Breakdown of distyly in a tetraploid variety of <Emphasis Type="Italic">Ophiorrhiza</Emphasis><Emphasis Type="Italic"> japonica</Emphasis> (Rubiaceae) and its phylogenetic analysis

We examined the floral morph of tetraploid Ophiorrhiza japonica Blume var. amamiana Hatus. and diploid O. japonica var. japonica to elucidate the association of distyly and ploidy levels. Chloroplast DNA phylogeny was reconstructed to determine the number of tetraploidization events and floral morph shifts in O. japonica. All individuals of O. japonica var. amamiana proved to be long-homostylous, whereas O. japonica var. japonica was distylous with typical long- and short-styled flowers. Distyly is related to the ploidy level. The bagging treatment of flowers indicated that O. japonica var. amamiana is self-compatible and potentially automatically self-pollinating. In cpDNA sequencing analysis, no haplotype was shared between the two varieties. The cpDNA haplotype network displayed the monophyly of O. japonica var. amamiana, suggesting a single origin of this variety. Hence, both tetraploidization and the breakdown of distyly to homostyly in O. japonica var. amamiana likely occurred just once. Because O. japonica var. amamiana having the morphological and cytological entity is recognized as a single lineage and clearly separated from O. japonica var. japonica, this variety can be considered to be a distinct species. We therefore propose to raise O. japonica var. amamiana to the rank of species. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

New species of the genus <Emphasis Type="Italic">Scolecostigmina</Emphasis> and revision of <Emphasis Type="Italic">Cercospora cryptomeriicola</Emphasis> on conifers

Scolecostigmina chibaensis on Pinus parviflora is described as a new species. Cercospora cryptomeriicola Sawada on Cryptomeria japonica is transferred to the genus Pseudocercospora, based on the morphological characteristics of the type specimen and newly collected specimens.     

Phylogenetic analyses of <Emphasis Type="Italic">Uromyces viciae-fabae</Emphasis> and its varieties on <Emphasis Type="Italic">Vicia</Emphasis>, <Emphasis Type="Italic">Lathyrus</Emphasis>, and <Emphasis Type="Italic">Pisum</Emphasis> in Japan

A pea rust fungus, Uromyces viciae-fabae, has been classified into two varieties, var. viciae-fabae and var. orobi, based on differences in urediniospore wall thickness and putative host specificity in Japan. In principal component analyses, morphological features of urediniospores and teliospores of 94 rust specimens from Vicia, Lathyrus, and Pisum did not show definite host-specific morphological groups. In molecular analyses, 23 Uromyces specimens from Vicia, Lathyrus, and Pisum formed a single genetic clade based on D1/D2 and ITS regions. Four isolates of U. viciae-fabae from V. cracca and V. unijuga could infect and sporulate on P. sativum. These results suggest that U. viciae-fabae populations on different host plants are not biologically differentiated into groups that can be recognized as varieties.Contribution no. 184, Laboratory of Plant Parasitic Mycology, Institute of Agriculture and Forestry, University of Tsukuba, Japan     

Life cycle of <Emphasis Type="Italic">Uromyces appendiculatus</Emphasis> var. <Emphasis Type="Italic">azukicola</Emphasis> on <Emphasis Type="Italic">Vigna angularis</Emphasis>

Field observations and inoculation experiments revealed that Uromyces appendiculatus var. azukicola has an autoecious and macrocyclic life cycle and produces spermogonia, aecia, uredinia, and telia on Vigna angularis var. angularis and V. angularis var. nipponensis. From inoculation experiments, it was suggested that this rust fungus has different host relationships from other varieties. Morphological examinations revealed that the characteristics of urediniospores and teliospores are different among varieties, although aeciospores are morphologically similar to each other.Contribution no. 182, Laboratory of Plant Parasitic Mycology, Institute of Agriculture and Forestry, University of Tsukuba, Japan     

Two new species of <Emphasis Type="Italic">Lirula</Emphasis> on <Emphasis Type="Italic">Abies</Emphasis> from Japan

Two new species of Lirula (L. japonica and L. exigua) on Abies mariesii collected in subalpine areas of northern Japan are described as members of Rhytismatales, Discomycetes. Lirula japonica causes needle cast in fir, but L. exigua seems to occur on the needles of physically damaged twigs. Morphological characteristics of both species are discussed.     

Seasonal variation in allelopathic potential of<Emphasis Type="Italic">Artemisia princeps</Emphasis> var.<Emphasis Type="Italic">orientalis</Emphasis> on plants and microbes

We investigated seasonal variations in allelopathic potential ofArtemisia princeps var.orientalis. Aqueous and meth-anol extracts and volatile substances were prepared in the laboratory from samples collected monthly (April through October). Their impacts were then assessed on the germination and seedling growth ofLactuca sativa andAchyranthes japonica. The allelopathic potential varied with the time of sample collection and the concentration tested. For example, germination ofL. sativa was not inhibited by the aqueous extract but seedling growth (shoots and roots) was, with its seasonal effect being significant. ForA. japonica, seed germination was not inhibited at lower concentrations (except for August samples). However, at higher concentrations and in certain months (especially July), germination was more negatively affected. The degree of seedling growth inhibition also differed by month and by extract concentration, with roots being impacted more than shoots. Volatile substances also had a time-dependent influence on the germination and seedling elongation ofA. japonica. In a separate experiment, the ethyl-acetate and water fractions of a crude methanol extract were prepared monthly fromA. princeps var.orientalis. Here, we examined their antimicrobial activities against three gram-positive bacteria (Bacillus cereus, Bacillus subtilis, andStaphylococcus aureus), two gramnegative bacteria (Escherichia coli andPseudomonas fluorescens), and one lactic acid bacterium,Lactobacillus plantar urn. The ethyl-acetate fraction that was sampled in September was remarkably potent againstB. cereus andB. subtilis, whereas the water fraction collected in August and September showed great antimicrobial activity against the grampositive and -negative bacteria. In contrast,L. plantarum was not inhibited by the water fraction, regardless of the sampling month. Likewise, the ethyl-acetate and water fractions collected in April and October had the lowest levels of antimicrobial activity.     

Endophytic fungi from rhizomes of <Emphasis Type="Italic">Paris polyphylla</Emphasis> var. <Emphasis Type="Italic">yunnanensis</Emphasis>

About 63 fungal endophytic isolates were separated from rhizomes of Paris polyphylla var. yunnanensis, which is a traditional medicinal plant mainly distributed in China. The isolates were characterized and grouped based on the culture characteristics and the morphology of colony growth and conidia. Eleven representative ones were selected for further taxonomical identification. Five genera namely Fusarium, Gliocladiopsis, Gliomastix, Aspergillus and Cylindrocarpon were identified on the basis of their morphological characterizations. Of them, the most frequent genus was Fusarium (i.e. Ppf1, Ppf3 and Ppf14). Their ITS-rDNA sequences were compared with those available in the GeneBank databases to obtain the closest related species by BLAST analysis as well as to analyze their phylogenetic affiliation. The isolates were identified as Gliocladiopsis irregularis (Ppf2), Plectosphaerella cucumerina (Ppf4), Padospora sp. (Ppf6), Gliomastix murorum var. murorum (Ppf7), Aspergillus fumigatus (Ppf9), Pichia guilliermondii (Ppf10), Neonectria radicicola (anamorph: Cylindrocarpon) (Ppf12) and one uncultured mycorrhizal ascomycete (Ppf13) separately based on their morphological and molecular features. The molecular characters of the endophytic fungi were basically coincident with their morphology. The broad diversity and taxonomic spectrum were exhibited by the endophytic fungi from P. polyphylla var. yunnanensis.     

Regeneration of Three <Emphasis Type="Italic">Sphagnum</Emphasis> species

Sphagnum capillifolium var. tenellum, S. magellanicum, and S. recurvum var. brevifolium were regenerated from stem pieces grown in containers to assess their potential for use in peatland restoration projects. The effect of two water levels; peat, peat/sand or peat/clay substrates; and peat decomposition level on the species’ regeneration was evaluated. S. magellanicum attained the greatest cover on the peat or peat/sand mixture using decomposed peat when the growing surface was occasionally inundated. S. recurvum attained the greatest cover grown on the peat or peat/sand mixture using undecomposed peat when the water level was kept below the surface. S. capillifolium showed an affinity for the peat/clay mixture, and overall attained a greater total cover than the other species when grown under the lower water level on all substrate types, with total cover approximately three to five times that of the others. When developing management plans for restoration of mined peatlands, species-specific responses to water level, type and extent of mineral soil mixed with the peat surface, and peat decomposition level should be considered.     

New life cycles of <Emphasis Type="Italic">Porphyra katadae</Emphasis> var. <Emphasis Type="Italic">hemiphylla</Emphasis> in culture

Porphyra katadae Miura var. hemiphylla Tseng et T. J. Chang, a species distributed around the Liaodong and Shandong Peninsulas of China, produces gametophytes from late winter to early spring. These are monoecious with male and female reproductive tissues in distinct halves or sectors. Vegetative tissues from sectors expected to differentiate into sexual tissue were cultured in the laboratory. Male and female reproductive organs, as well as conchocelis and blades, were differentiated from these tissues. The male and female reproductive tissues were in patches and mixed on the cultured tissue pieces. This was quite different from the wild-type sectored individuals. The F₁ conchospore germlings also produced monospores, carposporangia, spermatangia and conchocelis. These carposporangia and spermatangia were in patches and were mixed on the F₁ fronds. The results imply that P. katadae var. hemiphylla is possibly sex-differentiated rather than sex-determined. This is the first report of such a dimorphic life history in the genus Porphyra.     

Isolation and characterization of melanin pigment from <Emphasis Type="Italic">Pleurotus cystidiosus</Emphasis> (telomorph of <Emphasis Type="Italic">Antromycopsis</Emphasis><Emphasis Type="Italic">macrocarpa</Emphasis>)

Melanins are enigmatic pigments that are produced by a wide variety of microorganisms including several species of bacteria and fungi. For more than 40 years, fungi have been known to produce pigments called melanins. Melanin pigment production by mushrooms was not intensively studied. The present study was carried out on isolation and characterization of melanin from an edible mushroom Pleurotus cystidiosus var. formosensis. The mushroom produced dark mucous mass of hyaline arthrospores on mycelium. The coremia exclusively produced dikaryotic arthrospores with the remnant of a clamp connection. Continuous cell extension and division in the coremium stipe supplied cells for arthroconidiation at the coremium apex, which is surrounded by a liquid droplet (coremioliquid). The black coloured coremea (conidia) were produced by Antromycopsis macrocarpa (anamorph of P. cystidiosus) when cultured on potato dextrose agar medium. The agar plate was incubated at continuous light illumination for high amount of pigment (coremea) production. The slimy layer of the coremea was extracted and partially purified by alkaline and acid treatment. The black pigment was confirmed as melanin based on UV, IR and EPR spectra apart from chemical analysis. This is the first report on characterization of melanin obtained from Pleurotus cystidiosus var. formosensis.     

A new species of <Emphasis Type="Italic">Brasiliomyces</Emphasis> (Erysiphaceae) on <Emphasis Type="Italic">Dalbergia cultrata</Emphasis> var. <Emphasis Type="Italic">cultrata</Emphasis> from Thailand

A powdery mildew fungus on leaves of Dalbergia cultrata var. cultrata (Fabaceae) collected at the Queen Sirikit Botanical Garden in northern Thailand is proven to be a new species of the genus Brasiliomyces and is described as B. chiangmaiensis sp. nov. with light and SEM micrographs. Differences in known Brasiliomyces species are discussed, and a key to species of this genus is provided.     

Two new records of <Emphasis Type="Italic">Mycena</Emphasis> sect. <Emphasis Type="Italic">Sacchariferae</Emphasis> from Japan and type study of <Emphasis Type="Italic">Mycena cryptomeriicola</Emphasis> (sect. <Emphasis Type="Italic">Sacchariferae</Emphasis>)

Mycena cupulicola sp. nov. and M. adscendens var. carpophila, new to Japan, are described and illustrated. The former is characterized by having lageniform caulocystidia with a slightly thick-walled broadened base and no cheilocystidia. The latter is characterized by having a white pileus up to 1mm in diameter and narrowly conical caulocystidia. Mycena cryptomeriicola was confirmed to have inamyloid basoidiospores.     

Revision of <Emphasis Type="Italic">Heterosavia</Emphasis>, stat. nov., with notes on <Emphasis Type="Italic">Gonatogyne</Emphasis> and <Emphasis Type="Italic">Savia</Emphasis> (Phyllanthaceae)

Heterosavia (Phyllanthaceae) is segregated from Savia (tribe Bridelieae), recognized at generic rank, and placed in tribe Phyllantheae. Floral, fruit, leaf anatomical, leaf venation, and pollen characters of the neotropical taxa previously united as Savia including Gonatogyne are discussed and illustrated. Keys to the three genera and to the species of Heterosavia are presented. Four species (all new combinations), Heterosavia bahamensis, H. erythroxyloides, H. laurifolia, and H. maculata, are recognized. The new combinations Heterosavia laurifolia var. intermedia and H. maculata var. clementis are proposed. The names Heterosavia, H. erythroxyloides, H. laurifolia, Savia clementis, S. clusiifolia, S. clusiifolia var. fallax, and S. longipes are lectotypified. Distribution maps and conservation assessments (IUCN ratings) of Heterosavia species and varieties are provided.     

Four new species of <Emphasis Type="Italic">Phyllosticta</Emphasis>, one new species of <Emphasis Type="Italic">Pseudocercospora</Emphasis>, and one new combination in <Emphasis Type="Italic">Passalora</Emphasis> from Japan

During a survey of plant-inhabiting fungi in a botanical garden in Japan, some noteworthy fungi were collected from leaf spots of some herbal and arboreal plants. Among them, five new species are described, namely: Phyllosticta. ardisiicola on Ardisia crenata, Phy. aspidistricola on Aspidistra elatior, Phy. kerriae on Kerria japonica, Phy. fallopiae on Fallopia japonica, and Pseudocercospora davidiicola on Davidia involucrata. Passalora pyrrosiae, a new combination for Pseudocercospora pyrrosiae on Pyrrosia lingua, is proposed based on its morphological characteristics designating the neotype specimen.     

Ploidy chimeras induced in haploid sporophytes of <Emphasis Type="Italic">Osmunda claytoniana</Emphasis> and <Emphasis Type="Italic">Osmunda japonica</Emphasis>

Haploid sporophytes of Osmunda claytoniana (2n = x = 22) were apogamously produced from calli when cultivated on a hormone-free medium. Flow cytometric analysis showed that ploidy chimeras were spontaneously produced in a haploid sporophyte of O. claytoniana and those of O. japonica that were obtained in the previous study. In the haploid sporophyte of O. claytoniana, a diploid pinnule and a partially diploid terminal segment were produced in a haploid pinna. In O. japonica, a haploid sporophyte yielded a diploid pinna in a haploid frond, and another haploid sporophyte yielded a diploid pinnule in a haploid pinna. Diploid chimeras were large in size and could be readily distinguished from other haploid parts of the fronds. It is likely that the chimeras were produced clonally from a single diploid cell that established chromosome doubling.     

A new variety of <Emphasis Type="Italic">Cortinarius subalboviolaceus</Emphasis> (Sect. <Emphasis Type="Italic">Sericeocybe</Emphasis>) from Japan

Cortinarius subalboviolaceus var. niigatensis var. nov., which grows in deciduous forests, is described and illustrated from Niigata and Gunma prefectures, Japan. It differs from C. subalboviolaceus var. subalboviolaceus mainly in its more distinct violet coloration and somewhat larger size of the basidiocarp.     

Ecological niche of <Emphasis Type="Italic">Cryptococcus neoformans</Emphasis> var. <Emphasis Type="Italic">grubii</Emphasis> and <Emphasis Type="Italic">Cryptococcus gattii</Emphasis> in decaying wood of trunk hollows of living trees in Jabalpur City of Central India

Cryptococcus neoformans var. grubii and C. gattii were repeatedly isolated from decaying wood of trunk hollows in living trees growing in Jabalpur City in Central India. The isolation of C.␣gattii has been reported from decayed wood inside trunk hollow of Tamarindus indica (15.6%), Mangifera indica (2.2%), Pithecolobium dulce (12.5%), Syzygium cumini (14%), and one from bark of S.␣cumini. C. n. var. grubii was isolated from decaying wood debris of T. indica (4.4%), M. indica (13.3%), Terminalia arjuna (25%), S. cumini (2%), Cassia fistula (4.5%), and two from bark of S. cumini. The two varieties never co-occurred in the same hollow. C. gattii and C. n. var. grubii isolates belonged to serotype B and serotype A respectively. The data strongly supported the colonization of the pathogen in␣decaying wood hollow of all six-tree species. Evidence of this was found by repeated isolation up to 820 days. P. dulce is being reported for the first time as natural habitat of C. gattii and T.␣arjuna and C. fistula as natural habitat for C. n. var. grubii. M. indica is being reported for the second time as the natural habitat of both varieties (C. n. var. grubii and C. gattii). The population density of these pathogens from decaying wood debris of various tree species ranged between 0.5 × 10³ cells/g and 6 × 10⁵ cells/g. The seasonal variation has been seen in isolation of this yeast. Our result further reinforce the recently emerging evidence that the natural habitat of C. n. var. grubii and C. gattii is more generalized.     

The <Emphasis Type="Italic">Caenorhabditis</Emphasis><Emphasis Type="Italic">briggsae</Emphasis> genome contains active <Emphasis Type="Italic">CbmaT1</Emphasis> and <Emphasis Type="Italic">Tcb1</Emphasis> transposons

The maT clade of transposons is a group of transposable elements intermediate in sequence and predicted protein structure to mariner and Tc transposons, with a distribution thus far limited to a few invertebrate species. We present evidence, based on searches of publicly available databases, that the nematode Caenorhabditis briggsae has several maT-like transposons, which we have designated as CbmaT elements, dispersed throughout its genome. We also describe two additional transposon sequences that probably share their evolutionary history with the CbmaT transposons. One resembles a fold back variant of a CbmaT element, with long (380-bp) inverted terminal repeats (ITRs) that show a high degree (71%) of identity to CbmaT1. The other, which shares only the 26-bp ITR sequences with one of the CbmaT variants, is present in eight nearly identical copies, but does not have a transposase gene and may therefore be cross mobilised by a CbmaT transposase. Using PCR-based mobility assays, we show that CbmaT1 transposons are capable of excising from the C. briggsae genome. CbmaT1 excised approximately 500 times less frequently than Tcb1 in the reference strain AF16, but both CbmaT1 and Tcb1 excised at extremely high frequencies in the HK105 strain. The HK105 strain also exhibited a high frequency of spontaneous induction of unc-22 mutants, suggesting that it may be a mutator strain of C. briggsae.