Multicellular Conidia Research Articles

First report of Epicoccum nigrum causing leaf spots on Campomanesia guazumifolia (Camb.) Berg in Brazil.

Campomanesia guazumifolia is a native tree that produces fruit that can be consumed fresh or used by industry (Donadio et al., 2002). In February 2022, in the experimental area of the Universidade Tecnológica Federal do Paraná - Brazil, disease was observed in 22 trees, with 50% to 80% severity in crown leaves. Symptoms were small, irregular, or circular-shaped, dark-brown lesions with yellow halos (Figure S1). As the disease progressed, the lesions increased in size, without distinction between mature and young tissues, causing complete leaf wilting. Twenty symptomatic leaves from 11 trees grown in the same orchard line were collected. For fungal isolation, the leaf surfaces were disinfected with 0.5% NaOCl solution for 1 min, rinsed in sterile distilled water, and dried on sterile filter paper. Five fragments of diseased leaf tissue were placed on a potato dextrose agar medium. The morphological characteristics of the colony, such as filamentous mycelium and golden yellow on the upper part, with the presence of circular to ovoid and multicellular conidia (mean 21.00 µm x 24.45 µm, n = 30) of the nine isolates, coincided with the description of the fungus of the genus Epicoccum (Valenzuela-Lopez et al., 2018). Further identification of one of these nine isolates was confirmed by amplifying and sequencing three loci (ITS, β-tubulin, and RPB2) using the ITS1/ITS4, Bt2a/Bt2b, and 5F2/7cR primer pairs, respectively (White et al., 1990, Glass and Donaldson, 1995, O'Donnell et al., 2007). A single representative isolate (Cgen01) was analyzed and submitted to GenBank (OR020968, OR079879, and OR079878). The Bayesian Inference was used to reconstruct the phylogenetic trees (Figure S2), starting from random trees for 5,000,000 generations, using MrBayes v. 3.2.1 (Ronquist et al., 2012). The isolate clustered together with the isolate of Epicoccum nigrum (Chen et al., 2017) with a high posterior probability (0.98). For the pathogenicity tests, four young, healthy branches containing 20 leaves were spray-inoculated with 1.5 mL of conidia suspension of Cgen01 (106 conidia mL-1), covered with perforated transparent plastic bags, and moistened with distilled water in the orchard. The air temperature ranged from 14ºC to 25ºC. Sterile distilled water was used as a control. Three replicates (pathogen and control) on different trees were evaluated. After five days, the fungus was re-isolated from the symptomatic lesion, showing morphological characteristics similar to those of Cgen01. Control branches did not show fungal growth. The inoculation test was conducted twice and similar symptoms were observed. This is the first report of leaf spots caused by E. nigrum on C. guazumifolia in Brazil. E. nigrum, an endophytic fungus described as a mycoparasite, showed phytopathogenic behavior in this study, causing spots and loss of leaves in C. guazumifolia, drastically reducing the production of photoassimilates and affecting the quality of the fruits.

Exploring Factors Influencing the Inhibitory Effect of Volume Dielectric Barrier Discharge on Phytopathogenic Fungi

The application of Low-Temperature Plasmas has shown great potential as an effective and alternative tool for microbial inactivation in recent years. Nevertheless, further investigations are required to fully understand the possible factors influencing these processes. The present study aims to investigate the effectiveness of square wave modulated Volume Dielectric Barrier Discharge on the direct inhibition of conidial germination in five different fungal species, various discharge conditions, and medium composition. The five different fungal species used were Botrytis cinerea, Monilinia fructicola, Aspergillus carbonarius, Fusarium graminearum, and Alternaria alternata. On water agar, the inhibition of Botrytis cinerea was influenced by the applied voltage, which mainly reflects the uniformity of the treatment. Under the selected voltage condition, the inhibition increased with treatment duration and decreased with fungal spore complexity. B. cinerea and M. fructicola, with unicellular conidia and low melanin content, showed similar behaviour and high sensitivity to the treatment. F. graminearum and A. alternata, both having multicellular conidia, were more resistant to the plasma treatment and showed different sensitivity likely due to different content in melanin. However, after 1 min of treatment, complete inhibition of conidial germination was achieved for all the tested species. Inhibition of A. carbonarius conidia on different agarized media containing dextrose or malt extract was influenced by the complexity and composition of the medium, being potato dextrose agar that more hindered the plasma efficacy. Each medium exhibited a different electrical response studied by Electrical Impedance Spectroscopy and morphology observed by Scanning Electron Microscopy images. These differences translated into a different response to the applied electrical field, influencing plasma generation and uniformity.

Recent studies on aero-aquatic fungi, with special reference to diversity of conidial morphology and convergent evolution.

Aero-aquatic fungi compose an ecological group of saprophytes inhabiting the submerged decaying substrates in stagnant freshwater environment. They produce three-dimensional shaped, multi-cellular conidia, which float on water surface by holding air between conidial cells. Because the conidia show diverse morphology, genus and species level classification have been based on their features. They are mostly known as asexual morphs of Ascomycota or Basidiomycota. Recent phylogenetic study revealed the aero-aquatic fungi appeared mainly in the lineages of Leotiomycetes, Dothideomycetes, and Sordariomycetes. Furthermore, the phylogenetic tree showed the aero-aquatic fungi have polyphyletic origins and similar three-dimensional conidial morphology generated as a convergent evolution among different lineages of fungi by the selection pressure for inhabiting freshwater environment. Recent studies suggested the ancestors of the aero-aquatic fungi were terrestrial fungi.

First report of Alternaria alternata causing Alternaria leaf spot of Fig in Pakistan.

Fig (Ficus carica) is a species of flowering plants within the mulberry family. During June 2020, leaf spots were observed on several fig plants (31°26'15.0"N 73°04'25.6"E) at the University of Agriculture, Faisalabad, Pakistan. Early symptoms were small, oval to circular, light brown, sunken spots that were uniformly distributed on the leaves. Spots gradually enlarged and coalesced into circular to irregular dark brown to black spots that could be up to 3cm diam. with no or small sized fruit. Disease incidence was approximately 25%. To identify the causal agent of the disease, 15 symptomatic leaves were collected. Small pieces from all diseased samples were removed from the margin between healthy and diseased tissues were surface disinfested in 70% ethanol for 2 min, rinsed three times with sterile distilled water, plated on Potato dextrose agar and incubated at 25 ± 2°C with a 12-h photoperiod. Fungal isolation on PDA medium frequency was 95% from diseases leaves. Morphological observations were made on 7- day- old single-spore cultures. The colonies initially appeared light grayish which turned sooty black in color. All fungal isolates were characterized by small, short-beaked, multicellular conidia. The conidia were ellipsoidal or ovoid and measured 9 to 25 μm × 5 to 10 μm (n = 40) with longitudinal and transverse septa. The morphological characters matched those of Alternaria alternata (Simmons et al. 2007). Genomic DNA of a representative isolate (FG01-FG03) was extracted using DNAzol reagent (Thermo Fisher Scientific MA, USA) and PCR amplification of the internal transcribed spacer (ITS) rDNA region, was performed with primers ITS1/ITS4 (White et al. 1990), partial RNA polymerase II largest subunit (RPB2) with RPB2-5F/RPB2-7cR (Liu et al. 1999) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene regions was performed with gpd1/gpd2 (Berbee et al. 1999). The obtained sequences were deposited in GenBank with accession numbers MW692903.1 to MW692905.1 for ITS-rDNA gene, MZ066731.1 to MZ066733.1 for RPB2 and MZ066728.1 to MZ066730.1 for GAPDH. BLASTn analysis showed 100% identity with the submitted sequences of A. alternata for ITS rDNA, RPB2, and GAPDH. To confirm pathogenicity, 2-month-old 15 healthy potted F. carica plants were sprayed at true leaf stage with conidial suspension by using an atomizer in a greenhouse. Each representative A. alternata isolate (FG01-FG03) was inoculated on every three plants with conidial suspensions (106 conidia/ml; obtained from 1-week-old cultures) amended with 0.1% (vol/vol) of Tween 20 until runoff (1.5 to 2 ml per plant) whereas, three control plants were sprayed with sterile distilled water amended with 0.1% Tween 20. All plants were incubated at 25 ± 2°C in a greenhouse, and the experiment was conducted twice. After 10 days of inoculation, each isolate induced leaf spots similar to typical spots observed in the field, whereas the control plants remained symptomless. The fungus was re-isolated from symptomatic tissues and reisolation frequency was 100%. Re-isolated fungal cultures were again morphologically and molecularly identical to A. alternata, thus fulfilling Koch's postulates. Previously, A. alternata has been reported cause fruit disease of fig in Pakistan and California, USA (Alam et al. 2021; Latinović et al. 2014). To our knowledge, this is the first report of A. alternata causing leaf spot on common fig in Pakistan. In Pakistan, fig is widely grown for drying, and this disease may represent a threat to fig cultivation.

Alternaria alternata causing Alternaria Leaf Spot of Cucumis melo (Muskmelon) in Pakistan.

In July 2019, leaf spot symptoms were observed on muskmelon (Cucumis melo L.) cv. Jackball-1 plants in an experimental field of 2.02 ha with a disease incidence of 30% (31°26'05.4"N 73°04'30.3"E) at the University of Agriculture, Faisalabad, Pakistan. Early symptoms consisted of small, circular, brown, necrotic spots 1 to 2 mm in size covering 10 to 30% of the leaf blade, which gradually enlarged and developed concentric rings. To identify the causal agent of the disease, a total of 20 symptomatic leaves were collected. Small pieces removed from the margin between healthy and diseased tissues were surface disinfected in 70% ethanol for 2 min, rinsed three times with sterile distilled water, plated on Potato dextrose agar and incubated at 25 ± 2°C with a 12-h photoperiod. Morphological observations were made on 7-day-old single-spore cultures. The colonies initially appeared white and then turned olive-green. All 20 fungal isolates were characterized by small, short-beaked, multicellular conidia. The conidia were ellipsoidal or ovoid and measured 11.5 to 30 μm × 7.5 to 15 μm (n = 50) with longitudinal and transverse septa. Conidia were produced on short conidiophores in chains. The beaks were short (often less than one-third the body length) and conical or cylindrical. These morphological features concur with the description of Alternaria alternata (Fr.) Keissler (Woudenberg et al. 2013). For molecular identification, genomic DNA of four representative isolates (HMSMZA 07, 08, 09, 10) were extracted and PCR amplification of the internal transcribed spacer (ITS)-rDNA, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and translation elongation factor-1 alpha (TEF-1α) gene regions were performed (White et al. 1990, Berbee et al. 1999, Carbone & Kohn, 1999) respectively. The obtained sequences were deposited in GenBank with accession numbers MT253643.1-MT253646.1 (ITS-rDNA), MT318260.1-MT318263.1 (GAPDH), and MT318280.1-MT318283.1 (TEF-1α). BLASTn analysis of HMSMZA 07 sequences showed 100% identity with ITS rDNA (MN615420.1), GAPDH (MK637438.1) and TEF-1α (MN807795.1) sequences of A. alternata. To confirm pathogenicity, 5-6 weeks-old Muskmelon (Cucumis melo L.) cv. Jackball-1 plants (true leaf stage) were sprayed until runoff (1.5 to 2 ml per plant) with A. alternata conidial suspension (106 conidia/ml; obtained from 1 week-old cultures) amended with 0.1% (vol/vol) of Tween 20 using an atomizer in the green house. The experiment included four A. alternata isolates inoculated onto three muskmelon plants per each isolate, whereas control plants (n = 3) were sprayed with sterile distilled water amended with 0.1% Tween 20. The plants were incubated at 25 ± 2°C in a greenhouse and the experiment was conducted twice. After 5 to 7 days post inoculation, necrotic leaf spots were observed on the inoculated plants and A. alternata was reisolated and confirmed by morphological and molecular (ITS) features. No disease was observed on control plants. Previously, A. alternata on muskmelon has been reported in Pakistan (Ahmad et al. 1997), however this study provides a detailed description of disease symptoms, morphological and molecular identity of the causal agent including completion of Koch's postulates. The disease could represent a threat for muskmelon crop in Pakistan due to its increasing cultivation and therefore warrants the need to develop disease management strategies.

First Report of Leaf Spot Caused by Alternaria alternata on Phlox maculata in Italy

Phlox maculata (common name spotted phlox), Polemoniaceae family, is an herbaceous perennial plant with showy panicle inflorescences, commonly grown in mixed borders. During summer 2019, about twenty 5-year-old plants of P. maculata growing in a garden located in the Biella province (northern Italy) showed symptoms of a leaf spot, starting from the basal part of the foliage. Symptoms consisted in brown spots with not well-defined margins, irregularly circular to polygonal in shape, up to 2 to 3 mm in diameter, sometimes confluent. The internal part of spots dried. Chlorosis and yellowing surrounded the spots, involving the most part of affected leaf surfaces, due to the spread of the disease. The ornamental value of affected plants was significantly reduced. Several symptomatic leaves were washed in sterile water for 1 min. Isolations were carried out, taking several small fragments of tissues from the margins of the spots and placing them on potato dextrose agar. Fungal colonies that developed were dark green at the end of development. Successively, single-spore colonies were obtained, and pure cultures were transferred on potato carrot agar (PCA, Simmons 2007) under a light/dark regime of 11 h/13 h. On this medium, colonies formed branched conidiophores with brown, multicellular conidia, ovoid to obclavate in shape, containing one to four (average 2.4, n = 50) transverse septa and zero to one (average 0.1, n = 50) longitudinal septa. Conidia measured 10.8 to 29.3 × 5.3 to 11.2 µm (average 18.7 × 7.6 µm, n = 50). If present, the beak was 2.1 to 11.0 µm (average 5.5 µm) long. These morphological characteristics permitted identifying the fungus isolated from P. maculata as Alternaria section alternata (Woudenberg et al. 2015). The fungal DNA from the 19/33 isolate was extracted using the E.Z.N.A. Fungal DNA Mini Kit (Omega Bio-Tek, Darmstadt, Germany), and PCR was performed using the ITS (White et al. 1990), the rpb2, and the endoPG primers (Woudenberg et al. 2015). Three sequences with 529 bp (ITS), 889 bp (rpb2), and 452 bp (endoPG) (GenBank accession nos. MN871825, MN871979, and MN871978, respectively) were produced. A BLASTn analysis of these sequences exhibited 100% identity with the reference strain CBS 117.44 of A. alternata (Fries) Keissler in rpb2 and endoPG portions (accession nos. KP124772 and KP124001, respectively) and 99.6% identity in ITS region (accession no. KP124303). One P. maculata fungal isolate (19/33) was grown on PCA. A spore suspension obtained from 15-day-old cultures was sprayed at the concentration of 10⁵ CFU/ml onto leaves of three healthy plants of P. maculata. Three control plants were treated with sterile water. All plants were closed in moist plastic bags for 7 days and maintained at temperatures ranging from 15 to 28°C. The first symptoms of leaf spot appeared 10 days after the inoculation, and A. alternata was reisolated from affected leaves. Control plants remained symptomless. The pathogenicity test was repeated once using the isolate 19/40, obtaining the same results. To our knowledge, this is the first report of A. alternata on P. maculata in Italy as well as worldwide. The spread of A. alternata on P. maculata and the difficulty to manage this disease, particularly in public gardens, may discourage the use of this ornamental in Italy.

Fungal Record in a Legume Wood from the Chiquimil Formation (Upper Miocene), Catamarca, Argentina

Herein, we described three types of fossil fungi associated with permineralized legume wood from Upper Miocene deposits in the Rio de Agua Verde locality (Chiquimil Formation, Catamarca Province, Argentina). These are represented by diverse fungal remains, including septate hyphae bearing unicellular and multicellular conidia, distributed inside the vessel elements. Vegetative and asexual reproductive structures preserved were compared to extant mitosporic fungi and related to three different genera. The fossils are particularly similar to extant Gilmaniella Barron, Brachysporium Saccardo, and Ramichloridium Stahel ex de Hoog, but display some differences that preclude unequivocal assignment. Due to the absence of host reactions signs in the infected wood, we suggest that the fungi were probably saprotroph and invaded the fossil wood after its death.

FUNGAL RECORD IN A LEGUME WOOD FROM THE CHIQUIMIL FORMATION (UPPER MIOCENE), CATAMARCA, ARGENTINA

Herein, we described three types of fossil fungi associated with permineralized legume wood from Upper Miocene deposits in the Río de Agua Verde locality (Chiquimil Formation, Catamarca Province, Argentina). These are represented by diverse fungal remains, including septate hyphae bearing unicellular and multicellular conidia, distributed inside the vessel elements. Vegetative and asexual reproductive structures preserved were compared to extant mitosporic fungi and related to three different genera. The fossils are particularly similar to extant Gilmaniella Barron, Brachysporium Saccardo, and Ramichloridium Stahel ex de Hoog, but display some differences that preclude unequivocal assignment. Due to the absence of host reactions signs in the infected wood, we suggest that the fungi were probably saprotroph and invaded the fossil wood after its death.

Vertical Divergence of Cultured Microfungal Communities at the Hallamish Dune Field, Western Negev Desert, Israel

ABSTRACTWe examined the depthwise distribution of microfungi through 0–40 cm of sandy profiles under bare surface and different shrub species at the Nizzana research site (NRS) in the Negev Desert, Israel. A total of 125 species from 61 genera were isolated using the soil dilution plate method. At the topsoil of all profiles melanin-containing species with large multicellular conidia predominated. In the middle depths at the majority of the profiles, mesic group of microfungi, Penicillium spp. substantially increased in abundance. Density of microfungal isolates decreased with depth under shrubs and did not substantially vary in the nonshaded profiles, highly positively correlating with electrical conductivity and organic matter (OM) content. Substantial increase in isolate densities at the undercanopy habitats, especially in the interdune depression with higher substrate stability raising the accumulation of OM, highlights the role of shrubs as “islands of fertility” for microfungal communities at NRS.

Vertical Divergence of Cultured Microfungal Communities Through the Depth in Different Soil Formations at Nahal Nizzana, Western Negev Desert, Israel

ABSTRACTWe examined the depth-wise distribution of microfungi through 0–50-cm crusted sandy and playa profiles at the Nizzana research site, the Negev desert, Israel. A total of 188 species from 77 genera was isolated using the soil dilution plate method. Density of microfungal isolates sharply decreased with depth highly positively correlating with organic matter content at the sandy profiles. High load of solar radiation (at the surface) as well as strongly limited aeration, water infiltration, and increased salinity (at the playa depth) led to dominance of melanin-containing species with large, multicellular conidia both in the topsoil of all profiles and in the deep playa layers, while at 1–30 cm, species producing light-colored small one-celled conidia mostly prevailed. In that way, the above vertical variations resemble differences in the composition of topsoil microfungal communities found between the sites located in the Negev desert and in the Mediterranean region of Israel.

First Report of Pyrenophora seminiperda in Turkey and Greece.

Pyrenophora seminiperda (Brittleb. & D.B. Adam) Shoemaker (anamorph Drechslera campanulata (Lév.) B. Sutton) is a generalist seed pathogen that can cause high mortality in the seed banks of annual and perennial grasses and a minor leaf spot disease. Its current reported distribution is mainly temperate grasslands, deserts, and winter cereal-growing regions in Argentina, Australia, Canada, Egypt, New Zealand, South Africa, and the United States (2). P. seminiperda was originally described in France in the mid-1800s, but there are no recent reports from eastern Europe or Asia (2). In May of 2008, we observed what appeared to be P. seminiperda on seeds from seed bank samples collected in Turkey. Evidence of disease was observed as macroscopic black stromata protruding from the seed. The characteristic club-shaped stromata were collected from a Taeniatherum caput-medusae seed near Pamukkale, Turkey and six Bromus tectorum seeds in Love Valley near Goreme, Turkey. An additional collection from a single undispersed B. tectorum seed was obtained from Perissa, Greece. Identity of the pathogen was tentatively established by evaluating morphological characteristics for nine isolates in V8 agar culture. After 4 days of incubation at 20°C with a 12-h photoperiod, the cultures produced white mycelium. Following wounding, the mycelium produced black, club-shaped stromata (2 to 8 × 0.4 to 0.9 mm) in a radial pattern. These produced branched conidiophores bearing crescent-shaped, multicellular conidia (79 to 125 μm long). These attributes are consistent with those of the anamorph of P. seminiperda as described by Shoemaker (4) and Campbell et al. (1). The teleomorph was not observed. The identity of the isolates as P. seminiperda was confirmed with ribosomal DNA internal transcribed spacer (ITS) genetic sequencing analysis. ITS sequences obtained were identical to sequences for North American haplotypes of this species. Four of the Love Valley isolates, (representative isolate: GQ168725, BPI 879142, NRRL 54032) matched the HTA haplotype (GQ168724), while the other four (representative isolate: GQ168736, BPI 879143, NRRL 54033) matched the HTJ haplotype (GQ168735). The isolate from Perissa, Greece (GQ168728, BPI 879144, NRRL 54034) matched the HTC haplotype (GQ168727). Pathogenicity of several Love Valley isolates was confirmed by producing conidia in culture, dusting nondormant B. tectorum seeds with 0.003 g of conidial inoculum per 50 seeds, and incubating for 14 days at 10/20°C with a 12-h photoperiod. Stromata developed on >90% of inoculated seeds and mortality as high as 34% was observed. Morphological similarities combined with ITS sequence data provide conclusive evidence that P. seminiperda occurs in Turkey and Greece. The discovery of this pathogen in these countries indicates that it may be widespread in Eurasia and that it could have arrived in North America on seeds of B. tectorum (3) rather than representing a novel pathogen for this important weed in its North American range.

A new species of Petractis (Ostropales s. lat., lichenized Ascomycota) from Wales

AbstractPetractis nodispora is described as a new species, characterized by an endolithic thallus with Trentepohlia as photobiont, immersed apothecia, 3-septate, halonate ascospores, and very distinctive multicellular conidia. Nuclear ribosomal SSU and LSU sequences show that it is closely related to P. luetkemuelleri, but its relationship to the type species of Petractis, P. clausa, is unclear.

First Report of Alternaria alternata f. sp. cucurbitae Causing Alternaria Leaf Spot of Melon in the Mid-Atlantic Region of the United States.

Alternaria alternata f. sp. cucurbitae, the casual agent of Alternaria leaf spot, was first described in Greece where it caused severe losses to greenhouse-grown cucumbers (Cucumis sativus) (3,4). The fungus also attacks melon (C. melo) and watermelon (Citrullus lanatus) (1-3). In late June of 2006, following a period of windy and rainy days, numerous dark brown, circular lesions, 0.5 to 1 mm in diameter, were observed on leaves of melons in a field in Wicomico County, Maryland. The lesions gradually enlarged and coalesced into large, nearly circular, or irregularly shaped lesions that could be as long as 3 cm. The center of the lesions was light tan, surrounded by a dark brown ring and a chlorotic halo, and tended to split in the later development stages. Most of the lesions appeared on the edge of the leaves and no lesions developed on the stems and fruit. Lesions first started on old leaves and then developed on leaves in the middle part of the canopy. Leaf lesions were observed on melon cvs. Ananas, Honeydew Greenflesh, and Israeli. Disease severity ranged from 3 to 20% of the leaf area affected. Small pieces (3 × 3 mm) of tissue removed from the margin between healthy and diseased tissue were surface disinfected in 0.5% NaOCl for 2 min and plated on acidified, ¼-strength potato dextrose agar. Isolations made from diseased tissue frequently (61%) yielded fungal colonies with morphological features and spore dimensions that were consistent with the description of A. alternata f. sp. cucurbitae (1,3). Fungal isolates were characterized by small, short-beaked, multicellular conidia. Conidia were ovoid, obclavate, and sometimes ellipsoidal with the average overall body length of 39 μm (range, 17 to 80 μm) and width of 14 μm (range, 7 to 20 μm). Conidia were produced on short conidiophores in chains. The beaks were short (often less than one-third the body length) and conical or cylindrical. Pathogenicity of six single-spore isolates was determined on four melon cultivars (Honeydew Greenflesh, Israeli, Tam Dew, and Topmark) and one watermelon cultivar (Sugar Baby) in a greenhouse. Twenty plants of each cultivar at the one-true-leaf stage were sprayed with a conidial suspension (106 conidia/ml) of each isolate amended with 0.1% (vol/vol) of Tween 20 until runoff (1.5 to 2 ml per plant). Inoculation with sterile distilled water amended with 0.1% Tween 20 served as controls. The plants were placed in a dew growth chamber for 48 h at 24°C and subsequently maintained in a greenhouse at 21 to 29°C. At 4 to 5 days after inoculation, each isolate induced leaf lesions on each inoculated cultivar similar to typical lesions observed in the field. There was no significant difference in disease severity among the cultivars tested or between melon and watermelon. Control plants remained symptomless. The fungus was readily reisolated from symptomatic tissues. To our knowledge, this is the first report of A. alternata f. sp. cucurbitae causing Alternaria leaf spot of melon in the Mid-Atlantic United States and the only report outside Georgia in the southern region of the United States (D. B. Langston, personal communication) and Greece.

Trichocladium ismailiense, a new dematiaceous hyphomycete from Egyptian soils

Trichocladium ismailiense is described and illustrated from Egyptian soils. It is characterized by deeply constricted, multicellular conidia with the top cells showing conspicuously reticulate walls and 1-3 germ pores

Cell wall structures of Epicoccum nigrum (Hyphomycetes)

Two structurally distinct types of secondary wall layers are present in older hyphae in addition to the primary wall. A coarsely fibrous outer wall layer often becomes quite massive and frequently fuses with the outer wall layers of adjacent cells in the formation of hyphal strands. The uneven deposition of this outer layer often produces large verrucosities. The inner secondary wall layer is relatively electron transparent and contains a reticulum of electron-dense lines. The interface of the inner secondary wall with the cytoplasm is often very irregular, and sections of the plasma membrane are frequently overlain by wall material. The outer secondary wall of conidia is composed of an electron-dense material different from that of the outer wall of hyphae. Cells in the multicellular conidia tend to be polyhedral in shape with either very thick primary walls or thin primary walls having a thick inner wall deposit.

Two New Genera of Stilbellaceous Fungi

Two new synnematous genera of the Fungi Imperfecti are described. Dictyocatenulata, with the type species D. alba, is pro- posed for the Dictyosporae group and Hyalotrocophora, with the type species H. lignitalis, is proposed for the Helicosporae group of the Stil- bellaceae. During our investigations on the stilbellaceous Fungi Imperfecti, a fungus possessing hyaline, muriform conidia which occur in long chains has been encountered frequently. Herbarium specimens of this fungus have been identified as Stilbum glomerulisporum Ell. & Everh. Hughes (1953) considered the name to be a nomen nudum. We are unable to find any other record of it. Hughes did not consider the species to be a Stilbum/Stilbella and we concur. The fungus was collected in the Panam'a Canal Zone by E. F. Morris and J. W. Strain in 1964. The only difference between this specimen and the older ones is in the length of the synnemata. The tropical specimens have longer synnemata. The hyaline, muriform conidia of this fungus are quite distinctive. As stated by Hughes, conidia are composed of adpressed and coiled outgrowths of the initial cells . Hughes placed this fungus in Section V of his classification scheme. Section V is characterized by Conidia developing in gradually maturing basipetal series and originating by the meristematic growth of the apical region of the conidiophore in such a way that the chain of conidia merges im- perceptibly with the conidiophore that gives rise to the chain. Although the fungus has not been obtained in culture, this is probably a good conjecture. However, it is entirely possible that these conidia arise as arthrospores which, after formation, continue their maturation by production of the coiled outgrowths which result in multicellular conidia. It is somewhat difficult to ascertain the presence or absence of a meristematic zone from herbarium materials. Hughes' Section VII is characterized by Conidia developing by the basipetal fragmentation of conidiophores of determinate length, and which do, not possess a meristematic zone. Regardless of the production of meristem arthro- spores or arthrospores, the fungus cannot be satisfactorily placed in a known genus and is here described as new.