Development and diversity of microsatellite markers for endangered species,Morus boninensis Koidz., to establish conservation program

We have developed seven microsatellite markers from an enrichment library of genomic DNA for an endangered species, Morus boninensis. A total of 112 of the 320 clones were found to have unique sequences with microsatellite repeats. Seven of 54 primer pairs revealed clear chromatograms and polymorphisms among 36 individuals sampled from three of the Bonin Islands. Seven to 17 alleles per locus were detected, and the expected heterozygosity without considering double reduction ranged from 0.429 to 0.819. These findings should be useful for those studying the conservation genetics of M. boninensis.     

Oviposition site selection by a lepidopteran leafminer in response to heterogeneity of leaf surface conditions: structural traits and microclimates

1. Leafminer larvae are sedentary and make feeding tracks called mines. Their spatial distribution in trees affects their growth and survival through interaction with the heterogeneity of environments, such as leaf traits and microclimate. Lepidopteran leafminers that mine lower leaf surfaces have shown evolutionary radiation, suggesting that lower surfaces improve leafminer performance. 2. The lepidopteran multivoltine leafminer Phyllocnistis sp. Zeller (Gracirallidae: Phyllocnistinae) uses the Japanese privet Ligustrum japonicum Thunb. (Oleaceae). It mines only the lower‐surface epidermal layer of primary shoot leaves early in the occurrence season, but once lammas shoots appear, which happens in seasons other than spring, it preferentially uses the lower surface, but also uses the upper surface of the leaves. This study examined whether selection of oviposition sites was associated with the structural traits and microclimate of the leaf surface. 3. The shift of oviposition site from primary to lammas shoot leaves followed increasing hardness and epidermal cell wall thickness of primary shoot leaves during leaf development, and mine initiation rates decreased below 20% after oviposition on mature primary shoot leaves. Preference for the lower surface was related to the thinner cuticle. However, the thinner cuticle of the upper surface on lammas shoot leaves allowed Phyllocnistis sp. to expand its mining sites to both surfaces with no decrease in mine initiation and emergence rates. 4. Microclimates (leaf surface and mine temperatures) did not differ between upper and lower surfaces, suggesting that microclimate did not affect oviposition site selection by Phyllocnistis sp. These results suggest that the adaptive radiation of lower‐surface mining may have been influenced by the leaf surface characteristics.     

Maternal and paternal effects on the germination time of non‐dormant seeds of a monocarpic perennial species,Aster kantoensis (Compositae)

In plant species producing non‐dormant seeds, the germination time (from the start of imbibition to radicle emergence) is the main factor determining the timing of seedling emergence. We investigated maternal and paternal genetic effects on the germination time of non‐dormant seeds of a monocarpic perennial, Aster kantoensis Kitamura (Compositae). Three sets of reciprocal diallel crosses among five plants were conducted to produce genetic variation in seeds, and the germination time of the progeny of each parent was determined. The effects of the maternal parent and the interaction of maternal and paternal parents on the germination time of progeny were significant in all sets, and the effect of the paternal parent was significant in two of the three sets. This result means that the germination time of the progeny of a maternal or paternal parent can vary with the genotype of its mating partners. Because variation in the emergence time of seedlings contributes to avoiding seedling loss owing to unpredictable environmental changes, genetic variation in the germination time among the progeny of each parent mating with multiple partners could contribute to the establishment of the parent's seedlings in species producing non‐dormant seeds in the field.     

Encephalitozoon-Like Organisms in Patients with Alveolar Hydatid Disease: Cell Culture, Ultrastructure, Histoimmunochemical Localization and Seroprevalence

ABSTRACT. We found Encephalitozoon -like organisms in an in vitro culture of a human liver lesion which was due to larval Echinococcus multilocularis. The organisms developed in the same fashion as an Encephalitozoon cunculi. The spores that developed in parasitophorous vacuoles were 2.0–2.6 × 1.1–1.5 μm: each contained a single nucleus and 4–5 polar tubule coils, closely resembling E. cuniculi in its ultrastructure. Mature spores were collected from the supernatants by the use of Percoll centrifugation resulting in the banding of the spores on continuous gradients. We prepared three sorts of spores which were different in buoyant density in 0.15 M NaCl: 1.05–1.07 g/ml spores, 1.12 g/ml spores, and spores of over 1.14 g/ml. Polyclonal antibodies to a pool of each spore preparation were produced in a rabbit and applied to the detection of microsporidian antigen in situ. The histoimmunoperoxidase (HIP) procedure was used to detect the microsporidian antigen in echinococcal liver lesions from patients with alveolar hydatid disease (AHD). Ten echinococcal liver lesions from different AHD patients were examined and four were found to be positive in the HIP test. The Percoll-separated spores were also used as an antigen to detect for antibodies in the sera from the patients with AHD by Western blotting. Antibodies were detected in 62 (52%) of the 119 AHD patients and in only 8 (5%) of the 159 normal healthy individuals.     

Central Die-back of Monoclonal Stands of Reynoutria japonica in an Early Stage of Primary Succession on Mount Fuji

Reynoutria japonica is a common perennial pioneer species onJapanese volcanoes. In a volcanic desert (1500m above sea level)on Mount Fuji (3776m), central Japan, this species forms circularstands (patches). As a patch develops, shoot density decreasesin its centre (‘central die-back’). The centraldie-back has been considered a key process in the early stagesof primary succession, though its mechanism has been unknown. The pattern of patch development, population dynamics of aerialshoots, and growth patterns of below-ground organs were analysedin order to investigate the mechanism of die-back, and the followingtraits are clarified: (1) central die-back areas occur in mostsmall patches (approx. 1m²) without later successional species;(2) shoot characteristics are dependent both on their positionwithin a patch and on patch size; (3) despite the large differencesin shoot density, neither time course of shoot growth nor theirmortality differs between the centre and periphery of patches;and (4) rhizomes ofR. japonicagrow outwards with regular sympodialbranching. From these results, it is concluded that neither interspecificnor intraspecific competition is likely to be a primary causeof the die-back phenomenon, but that central die-back is broughtabout intrinsically by the growth pattern of the rhizome systems.We also discuss the importance of the central die-back in facilitatingestablishment of later successional species in the early stagesof primary succession. Clonal plant; central die-back; competition; facilitation; Japanese knotweed; Mount Fuji; primary succession;Reynoutria japonica ; rhizome growth; volcanic desert     

Seedling Establishment of a Canopy Tree Species in Malaysian Tropical Rain Forests

Abstract Unlike many other canopy tree species in tropical rain forests, Dryobalanops aromatica (Dipterocar-paceae, local name: Kapur) establishes monospecific dominant (monodominant) forests in Peninsular Malaysia. In natural conditions, monodominance of Kapur does not necessarily mean low species diversity of the Kapur forests. While the emergent canopy layer is occupied by Kapur, many other plant species, which are common to lowland dipterocarp primary forests in the same region, are found in lower canopy layer and understory.
To understand the ecological implications of the monodominance of Kapur, we monitored post dispersal survival and seedling establishment in a pure stand of Kapur in a plantation in Kepong, near Kuala Lumpur. Immediately after seed fall, seeds and cotyledon-stage seedlings suffered high predation by vertebrates such as rodents. The predation pressure was higher in a more general fruiting year (1991) than in a sporadic fruiting year (1992). In contrast to the high mortality of seeds and newly emerged seedlings, seedlings surviving to the six-leaf stage showed low mortality, which allowed the establishment of a sapling bank.
The occurrence of saplings of Kapur with a wide range of size classes in natural forests indicates that this species is more shade tolerant than other dipterocarp species such as the Shorea group and that it could well respond to enhanced light conditions caused by canopy opening. These characteristics may partly contribute to maintaining monodominance of Kapur.     

Ancient genome‐wide admixture extends beyond the current hybrid zone between Macaca fascicularis and M. mulatta

Macaca fascicularis and Macaca mulatta are two of the most commonly used laboratory macaques, yet their genetic differences at a genome‐wide level remain unclear. We analysed the multilocus DNA sequence data of 54 autosomal loci obtained from M. fascicularis samples from three different geographic origins and M. mulatta samples of Burmese origin. M. fascicularis shows high nucleotide diversity, four to five times higher than humans, and a strong geographic population structure between Indonesian‐Malaysian and Philippine macaques. The pattern of divergence and polymorphism between M. fascicularis and M. mulatta shows a footprint of genetic exchange not only within their current hybrid zone but also across a wider range for more than 1 million years. However, genetic admixture may not be a random event in the genome. Whereas randomly selected genic and intergenic regions have the same evolutionary dynamics between the species, some cytochrome oxidase P450 (CYP) genes (major chemical metabolizing genes and potential target genes for local adaptation) have a significantly larger species divergence than other genes. By surveying CYP3A5 gene sequences of more than a hundred macaques, we identified three nonsynonymous single nucleotide polymorphisms that were highly differentiated between the macaques. The mosaic pattern of species divergence in the genomes may be a consequence of genetic differentiation under ecological adaptation and may be a salient feature in the genomes of nascent species under parapatry.