<Emphasis Type="Italic">Erysiphe fimbriata</Emphasis> sp. nov.: a powdery mildew fungus found on <Emphasis Type="Italic">Carpinus laxiflora</Emphasis>

Ascomata of a powdery mildew-like fungus have been found on Carpinus laxiflora in Tochigi Prefecture of Japan since 2003. The morphological and molecular characteristics of this fungus are reported, and a new species, Erysiphe fimbriata, is proposed. It has large chasmothecia (200–250 μm in diameter) with long (up to 4–5 mm in length), fimbriate appendages arising from the upper half of the chasmothecia and turning upward, and numerous asci (22–38 per chasmothecium). Erysiphe fimbriata is a unique fungus both genetically and morphologically.     

New species of <Emphasis Type="Italic">Clohiesia</Emphasis> and <Emphasis Type="Italic">Paraniesslia</Emphasis> collected from freshwater habitats in China

Two fungi collected from submerged woody debris were found to represent hitherto undescribed species of the ascomycete genera Clohiesia and Paraniesslia. They are described as Clohiesia curvispora sp. nov. and Paraniesslia aquatica sp. nov. based on morphological characters. Clohiesia curvispora is characterized by immersed ascomata under a clypeus, and unitunicate, cylindrical asci containing one-celled, curved, elongate-fusiform ascospores. Paraniesslia aquatica is characterized by small, superficial, setose ascomata, and unitunicate, clavate asci containing verrucose, brown ascospores. Each species is illustrated with light micrographs and compared with similar taxa in this article.     

Identification,distribution and occurrence of the ascomycete <Emphasis Type="Italic">Metschnikowia typographi</Emphasis> in the great spruce bark beetle, <Emphasis Type="Italic">Dendroctonus micans</Emphasis>

This is the first report on the ascomycete Metschnikowia typographi from the adults and larvae of the great spruce bark beetle Dendroctonus micans in Turkey. In total, 910 of 1928 adults and 44 of 149 larvae investigated during the two years were infected by the pathogen. In a fresh smear the asci of the pathogen measure 18.5 ± 2.05 μm (14.7–22.3) in length and 2.1 ± 0.4 μm in width (n = 35). The ascospores are about 2 μm shorter than asci, having an average length of 16.4 ± 1.5 μm (14.2–18.0). The total infection rate of D. micans was 47.2 %. The prevalence of M. typographi infections differed between localities and years. Different infection rates of male and female beetles of D. micans were not recognized.     

Reclassification of Two <Emphasis Type="Italic">Peronospora</Emphasis> Species Parasitic on <Emphasis Type="Italic">Draba</Emphasis> in <Emphasis Type="Italic">Hyaloperonospora</Emphasis> Based on Morphological and Molecular Phylogenetic Data

On the family Brassicaceae, the causal agent responsible for downy mildew disease was originally regarded as a single species, Peronospora parasitica (now under Hyaloperonospora), but it was recently reconsidered to consist of many distinct species. In this study, 11 specimens of Peronospora drabae and P. norvegica parasitic on the genus Draba were investigated morphologically and molecularly. Pronounced differences in conidial sizes (P. drabae: 14–20 × 12.5–15.5 μm; P. norvegica: 20–29 × 15.5–22 μm) and 7.8% sequence distance between their ITS1-5.8S-ITS2 rDNA sequences confirmed their status as distinct species. Based on ITS phylogeny and morphology (monopodially branching conidiophores, flexuous to sigmoid ultimate branchlets, hyaline conidia and lobate haustoria), the two species unequivocally belong to the genus Hyaloperonospora and not to Peronospora to which they were previously assigned. Therefore, two new combinations, Hyaloperonospora drabae and H. norvegica, are proposed. The two taxa are illustrated and compared using the type specimen for H. norvegica and authentic specimens for H. drabae, which is lectotypified.     

<Emphasis Type="Italic">In vitro</Emphasis> propagation of <Emphasis Type="Italic">Decalepis arayalpathra</Emphasis>, a critically endangered ethnomedicinal plant

Summary This study reports a protocol for successful micropropagation of Decalepis arayalpathra (Joseph and Chandras) Venter. (Janakia arayalpathra Joseph and Chandrasekhran; Periplocaceae), a critically endangered and endemic ethnomedicinal plant in the southern forests of the Western Ghats which is overexploited for its tuberous medicinal roots by the local Kani tribes. Natural regeneration is rare and conventional propagation is difficult. Conservation of the species through micropropagation was attempted. The nodal explants of greenhouse-raised plants, were more desirable than cotyledonary nodal explants of aseptic seedlings. The basal nodes (73%) of 12–16-wk-old greenhouse-grown plants cultured in Murashige and Skoog (MS) medium containing 12.96 μM 6-benzyladenine (BA), 2.48 μM 2-isopentenyladenine (2-ip) and 2.68 μM α-naphthaleneacetic acid (NAA) formed 16–17 cm long unbranched robust solitary shoots in 8 wk. Cotyledonary nodal explants cultured in the same medium showed multiple shoot formation and axillary branching. But the shoots were thin, fragile and not suitable for mass propagation. Single nodes of a solitary shoot subcultured on MS medium containing 2.22 μM BA and 0.24 μM 2-ip together produced 9.8±0.3 nodes from 18.0±0.6 cm long shoots within 5–6 wk. The basal nodes of the shoots so formed were repeatedly subcultured to increase the stock of propagules while the 2.5–3.0 cm terminal cuttings were used for rooting. The best root induction (68%) and survival (86%) was achieved on half-strength MS medium supplemented with 1.07 μM NAA. Field-established plants showed uniform growth and phenotypic similarity to parental stock.     

<Emphasis Type="Italic">Hirsutella proturicola</Emphasis> sp. nov. isolated from a proturan, <Emphasis Type="Italic">Baculentulus</Emphasis> densus (Protura,Hexapoda)

A new species of Hirsutella, H. proturicola, isolated from a subterranean proturan (Baculentulus densus; Protura, Hexapoda), is described and illustrated. Hirsutella proturicola is characterized by producing monoblastic phialides of 24–51.5 × 2.5–5 μm with a slightly roughened neck, fusiform and curved conidia of 9–18 × 2.5–4 μm that have a truncate base and a papillate projection often capped with sheath-like mucilage, and pluricellular, globose to subglobose chlamydospores of 21–48 × 21–41.5 μm. This species is morphologically and phylogenetically close to H. rostrata, an acaropathogenic species, but can be distinguished from the size of the phialides and the size and shape of the conidia.     

<Emphasis Type="Italic">In vitro</Emphasis> organogenesis of <Emphasis Type="Italic">Pelecyphora aselliformis</Emphasis> erhenberg (cactaceae)

Summary In vitro propagation of Pelecyphora aselliformis, a Mexican cactus which is considered rare and is highly valued in the commercial market, was initiated using seeds as explants. The longitudinal explants from seedlings germinated in vitro were cultivated on Murashige and Skoog medium containing 8.8 μM benzyladenine (BA) or 4.6 μM kinetin at pH 7.0. After 120 d, each explant gave rise to five shoots and this number of shoots increased 20–25% after subculture. The hyperhydricity was similar in both media, but callus formation was lower on the medium with BA. The shoot development, in terms of epicotyl length, and fresh and dry weight after 6 wk, was also recorded. The epicotyl length was similar on shoot-forming media but the quality of shoots was better on media containing BA. In about 1 yr, 500–600 well-defined shoots were obtained. The rooting of shoots was very slow and a vigorous radical system was observed after 1 yr of culture.     

Pleosporales in Japan (2): the genus <Emphasis Type="Italic">Lophiotrema</Emphasis>

 Five species of the genus Lophiotrema are reported in this article. Of these, Lophiotrema vitigenum is described and illustrated as a new species. Three species, L. nucula, L. neohysterioides, and L. vagabundum, are newly added to the Japanease mycoflora. Lophiosphaera orientalis, Nodulosphaeria araucariae, and N. rosae, which had been recorded in Japan, are treated as synonyms of Lophiotrema fuckelii. A key to the species of Lophiotrema in Japan is given. Received: October 9, 2002 / Accepted: January 20, 2003 Acknowledgments We are grateful to Drs. Shuhei Tanaka (curator of YAM) and Ken Katumoto for the loan of fungal material. Correspondence to:K. Tanaka     

Pleosporales in Japan (1): the genus <Emphasis Type="Italic">Lophiostoma</Emphasis>

 Seven species of the genus Lophiostoma were the subject of this study. Among these, Lophiostoma mucosum is described and illustrated as a new species. All other species, L. macrostomum, L. semiliberum, L. arundinis, L. caulium, L. caudatum, and L. winteri, are reported for the first time in Japan. A key to the species of Lophiostoma in Japan is given. Received: August 5, 2002 / Accepted: November 28, 2002 Acknowledgments We are grateful to Dr. Hideki Takahashi (curator of SAPA) for the loan of fungal material. Correspondence to:Y. Harada     

<Emphasis Type="Italic">In vitro</Emphasis> propagation of eight species or subspecies of <Emphasis Type="Italic">Turbinicarpus (Cactaceae)</Emphasis>

Summary In vitro propagation systems by means of areole activation were developed for Turbinicarpus laui, T. lophophoroides, T. pseudopectinatus, T. schmiedickeanus subsp. flaviflorus, T. schmiedickeanus subsp. klinkerianus, T. schmiedickeanus subsp. schmiedickeanus, T. subterraneus, and T. valdezianus. In vitro-germinated seedlings were used as a primary source of explants. Multiple shoot formation from areoles was achieved for three explant types (apical, lateral, and transverse), cultured on Murashige and Skoog (MS) basal medium supplemented with 3% sucrose, 10 gl⁻¹ agar and several treatments with cytokinins. Efficiencies were in the range from 7.8 shoots per explant in T. valdezianus up to 19.7 shoots per explant in T. pseudopectinatus, using the best treatment for each species and in a single proliferation cycle. Four of the studied species responded best when 6-benzylaminopurine (3.3–8.8μM) was used, while 6-(γ,γ-dimethylallylamino)purine (19.7–24.6μM) showed better results in two species. The two remaining species showed no significant differences in their response to both cytokinins. Regarding explant type, the best results were obtained with transverse cuts for five species, with apical explants for one species, and the two remaining species showed no significant differences among the explants tested. Rooting of the in vitro-generated shoots was achieved most efficiently on half- or full-strength MS basal medium. Rooting frequencies were in the range from 54.2 to 94.2%, and the frequency of survival of the plants once transferred to soil was 91.6% on average.     

<Emphasis Type="Italic">Subsessila turbinata</Emphasis> gen. et. sp. nov. (Beltraniaceae), a <Emphasis Type="Italic">Beltrania</Emphasis>-like fungus from Thailand

A new monotypic Beltrania-like genus, Subsessila, with its type species S. turbinata, is described, illustrated and compared with similar genera. The new genus is introduced in the family Beltraniaceae based on phylogenetic analysis and morphological characters. Subsessila can be easily distinguished from other Beltrania-like genera by dark setae arising from radially lobed basal cells, mostly lacking macronematous conidiophores. Conidiogenous cells are ampulliform or doliiform and produce turbinate to clavate conidia with rostrate proximal end and rounded distal end. Evidence for establishment of the new genus is provided based on morphological comparison and DNA sequence data analyses.     

Somatic embryogenesis and <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation in <Emphasis Type="Italic">Bixa orellana</Emphasis> L.

Establishment, maintenance, regeneration, and transformation of somatic embryos by both direct and indirect means (callus-mediated) was achieved for Bixa orellana, a tropical plant whose seeds produce commercially edible ‘annatto pigment,’ which mainly constitutes an apocarotenoid called bixin. Callus-mediated methodology was found to be efficient in producing a greater number of embryos in a short time. The maximum of 28 somatic embryos were produced in 16–18 weeks when immature zygotic embryonic stalks were inoculated onto Murashige and Skoog (MS) medium containing B5 vitamins supplemented with 0.44 μM benzyladenine (BA), 0.054 μM α-naphthaleneacetic acid (NAA), 2.89 μM gibberellic acid (GA₃), 0.02 μM triiodobenzoic acid (TIBA), and 0.011 μM triacontanol (TRIA). Callus initiation from hypocotyl explants was obtained on MS medium supplemented with 1.07–2.14 μM NAA and 10.2 μM BA. In 3 months, somatic embryos were produced when callus was inoculated onto MS medium supplemented with 4.44 μM BA, 40 μM AgNO₃, and 0.011 μM TRIA. Somatic embryos were efficiently regenerated on MS basal solid and liquid media supplemented with 0.44–4.4 μM BA, 0.54–2.69 μM NAA, 4.92 μM 2iP, 2.1 μM calcium d-pantothenate, 0.21 μM biotin, 227.7 μM cysteine HCl monohydrate, and 108.6 μM adenine sulfate. Agrobacterium tumefaciens GV 3101 harboring pCAMBIA 1305.2 binary vector-mediated stable transformation of somatic embryos exhibited a transformation frequency of 2.56%. As somatic embryogenesis in any perennial system is useful in terms of both commercial and scientific nature, this somatic embryo-based transformation protocol for the commercially important dye-yielding tropical plant B. orellana is useful for its improvement through genetic engineering.     

<Emphasis Type="Italic">Erysiphe patagoniaca</Emphasis>: a new species of <Emphasis Type="Italic">Erysiphe </Emphasis>sect. <Emphasis Type="Italic">Uncinula </Emphasis>from Patagonia,Argentina

 A new species of Erysiphe sect. Uncinula is described and illustrated from Patagonia, Argentina. Erysiphe patagoniaca sp. nov., found on leaves of Nothofagus × antarctica, is similar to E. nothofagi and E. kenjiana, but differs in its appendages being twisted throughout their length and the number of appendages, asci, and ascospores. The two endemic species of Erysiphe sect. Uncinula, E. magellanica and E. nothofagi, coexisted on the same leaves together with Erysiphe patagoniaca. Received: September 19, 2002 / Accepted: November 28, 2002 Acknowledgments The authors are grateful to Ms. Seiko Niinomi for providing the micrographs of ascomata of Erysiphe spp. on Nothofagus. Correspondence to:S. Takamatsu     

Mercury Tolerance of Thermophilic <Emphasis Type="Italic">Bacillus</Emphasis> sp. and <Emphasis Type="Italic">Ureibacillus</Emphasis> sp

Although resistance of microorganisms to Hg(II) salts has been widely investigated and resistant strains have been reported from many eubacterial genera, there are few reports of mercuric ion resistance in extremophilic microorganisms. Moderately thermophilic mercury resistant bacteria were selected by growth at 62 °C on Luria agar containing HgCl₂. Sequence analysis of 16S rRNA genes of two isolates showed the closest matches to be with Bacillus pallidus and Ureibacillus thermosphaericus. Minimum inhibitory concentration (MIC) values for HgCl₂ were 80 μg/ml and 30 μg/ml for these isolates, respectively, compared to 10 μg/ml for B. pallidus H12 DSM3670, a mercury-sensitive control. The best-characterised mercury-resistant Bacillus strain, B. cereus RC607, had an MIC of 60 μg/ml. The new isolates had negligible mercuric reductase activity but removed Hg from the medium by the formation of a black precipitate, identified as HgS by X-ray powder diffraction analysis. No volatile H₂S was detected in the headspace of cultures in the absence or presence of Hg²⁺, and it is suggested that a new mechanism of Hg tolerance, based on the production of non-volatile thiol species, may have potential for decontamination of solutions containing Hg²⁺ without production of toxic volatile H₂S.     

<Emphasis Type="Italic">Neoskrjabinolepis nuda</Emphasis> n. sp. from shrews on Sakhalin Island,Russia, with a taxonomic review of <Emphasis Type="Italic">Neoskrjabinolepis</Emphasis> Spasskii, 1947 (Cestoda: Cyclophyllidea: Hymenolepididae)

Neoskrjabinolepis (Neoskrjabinolepidoides) nuda n. sp. is described from the shrews Sorex unguiculatus (type-host), S. gracillimus, S. isodon and S. caecutiens on Sakhalin Island, Russia. The new species is characterised by: rostellar hooks 40–44 μm long and provided with small epiphyseal thickening of the handle; a long (95–100 μm) cirrus consisting of basal region with claw-shaped spines, a parabasal region with thin needle-shaped spines and an unarmed distal region; a cirrus-sac extending well into the median field; and 15–22 eggs per gravid uterus. A review of the species of Neoskrjabinolepis Spasskii, 1947 is presented. Currently, this genus includes nine species and is divided in two subgenera on the basis of strobilar development, which is gradual in the subgenus Neoskrjabinolepis (four species) and serial in the subgenus Neoskrjabinolepidoides Kornienko, Gulyaev & Mel’nikova, 2006 (five species). An amended generic diagnosis and an identification key to Neoskrjabinolepis spp. are presented.     

Shooting of sporidium by "gun" cells in <Emphasis Type="Italic">Haptoglossa heterospora</Emphasis> and <Emphasis Type="Italic">H. zoospora</Emphasis> and secondary zoospore formation in <Emphasis Type="Italic">H. zoospora</Emphasis>

Haptoglossa spp. (Lagenidiales, Oomycetes) have been known to parasitize microscopic animals by means of a "gun" cell that shoots an infection cell, named the sporidium, into the body of the animal. A thallus grown from the sporidium changes into a zoosporangium at maturation to produce a number of zoospores that encyst after a swarming period, and the resulting cysts germinate to produce gun cells. In Haptoglossa zoospora, endoparasitic in nematodes, the cysts of primary zoospores that swam for about 5 min did not develop gun cells but produced secondary zoospores that swam for about 3 h. After encystment of the secondary zoospores, each secondary cyst germinated to produce a gun cell. In the present study, the secondary zoospores of the genus Haptoglossa could be recorded with a videocassette recorder for the first time. The videocassette recording also revealed the infection of a nematodes by H. zoospora and H. heterospora to be composed of two steps of injection of a sporidium by the gun cell, in which the gun cell came in contact with the cuticle of a nematode and produced a spherical adhesorium on the tip of the cell in 0.07–0.1 s in both species. The adhesorium was ∼2 μm in H. zoospora and ∼4 μm in H. heterospora. When the adhesorium infiated to full size, it shot the sporidium into the nematode's body in 0.5–0.65 s and in 0.2–0.5 (or rarely 1.0) s in H. zoospora and H. heterospora, respectively. After shooting, the empty gun cell with an empty cyst case was separated from the cuticle immediately in both species. Received: October 3, 2001 / Accepted: December 13, 2001     

<Emphasis Type="Italic">Kazachstania intestinalis</Emphasis> sp. nov., an ascosporogenous yeast from the gut of passalid beetle <Emphasis Type="Italic">Odontotaenius disjunctus</Emphasis>

Three ascosporogenous yeast strains were isolated from the gut of the passalid beetle, Odontotaenius disjunctus, inhabiting on rotten oak trees. DNA sequence comparison and other taxonomic characteristics identified the strains as a novel species in the genus Kazachstania. The name Kazachstania intestinalis sp. nov. (type strain EH085^T = ATCC MYA-4658^T = CBS 11839^T) is proposed for the strains. The yeast is homothallic, producing persistent asci with 1–4 spheroidal ascospores. Molecular phylogeny from ribosomal RNA gene sequences placed this novel species on the basal lineage of a clade including Kazachstania lodderae, Kazachstania exigua, Kazachstania martiniae, and other related Kazachstania spp., but none of those species was a close sister to K. intestinalis.     

Three new <Emphasis Type="Italic">Tomentella</Emphasis> species from West Africa identified by anatomical and molecular data

We used a combination of molecular-phylogenetic inference of 82 ITS rDNA sequences and anatomical approach to describe three new west African thelephoroid species, namely Tomentella afrostuposa, T. guineensis and T. guinkoi. Anatomically, T. afrostuposa is reminiscent of T. stuposa with globose to broadly ellipsoid large basidiospores of 8–14 μm, long aculei of up to 3 μm and prominent apiculi of 2 μm width. Molecular-phylogenetically, it falls within the T. stuposa complex. However, T. afrostuposa deviates by at least 7.80–10.74% from T. stuposa in regard with the ITS rDNA sequences. Tomentella guineensis is characterised by long (up to 85 μm) utriform basidia, the presence of reniform basidiospores in lateral view (up to 9 μm) with aculei not exceeding 1 μm and a strong cyanescent reaction of the subhymenial hyphae and basidia in 2.5% KOH. It forms a sister species of the newly described species Tomentella maroana; however, deviating from the last species by at least 9.75–10.04%. The very short, inflated (up to 14 μm) and thick-walled septate (septa up to 1.5 μm) subhymenial hyphae combined with ellipsoid basidiospores (up to 8 μm) and short aculei not exceeding 0.5 μm characterise Tomentella guinkoi. Anatomically, T. guinkoi recalls T. ellisii. Genetic distance between both species ranges from 12.67 to 13.73% according to ITS rDNA sequences analyses. Tomentella guinkoi forms a sister species of the group composed of T. ellisii, T. hjortstamiana and T. pisoniae. Detailed anatomical comparisons between the newly described species and their close relatives are given.     

Revision of <Emphasis Type="Italic">Trimmatothele</Emphasis> (Verrucariaceae), and description of <Emphasis Type="Italic">Oevstedalia</Emphasis> for <Emphasis Type="Italic">Trimmatothelopsis antarctica</Emphasis>, a new lichen genus with true ascoconidia

The type species of Trimmatothele is shown to differ from Verrucaria only by the multispored asci. Trimmatothele is consequently regarded as a younger synonym of Verrucaria, and the new combination Verrucaria perquisita proposed. The species is just known from Europe, where it is very rare on calcareous rocks and has a boreo-alpine distribution. Trimmatothelopsis antarctica is a very distinct lichen, not related with Verrucaria, and the new genus Oevstedalia is described to accommodate it. Oevstedalia is characterized by relatively pale perithecia with a hyphal wall, abundant pseudoparaphyses, sparse, thin, basally branched and anastomosed interascal filaments, an I− and K/I− centre, and very large, subcylindrical, entirely thin-walled, K/I+ blue asci containing in the living state 8 conidial balls filled with numerous ascoconidia. Oevstedalia might be related to the Epigloeaceae (family inc. sedis) and represents the first example of a lichenized ascomycete forming true ascoconidia. O. antarctica appears to be endemic in Antarctica, where it seems to be widespread and relatively common. No material is available for Trimmatothele umbellulariae and Lesdainea maritima, and no final conclusion can be given for the taxonomic status of these two species.