Apothecia Research Articles

Mating system and role of pycnidiospores in biology of Polystigma amygdalinum , the causal agent of almond red leaf blotch

Polystigma amygdalinum is a serious leaf pathogen of almonds. Ascospores are believed to be the only inoculum of this pathogen. Ascocarp initials develop in infected leaves in contact with, or in close proximity to, filimentous spore-bearing bodies, which are the first to appear. While the aseptate filiform spores (scolecospores), are suspected of involvement in the sexual cycle of P. amygdalinum , there is no direct evidence of this. In order to study the role of the scolecospores, infected almond leaves, collected during the period from first appearance of symptoms to perithecium development, were sectioned by freezing microtome and the developmental anatomy was examined. Since ascogonia were not evident prior to the appearance of scolecospores, it was concluded that these were part of the sexual cycle of P. amygdalinum. This is supported by observations that perithecium initials always developed beside scolecospore fruit bodies, scolecospores failed to germinate in vitro , and almond leaves inoculated with them never became infected. We conclude that the filiform spores are spermatia essential to the sexual process and not asexual spores. Moreover, P. amygdalinum is likely to be homothallic, because single lesions in leaves infected by P. amygdalinum , assumed to have arisen from single ascospore infections, produced perithecia as abundantly as occurred in coalescing lesions caused by multiple ascospore infections.

Growth and development of ascomata in two members of the British Graphidaceae (lichenized Ascomycota; Ostropales)

Growth and development of ascomata in two members of the British <i>Graphidaceae</i> (lichenized Ascomycota; <i>Ostropales</i>)

FIRST REPORT OF PLEOSPORA TARDA CAUSING LEAF SPOT OF LEPIDIUM DRABA IN IRAN

Since 2009, leaf spot symptoms were observed on Lepidium draba (whitetop) an invasive weed collected in wheat plots (Triticum sp.) in the Hamadan Province of Iran. Typical symptoms included chlorosis of the basal leaves with black discrete ascomata protruding from the leaf surface. Colonies of a fungus recovered from diseases leaves were brown to olivaceous with a white aerial mycelium in the center. Since the morphological characters of this mycete were consistent with the description of Pleospora tarda (Simmons, 1985), its identification was confirmed by amplifying and sequencing the internal transcribed spacer region (ITS) of a representative isolate (GenBank accession No. KC660992). To fulfill Koch’s postulates, a spore suspension (105 ascospores/ml) of the fungus was sprayed onto leaves of whitetop seedlings at the 4-6 leaf stage. Inoculated seedlings were maintained in a growth chamber at 25°C, with more than 75% relative humidity and a 12 h light photoperiod. Inoculated plants showed symptoms like those observed in the field 14 days post inoculation. P. tarda was reisolated from inoculated plants and a culture was deposited in the Iranian Research Institute of Plant Protection with the accession code IRAN 2184C. To our knowledge, this is the first report of P. tarda infecting whitetop in Iran and in the world.

FIRST REPORT OF WHITE MOULD CAUSED BY SCLEROTINIA SCLEROTIORUM ON MARIGOLD (TAGETES ERECTA) IN BANGLADESH

Marigold (Tagetes erecta) is one of the commercial ornamental flowers commonly grown in Bangladesh. In January 2011, rotten flowers of marigold covered with fluffy white mycelia were found in Rangpur, Bangladesh. Symptoms began from petals and subsequently expanded to the whole flower and the lower part of the plant. Dark brown lesions with necrotic tissues appeared on the infected leaves and stems. Seriously infected plants showed flower dropping and branch wilting. A fungus showing hyaline and septate mycelia was isolated from the infected tissue using potato dextrose agar. The fungus formed round to irregular-shaped sclerotia. Apothecia development from sclerotia were also observed on moist sand. The partial sequence (531 bp, GenBank accession No. AB937102) of the internal transcribed spacer region of rDNA was determined by the method described in Rahman et al. (2015). The result showed a 100% identity with those of Sclerotinia sclerotiorum, which is known as a pathogen to various plants (Purdy, 1979). Pathogenicity of the isolate to marigold was examined by detached leave and flower assays using the mycelial block. After incubation at 20°C with 90% relative humidity in 12 h light/12 h dark conditions, all inoculated leaves and flowers were rotten, while no symptom developed on the controls. The fungus was re-isolated from the symptomatic tissues and Koch’s postulates were confirmed. In Bangladesh, the fungus is becoming an emerging threat to various hosts (e.g. Rahman et al., 2015). To our best knowledge, this is the first report of S. sclerotiorum infection on marigold in the country.

First report of stem and pod blight of hyacinth bean caused by Sclerotinia sclerotiorum in Bangladesh.

Hyacinth bean (Lablab purpureus) is an almost year round vegetable crop in Bangladesh. During January 2011, while surveying hyacinth bean fields in the Gazipur district, plants with light tan to brown blighted stems and pods were observed. Within the pod pith, large dark irregular sclerotia embedded in a white fluffy mycelium were found. Symptomatic tissues were excised, surface-sterilized in 0.5% sodium hypochlorite and placed on potato dextrose agar (PDA). Isolated fungal colonies consistently yielded white mycelium and produced a ring of large sclerotia near the edge of PDA plates. Under microscope, the hyphae were hyaline, branched and multinucleate. Sclerotia were induced to produce apothecia, asci and ascospores following a conditioning process (Smith and Boland, 1989). Based on morphology, the fungus was assumed to be Sclerotinia sclerotiorum. Pathogenicity of the fungal isolate was proven by placing seven-day-old PDA mycelial plugs (5 mm) on hyacinth bean stems. Seven days post inoculation, all inoculated plants started wilting and stem breakage were also observed for some dead plants. Dark sclerotia were found after splitting the dead stems of all inoculated plants. Identification of the fungus was confirmed by comparing the sequences generated from the internal transcribed spacer (ITS) region of the ribosomal DNA (ITS1 and ITS4 primers) (White et al., 1990). The nucleotide sequence was deposited in GenBank as accession No. KF791510. These sequences shared 100% nucleotide similarity with those of S. sclerotiorum. This pathogen has previously been reported on different crops (Huang et al., 2005). To our knowledge, this is the first report of S. sclerotiorum on hyacinth bean in Bangladesh.

Non-developing ascospores in apothecia of asexually reproducing lichen-forming fungi.

The presence of apothecia in mixed species (vegetatively reproducing lichens, occasionally producing ascomata) has been interpreted as a mechanism to increase genetic variability in mostly clonal populations. However, spore viability from these apothecia has not been studied. We asked whether ascospores of the mixed species Physconia grisea are viable and thereby contribute to increasing the genetic diversity within populations of this species. An ontogenetic study of spores in cultures of P. grisea and a related sexual species (P. distorta), showed that although mature apothecia from both species produced and discharged meiospores capable of germination, spores from P. grisea were only rarely (0.43%) able to continue development whereas those from P. distorta germinated and developed successfully. The strongly reduced viability of P. grisea spores suggested that they do not have a strong reproductive function, at least in the two local populations analyzed. Additionally, we show that the segregation of Physconia grisea ssp. lilacina does not have molecular support.

Reducing Primary Inoculum of Apple Scab Using Foliar Application of Urea in Autumn

The effect of applications of urea 5% after harvest but before leaf-fall, as foliarapplication, in order to restrict perithecial production by Venturia inaequalis in a commercial superintensive apple orchard situated near Cluj-Napoca, Romania was studied. The urea works in two ways:directly by inhibiting the development of ascospores, and by stimulating the development ofantagonistic microorganisms from against V. inaequalis. There are several possibilities to reduce theamount of overwintering apple scab, such as removing fallen leaves by raking, vacuuming, or usingleaf blowers, chopping or shredding the leaf litter in the spring. Primary infection of scab conidia isproduced by infected leaf litter on the orchard floor. Any modalities that reduce the amount of leaflitter will help to reduce the amount of inoculum for primary infection with scab. Urea-treated leavesdecomposed rapidly infected leaf litter, thus destroying the overwintering substrate for the V.inaequalis fungus. The results have shown large reductions in spore production, often as high as 70 to80%, following application of 5% urea. Spraying the surface of the leaves on the ground with urea 5%reduced primary infection by about 60%. In conclusion, urea applications proved to be effective toreduce apple scab inoculum. Combining the urea with other modalities of leaf shredding provides thegreatest reductions in ascospores.

Five Apothecium-producing Lichenized Fungi of the Genus Usnea in Taiwan

Most species of the genus Usnea that commonly produce apothecia in Taiwan have not been collected for a long time. We rediscovered five species, including Usnea masudana, U. orientalis, U. pseudogatae, U. shimadae, and U. sinensis, by investigating their morphology, chemistry, and molecular data. Ten internal transcribed spacer (ITS) sequences of the five species were first reported. Distributions of the species were also provided. Although no latitudinal tendency was observed among the Usnea species in Taiwan, we consider high elevation places of the subtropics and tropics to contain plenty of Usnea species that produce apothecia.

AN OUTBREAK OF CYCLANEUSMA MINUS NEEDLE CAST ON SWISS MOUNTAIN PINE (PINUS UNCINATA) IN ITALY

In September 2010, a significant decline of Pinus uncinata was observed in a mixed forest of Larix decidua and P. cembra in Claviere (western Italian Alps). In the forest, which covered about 32 ha and was located between 1800 and 1900 m of elevation (coordinates: 44°56’05.19’’N 6°45’28.46’’E), P. uncinata accounted for 12% of the trees. The disease affected 85-90% of P. uncinata trees belonging to all diameter classes. More than 70% of the trees showed needle yellowing, but the remaining trees showed thin and transparent crowns, and needle cast that progressed from the inner to the outer parts of the twigs. No symptoms were observed on L. decidua and P. cembra. Twenty symptomatic twigs were collected from 10 randomly selected trees. Needles were placed in Petri dishes containing a sterile piece of filter paper dampened with sterile water and incubated at room temperature for 10-12 days to induce sporulation. After incubation, white to creamy, scattered, elliptical apothecia embedded in yellowed needles were observed. Apothecia produced hyaline, filiform, 2septate, smooth ascospores 80-90×2.5-3.0 μm in size. Based on these macroand micro-morphological characters, the fungus was identified as Cyclaneusma minus (Butin) Di Cosmo, Peredo et Minter (syn. Naemacyclus minor Butin). C. minus has been reported both as an endophyte on symptomless P. uncinata needles (Sieber et al., 1999) and as a pathogen on needles of several pine species (Millar and Minter, 1980). In Italy, the fungus has previously been observed on Pinus sylvestris (Gonthier et al., 2010) but, to our knowledge, this is its first report on P. uncinata in this country.

Complete genome sequence of the first endornavirus from the ascocarp of the ectomycorrhizal fungus Tuber aestivum Vittad.

The genus Endornavirus was recently approved by the International Committee on Taxonomy of Viruses (ICTV) [1, 10] to be separated from the family Partitiviridae, and endornaviruses have only been described in plants and fungi [4, 15]. They share common properties such as symptom-free infection of their host, occurrence in lowcopy number in host cells, lack of true virions [7], and an efficient vertical transmission in their hosts [16]. Endornaviruses have unique plasmid-like properties that differ markedly from those of other conventionally encapsidated viruses [5, 12, 16]. The genomes of endornaviruses consist of linear double-stranded RNA (dsRNA) with a characteristic, single open reading frame (ORF) of up to 18 kb in length, often preceded by a site-specific nick at the 5’ end [5, 7, 8, 12, 18]. The predicted protein from this ORF harbours in its N-terminal part characteristic amino acid (aa) sequence motifs for viral RNA helicases (VHel) or viral methyltransferases (VMet), while aa sequence motifs for UDP-glycosyltransferases (UGT) and RNA-dependent RNA polymerases (RdRp) are located at close proximity to the 30 end [7]. The remaining polyprotein is unrelated to any known functional proteins in the database. Currently, the INSDC databases contain complete sequences of seven endornaviruses. Three are plant-associated viruses infecting cultivated and wild rice (Oryza sativa endornavirus [OsEV] and Oryza rufipogon endornavirus [OrEV]) [5, 11], respectively), as well as broad bean (Vicia faba endornavirus [VFV]) [13]. In addition, four fungal isolates are known. One isolate each was reported from the mycelia of the violet root rot fungus (Helicobasidium mompa endornavirus 1-670 [HmEV1-670]) [12] and from an informally designated plant-pathogenic oomycete Phytophtora spp. (Phytophtora endornavirus 1 [PEV1]) [6], while two isolates originate from the pathogenic ascomycete Gremmeniella abietina (Gremmeniella abietina type B RNA virus) [15]. Here, we report the complete nucleotide sequence of an isolate of a putatively novel species in the genus Endornavirus from an ectomycorrhizal fungus, isolated from the hypogeous ascomata of a black truffle (Tuber aestivum Vittad.). Accordingly, we propose the name Tuber aestivum endornavirus (TaEV). The TaEV genome structure is highly similar to that of other Ascomycetes-infecting endornaviruses, such as Gremmeniella abietina type B RNA virus [15], containing characteristic motifs for the DEAD-like helicase (DEXDc), VMet and RdRp.

Independent Origins of Cleistocarpy in the Splachnaceae: Analyses of cpDNA Sequences and Polyphyly of the Voitioideae (Bryophyta)

Many dung mosses (Splachnaceae) are characterized by insect-mediated spore dispersal. All of the ento- mophilous species are coprophilous, whereas anemophilous species are humicolous or epiphytic. The three species of the Voitioideae are coprophilous but are distinguished from other members of the family by sporangia that remain closed (cleistocarpous) and lack a peristome. Spores are released when the sporangial wall disintegrates. Phylogenetic analyses of nucleotide sequences of the trnL-trnF region and the rps4 locus (cpDNA) for 25 species of Splachnaceae suggest that this combination of characters arose twice within the Splachnaceae and that Voitia grandis is more closely related to species of Tayloria subgenus Tayloria rather to the other species of Voitia, which are nested within the genus Tetraplodon. Hence the new combination Tayloria grandis (Long) Goffinet & Shaw is made. Although the optimal trees were left unrooted, our results resolve the Voitioideae (i.e., the genus Voitia ) as nested within the Splachnoideae. The phylogenetic significance of sporophytic characters within the family Splachnaceae is briefly discussed. Some members of the moss family Splachnaceae are unique among seedless land plants: they display fea- tures considered linked with insect-mediated spore dispersal. Such putative adaptations include an en- larged, brightly colored hypophysis, emission of vola- tile compounds that attract insects, sticky spores, and a pseudocolumella (Koponen 1990; Demidova and Fi- lin 1994). The systematics of the family is based pri- marily sporophytic features, and in particular of com- binations of entomophily related characters and the ar- chitecture of the peristome (Koponen 1982). The

New Insights into Cerebral Malaria

Cerebral malaria, defined as coma not attributable to any other cause in an individual with Plasmodium falciparum malaria, has a fatality rate ≥15%.

Mycocaliciales, a New Order for Nonlichenized Calicioid Fungi

The new order Mycocaliciales is described. It includes the nonlichenized Mycocaliciaceae and Sphinctrinaceae, which occur as saprobes or commensals on lichens. They are further characterized by having stalked or sessile ascomata consisting of sclerotized,

Two Inoperculate Discomycetes with White Hairs from Taiwan

Two inoperculate discomycetes in the genus Lachnum (Hyaloscyphaceae) with white-haired apothecia are newly reported from Taiwan. These two species are L. albidulum and L. virgineum which occurred either on leaves of broadleaf trees, stem of herbaceous plants or decayed wood. The microscopic characteristics and habitats are described and discussed in detail in order to distinguish them from each other for their morphological similarity.

Chaetosphaeria ampulliformis sp. nov. associated with a Hemicorynespora anamorph, and a key to Hemicorynespora species

The new ascomycete Chaetosphaeria ampulliformis sp. nov. found on decaying wood of an unidentified angiosperm in Taiwan is described and illustrated. It is closely associated with an Hemicorynespora anamorph which grows mostly on the necks of the flask-shaped ascomata. This anamorph is described as Hemicorynespora multiseptata sp. nov. The relationships of these two species to others in the genera are discussed, and a key to the recognized species of Hemicorynespora is provided.

Yenidoğanda Tizanidin Zehirlenmesi

Tizanidine an imidazole derivative is a novel centrally acting muscle relaxant Adult case of tizanidine over dosage is expressed as coma miosis nistagmus and respiratory depression with Cheyne Stokes respiration There is limited experience with the drug in children and there have been no reported cases of tizanidine toxicity in a neonate In this paper a 25 day old neonate with tizanidine intoxication is presented to point out the hazard of improper use of drugs in the neonatal period Key words: Neonate Intoxication Tizanidine

Coprophilous fungi in New Zealand. I. Podospora species with swollen agglutinated perithecial hairs

New Zealand coprophilous Podospora species with swollen agglutinated perithecial hairs include P. aloides, P. conica, P. curvuloides, P. dakotensis, P. glutinans, P. miniglutinans, P. tetraspora an...

A new species of Tuberales from China

A new species of Tuberales, Hydnobolites baodingensis , from Baoding, China, is described and compared with allied species. The large ascocarps of the new species are the principal distinction from other species in the genus. This is the first record of Hydnobolites from China.

Placoparmelia patagonica, a new lichen genus and species from Argentina (Parmeliaceae)

Placoparmelia patagonica Henssen gen. et sp. nov. is described from two localities in Argentina. The placodioid thallus of P. patagonica has a primitive paraplectenchymatous upper cortex in parts but is devoid of a clearly defined lower cortex, true rhizines or tufts of rhizoidal hyphae. The upper surface is minutely porous and reacts HNO, + blue-green indicating the presence of glomellifera brown. Norstictic acid with small amounts of connorstictic and cryptostictic acid, and a trace of an unknown substance were detected by TLC. The ontogeny and structure of the apothecia and pycnidia correspond to those of the Parmeliaceae. The anatomy of the thallus in P. patagonica is compared with that in Neofuscelia squamariata, N. pulla and Parmelia saxatilis.

A Method for Sectioning Ascomycete Herbarium Specimens for Light Microscopy

(1991). A Method for Sectioning Ascomycete Herbarium Specimens for Light Microscopy. Mycologia: Vol. 83, No. 4, pp. 520-524.