Conidial Morphology Research Articles

Powdery Mildew Caused by Golovinomyces cichoracearum on Moth Mullein (Verbascum blattaria) in Italy.

Moth mullein (Verbascum blattaria) is an herbaceous plant belonging to the Scrophulariaceae family. It has alternate, simple leaves on stiffly, erect, green stems. Flowers are yellow or white, borne in summer through fall, and is increasingly used in gardens in low-maintenance borders. During the fall of 2009, 4-month-old plants grown in a greenhouse near Torino (northern Italy) showed signs and symptoms of an unknown powdery mildew. The adaxial leaf surfaces were covered with white mycelia and conidia, while the abaxial surfaces were less infected. As the disease progressed, infected leaves turned yellow and wilted. Mycelia were also observed on stems, petioles, and flower calyxes of inflorescences. Powdery mildew was observed on moth mullein naturally diffused in Italian flora and on V. blattaria var albiflorum cv. White Blush. The same symptoms and signs were observed in summer 2010 on V. blattaria plants grown in a garden near Biella. Conidia were hyaline, elliptical, borne in short chains (as many as five conidia per chain), and measured 35 × 22 (29 to 42 × 19 to 24) μm. Conidiophores were erect with a cylindrical foot cell measuring 147 × 11 (93 to 177 × 10 to 12) μm, followed by one to two shorter cells measuring 23 × 11 (15 to 33 × 10 to 12) μm. Fibrosin bodies were absent. Chasmothecia were not observed in the collected samples. The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS4/ITS6 and sequenced (1) (GenBank Accession No. HQ316555). The 542-bp amplicon had 99% homology with the sequence of Golovinomyces cichoracearum (GenBank Accession No. EU819552. Pathogenicity was confirmed through inoculation by gently pressing diseased leaves onto leaves of healthy V. blattaria plants. Five plants were inoculated, while the same number of noninoculated plants served as a control. Plants were maintained at temperatures from 19 to 25°C. Fifteen days after inoculation, symptoms and signs of powdery mildew developed on inoculated plants. The conidial morphology of the powdery mildew fungus that developed on inoculated plants was identical to the conidial morphology observed in the original fungus. Noninoculated plants remained healthy. The pathogenicity test was carried out twice. G. cichoracearum, formerly Erysiphe verbasci (synonym E. cichoracearum), has been reported on V. blattaria in Hungary, Romania, and the former USSR (2,3). In conclusion, to our knowledge, it is the first report of G. cichoracearum affecting moth mullein in northern Italy. The economic importance of this disease is at present limited in Italy because of limited planting of this host. However, the ecological characteristics and flowering of V. blattaria make it interesting for low-maintenance gardens.

Scleroramularia gen. nov. associated with sooty blotch and flyspeck of apple and pawpaw from the Northern Hemisphere

Scleroramularia is proposed as a new hyphomycetous fungal genus associated with sooty blotch and flyspeck (SBFS) blemishes of apple and pawpaw fruit in the Northern Hemisphere. Morphologically the genus closely resembles Ramularia (Mycosphaerellaceae), based on its chains of hyaline conidia, with thickened, darkened, somewhat refractive conidiogenous loci. Scleroramularia is distinguished by forming black sclerotial bodies in culture, and having conidial chains that do not quickly disarticulate as observed in Ramularia. Based on the nuclear ribosomal DNA phylogeny (LSU), Scleroramularia represents an undescribed order in the Dothideomycetes, clustering between the Pleosporales and the Botryosphaeriales. Further analysis of morphology in combination with DNA phylogeny of the nuclear ribosomal ITS region and partial translation elongation factor 1-alpha (TEF) gene sequences delimited five species. These include S. asiminae on Asimina triloba (pawpaw fruit) in the U.S.A., and four other species occurring on apple fruit, namely S. abundans (on a local cultivar in Ardesen, Rize, Turkey), S. shaanxiensis (on ‘Fuji’ in China), S. pomigena (on ‘Golden Delicious’ in the U.S.A.), and S. henaniensis (on ‘Fuji’ in China, and ‘Golden Delicious’ and ‘Gold Rush’ in the U.S.A.). Morphologically these taxa can be distinguished based on a combination of culture characteristics and conidial morphology in vitro, which is reflected in a key to the species treated.

Genetic Stability of Magnaporthe oryzae during Successive Passages through Rice Plants and on Artificial Medium

Genetic instability of the rice blast fungus Magnaporthe oryzae has been suggested as a major factor underlying the rapid breakdown of host resistance in the field. However, little information is available on the mechanism of genetic instability. In this study, we assessed the stability of repetitive DNA elements and several key phenotypic traits important for pathogenesis after serially transferring two isolates though rice plants and an artificial medium. Using isolate 70-15, we obtained a total of 176 single-spore isolates from 10 successive rounds of culturing on artificial medium. Another 20 isolates were obtained from germ tubes formed at the basal and apical cells of 10 three-celled conidia. Additionally, 60 isolates were obtained from isolate KJ201 after serial transfers through rice plants and an artificial medium. No apparent differences in phenotypes, including mycelial growth, conidial morphologies, conidiation, conidial germination, appressorium formation, and virulence, or in DNA fingerprints using MGR586, MAGGY, Pot2, LINE, MG-SINE and PWL2 as probes were observed among isolates from the same parent isolate. Southern hybridization and sequence analysis of two avirulence genes, AVR-Pita1 and AVR-Pikm, showed that both genes were also maintained stably during 10 successive generations on medium and plants. However, one reversible loss of restriction fragments was found in the telomere-linked helicase gene (TLH1) family, suggesting some telomere regions may be more unstable than the rest of the genome. Taken together, our results suggest that phenotype and genotype of M. oryzae isolates do not noticeably change, at least up to 10 successive generations on a cultural medium and in host plants.

Genetic Diversity and Pathogenicity Analysis of Sphaceloma ampelinum Causing Grape Anthracnose in Thailand

AbstractAnthracnose is one of the major diseases affecting grape (Vitis vinifera L.) cultivars in Thailand. Isolates of Sphaceloma ampelinum, the anamorph stage of Elsinoe ampelina, were collected from various regions of Thailand. Nineteen single‐conidial isolates were evaluated for differences in conidial morphology, DNA patterns and pathogenicity. These isolates could not be unambiguously distinguished based on conidial morphology; however, they were genetically differentiated using random amplified polymorphic DNA markers. Cluster analysis by the unweighted paired grouped mean arithmetic average classified these isolates into four groups. Pathogenicity analysis using nine grape genotypes and five S. ampelinum isolates showed that ‘Wilcox321’ and ‘Illinois547‐1 were highly resistant to all isolates, suggesting their usefulness as resistant sources in future breeding programmes.

Proteomic changes associated with deletion of the Magnaporthe oryzae conidial morphology-regulating gene COM1

BackgroundThe rice blast disease caused by Magnaporthe oryzae is a major constraint on world rice production. The conidia produced by this fungal pathogen are the main source of disease dissemination. The morphology of conidia may be a critical factor in the spore dispersal and virulence of M. oryzae in the field. Deletion of a conidial morphology regulating gene encoding putative transcriptional regulator COM1 in M. oryzae resulted in aberrant conidial shape, reduced conidiation and attenuated virulence.ResultsIn this study, a two-dimensional gel electrophoresis/matrix assisted laser desorption ionization- time of flight mass spectrometry (2-DE/MALDI-TOF MS) based proteomics approach was employed to identify the cellular and molecular components regulated by the COM1 protein (COM1p) that might contribute to the aberrant phenotypes in M. oryzae. By comparing the conidial proteomes of COM1 deletion mutant and its isogenic wild-type strain P131, we identified a potpourri of 31 proteins that exhibited statistically significant alterations in their abundance levels. Of these differentially regulated proteins, the abundance levels of nine proteins were elevated and twelve were reduced in the Δcom1 mutant. Three proteins were detected only in the Δcom1 conidial proteome, whereas seven proteins were apparently undetectable. The data obtained in the study suggest that the COM1p plays a key role in transcriptional reprogramming of genes implicated in melanin biosynthesis, carbon and energy metabolism, structural organization of cell, lipid metabolism, amino acid metabolism, etc. Semi-quantitative RT-PCR analysis revealed the down-regulation of genes encoding enzymes involved in melanin biosynthesis in the COM1 mutant.ConclusionsOur results suggest that the COM1p may regulate the transcription of genes involved in various cellular processes indispensable for conidial development and appressorial penetration. These functions are likely to contribute to the effects of COM1p upon the aberrant phenotypes of M. oryzae.ReviewersThis article is reviewed by George V. Shpakovski, Karthikeyan Sivaraman (nominated by M. Madan Babu) and Lakshminarayan M. Iyer.

Effects of Cadmium and Phenanthrene Mixtures on Aquatic Fungi and Microbially Mediated Leaf Litter Decomposition

Urbanization and industrial activities have contributed to widespread contamination by metals and polycyclic aromatic hydrocarbons, but the combined effects of these toxics on aquatic biota and processes are poorly understood. We examined the effects of cadmium (Cd) and phenanthrene on the activity and diversity of fungi associated with decomposing leaf litter in streams. Leaves of Alnus glutinosa were immersed for 10days in an unpolluted low-order stream in northwest Portugal to allow microbial colonization. Leaves were then exposed in microcosms for 14days to Cd (0.06-4.5mgL(-1)) and phenanthrene (0.2mgL(-1)) either alone or in mixture. A total of 19 aquatic hyphomycete species were found sporulating on leaves during the whole study. The dominant species was Articulospora tetracladia, followed by Alatospora pulchella, Clavatospora longibrachiata, and Tetrachaetum elegans. Exposure to Cd and phenanthrene decreased the contribution of A. tetracladia to the total conidial production, whereas it increased that of A. pulchella. Fungal diversity, assessed as denaturing gradient gel electrophoresis fingerprinting or conidial morphology, was decreased by the exposure to Cd and/or phenanthrene. Moreover, increased Cd concentrations decreased leaf decomposition and fungal reproduction but did not inhibit fungal biomass production. Exposure to phenanthrene potentiated the negative effects of Cd on fungal diversity and activity, suggesting that the co-occurrence of these stressors may pose additional risk to aquatic biodiversity and stream ecosystem functioning.

Potential of chitosan coating in delaying the postharvest anthracnose (Colletotrichum gloeosporioides Penz.) of Eksotika II papaya

SummaryThe in vitro and in vivo fungicidal activity of chitosan was studied against Colletotrichum gloeosporioides, the causal agent of anthracnose in papaya fruits. Chitosan at 1.5% and 2.0% concentrations showed a fungistatic effect with 90–100% inhibition (significant at P ≤ 0.05) of the fungal mycelial growth. Changes in the conidial morphology were also observed with the higher chitosan concentrations after 7‐ h incubation. In vivo studies showed that 1.5% and 2.0% chitosan coatings on papaya not only controlled the fruit decay but also delayed the onset of disease symptoms by 3–4 weeks during 5 weeks storage at 12 ± 1 °C and slowed down the subsequent disease development. However, when leaving the fruits to ripen at ambient temperature (28 ± 2 °C), 2.0% chitosan was less effective than 1.5% in controlling the disease development. Chitosan coatings also delayed the ripening process by maintaining the firmness levels, soluble solids concentration and titratable acidity values during and after storage.

Species confirmation of fungal isolates by molecular analysis

Traditional taxonomy of hyphomycetes has been based on conidial morphology and development. In order to confirm species level for the detection and identification of the entomopathogenic fungus, we analysed the species-specific fingerprints to investigate molecular characteristics within isolates of six species and to resolve morphologically atypical isolates. The extent of fingerprint profile observed by RAPD was sufficient to confirm the species level of all the isolates. The genetic similarity among morphologically identified isolates of each species was considerably higher, allowing us to conclude that all the isolates are of same species. These results establish a molecular framework for further taxonomic, phylogenetic and comparative biological investigations.

Description of Pestalotiopsis pallidotheae: a new species from Japan

A new coelomycetes fungal species, Pestalotiopsis pallidotheae, is described. This endophytic fungus, isolated from a leaf of Japanese andromeda (Pieris japonica), has knob-tipped appendages on the apical and basal cells of the conidia. The conidial morphology is similar to that of Pestalotiopsis theae except that the color of the median cells is paler in P. pallidotheae. Molecular analyses of the ITS1-5.8S rRNA-ITS2 site placed P. pallidotheae in a group different from P. theae.

First Report of Occurrence of Benomyl Resistance in Botrytis cinerea Isolates on Raspberry in Serbia.

Botrytis fruit rot, caused by Botrytis cinerea, is one of the major diseases limiting production of raspberries (Rubus idaeus) in Serbia. Yield losses in commercial fields can exceed 50%, especially during periods of rainy, wet weather before harvest. Development of resistance to fungicides with site-specific modes of action is a serious problem in the control of B. cinerea worldwide. To insure the longest possible useful life of a fungicide, an early detection of shifts of sensitivity in pathogen population is crucial (1). The goal of this study was to evaluate sensitivity of B. cinerea isolates from commercial raspberry fields in Serbia to several fungicides that are frequently used: vinclozolin, benomyl, pyrimethanil, and fenhexamid. Initial isolation was done from sporulating berries during harvest. Single-spore isolates were identified based on colony and conidial morphology and by PCR amplification of an expected 0.7-kbp DNA fragment using B. cinerea-specific primer pair C729+/729- (3). Sensitivity of 130 isolates from six localities (20 to 30 isolates per locality) was determined on potato dextrose agar (PDA) amended with fungicides at discriminatory concentrations (1 and 10 mg/liter). Fungicides were suspended in sterile distilled water and added to autoclaved media that had cooled to 50°C. Inverted mycelial plugs (10-mm diameter), which had been cut from the edge of 4-day-old colonies on PDA, were placed on fungicide amended media and incubated for 48 h at 20°C. Treatments were replicated four times and the experiment repeated once. Strain SAS 56, which is sensitive to benzimidazoles and dicarboximides, and strain SAS 405, which is resistant to these fungicide classes, originating from German Collection of Microorganisms and Cell Cultures, were used as standards in the experiment. Isolates that did not grow at 1 mg/liter were designated as sensitive, those that grew at 10 mg/liter were considered highly resistant, and those that grew at 1 mg/liter but not at 10 mg/liter were classified as weakly resistant to all fungicides tested. Values of EC50 for all highly resistant strains were determined in radial growth experiments on PDA supplemented with a range of concentrations (5,000, 2,500, 1,000, and 500 mg/liter) of benomyl or thiophanate-methyl, according to the method described by Leroux and Gredt (2). All tested isolates were sensitive to vinclozolin, pyrimethanil, and fenhexamid. Nine of 130 isolates were highly resistant to benomyl with EC50 values between 1,056 and 1,523 mg/liter. The reference strain SAS 56 had an EC50 value of 0.17 mg/liter, compared to an EC50 value for SAS 405 strain of 1,548 mg/liter. All benomyl resistant isolates were also resistant to thiophanate-methyl and EC50 values ranged from 2,328 to 7,699 mg/liter. To our knowledge, this is the first report of benomyl resistance in isolates of B. cinerea on raspberry in Serbia.

Identification of Pyrenophora teres f. maculata, Causal Agent of Spot Type Net Blotch of Barley in North Dakota.

Net blotch of barley (Hordeum vulgare L.) caused by the fungus Pyrenophora teres (anamorph Drechslera teres) is found in two forms, net form net blotch (NFNB) and spot form net blotch (SFNB). When inoculated on susceptible varieties, P. teres f. teres produces lesions with a characteristic net-like pattern surrounded by necrosis or chlorosis (NFNB), whereas P. teres f. maculata produces lesions consisting of spots surrounded by necrosis or chlorosis (SFNB). Recently, epidemics of SFNB have occurred throughout the world (4). Currently, net blotch is a significant foliar disease of barley in the North Dakota-Northwestern Minnesota agricultural region, a leading barley-production area. Diseased barley leaf tissue was collected annually from 2004 to 2008 in Fargo and Langdon, ND. Diseased leaves were incubated to promote sporulation. Ten single-spore isolates of P. teres collected from each location each year were tested for virulence by inoculation on 20 commonly used barley net blotch differential lines. Among the 100 isolates collected, one isolate collected in Fargo in 2006 (FGOH06Pt-8) and one isolate collected in Langdon in 2008 (LDNH08Pt-4) were identified as P. teres f. maculata due to their induction of spot-type lesions across the differential set. Conidial morphology of the two isolates was similar to P. teres f. teres isolates. A pathogenicity test of all isolates was performed on regional barley cvs. Tradition, Robust, and Lacey as well as barley lines Rika and Kombar (1) as previously described (3). The net form isolate 0-1 and spot form isolate DEN2.6 (obtained from B. Steffenson, University of Minnesota) were used as controls. The P. teres f. teres isolate 0-1 produced typical net type symptoms on all barley lines except the resistant line Rika, in which only small, dark spots were observed. DEN2.6 produced pin-point spot-like lesions with an extensive yellow halo on Robust, Lacey, Rika, and Kombar, but without chlorosis on Tradition. The two newly identified isolates induced elliptical spot-type lesions measuring 3 × 6 mm, larger than those produced by P. teres f. maculata isolate DEN 2.6, suggesting a higher level of virulence. We constructed a neighbor-joining phylogenetic tree using ClustalW2 ( http://www.ebi.ac.uk/ ) based on sequence identity of the internal transcribed spacer (ITS) region from 0-1 (GenBank No. GU014819), DEN2.6 (GenBank No. GU014820), FGOH06Pt-8 (GenBank No. GU014821), and LDNH08Pt-4 (GenBank No. GU014822) as well as P. teres f. maculata, P. teres f. teres, and P. tritici-repentis (causal agent of tan spot of wheat) accessions obtained from GenBank (2). All P. teres isolates clustered together and were clearly separated from the P. tritici-repentis cluster. Isolates FGOH06Pt-8 and LDNH08Pt-4 had identical ITS sequences and differed from DEN2.6 by only a single nucleotide. To our knowledge, this is the first report of P. teres f. maculata in North Dakota. Resistance to SFNB should now be considered in local barley breeding programs and cultivar releases. Reference: (1) M. Abu Qamar. Theor. Appl. Genet. 117:1261, 2008. (2) R. M. Andrie et al. Fungal Genet. Biol. 45:363, 2008. (3) Z. Lai et al. Fungal Genet. Biol. 44:323, 2007. (4) M. S. McLean et al. Crop Pasture Sci. 60:303, 2009.

Endophytic and canker-associated Botryosphaeriaceae occurring on non-native Eucalyptus and native Myrtaceae trees in Uruguay

Species of the Botryosphaeriaceae are important pathogens causing cankers and die-back on many woody plants. In Uruguay, Neofusicoccum eucalyptorum, N. ribis and B. dothidea have previously been associated with stem cankers on plantation-grown Eucalyptus globulus. However, very little is known regarding the occurrence and species diversity of Botryosphaeriaceae in native Myrtaceae forests or what their relationship is to those species infecting Eucalyptus in plantations. The objectives of this study were to identify the Botryosphaeriaceae species present as endophytes or associated with cankers in both introduced and native tree hosts in Uruguay, and to test the pathogenicity of selected isolates obtained from native trees on Eucalyptus. Symptomatic and asymptomatic material was collected countrywide from Eucalyptus plantations and native Myrtaceae trees. Single spore cultures were identified based on conidial morphology and comparisons of DNA sequences of the ITS and EF1-α regions. Six Botryosphaeriaceae species were identified. Botryosphaeria dothidea, N. eucalyptorum and specimens residing in the N. parvum-N. ribis complex were isolated from both introduced Eucalyptus and native Myrtaceae trees, whereas Lasiodiplodia pseudotheobromae was found only on Myrcianthes pungens. Diplodia pseudoseriata sp. nov. and Spencermartinsia uruguayensis sp. nov. are novel species found only on native myrtaceous hosts. Pathogenicity tests showed that isolates obtained from native trees and identified as L. pseudotheobromae, N. eucalyptorum and the N. parvum-N.ribis complex are pathogenic to E. grandis. Interestingly, Lasiodiplodia pseudotheobromae has not previously been found on Eucalyptus in Uruguay and represents a potential threat to this host.

Description of <i>Pestalotiopsis pallidotheae</i>: a new species from Japan

A new coelomycetes fungal species, Pestalotiopsis pallidotheae, is described. This endophytic fungus, isolated from a leaf of Japanese andromeda (Pieris japonica), has knob-tipped appendages on the apical and basal cells of the conidia. The conidial morphology is similar to that of Pestalotiopsis theae except that the color of the median cells is paler in P. pallidotheae. Molecular analyses of the ITS1-5.8S rRNA-ITS2 site placed P. pallidotheae in a group different from P. theae.

A Novel Protein Com1 Is Required for Normal Conidium Morphology and Full Virulence in Magnaporthe oryzae

In Magnaporthe oryzae, pyriform conidia are the primary inoculum and the main source for disease dissemination in the field. In this study, we identified and characterized the COM1 gene that was disrupted in three insertional mutants producing slender conidia. COM1 encodes a putative transcription regulator unique to filamentous ascomycetes. The com1 disruption and deletion mutants had similar defects in conidium morphology and were significantly reduced in virulence on rice and barley seedlings. Microscopic examination revealed that the Deltacom1 mutants were defective in appressorium turgor generation, penetration, and infectious growth. COM1 was expressed constitutively in M. oryzae. The Com1 protein had putative helix-loop-helix structures and three predicted nuclear localization signal sequences. In transformants expressing COM1(335-613)-enhanced green fluorescent protein fusion constructs, fluorescence signals were observed in the nucleus. Our data indicated that the COM1 gene may encode a novel transcription regulator that regulates conidial development and invasive growth in M. oryzae.

Ambient pH intrinsically influences Trichoderma conidiation and colony morphology

Conidiation in Trichoderma has been demonstrated to be favoured by a low ambient pH and more recently PacC (Pac1) mediated pH-regulation has been implicated in the control of conidiation. In this study, ambient pH effects on conidiation were investigated in three isolates ( Trichoderma hamatum, Trichoderma atroviride and Trichoderma pleuroticola) exposed to a single blue-light burst or to mycelial injury. Disks of conidiation were observed for T. atroviride in response to a single light exposure, which clearly demonstrates that all cells are potentially competent for photoconidiation. Previous studies have suggested T. hamatum does not conidiate in response to mycelial injury, however, in this study a clear injury response was observed from pH 2.8 to 3.2. T. pleuroticola displayed three distinct pH-dependent colony morphologies from pH 2.8 to 5.2. Conidiation was strictly low pH-dependent on buffered media and observed at all pH values on unbuffered media. The dependence of the conidial phenotype on the buffering state of the medium rather than the pH per se, was unexpected as it has been suggested that conidiation is PacC regulated. Conversely, excretion of an anthraquinone was strictly pH-dependent regardless of the buffering state. These studies highlight the complexity of ambient pH effects on Trichoderma spp. and demonstrate a need to widen the scope of research to multiple species.

New record of fungus Mycoleptodiscus terrestris from New Zealand

During a survey of submersed aquatic macrophytes and their associated fungi from lakesand waterways in New Zealand’s North Island, an isolate of Mycoleptodiscus terrestris (Gerd.) Ostaz. was confirmed on the basis of colony characteristics, sporodochia, and conidial morphology. This is a new record of M. terrestris in New Zealand and a new host species, the introduced aquatic macrophyte, Ceratophyllum demersum L.

Sequencing DNA extracted from single conidia of aquatic hyphomycetes

Abstract Taxonomy and identification of aquatic hyphomycetes have traditionally been based on the morphology and development of conidia. Conidium morphology has been shaped by natural selection to facilitate attachment to substrata in running water, and convergent evolution has favoured two major types, scolecoid and multiradiate spores. As a result, phylogenetically distinct species often produce morphologically very similar spores. This raises the danger of misidentification when relying on spores, and pure cultures are often needed. An alternative is the use of unique DNA sequences (barcodes) that allow identification regardless of the presence of spores or their shape. We successfully extracted DNA from single, stained spores (cotton blue in lactophenol) and amplified a partial ITS region with a sequence similarity of up to 99.6 % to deposited GenBank sequences. With single spores released from pure culture, the success rate (DNA extraction, PCR, sequence similarity ≥99 %) was close to 50 % after optimizing the initial steps of spore recovery. Magnetic beads and Whatman FTA cards were equally successful, but FTA cards potentially allow successive amplifications of different sequences from a single extraction. A simple modification of the foam sampling technique may allow us to take a much deeper and broader look at the taxonomy and biogeography of aquatic hyphomycetes.

Neofusicoccum spp. Associated with Stem Canker and Dieback of Blueberry in Chile

Blueberry (Vaccinium spp.) plantings have significantly increased in Chile during the last decade and, currently, over 10,700 ha are cultivated throughout the country. Among other diseases, stem canker and dieback has been frequently observed in commercial plantations with incidences between 15 and 45%. The aim of this study was to identify and characterize Neofusicoccum spp. causing stem canker and dieback of blueberry in Chile. Three species, N. arbuti, N. australe, and N. parvum, were identified based on colony and conidia morphology, and nucleotide sequence analysis of the internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2). These Neofusicoccum spp. were found alone or coexisting with Pestalotiopsis spp., Truncatella spp., or Phomopsis spp. Koch's postulates showed all Neofusicoccum spp. isolated from infected plants to be pathogenic when inoculated on blueberry fruit and twigs using both mycelia and conidia suspension. All blueberry cultivars tested, including, Brigitta, Bluecrop, Brightwell, Duke, Elliott, Misty, and O'Neal, were susceptible to Neofusicoccum spp. infection. Pathogenicity tests showed N. parvum to be the most virulent species and Elliott to be the most susceptible cultivar. This report represents the first description of N. arbuti, N. australe, and N. parvum as canker-causing agents on blueberry in Chile.

Systematic analysis of the falcate-spored graminicolous Colletotrichum and a description of six new species from warm-season grasses

Species limits in the fungal genus Colletotrichum are traditionally distinguished by appressorial and/or conidial morphology or through host plant association, but both criteria are criticized for their inability to resolve distinct taxa. In previous research eight novel falcate-spored Colletotrichum species were identified from graminicolous hosts using multilocus molecular phylogenetic analysis. In the present work formal descriptions and illustrations are provided for six of the new taxa: C. hanaui sp. nov., C. nicholsonii sp. nov., C. paspali sp. nov., C. jacksonii sp. nov., C. miscanthi sp. nov. and C. axonopodi sp. nov.; and an emended description with epitypification is provided for C. eleusines. Comparison of hyphopodial appressoria and host association against phylogenetic species boundaries and evolutionary relationships in the graminicolous Colletotrichum group demonstrate that, while these characters can be useful in combination for the purpose of species diagnosis, erroneous identification is possible and species boundaries might be underestimated if these characters are used independently, as exemplified by the polyphyletic taxa C. falcatum. Appressoria have been subject to convergent evolution and were not predictive of phylogenetic relationships. Despite these limitations, the results of this work establish that in combination appressorial and host range characters could be used to generate informative dichotomous identification keys for Colletotrichum species groups when an underlying framework of evolutionary relationships, taxonomic criteria and nomenclature have been satisfactorily derived from molecular systematic treatments.

Three new species of Piricaudiopsis from southern China

Piricaudiopsis punicae sp. nov., P. rosae sp. nov. and P. rhaphidophorae sp. nov., occurring respectively on dead branches of Punica granatum, Rosa chinensis and Rhaphidophora decursiva were collected from tropical forest in southern China. They differ from the three previously described Piricaudiopsis species in conidial morphology and proliferation of the conidiogenous cell. The presence or absence of percurrent proliferation of the conidiogenous cells and conidial appendages and the height of conidia are recognized as important characters in the delimitation of species of Piricaudiopsis. A key to Piricaudiopsis species is provided.