal part of assimilatory filaments. DNA sequences of the Rubisco‐spacer (rbc‐spacer) region and the nuclear rDNA ITS region (ITS1, 5.8S and ITS2) supported the distinctness of the species.     

Morphological discrimination of two genetic groups of a Japanese salamander, Hynobius naevius (Amphibia, Caudata)

Hynobius naevius, distributed on western Honshu, Shikoku, and Kyushu Islands of Japan, includes two genetically distinct groups (Groups A and B) that have never been delimited morphologically. Using specimens from the entire species range, we investigated the possibility of distinguishing these groups morphologically. Multivariate analyses of morphometric characters resulted in recognition of two groups that corresponded well to the two genetic groups. One (Group A) was characterized by larger body, compressed tail, shallower vomerine tooth series, bluish- or reddish-purple ground color, and pale-white lateral markings. In contrast, another (Group B) was characterized by smaller body, cylindrical tail, longer vomerine tooth series, reddish-brown ground color, and white lateral markings. Group A was composed of populations from the Chugoku District of Honshu and northern Kyushu, and could not be divided into subgroups, while Group B encompassed populations from the Chubu and Kinki Districts of Honshu, Shikoku and Kyushu, and was subdivided into three local subgroups that are geographically separated by marine straits. Morphometric differentiation in Group A is presumed to have been less affected by genetic factors than by other factors, such as ecological relationships with other, coexisting species. Differentiation in Group B is assumed to have been enhanced not only by genetic but also by climatological factors.     

Karyotypic analysis of <Emphasis Type="Italic">Skimmia japonica</Emphasis> (Rutaceae) and related species

A karyotypic analysis of three species of Skimmia (Rutaceae) in East Asia was performed that examined 88 individuals from 53 localities. Chromosome numbers of S. japonica, S. reevesiana and S. arisanensis were 2n=30, 31, 32 (=30+0−2B), 2n=60 and 2n=60, respectively. The chromosome number of S. arisanensis was reported for the first time. All species had a large chromosome pair or quartet (the first pair or quartet) with a median–submedian centromere in the karyotype. In S. japonica the arm ratio of this first pair was considerably variable and showed a geographical pattern. In the northern half of the distribution range, Sakhalin, Hokkaido, Honshu, Shikoku and part of Kyushu, the arm ratio was 1–1.2, while in the southern half, part of Kyushu, Ryukyu and Taiwan, the arm ratio was very variable and ranged from 1.2 to 2.4. In S. japonica the first pair was sometimes rather heteromorphic; however, the heteromorphism was not related to sex of the plant.     

Combined effects of tidal and diurnal cycles on spawning of the puffer,Takifugu niphobles (Tetraodontidae)

Synopsis Spawning in the pufferTakifugu niphobles was observed at Tomioka in western Kyushu, Japan, and the factors affecting the spawning time were estimated using a multiple regression analysis. Spawning in this puffer occurred in the intertidal beach during evening rising tides around the full and new moons. Both the time of commencement and ending of a day's spawning were related mainly to the tidal cycle, and occurred later when the high tides occurred later. However, the termination of a day's spawning was also affected by the diurnal cycle; spawning ended earlier when sunset was earlier. Besides, a day's spawning tended to be concentrated within a shorter time span when the interval between the commencement of spawning and the sunset was shorter. Thus the spawning time of the puffer is regulated not only by the tidal but also by the diel cycle, and it is suggested that the adaptive significance of the spawning reactions to both cycles should be examined separately.     

Origins of recently re‐established and newly discovered populations of the endangered butterfly Shijimiaeoides divinus (Lepidoptera: Lycaenidae) in Oita Prefecture,Japan

The endangered butterfly Shijimiaeoides divinus was believed to have been extirpated from Oita Prefecture, Kyushu, Japan, but was rediscovered in Taketa in recent years. This population is considered to have re‐established as a result of natural dispersal from Kumamoto, a neighboring prefecture located to the west of Oita. Furthermore, another population was recently found in Yufu, Oita Prefecture, which is an area where the species had never been recorded. To elucidate the origins of these two populations newly found from Oita Prefecture, their DNA sequences of the mitochondrial cytochrome c oxidase subunit I (COI) gene were compared with those of other S. divinus populations from Kumamoto Prefecture, Honshu and Korea. The results supported the hypothesis that the Taketa population originated from Kumamoto Prefecture. However, it was not clear whether this population originated from the natural dispersal or deliberate release of individuals. It was also found that the Yufu population was not established by the deliberate release of individuals from Honshu or Korea; however, it remained unclear whether the population of S. divinus was native to Yufu, or originated from other localities in Kyushu.     

Living in a tilted world: climate change and geography limit speciation in Old World anchovies (Engraulis; Engraulidae)

Historical changes in the distributions of temperate species in response to Milankovitch climate cycles have been well documented in palaeontological studies and recently evaluated with phylogeographical methods. How these cycles influence biological diversity remains a matter of debate. Molecular surveys of terrestrial and freshwater fauna demonstrate glacial refugia in low latitudes and range expansions into high latitudes, but few genetic studies have assessed the corresponding impact on marine fauna. In the present study, mtDNA sequences (N = 84) are surveyed to understand the impact of long‐term climate oscillations on ‘Old World’ anchovies (genus Engraulis), a monophyletic group occurring in north and south temperate zones of the eastern Atlantic and the western Pacific. The analysis of a 521‐bp sequence of mtDNA cytochrome b indicates a late Miocene or Pliocene dispersal from the north‐eastern Pacific (California–Mexico) to the north‐western Pacific (Japan), followed by Pleistocene dispersal from the north‐western Pacific to Europe. Geography mandates that populations in southern Africa and Australia were stepping‐stones for this dispersal. However, neither population occupies an intermediate position in the mtDNA genealogy; both populations are more recently derived from their northern neighbours. Haplotype diversity is high (h = 0.93–0.97) in European, Australian, and Japanese anchovies, but low (h = 0.22) in the southern African population, where all haplotypes are more closely related to European specimens than to each other. These southern populations occupy a precarious position, lacking north–south coastlines that allow range shifts during climatic extremes. Recurring extinctions and episodic recolonizations from northern hemisphere populations are the likely results. In this case, ocean‐climatic changes retard rather than enhance opportunities for evolutionary radiations. © 2006 The Linnean Society of London, Biological Journal of the Linnean Society, 2006, 88 , 673–689.     

AGE, GROWTH, AND REPRODUCTION OF THE FINLESS PORPOISE, NEOPHOCAENA PHOCAENOIDES, IN THE COASTAL WATERS OF WESTERN KYUSHU, JAPAN

Abstract: In the coastal waters of western Kyushu, Japan, a total of 97 incidentally taken or stranded finless porpoises, Neophocaena phocaenoides , was collected for studying age, growth and reproduction. An additional 17 specimens from the Inland Sea were used for a comparison of life history. Mean neonatal body length was 78.2 cm. Both males and females grew to around 140 cm by 5 yr of age. The maximum body lengths of males and females in western Kyushu were 174.5 cm and 165.0 cm, respectively, which were smaller than those recorded in other Japanese waters. Females probably attain sexual maturity at ages of 6–9 yr and at body lengths of 135–145 cm. Males probably mature sexually at ages of 4–6 yr, at body lengths of 135–140 cm and at weight of testis of 40–150 g. The lack of females aged 5–6 yr and males aged 4–5 yr precluded firm conclusions on ages at sexual maturity. Parturition in western Kyushu was estimated to be prolonged from autumn to spring, whereas in the Inland Sea and Pacific waters it was restricted from spring to summer with a peak in April. These geographical differences and available information on distribution implies that the finless porpoises in western Kyushu constitute a local population.     

Concentric leaf spot of yam (Dioscorea spp) in South-western Nigeria

The etiology and epidemiology of concentric leaf spot and seedling blight diseases of yam (Dioscorea spp) was investigated at Jbadan, south western Nigeria, in the low land humid tropics. Sclerotium rolfsii was associated with the concentric leaf spot and seedling blight of yam. The pathogen was also harbored by weeds namely Mucuna pruriensis, Commelina erecta, ipomea triloba, I. involucrataand Ipomoea sp found growing within the yam plots. A high mean inoculum densities of 3.0 ± 0.5 × 10⁶ colony forming unit/g of soil was estimated in the soil samples obtained from the yam plots This revised version was published online in June 2006 with corrections to the Cover Date.     

Parasitism of three attelabid weevils by Bracon (Uncobracon) apoderi Watanabe (Hymenoptera: Braconidae) in Kyushu,Japan

The larval parasitoid Bracon (Uncobracon) apoderi Watanabe (Hymenoptera: Braconidae) is reported from Kyushu for the first time. Two host weevils, Phymatapoderus latipennis and Paroplapoderus pardalis (Coleoptera: Attelabidae), are newly recorded. Bracon apoderi parasitized three attelabid species, including Apoderus balteatus, but did not parasitize Cycnotrachelus roelofsi even though it is a dominant attelabid species in the study site. In addition, B. apoderi mainly parasitized suspended cradles rather than cut‐off cradles.