Scanning Electron Microscopic Observation of Differentiation from the Reproductive Short Form to the Infectious Long Form of Holospora obtusa

To identify the surface features of Holospora obtusa during its differentiation from the reproductive short form to the infectious long form, bacteria of four different buoyant densities were isolated by Percoll density gradient centrifugation of homogenates of host cells or isolated macronuclei, and examined with a scanning electron microscope. Bacteria of buoyant density 1.09 g/ml were reproductive short forms as well as cells at various stages in the elongation process including fully elongated ones. Bacteria of buoyant densities 1.11 g/ml and 1.13 g/ml were premature long forms and those of 1.16 g/ml were mature infectious long forms. Bacteria of buoyant density 1.09 g/ml had an entirely rough surface while those of buoyant densities 1.11 g/ml and 1.13 g/ml were smooth and had wale-like stripes on their surface. A small tapered tip was observed at one end of the bacteria of buoyant density 1.13 g/ml. Bacteria of buoyant density 1.16 g/ml had an entirely smooth surface, but one end always showed a rough surface; this locally differentiated surface of the special tip of the infectious long form may be responsible for both the nuclear and species specificities of the infectivity of H. obtusa. These observations indicate that the surface of H. obtusa changes during differentiation and the special tip develops in bacteria of buoyant density 1.13 g/ml.     

Developmental morphology of foliose shoots and seedlings of Dalzellia zeylanica (Podostemaceae) with special reference to their meristems

The developmental morphology of seedlings and shoots of Dalzellia zeylanica was examined with reference to the meristem in order to understand the dorsiventral, foliose shoot. In seedlings, no obvious primary shoot and no root are formed. Subsequent to disappearance of the vestigial primary shoot meristem, two shoot meristems are established in the axils of the cotyledons, one of which grows into a secondary shoot. Microtome and SEM examinations of mature plants show that the shoot meristem is complex, comprising three zones along the shoot margin. The organogenetic zone, equivalent to the shoot apical meristem, produces dorsal leaves proximally and much fewer marginal leaves distally. During development, the zone repeatedly changes into a dorsal zone, while a new organogenetic zone is formed in an area between developing marginal leaves, resulting in the alternation of the organogenetic and dorsal zones, which allowed development of the coenosomic structure of the shoot. The dorsal and ventral zones do not produce leaves, but contribute to shoot expansion. The ventral zone also forces the marginal leaves to shift to the lateral side of the shoot. The rosette with tufted leaves might be a modification of the short shoot (ramulus) of other Tristichoideae.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 144 , 289–302.     

A nocturnal Provespa wasp species as the probable pollinator of epiphytic orchid Coelogyne fimbriata

Many vespid wasps visit flowers to forage nectar. These hymenopterans sometimes contribute to flower pollination. However, none of the nocturnal wasp species is a known pollinator. We collected individuals of light‐attracted Provespa nocturna workers in a montane rainforest on Peninsular Malaysia: some wasps collected bore orchid pollinia on their thoraxes. Among 114 trapped individuals, four bore pollinaria and nine bore only viscidia, suggesting that pollinia had been successfully transported. Molecular barcoding of the pollinia (based on their ITS sequences) assigned the orchid to a species in Coelogyne fimbriata complex. These findings and our other analyses suggest that this nocturnal wasp contributes to pollination of an epiphytic nectarless orchid that probably releases olfactory attractants. This discovery sheds light on the importance of mutualistic relationships between the nocturnal social wasps and epiphytic orchids in Southeast Asian tropical rainforest canopies.     

Assembly,Target‐Signaling and Intracellular Transport of Tyrosinase Gene Family Proteins in the Initial Stage of Melanosome Biogenesis

Assembly, target‐signaling and transport of tyrosinase gene family proteins at the initial stage of melanosome biogenesis are reviewed based on our own discoveries. Melanosome biogenesis involves four stages of maturation with distinct morphological and biochemical characteristics that reflect distinct processes of the biosynthesis of structural and enzymatic proteins, subsequent structural organization and melanin deposition occurring in these particular cellular compartments. The melanosomes share many common biological properties with the lysosomes. The stage I melanosomes appear to be linked to the late endosomes. Most of melanosomal proteins are glycoproteins that should be folded or assembled correctly in the ER through interaction with calnexin, a chaperone associated with melanogenesis. These melanosomal glycoproteins are then accumulated in the trans Golgi network (TGN) and transported to the melanosomal compartment. During the formation of transport vesicles, coat proteins assemble on the cytoplasmic face of TGN to select their cargos by interacting directly or indirectly with melanosomal glycoproteins to be transported. Adapter protein‐3 (AP‐3) is important for intracellular transport of tyrosinase gene family proteins from TGN to melanosomes. Tyrosinase gene family proteins possess a di‐leucine motif in their cytoplasmic tail, to which AP‐3 appears to bind. Thus, the initial cascade of melanosome biogenesis is regulated by several factors including: 1) glycosylation of tyrosinase gene family proteins and their correct folding and assembly within ER and Golgi, and 2) supply of specific signals necessary for intracellular transport of these glycoproteins by vesicles from Golgi to melanosomes.     

Current progress in DNA barcoding and future implications for entomology

DNA barcoding is a technique for identifying organisms based on a short, standardized fragment of genomic DNA. The standardized sequence region is called a DNA barcode because it is like a barcode tag for each taxon. Since the proposition of this concept and the launch of a large project named the Barcode of Life, this simple technique has attracted attention from taxonomists, ecologists, conservation biologists, agriculturists, plant‐quarantine officers and others, and the number of studies using the DNA barcode has rapidly increased. The extreme diversity of insects and their economical, epidemiological and agricultural importance have made this group a major target of DNA barcoding. However, there is some controversy about the utility of DNA barcoding. In this review, we present an overview of DNA barcoding and its application to entomology. We also introduce current advances and future implications of this promising technique.     

Effects of forest loss and fragmentation on pollen diets and provision mass of the mason bee,Osmia cornifrons,in central Japan

1. Habitat loss and fragmentation potentially affect the performance of bees that forage nectar and pollen of plants in their habitats. In forest landscapes, silvicultural conifer plantations often have reduced and fragmented natural broadleaf forests, which seem to provide more floral resources for bees than do the plantations. 2. This study evaluated the effects of forest characteristics (i.e. elevation, area, edge length, and tree size of natural forests) on pollen diets (plant taxa assemblages of pollen grains in provisions) and total provision mass in oviposited chambers in nests made by a standardised number of Osmia cornifrons bees at 14 sites in a forestry area in central Japan. 3. From April to May, the numbers of nests and chambers per nest increased, and the provision mass per chamber decreased. Main pollen sources were Prunus at higher elevations in April and Wisteria at lower elevations in May, foraging on which increased the numbers of nests and chambers per nest. The provision mass per chamber was smaller at higher elevations in more fragmented natural forests. Decreases in the area of natural forests within the foraging range (400‐m radii) of O. cornifrons increased the utilisation of Rubus pollen and decreased the total provision mass. 4. These findings suggest that the loss and fragmentation of natural broadleaf forests change pollen diets and reduce the provision mass of mason bees, which may reduce the number and size of their offspring.     

RELATION BETWEEN THE ACROSOME REACTION AND FERTILIZATION IN THE SEA URCHIN. II. FERTILIZATION IN ACIDIFIED SEA WATER WITH EGG-WATER-TREATED SPERMATOZOA*

Fertilization of sea urchin eggs fails to occur at a pH lower than 6.5. Analytical studies on this problem were made with Hemicentrotus pulcherrimus, Anthocidaris crassispina and Pseudocentrotus depressus. If the spermatozoa have been pretreated with egg water, eggs can be fertilized even at pH 6.5 and 6.0. The acrosome reaction is inhibited at a pH lower than 6.5. Intact spermatozoa fail to adhere to the fixed eggs in acidified sea water, whereas egg-water-treated spermatozoa adhere even at pH 6.5 and 6.0. From these results we infer that the failure of fertilization at pH 6.5–6.0 is caused by non-occurrence of the acrosome reaction, and that fertilization reactions other than the acrosome reaction, such as the binding and fusion of the gametes, are not inhibited in this range of pH. At pH 5.5, the spermatozoa become inert and fertilization is inhibited or suppressed, even though egg-water-treated spermatozoa are employed.