Occurrence and characterization of Colletotrichum dematium (Fr.) grove

Colletotrichum dematium was isolated from caraway for the first time in Poland in 2005. Isolations of this fungus were repeated in 2006 and 2007. The cultures of fungus were obtained from superficially disinfected leaves, root necks, roots, stems and umbels. The isolates were identified on culture media: PDA and malt agar with addition of pieces of caraway stems and on the base of macro and microscopic structures. Studies on the biotic effect between C. dematium and other species of phyllosphere fungi of caraway showed that the majority of the studied species limited the growth and development of C. dematium, but the size of the limiting effect was different. The species from Trichoderma and Gliocladium genera were the most effective against C. dematium, causing degeneration and lysis of hyphae and precluded the formation of the pathogen's acervuli and conidia. C. dematium in dual culture with E. purpurascens, A. radicina, S. sclerotiorum, B. cinerea and R. solani produced an inhibition zone which indicated its capacity for antibiosis.     

Tympanosporium parasiticum gen. et sp. nov., a hyperparasitic Hyphomycete on Tubercularia vulgaris with pleomorphic conidiogenesis

Tympanosporium parasiticum forms annellated conidiogenous cells with short cylindric (tympaniform) conidia in white sporodochia on Tubercularia vulgaris Tode ex Fr. On agar media the conidia form short necks and blow out narrow secondary conidia in a phialidic manner producing very restricted yeast-like colonies. Only in the vicinity of the host hyphae (without tropic growth or hyphal connections) mycelia develop which form phialidic or annellate conidiogenous cells with narrowly cylindrical conidia in the agar, and sporodochia with short truncate conidia like those on the natural substrate in the aerial mycelium. No other host fungus was found to support this kind of growth. The mode of parasitism is compared with that of other hyperparasitic fungi.For Tympanosporium parasiticum the name Oospora candidula Sacc. was incorrectly used by Grove (1885) and the name Oospora filamentosa was proposed by Arnaud but never published.     

Infection of Subterranean Clover (Trifolium subterraneum) by Kabatiella caulivora

Kabatiella caulivora is a serious pathogen of clover ( Trifolium ) spp. Subterranean clover ( T. subterraneum ) cv. Woogenellup was inoculated with K. caulivora , to study the attachment and germination of conidia, germ-tube penetration of the plant surface, and histochemistry and ultrastructure of changes in the host associated with lesion development. The foliar architecture caused the conidia to concentrate at the base of leaflets and on the petiolules (between the leaflets and petioles). Epidermal cells immediately beneath conidia and, occasionally, also adjacent cells developed a yellow-brown discoloration 1 day post-inoculation. Penetration appeared to be directly through the cuticle, characterized by constricted hyphae at the point of entry. No appressoria were observed. In leaves, invasion was restricted to the area proximal to the petiolule and leaf mid-rib. In petioles and petiolules, the hyphae initially remained between the epidermal cells and first layer of mesophyll cells before moving intercellularly through the mesophyll tissue towards phloem tissues. The cuticle was occasionally degraded in petiole and petiolule infections, the loss of epidermal and mesophyll cell wall components was detected, and chloroplasts and starch grains were disrupted. Plants developed macroscopic symptoms 10–11 days post-inoculation with necrotic lesions occurring on leaves, petioles and petiolules. Sporulation occurred approximately 15–18 days post-inoculation when affected plants collapsed. This information may be useful for breeding programmes aimed at selecting varieties with improved resistance to the clover scorch disease.     

Surface Ultrastructural Studies on Penetration and Infection Process of Colletotrichum gloeosporioides on Mulberry Leaf Causing Black Spot Disease

Unicelled conidia of Colletotrichum gloeosporioides germinated 3 h after inoculation producing single germ tubes. The orientation of the germ tubes and their lateral branches as they grew was towards the open stomata and away from closed stomata. By 24 h post-inoculation, the lateral branches had developed specialized infection vesicles either over the stomata or within the stomatal cavities. The infection vesicles produced primary infection hyphae which entered the leaves through stomatal openings. The disease symptoms became apparent by 6 days post-inoculation when clusters of abundant conidiophores emerged by rupturing the leaf epidermal layer, forming acervuli mostly near the base of idioblasts. The pressure exerted by the emerging conidiophores caused stretching of epidermal layer leading to the widening of the acervuli. The conidia are borne on the tips of the erect conidiophores.     

Dendritic cells transport conidia and hyphae of Aspergillus fumigatus from the airways to the draining lymph nodes and initiate disparate Th responses to the fungus.

Aspergilli are respiratory pathogens and pulmonary infections are usually acquired through the inhalation of conidia, able to reach small airways and the alveolar space where the impaired host defense mechanisms allow hyphal germination and subsequent tissue invasion. The invasive pulmonary aspergillosis is the most common manifestation of Aspergillus fumigatus infection in immunocompromised patients and is characterized by hyphal invasion and destruction of pulmonary tissue. A Th1/Th2 dysregulation and a switch to a Th2 immune response may contribute to the development and unfavorable outcome of invasive pulmonary aspergillosis. Dendritic cells (DC) have a primary role in surveillance for pathogens at the mucosal surfaces and are recognized as the initiators of immune responses to them. In the present study, we assessed the functional activity of pulmonary DC in response to A. fumigatus conidia and hyphae, both in vitro and in vivo. We analyzed mechanisms and receptors for phagocytosis by DC as well as DC migration, maturation, and Th priming in vivo upon exposure to either form of the fungus. We found a remarkable functional plasticity of DC in response to the different forms of the fungus, as pulmonary DC were able to: 1) internalize conidia and hyphae of A. fumigatus through distinct phagocytic mechanisms and recognition receptors; 2) discriminate between the different forms in terms of cytokine production; 3) undergo functional maturation upon migration to the draining lymph nodes and spleens; and 4) instruct local and peripheral Th cell reactivity to the fungus.     

Root Infection and Systemic Colonization of Maize by Colletotrichum graminicola

Colletotrichum graminicola is a filamentous ascomycete that causes anthracnose disease of maize. While the fungus can cause devastating foliar leaf blight and stalk rot diseases, little is known about its ability to infect roots. Previously published reports suggest that C. graminicola may infect maize roots and that root infections may contribute to the colonization of aboveground plant tissues, leading to disease. To determine whether C. graminicola can infect maize roots and whether root infections can result in the colonization of aboveground plant tissues, we developed a green fluorescent protein-tagged strain and used it to study the plant root colonization and infection process in vivo. We observed structures produced by other root pathogenic fungi, including runner hyphae, hyphopodia, and microsclerotia. A mosaic pattern of infection resulted from specific epidermal and cortical cells becoming infected by intercellular hyphae while surrounding cells were uninfected, a pattern that is distinctly different from that described for leaves. Interestingly, falcate conidia, normally restricted to acervuli, were also found filling epidermal cells and root hairs. Twenty-eight percent of plants challenged with soilborne inoculum became infected in aboveground plant parts (stem and/or leaves), indicating that root infection can lead to asymptomatic systemic colonization of the plants. Many of the traits observed for C. graminicola have been previously reported for other root-pathogenic fungi, suggesting that these traits are evolutionally conserved in multiple fungal lineages. These observations suggest that root infection may be an important component of the maize anthracnose disease cycle.     

CYTOLOGICAL INVESTIGATIONS OF KABATIELLA CAULIVORA

Cole , Herbert , Jr ., and Houston B. Couch . (Penn. State U., University Park.) Cytological investigations of Kabatiella caulivora. Amer. Jour. Bot. 46(1) : 12-16. Illus. 1959.—Initial growth of K. caulivora on artificial media is characterized by budding, yeast-like, conidia, exclusive of mycelia. After 14 days growth, at 20°C., mycelial growth becomes macroscopically evident. A study of the germination behavior of a total of 5500 conidia from the initial yeast-like growth stage showed 59 to germinate by the production of mycelia, while the balance germinated by budding. Five thousand conidia of the mycelial growth type were studied in a successive, single-spore transfer series, and, in all cases, conidial germination continued to be by means of germ tubes. Conidia of both growth forms were studied by means of bright field and phase contrast microscopy, and found to be multinucleate—possessing variable nuclear numbers, ranging from 1-8 per cell. Mean nuclear number for both the mycelial and conidial types was 2.8. All nuclei of both growth types appeared to contain the same chromosome complement. The cultural variability exhibited by K. caulivora cannot be reconciled with the concept of dual phenomenon. It is suggested, rather, that the mycelial homotype probably arises as the result of a unidirectional mutation within the conidial growth form.     

<Emphasis Type="Italic">Colletotrichum kakivorum</Emphasis> sp. nov., a new leaf spot pathogen of persimmon in Korea

In this study, a new species within the genus Colletotrichum was isolated from diseased persimmon leaves collected from Sangju and Cheongdo in the Gyeongbuk province of Korea. The disease was characterized by leaf spots with a brown to gray center surrounded by a black ring with a green halo. Black acervuli with setae and numerous curved conidia were observed on the leaf spots. Phylogenetic analysis based on a combination of multiple loci, including sequences of the internal transcribed spacers, actin, beta-tubulin, chitin synthase-1, and glyceraldehyde-3-phosphate dehydrogenase genes, revealed that the isolates were clearly distinct from other species of the Colletotrichum dematium complex, with the closest species being C. anthrisci. Cultural and morphological characteristics and pathogenicity on persimmon leaves were examined. The isolates differed from C. anthrisci in their cultural and morphological characteristics, such as the conidia and appressoria. Based on molecular phylogenetic, morphological, and cultural characteristics and pathogenicity, this fungus was demonstrated to be a new species, Colletotrichum kakivorum, and the causal agent of a leaf spot disease on persimmon in Korea.     

Beauveria bassiana yeast phase on agar medium and its pathogenicity against Diatraea saccharalis (Lepidoptera: Crambidae) and Tetranychus urticae (Acari: Tetranychidae)

Beauveria bassiana colonizes insect hosts initially through a yeast phase, which is common in some artificial liquid cultures, but not reported on artificial solid media. We describe a yeast-like phase for B. bassiana isolate 447 (ATCC 20872) on MacConkey agar and its virulence toward Diatraea saccharalis and Tetranychus urticae. The yeast-like cells of B. bassiana developed by budding from germinating conidia after 24-h incubation. Cells were typically 5-10 microm and fungal colonies were initially circular and mucoid, but later were covered with mycelia and conidia. Ability to produce yeast-like cells on MacConkey medium was relatively common among different B. bassiana isolates, but growth rate and timing of yeast-like cell production also varied. Metarhizium anisopliae and Paecilomyces spp. isolates did not grow as yeast-like cells on MacConkey medium. Yeast-like cells of B. bassiana 447 were more virulent against D. saccharalis than conidia when 10(7)cells/ml were used. At 10(8)cells/ml, the estimated mean survival time was 5.4 days for the yeast suspension and 7.7 days for the conidial suspension, perhaps due to faster germination. The LC(50) was also lower for yeast than conidial suspensions. Yeast-like cells and conidia had similar virulence against T. urticae; the average mortalities with yeast-like cells and conidia were, respectively, 42.8 and 45.0%, with 10(7)cells/ml, and 77.8 and 74.4%, with 10(8)cells/ml. The estimated mean survival times were 3.6 and 3.9 for yeast and conidial suspensions, respectively. The bioassay results demonstrate the yeast-like structures produced on MacConkey agar are effective as inoculum for B. bassiana applications against arthropod pests, and possibly superior to conidia against some species. Obtaining well-defined yeast phase cultures of entomopathogenic hyphomycetes may be an important step in studies of the biology and nutrition, pathogenesis, and the genetic manipulation of these fungi.     

On an undescribed, pleomorphic hyphomycete from litter

Pseudoyima prolifica is described from litter of Scirpus microcarpus Presl. The species produces dark hyphal colonies with few or no conidia, and pallid conidial colonies in which hyphae are sparse or absent. Conidial colonies, macroscopically yeast-like, consist of proliferating scolecoconidia, stauroconidia, and arthroconidia.     

Phylogeny and morphology of Hirsutella tunicata sp. nov. (Ophiocordycipitaceae), a novel mite parasite from Peru

A new species of Hirsutella was isolated from unidentified mites on Petri plates inoculated with soil and root fragments collected from asparagus rhizosphere at Virú, Northern Peru. The fungus differs from other Hirsutella species by an envelope surrounding the conidium, conidia dimension and DNA sequences. In PDA cultures, the mycelium produced aerial hyphae with conidiogenous cells mainly at right angles, occasionally showing a secondary conidiophore. The solitary conidia are cymbiform, slightly apiculate, 5.0–6.0 × 3.0–4.0 μm. Phylogenetic analyses with partial rRNA and β-tubulin gene sequences confirmed the fungus as an Hirsutella (Ophiocordycipitaceae). Closest species shown by maximum likelihood and neighbor-joining trees were H. nodulosa and H. aphidis, from which the new species differs for conidium or conidiogenous cells dimensions, lack of synnemata and host type. A recombination event was also detected in the rRNA of the holotype strain, involving Ophiocordyceps sinensis as major parent and O. cochlidiicola as minor parent. A complement, inverted insertion was also found in its rRNA, involving part of the ITS2 and 5.8S regions, flanked by two short nucleotide arrays. Due to conidia dimension and phylogenetic position, the fungus is described as Hirsutella tunicata sp. nov. A review of mononematous Hirsutella species is provided.     

MripacC regulates blastosphere budding and influences virulence of the pathogenic fungus Metarhizium rileyi

Fungal dimorphism is the ability of certain fungi to switch between two different cellular forms, yeast and mycelial forms, in response to external environmental factors. The pacC/Pal signal transduction pathway responds to neutral and alkaline environments and is also involved in the fungal dimorphic transition. In this study, we investigated the function of the pacC homolog, MripacC, which regulates the dimorphic transition and modulates virulence of the insect pathogenic fungus Metarhizium rileyi. MripacC expression was upregulated under alkaline condition, with increased number of yeast-like cells compared to the number of hyphae cells. A MripacC deletion mutant (ΔMripacC) was obtained by homologous replacement and exhibited decreased blastospore budding, with direct development of conidia into hyphae without entering the yeast-like stage when cultured on alkaline medium. Observation of host hemolymph morphology and analysis of samples to detect the main immune factors revealed a decreased ability of ΔMripacC to evade the host immune system. The results of insect bioassays showed that ΔMripacC had decreased virulence with extended median lethality time. Together, the results suggested that MripacC not only regulated adaptation to acidic and alkaline environments, but also influenced virulence by budding blastospores. This elucidation of the function of MripacC adds to our understanding of blastospore budding and virulence of this fungal pathogen.     

Phaeomoniella zymoides and Phaeomoniella pinifoliorum spp. nov., new acid-tolerant epiphytic fungi isolated from pine needles in Korea

Two new epiphytes of anamorphic ascomycetes, Phaeomoniella zymoides sp. nov. and Phaeomoniella pinifoliorum sp. nov., were isolated from the needle surface of Pinus densiflora in Korea. The new taxa were characterized by acid-tolerant, slow, partially yeast-like growth and extensive production of emerging cells on convex wrinkled mycelial colonies. Phaeomoniella zymoides produced mycelium with large numbers of intercalary and lateral or terminal vesicles or swollen cells. Large conidiogenous cells had a swollen base and appeared to be phialidic, and many phialoconidia also were produced from lateral hyphal apertures. Maturing colonies of Ph. zymoides were made up of dark green to blackish areas and produced a Phoma-like synanamorph. Primary conidia became elongate mother cells giving rise to polar or lateral secondary conidia. Phaeomoniella pinifoliorum was characterized by reduced, swollen, phialide-like cells, lateral production of conidia from hyphae and terminal or subterminal, or less commonly lateral, secondary production of conidia from yeast-like primary conidia. When ITS and 28S rDNA sequences were compared and analyzed with those of best matching GenBank taxa, the Phaeomoniella group consisted of three lineages, "zymoides", "pinifoliorum" and "chlamydospora" clades, which again showed a complete sister relationship to Moristroma quercinum ined.     

嗜冷和冷育微生物的研究 Ⅲ.地丝霉属一新种,光滑地丝霉

在研究食品冷库中嗜冷微生物区系时,获得很多冷育型真菌。地丝霉和金孢菌类在这些菌中占优势。其中一菌在PYE上于18℃培养14天,菌落白色,厚毡状,中央常有淡黄色渗出液滴,菌落直径33mm,背面浅黄褐色,有辐射沟纹,老后仍为白色。菌丝分隔,直径1.0—2.0μm。该菌有明显的分生孢子梗,直立,宽度1.0—2.0μm,长10—25(—35)μm,无色或淡黄色,分生孢子梗具分枝,由分枝产分生孢子。分生孢子无色或淡黄色,顶端者为梨形、倒卵形或楔状,间生者矩椭圆形或桶状,1.5—2.5×2.5—3.5(—4.0)μm,光滑、1—3成链。该菌生长温度范围为0—25℃,最适温度为18℃,30℃不生长。此菌产生果胶酶,但是不分解角朊。该菌从冷库中生霉的蒜苗上分离得。其特征与Carmichael(1962),Domsch等(1980)所订正的金孢菌属(Chrysosporium Corda)中各种,以及Sigler和Carmichael(1976),Oorschot(1980)所订正的地丝霉属(Geomyces Traaen)中各种均有显著的差异,而其性状符合地丝霉属的特征集要。故命名为光滑地丝霉Geomyces laevis Z.Q.Li et Cui。模式:菌株AS 3.4605为模式菌株,保藏在中国科学院微生物研究所菌种保藏研究室;模式标本HMAS No.54875保存在该研究所真菌标本室。     

Dimorphism of Scopulariopsis brevicaulis: Morphogenesis of the mould to yeast phase

The dimorphism of Scopulariopsis brevicaulus was induced in malt agar (Oxoid), at 37°C and 5–10% rate of CO₂. Six cultures of S. brevicaulus were converted from hyphae to yeast-like cells by the following processes: One was that the hyphae formed intercalar and terminal chlamydoconidia-like cells. These cells increased in size, gained double contour and gave rise to budding cells. The other process was via conidia that assumed yeast characteristics. So, a transformation cycle M Y was proposed. The yeast-like form presented slow reversion to the mould phase, standing, many times, in transition forms. A high concentration of sugars, peptone, temperature of cultivation of 37°C and 5–10% rate of CO₂ seem to be favourable factors for the maintenance of the yeast phase. Ethanol, E.D.T.A., potassium nitrate, temperature of 25°C and aerobiosis are considered favourable factors to conversion of the mould phase.