Ultrastructure of the ascus apical apparatus of Encoelia furfuracea (Helotiales)

The ultrastructural characteristics of the ascus apical apparatus of Encoelia furfuracea are described by means of transmission electron microscopy. The structure of its ascus apex is considerably different from that of E. tiliacea and E. fimbriata, and from other members of Helotiales in which these structures have been studied. Remarkable interspecific differences in the ascus apical apparatus provide additional data indicating heterogeneity of the genus.     

On the ascus types in thePhysciaceae (Lecanorales)

Comparative studies in thePhysciaceae revealed that characters of the apical apparatus of asci differ between the various genera. Two major ascus types are observable in this family. They are found to correspond with certain ascospore types.     

New fungal genera, Tectonidula gen. nov. for Calosphaeria-like fungi with holoblastic-denticulate conidiogenesis and Natantiella gen. nov. for three species segregated from Ceratostomella

Two morphologically similar groups of ascomycetes with globose to subglobose perithecia, elongate necks, unitunicate asci floating freely at maturity, and hyaline ascospores currently placed in Calosphaeria s. lat. and Ceratostomella s. lat., respectively, are studied. The Calosphaeria-like fungi have groups of perithecia growing between cortex and wood, arranged in circular groups with converging necks and piercing the cortex in a common point; the asci with a shallow apical ring and U- to horseshoe-shaped hyaline ascospores are compared with Calosphaeria pulchella, the type species of the genus. Conidiogenesis of the investigated Calosphaeria-like fungi is holoblastic-denticulate; ramichloridium-like and sporothrix-like conidiophores and conidia were formed in vitro. Ascospore and ascus morphology, structure of the ascal apex, ascogenous system, mode of conidiogenesis and the large subunit rRNA sequences of this group differ considerably from C. pulchella and both groups are unrelated. Thus a new genus, Tectonidula, is described with two accepted species, T. hippocrepida and T. fagi; they are separated by ascospore and ascus morphology and holoblastic-denticulate conidiogenesis from the core species of Calosphaeria. The placement of Tectonidula among perithecial ascomycetes is discussed. The relationship of Tectonidula with Barbatosphaeria and two ramichloridium-like hyphomycetous genera Rhodoveronaea and Myrmecridium is investigated. Three species formerly attributed to Ceratostomella are studied. The revision of the herbarium type specimen and fresh material of Ceratostomella ligneola revealed that it is conspecific with Ceratostomella ampullasca and Ceratostomella similis. The LSU phylogeny clearly separated C. ligneola from Ceratostomella s. str. and morphologically similar Lentomitella. On the basis of molecular sequence data and detailed comparison of morphology of asci, ascospores and ascogenous system the genus Natantiella is described for C. ligneola with C. ampullasca and C. similis as its synonyms. Natantiella produced sterile mycelium in vitro.     

Division spindle and centrosomal plaques during mitosis and meiosis in some Ascomycetes

The behaviour of the division spindle and centrosomal plaques is described in four species of Ascomycetes (Ascobolus immersus, Ascobolus stercorarius, Podospora anserina and Podospora setosa) studied by light and electron microscopy. Two unique features of the kinetical apparatus were observed: presence of centrosomal plaques and intranuclear location of the spindle. In all types of mitoses (mycelium, crosier and postmeiotical mitosis) the apparatus is structurally identical to that found in meiosis. The centrosomal plaques, present in all divisions, are always contiguous with the nuclear envelope and never show centrioles similar to those commonly found in Metazoa and Protozoa. During metaphase and anaphase the plaque is constituted of two zones situated on each side of the nuclear envelope: an electron opaque outer zone and inner one less opaque in which most of the microtubules end. In Podospora the outer zone appears in sections as consisting of two dark layers separated by a clear one. Two dispositions of plaques are possible: either they are entirely contiguous with the nuclear envelope (Ascobolus) or only partially so, the remainder being perpendicular to the nuclear envelope (Podospora). — The localisation of the plaques in the ascus was determined by light and electron microscopy. The nuclear envelope was shown to remain intact during division. It was possible to observe that the sporal wall of each spore originated from the same unique double membrane formed in the ascus during the meiotic second division and postmeiotical mitosis. This fact is of genetical interest for the study of morphological and physiological characters of the spores.     

Female germ unit in Genlisea and Utricularia, with remarks about the evolution of the extra-ovular female gametophyte in members of Lentibulariaceae

Lentibulariaceae is the largest family among carnivorous plants which displays not only an unusual morphology and anatomy but also the special evolution of its embryological characteristics. It has previously been reported by authors that Utricularia species lack a filiform apparatus in the synergids. The main purposes of this study were to determine whether a filiform apparatus occurs in the synergids of Utricularia and its sister genus Genlisea, and to compare the female germ unit in these genera. The present studies clearly show that synergids in both genera possess a filiform apparatus; however, it seems that Utricularia quelchii synergids have a simpler structure compared to Genlisea aurea and other typical angiosperms. The synergids are located at the terminal position in the embryo sacs of Pinguicula, Genlisea and were probably also located in that position in common Utricularia ancestor. This ancestral characteristic still occurs in some species from the Bivalvaria subgenus. An embryo sac, which grows out beyond the limit of the integument and has contact with nutritive tissue, appeared independently in different Utricularia lineages and as a consequence of this, the egg apparatus changes position from apical to lateral.     

Euopsis and Harpidium,Genera of the Lichinaceae (Lichenes) with Rostrate Asci*

Based on corresponding ascocarp ontogeny and thallus structure, the genera Euopsis and Harpidium are included in the family Lichinaceae. In the two species of Euopsis, E. granatina and E. pulvinata, the apothecia develop from ascogonia in generative tissue, while in H. rutilans they are pycnoascocarps. In thallus anatomy, the species of Euopsis resemble Pyrenopsis haematopsis and allied species, while H. rutilans corresponds in structure and development of the thallus and apothecia to Pyrenopsis haemaleella (syn. P. sphinctotricha). H. rutilans is the first member of Lichinaceae known to have only a green algal symbiont. In E. granatina, two phycobionts are always present, a species of Gloeocapsa and a chlorococcalean alga. In Euopsis and Harpidium, the ascus wall is composed of an outer, non-expansible and an inner, expansible layer; the latter surrounds the protoplast as an amyloid collar, which expands during spore release into a long, tapering rostrum. In Euopsis, the outer wall layer is strongly amyloid and the upper part separated from the expanded amyloid rostrum by a non-amyloid zone, appearing like a slit in LM studies. The ultrastructure and function of the ascus in E. granatina has been studied in TEM and is interpreted as functionally unitunicate-rostrate. Unitunicate asci with short rostrum are described for P. haemaleella and P. haematopsis.     

ASCI OF THE PEZIZALES III: THE APICAL APPARATUS OF EUGYMNOHYMENIAL REPRESENTATIVES

Morphological, developmental and cytochemical examinations were made with light and electron microscopy on the apical apparatuses of four eugymnohymenial species, Pyronema domesticum, Ascodesmis sphaerospora, Coprotus winteri and C. lacteus. Ascal tips in all four species were notably thinner walled than the rest of the ascus. Ultrastructurally, demarcation of the opercula was enhanced after staining with silver methenamine. Wide zones of dehiscence are formed in the outer layer of A. sphaerospora and C. winteri. In P. domesticum the outer layer of the operculum is differentially stained from the rest of the ascal wall. Wall dimensions in the multispored asci of C. winteri are approximately three times greater than those in the eight-spored asci of C. lacteus. The shape of the apical apparatus of C. lateus is almost identical to that of C. winteri. Morphological and cytochemical similarities of the apical apparatuses in Ascodesmis, Pyronema and Coprotus help demonstrate greater relationship between these taxa and support the belief that these taxa are most closely related to members of the Otideaceae and Aleuriaceae.     

Two new species of pyrenomycetous ascomycetes from New Caledonia

Two new species of Pyrenomycetes from forest soil in New Caledonia,Anthostomella pacifica andChaetomium novaecaledonicum, are described and illustrated.Anthostomella pacifica is characterized by non-ostiolate ascomata, cylindrical asci with an amyloid apical apparatus, and two-celled ascospores (dark apical cylindrical and hyaline basal dwarfed cells) with longitudinal germ slits.Chaetomium novae-caledonicum is characterized by ostiolate ascomata, straight terminal hairs, arcuate lateral hairs with a recurved tip, and very small, ovoid-flattened ascospores.This research was supported in part by Monbusho International Scientific Research Program: Field Research, No. 05041093.     

Cephalodiate Arten der GattungLecidea sensu lato (Ascomycetes lichenisati)

(1) The ability to produce cephalodia is usually a genus-specific character in lichens. (2)Lecidea shushanii Thoms., is a member of the genusTephromela, closely related toT. aglaea. It is not clear, whether or not the cephalodia of this taxon are true cephalodia or colonies of epiphytic cyanobacteria and whether or notLecidea shushanii is an independent species. (3)Lecidea dovrensis Nyl., is, in contrast to the traditional concept, not conspecific withLecidea alpestris Sommerf., but an earlier name forLecidea pallida Th. Fr. (4)Lecidea dovrensis is described in some detail. Chemically the species is characterized by the presence of isousnic acid (previously unknown in lecideoid lichens). It is restricted to areas north of the 60th parallel with an oceanic climate. (5) In connection with the attempt to clarify the taxonomic relationships ofLecidea dovrensis, figures of ascus apical structures of the following species are given (marked by an asterisk are genera where we found discrepancies with published data):Austrolecia antarctica, Catillaria chalybeia, Lecidea alpestris, L. caesioatra, L. limosa, Lecidoma demissum, Koerberiella wimmeriana, Micarea assimilata, M. crassipes, M. melaenida, M. prasina, Pilophorus robustus, Placodiella olivacea, Placolecis opaca, Porpidia trullisata, Protoblastenia rupestris, Psilolechia lucida, Psorula rufonigra, Squamarina gypsacea, Xanthopsorella texana. (6) Among crustaceous lichens we find no groups related toLecidea dovrensis. We supportTimal's concept of including this species in the genusPilophorus. Pilophorus, as well asLecidea dovrensis is characterized by the same ascus type, by a similar structure of thallus, cephalodia, paraphyses, and ascocarp (although there is no pseudopodetium developed inLecidea dovrensis), and the presence of isousnic acid. In addition, both taxa are restricted to cool oceanic climates and non-calciferous substrates. The following combination is proposed:Pilophorus dovrensis (Nyl.)Timdal, Hertel & Rambold, comb. nova. (7) The species of theLecidea alpestris-group form an independent genus, probably near toAustrolecia Hertel.

Frau Prof. Dr.Elisabeth Tschermak-Woess zu ihrem 70. Geburtstag gewidmet.     

Genetic variation in the orientation of nuclear spindles during the development of asci in Neurospora

The effect of a single mutant gene, peak-2, in Neurospora crassa is to alter the ascus from one in which the eight spores are linearly arranged to one in which the eight spores are arranged non-linearly. The same gene transferred to N. tetrasperma determines an ascus similar to that of the mutant N. crassa but in striking contrast to the normal ascus for N. tetrasperma, which includes four heterokaryotic spores, linearly arranged. A comparative cytological study has been made on asci from both species, including material from zygotes that are homozygous normal, heterozygous for peak-2, and homozygous for peak-2. The observed morphogenetic differences are closely related to differences in nuclear spindle orientation, which in turn are correlated with the geometry of the cell. The observations are shown to bear on the basis of pseudohomothallism in N. tetrasperma. Speculations on the importance of the cell wall in relation to spindle orientation are presented. Differences of expression of peak-2 in the different species are discussed.     

Clavispora,a new yeast genus of the Saccharomycetales

In Candida lusitaniae van Uden et do Carmo-Sousa (1959), strains of opposite sex have been found. Cells of the opposite mating types conjugate and form asci with one to four clavate spores. These are easily liberated from the ascus. The type strain of Candida obtusa (Dietrichson) van Uden et do Carmo-Sousa ex van Uden et Buckley (1970) also produces ascospores after mating with one of the strains of Candida lusitaniae. As clavate ascospores are unknow in yeasts, a new genus, Clavispora, is proposed with Clavispora lusitaniae spec. nov. as type species.     

Ascospore formation and morphology in two species of the genusErysiphe remaining immature on the living host plant

Erysiphe cumminsiana andE. galeopsidis, which have immature asci in the current season, have been recorded from Japan, but the ascospores of both fungi have not been described. In the present experiments, some observations before and after overwintering were made on the cleistothecia ofE. cumminsiana on three species ofCacalia and two species ofLigularia, andE. galeopsidis onGeranium thunbergii. After overwintering, the former fungus developed six to eight, rarely four spores in an ascus and the latter fungus always four spores in an ascus. Their teleomorphic characteristics including those of ascospores are also described.     

Pleurosigma hinzianum sp. nov. and P. frenguellianum sp. nov. (Pleurosigmataceae,Bacillariophyta) from Argentinean coastal waters,in comparison with P. amara Stidolph and P. elongatum W. Smith

Two new marine diatom species from Argentinean coastal waters, Pleurosigma hinzianum Sterrenburg, Sunesen & Sar, sp. nov. and P. frenguellianum Sunesen, Sterrenburg & Sar, sp. nov., are described. The characters permitting their identification are specified, based on comparison with type material of the morphologically similar species P. amara Stidolph and P. elongatum W. Smith in the light (LM) and scanning electron microscope (SEM). New information on the type material of P. elongatum is presented and its taxonomic concept is emended. The main criteria for separation of the species discussed here are: valve outline, path of the raphe-sternum, raphe angle and angle of intersection of the oblique striae in LM; and external central and terminal raphe fissures, internal details of the central area, external and internal apical structure, and morphology of the hymen-occluded internal pores in SEM. The occurrence of these (and probably other) species in the genus Pleurosigma in net samples is adventitious and not indicative of a true planktonic mode of life.     

Further scanning electron microscope studies of lizard auditory papillae

The papillae basilares of 12 species of lizards from seven different families were studied by SEM. The iguanids, Sceloporus magister and S. occidentalis, have typical “iguanid type” papillae with central short-ciliated unidirectional hair cell segments and apical and basal long-ciliated bidirectional hair cell segments. These species of Sceloporus are unique among iguanids in that the bidirectional segments consist of but two rows of hair cells. The agamids, Agama agama and Calotes nigrolabius, have an “agamid-anguid type” papilla consisting of an apical short-ciliated unidirectional hair cell segment and a longer basal bidirectional segment. Agama agama is unusual in having a few long-ciliated hair cells at the apical end of the apical short-ciliated segment. The agamid, Uromastix sp., has an “iguanid type” papilla with a central short-ciliated unidirectional segment and apical and basal bidirectional segments. The anguid, Ophisaurus ventralis, has an “iguanid” papillar pattern with the short-ciliated segment centrally located. All the short-ciliated hair cells of the above species are covered by a limbus-attached tectorial network or cap and the long-ciliated hair cells, only by loose tectorial strands. The lacertids, Lacerta viridis and L. galloti, have papillae divided into two separate segments. The shorter apical segment consists of opposingly oriented, widely separated short-ciliated cells covered by a heavy tectorial membrane. The apical portion of the longer basal segment consists of unidirectionally oriented hair cells, while the greater part of the segment has opposingly oriented hair cells. The xantusiids, Xantusia vigilis and X. henshawi, have papillae made up of separate small apical segments and elongated basal segments. The apical hair cells are largely, but not exclusively, unidirectional and are covered by a heavy tectorial cap. The basal strip is bidirectional and the hair cells are covered by sallets. The kinocilial heads are arrowhead-shaped. The papilla of the cordylid, Cordylus jonesii, is very similar to that of Xantusia except that the apical segment is not completely separated from the basal strip. The papilla of the Varanus bengalensis is divided into a shorter apical and a longer basal segment. The hair cells of the entire apical and the basal three quarters of the basal segment are opposingly oriented, not with reference to the midpapillary axis but randomly to either the neural or abneural direction. The apical quarter of the basal segment contains unidirectional, abneurally oriented hair cells. The entire papilla is covered by a dense tectorial membrane. The functional correlations of the above structural variables are discussed.     

Comparative morphology of surface ultrastructure of diphyllobothriidean tapeworms (Cestoda: Diphyllobothriidea)

Microtriches on the scolices and adjacent strobila of seven species of diphyllobothriidean cestodes (Bothridium pithonis, Cephalochlamys namaquensis, Dibothriocephalus latus, Duthiersia expansa, D. fimbriata, Ligula intestinalis, and Schistocephalus solidus) from different hosts (frogs, snakes, lizards, birds, and mammals) and biogeographic areas were examined using scanning and transmission electron microscopy. The basic structure of the tegument of the seven species studied does not differ markedly from that found in other cestodes. The main characteristic is the presence of electron‐dense bodies and vesicles in the distal cytoplasm. However, this study has shown differences in the morphology of microtriches even among species of the same family. Two different types of microtriches were found, filitriches and spinitriches, with the latter represented by two forms. Our study reveals that capilliform filitriches are most commonly found in Diphyllobothriidea. They were observed mainly on the strobila and the scolices of all but one studied species; individuals of L. intestinalis bore only coniform spinitriches on their surface. The same type of microtriches was found on the cirrus in D. latus. Gladiate spinitriches covered the scolex in both species of Duthiersia, and gladiate spinitriches interspersed with capilliform filitriches were observed on the anterior part of the strobila in D. fimbriata and the posterior part of the scolex in B. pithonis. Individuals of C. namaquensis were covered only by small acicular filitriches. No obvious pattern in the type and distribution of microtriches was observed among species that belong to different families and parasitize distantly related definitive hosts.     

Host Associations Between Fungal Root Endophytes and Boreal Trees

Fungal root endophytes colonize root tissue concomitantly with mycorrhizal fungi, but their identities and host preferences are largely unknown. We cultured fungal endophytes from surface-sterilized Cenococcum geophilum ectomycorrhizae of Betula papyrifera, Abies balsamea, and Picea glauca from two boreal sites in eastern Canada. Isolates were initially grouped on the basis of cultural morphology and then identified by internal transcribed spacer ribosomal DNA sequencing or by PCR restriction fragment length polymorphism. Phylogenetic analysis of the sequence data revealed 31 distinct phylotypes among the isolates, comprising mainly members of the ascomycete families Helotiaceae, Dermateaceae, Myxotrichaceae, and Hyaloscyphaceae, although other fungi were also isolated. Multivariate analyses indicate a clear separation among the endophyte communities colonizing each host tree species. Some phylotypes were evenly distributed across the roots of all three host species, some were found preferentially on particular hosts, and others were isolated from single hosts only. The results indicate that fungal root endophytes of boreal trees are not randomly distributed, but instead form relatively distinct assemblages on different host tree species.     

Encystment Processes and the “Matrioshka-like Stage” in a Moss-dwelling and in a Limnic Species of Eutardigrades (Tardigrada)

Tardigrades have two forms of dormancy, namely cryptobiosis and encystment. The encystment is a form of diapause known for a limited number of species of tardigrades and still little studied. To increase the knowledge on encystment, two species of eutardigrades from Italy have been considered: the moss-dwelling Amphibolus volubilis (Eohypsibiidae), and the limnic Dactylobiotus parthenogeneticus (Murrayidae). Cysts have been collected in nature, or induced under laboratory conditions. In the latter case, it was possible to follow the several steps of encystment processes. Two different types of cyst (“type 1” and “type 2”) have been found in A. volubilis, while in D. parthenogeneticus only one type has been found. In general, the ovoid-shaped cysts are constituted by a series of cuticles surrounding the animals and resemble an onion or a Matrioshka Russian doll. In all three types of cyst, the encystment processes show both common and peculiar traits. Encystment begins with the discharging of the sclerified parts of the buccal-pharyngeal apparatus, as in the molting process, but without the loss of the old cuticle. Then, two or three new cuticles are serially synthesized, according to cyst type. In A. volubilis, the ultrastructure of these new cuticular involucra is similar to that of non-encysted animal cuticles, while in D. parthenogeneticus the ultrastructure of the new cuticular involucra differs from that of non-encysted animal cuticle. A modified buccal-pharyngeal apparatus has been observed both in “type 2” cyst of A. volubilis and in the D. parthenogeneticus cyst.