Fungal Trunk Pathogens Associated With Juglans regia in the Czech Republic.

Juglans regia L. (English walnut) trees with cankers and dieback symptoms were observed in two regions in the Czech Republic. Isolations were made from diseased branches. In total, 138 fungal isolates representing 10 fungal species were obtained from wood samples and identified based on morphological characteristics and molecular methods: Cadophora novi-eboraci, Cadophora spadicis, Cryptovalsa ampelina, Diaporthe eres, Diplodia seriata, Dothiorella omnivora, Eutypa lata, Eutypella sp., Peroneutypa scoparia, and Phaeoacremonium sicilianum. Pathogenicity tests conducted under field conditions with all species using the mycelium-plug method indicated that Eutypa lata and Cadophora spp. were highly virulent to woody stems of walnut. This is the first study to detect and identify fungal trunk pathogens associated with diseased walnut trees in Europe.

Role of cell cycle regulators in adipose tissue and whole body energy homeostasis.

In the course of the last decades, metabolism research has demonstrated that adipose tissue is not an inactive tissue. Rather, adipocytes are key actors of whole body energy homeostasis. Numerous novel regulators of adipose tissue differentiation and function have been identified. With the constant increase of obesity and associated disorders, the interest in adipose tissue function alterations in the XXIst century has become of paramount importance. Recent data suggest that adipocyte differentiation, adipose tissue browning and mitochondrial function, lipogenesis and lipolysis are strongly modulated by the cell division machinery. This review will focus on the function of cell cycle regulators in adipocyte differentiation, adipose tissue function and whole body energy homeostasis; with particular attention in mouse studies.

Diaporthe diversity and pathogenicity revealed from a broad survey of grapevine diseases in Europe.

Species of Diaporthe are considered important plant pathogens, saprobes, and endophytes on a wide range of plant hosts. Several species are well-known on grapevines, either as agents of pre- or post-harvest infections, including Phomopsis cane and leaf spot, cane bleaching, swelling arm and trunk cankers. In this study we explore the occurrence, diversity and pathogenicity of Diaporthe spp. associated with Vitis vinifera in major grape production areas of Europe and Israel, focusing on nurseries and vineyards. Surveys were conducted in Croatia, Czech Republic, France, Hungary, Israel, Italy, Spain and the UK. A total of 175 Diaporthe strains were isolated from asymptomatic and symptomatic shoots, branches and trunks. A multi-locus phylogeny was established based on five genomic loci (ITS, tef1, cal, his3 and tub2), and the morphological characters of the isolates were determined. Preliminary pathogenicity tests were performed on green grapevine shoots with representative isolates. The most commonly isolated species were D. eres and D. ampelina. Four new Diaporthe species described here as D. bohemiae, D. celeris, D. hispaniae and D. hungariae were found associated with affected vines. Pathogenicity tests revealed D. baccae, D. celeris, D. hispaniae and D. hungariae as pathogens of grapevines. No symptoms were caused by D. bohemiae. This study represents the first report of D. ambigua and D. baccae on grapevines in Europe. The present study improves our understanding of the species associated with several disease symptoms on V. vinifera plants, and provides useful information for effective disease management.

Magnetic resonance defecography versus videodefecography in the study of obstructed defecation syndrome: Is videodefecography still the test of choice after 50 years?

BACKGROUND: The aim of the present study was to evaluate the diagnostic accuracy of magnetic resonance (MR) defecography and compare it with videodefecography in the evaluation of obstructed defecation syndrome. METHODS: This was a prospective cohort test accuracy study conducted at one major tertiary referral center on patients with a diagnosis of obstructed defecation syndrome who were referred to the colorectal surgery clinic in a consecutive series from 2009 to 2012. All patients underwent a clinical examination, videodefecography, and MR defecography in the supine position. We analyzed diagnostic accuracy for MR defecography and performed an agreement analysis using Cohen's kappa index (κ) for each diagnostic imaging examination performed with videodefecography and MR defecography. RESULTS: We included 40 patients with Rome III diagnostic criteria of obstructed defecation syndrome. The degree of agreement between the two tests was as follows: almost perfect for anismus (κ = 0.88) and rectal prolapse (κ = 0.83), substantial for enterocele (κ = 0.80) and rectocele grade III (κ = 0.65), moderate for intussusception (κ = 0.50) and rectocele grade II (κ = 0.49), and slight for rectocele grade I (κ = 0.30) and excessive perineal descent (κ = 0.22). Eighteen cystoceles and 11 colpoceles were diagnosed only by MR defecography. Most patients (54%) stated that videodefecography was the more uncomfortable test. CONCLUSIONS: MR defecography could become the imaging test of choice for evaluating obstructed defecation syndrome.

The HERC1 E3 Ubiquitin Ligase is essential for normal development and for neurotransmission at the mouse neuromuscular junction.

The ubiquitin-proteasome system (UPS) plays a fundamental role in protein degradation in neurons, and there is strong evidence that it fulfills a key role in synaptic transmission. The aim of the present work was to study the implication of one component of the UPS, the HERC1 E3 Ubiquitin Ligase, in motor function and neuromuscular transmission. The tambaleante (tbl) mutant mouse carries a spontaneous mutation in HERC1 E3 Ubiquitin Ligase, provoking an ataxic phenotype that develops in the second month of life. Our results show that motor performance in mutant mice is altered at postnatal day 30, before the cerebellar neurodegeneration takes place. This defect is associated with by: (a) a reduction of the motor end-plate area, (b) less efficient neuromuscular activity in vivo, and (c) an impaired evoked neurotransmitter release. Together, these data suggest that the HERC1 E3 Ubiquitin Ligase is fundamental for normal muscle function and that it is essential for neurotransmitter release at the mouse neuromuscular junction.

Detection of Grapevine Fungal Trunk Pathogens on Pruning Shears and Evaluation of Their Potential for Spread of Infection.

Four vineyards visibly affected by trunk diseases were surveyed at pruning time in 2012 and 2013 in Spain, to determine whether pruning tools are capable of spreading grapevine trunk diseases from vine to vine. In each vineyard, pruning shears were regularly rinsed with sterile water, collecting liquid samples for analysis. Molecular detection of grapevine fungal trunk pathogens (GFTPs) was performed by nested polymerase chain reaction using specific primers to detect Botryosphaeriaceae spp. Eutypa lata, Cadophora luteo-olivacea, Phaeoacremonium spp., and Phaeomoniella chlamydospora. All of these GFTPs, with the exception of E. lata, were detected in samples from the four vineyards, C. luteo-olivacea and Phaeoacremonium spp. being the most prevalent. Co-occurrence of two, three, or four different GFTPs from the same sample were found, the simultaneous detection of C. luteo-olivacea and Phaeoacremonium spp. being the most prevalent. In addition, fungal isolation from liquid samples in semiselective culture medium for C. luteo-olivacea, Phaeoacremonium spp., and P. chlamydospora was also performed but only C. luteo-olivacea was recovered from samples collected in three of four vineyards evaluated. Pruning shears artificially infested with suspensions of conidia or mycelial fragments of C. luteo-olivacea, Diplodia seriata, E. lata, Phaeoacremonium aleophilum, and Phaeomoniella chlamydospora were used to prune 1-year-old grapevine cuttings of '110 Richter' rootstock. Successful fungal reisolation from the cuttings 4 months after pruning confirmed that infested pruning shears were able to infect them through pruning wounds. These results improve knowledge about the epidemiology of GFTPs and demonstrate the potential of inoculum present on pruning shears to infect grapevines.

Study of the frequency, localisation and morphology of supernumerary teeth in 1960 Spanish non-syndromic paediatric patients.

AIM: The main objective was to study supernumerary teeth diagnosed during the routine checkups at the Paediatric Dentistry Service of Hospital de Nens, Barcelona (Spain), for four months. MATERIALS AND METHODS: A transversal, descriptive study, was performed in 1,960 patients, aged 1 to 17 years, visited during routine oral checkups. An intraoral exploration (with intraoral mirror and probe) was performed to all patients, and subjects older than 5 years also underwent panoramic x-ray examination. RESULTS: A total of 33 patients showed supernumerary teeth (1.68%), 22 boys and 11 girls. A total of 10 patients (8 boys/2 girls) had supernumerary teeth in the temporary dentition, 20 patients (12 boys/8 girls) in the permanent dentition and 3 patients (2 boys/1girls) in both temporary and permanent dentition. A total of 46 supernumerary teeth were diagnosed. CONCLUSION: Any alteration in the number of teeth in patients younger than 5 years are difficult to diagnose, as x-rays are usually not taken. We believe that starting at 5 years old, a radiological exploration (panoramic x-ray) has to be carried out as a complement to the clinical examination.

Fungal Planet description sheets: 320-370.

Novel species of fungi described in the present study include the following from Malaysia: Castanediella eucalypti from Eucalyptus pellita, Codinaea acacia from Acacia mangium, Emarcea eucalyptigena from Eucalyptus brassiana, Myrtapenidiella eucalyptorum from Eucalyptus pellita, Pilidiella eucalyptigena from Eucalyptus brassiana and Strelitziana malaysiana from Acacia mangium. Furthermore, Stachybotrys sansevieriicola is described from Sansevieria ehrenbergii (Tanzania), Phacidium grevilleae from Grevillea robusta (Uganda), Graphium jumulu from Adansonia gregorii and Ophiostoma eucalyptigena from Eucalyptus marginata (Australia), Pleurophoma ossicola from bone and Plectosphaerella populi from Populus nigra (Germany), Colletotrichum neosansevieriae from Sansevieria trifasciata, Elsinoë othonnae from Othonna quinquedentata and Zeloasperisporium cliviae (Zeloasperisporiaceae fam. nov.) from Clivia sp. (South Africa), Neodevriesia pakbiae, Phaeophleospora hymenocallidis and Phaeophleospora hymenocallidicola on leaves of a fern (Thailand), Melanconium elaeidicola from Elaeis guineensis (Indonesia), Hormonema viticola from Vitis vinifera (Canary Islands), Chlorophyllum pseudoglobossum from a grassland (India), Triadelphia disseminata from an immunocompromised patient (Saudi Arabia), Colletotrichum abscissum from Citrus (Brazil), Polyschema sclerotigenum and Phialemonium limoniforme from human patients (USA), Cadophora vitícola from Vitis vinifera (Spain), Entoloma flavovelutinum and Bolbitius aurantiorugosus from soil (Vietnam), Rhizopogon granuloflavus from soil (Cape Verde Islands), Tulasnella eremophila from Euphorbia officinarum subsp. echinus (Morocco), Verrucostoma martinicensis from Danaea elliptica (French West Indies), Metschnikowia colchici from Colchicum autumnale (Bulgaria), Thelebolus microcarpus from soil (Argentina) and Ceratocystis adelpha from Theobroma cacao (Ecuador). Myrmecridium iridis (Myrmecridiales ord. nov., Myrmecridiaceae fam. nov.) is also described from Iris sp. (The Netherlands). Novel genera include (Ascomycetes): Budhanggurabania from Cynodon dactylon (Australia), Soloacrosporiella, Xenocamarosporium, Neostrelitziana and Castanediella from Acacia mangium and Sabahriopsis from Eucalyptus brassiana (Malaysia), Readerielliopsis from basidiomata of Fuscoporia wahlbergii (French Guyana), Neoplatysporoides from Aloe ferox (Tanzania), Wojnowiciella, Chrysofolia and Neoeriomycopsis from Eucalyptus (Colombia), Neophaeomoniella from Eucalyptus globulus (USA), Pseudophaeomoniella from Olea europaea (Italy), Paraphaeomoniella from Encephalartos altensteinii, Aequabiliella, Celerioriella and Minutiella from Prunus (South Africa). Tephrocybella (Basidiomycetes) represents a novel genus from wood (Italy). Morphological and culture characteristics along with ITS DNA barcodes are provided for all taxa.

First Report of Calosphaeria pulchella Causing Canker and Branch Dieback of Sweet Cherry Trees in Spain.

In autumn 2012, severe branch cankers and diebacks of sweet cherry trees (Prunus avium L.) were observed in orchards located in two different growing areas in Alicante Province (eastern Spain). In affected trees, leaves become dried without defoliation. Sectorial wood necrosis was also observed, occasionally associated with swollen bark and gum exudates. Isolations were made from diseased branches by surface-disinfecting small fragments of symptomatic tissue in 0.5% NaOCl, double-rinsing in sterile water, and plating them onto potato dextrose agar (PDA) amended with 0.5 g liter-1 of streptomycin sulfate. Plates were incubated at 25°C in the dark for 10 days, and all colonies were transferred to PDA. Pink to red colonies with white margins were consistently isolated. All isolates produced hyaline, allantoid to oblong-ellipsoidal conidia, 4 to 6 × 1.5 to 2 µm. The fungus was identified as Calosphaeria pulchella (Pers.: Fr.) J. Schröt (anamorph Calosphaeriophora pulchella Réblová, L. Mostert, W. Gams & Crous) based on morphology (1). Identification of C. pulchella isolates was confirmed by sequence comparison in GenBank database using the internal transcribed spacer region (ITS1-5.8S-ITS2) of the rDNA. Sequences showed 100% identity and 100% query coverage with C. pulchella reference isolate CBS 115999 (EU367451) (2). The ITS sequence of one of the isolates obtained in this study was deposited into GenBank (KJ396346). Two-year-old sweet cherry trees cv. Burlat were inoculated with two representative C. pulchella isolates from different orchards (1701 and 1702). A 5-mm cork borer was used to remove bark, and an agar plug from the growing margin of 20-day-old colonies was placed directly into the fresh wound, mycelium side down. Five trees were inoculated per isolate (five branches per tree) and 25 control branches were inoculated with non-colonized agar plugs. Inoculated tissue was covered with Vaseline and Parafilm to avoid the loss of water. Branches were taken to the laboratory 9 months after inoculation and thoroughly examined for canker development. The length of vascular discoloration was evaluated in each branch and resulting data were statistically analyzed. Length of vascular discoloration on the inoculated branches (6.6 ± 0.7) was significantly longer than in control plants (2.3 ± 0.3) at P < 0.001. Perithecia were neither observed on the artificially inoculated branches nor in the diseased sweet cherry trees from the sampled orchards. C. pulchella was re-isolated from the inoculated branches and no fungi were isolated from discolored tissue of the controls, confirming Koch's postulates. Canker of sweet cherry caused by C. pulchella is responsible for reducing yields and tree longevity in California and South Australia (3). Cultivated area of sweet cherry in Spain is around 25,000 ha. Hence, the potential economic loss from this pathogen could be substantial if left unchecked. To our knowledge, this is the first report of C. pulchella as a pathogen of sweet cherry trees in Spain. References: (1) M. E. Barr. Mycologia 77:549, 1985. (2) U. Damm et al. Persoonia 20:39, 2008. (3) F. P. Trouillas et al. Plant Dis. 96:648, 2012.

First Report of Phaeoacremonium venezuelense Associated with Wood Decay of Apricot Trees in Spain.

In July 2011, a survey was conducted to evaluate the phytosanitary status of apricot trees (Prunus armeniaca L.) in an orchard in Binissalem (Mallorca Island, Spain). Fungal isolation was performed on a 40-year-old apricot trees (cv. Galta Vermella, double-grafted onto bitter almond and Japanese plum) showing a collapse of branches, chlorosis of leaves, and shoot dieback. These symptoms appeared in approximately 10% of the trees. Black spots and dark streaking of the xylem vessels were observed in cross- or longitudinal sections of the branches. Symptomatic branches were collected and wood sections (10 cm long) were cut, washed under running tap water, surface-disinfested for 1 min in a 1.5% sodium hypochlorite solution, and washed twice with sterile distilled water. The sections were split longitudinally, and small pieces of discolored tissues were plated onto malt extract agar (MEA) supplemented with 0.5 g liter-1 of streptomycin sulfate. Dishes were incubated at 25°C in the dark for 14 to 21 days, and all colonies were transferred to potato dextrose agar (PDA). A Phaeoacremonium sp. was consistently isolated from necrotic tissues (more than 50% of the isolations). Single conidial isolates were obtained and grown on PDA and MEA in the dark at 25°C for 2 to 3 weeks until colonies produced spores (3). Colonies were brownish orange on PDA and MEA. Conidiophores were short and occasionally branched, and 26 to 35 (avg. 29) µm long. Phialides were terminal or lateral, mostly monophialidic. Conidia were hyaline, oblong-ellipsoidal or fusiform-ellipsoidal, 3 to 4 (avg. 3.9) µm long, and 1 to 1.5 (avg. 1.2) µm wide. Based on these characters, the isolates were identified as Phaeoacremonium venezuelense L. Mostert, Summerb. & Crous (2,3). DNA sequencing of a fragment of the beta-tubulin gene of the isolate 9.3 using primers T1 and Bt2b (GenBank Accession No. KF765487) matched P. venezuelense GenBank accession HQ605026. Pathogenicity tests were conducted using isolate 9.3. Ten 2-year-old apricot trees of cv. Galta Rotja grown in pots were wounded in two branches with a 8-mm cork borer. A 5-mm mycelium PDA plug from a 2-week-old culture was placed in the wound before being wrapped with Parafilm. Ten control plants were inoculated with 5-mm non-colonized PDA plugs. Plants were maintained in a greenhouse at 25 to 30°C. Within 5 months, shoots on all Phaeoacremonium-inoculated branches had weak growth with chlorosis of leaves and there were black streaks in the xylem vessels. The vascular necroses that developed on the inoculated plants were 5.5 ± 0.6 cm long, significantly greater than those on the control plants (P < 0.01). Control plants did not show any symptoms. The fungus was re-isolated from discolored tissue of all inoculated cuttings, completing Koch's postulates. P. venezuelense was reported as a pathogen of grapevines in Algeria (1) and South Africa (2) and, to our knowledge, this is the first report of P. venezuelense associated with wood decay of apricot trees in Spain or any country in the world. References: (1) A. Berraf-Tebbal et al. Phytopathol. Mediterr. 50:S86, 2011. (2) L. Mostert et al. J. Clin. Microbiol. 43:1752, 2005. (3) L. Mostert et al. Stud. Mycol. 54:1, 2006.

First Report of Alternaria Black Spot of Pomegranate Caused by Alternaria alternata in Spain.

Since 2010, a new foliar and fruit disease was observed in pomegranate (Punica granatum L.) orchards in Alicante Province (eastern Spain). Symptoms included black spots on leaves and fruits, as well as chlorosis and premature abscission of leaves. Fungal isolates were obtained by surface-disinfecting small fragments of symptomatic leaf and fruit tissues in 0.5% NaOCl, double-rinsing in sterile water, and plating them onto potato dextrose agar (PDA) amended with 0.5 g/liter of streptomycin sulfate. Gray-to-black colonies were obtained, which were identified as Alternaria sp. based on the dark, brown, obclavate to obpyriform catenulate conidia with longitudinal and transverse septa tapering to a prominent beak attached in chains on a simple and short conidiophore (4). Conidia (n = 100) measured (12.2-) 20.2 (-27.6) × (5.7-) 9.2 (-12.0) µm, and had 3 to 6 transverse and 0 to 5 longitudinal septa. Single spore cultures were obtained and their genomic DNA was extracted. The internal transcribed spacer (ITS) region of rDNA and partial sequences of the beta tubulin gene were amplified and sequenced with primers ITS1-ITS4 and Bt1a-Bt1b, respectively (3). BLAST analysis of the sequences showed that they were 100% identical to a pathogenic A. alternata (Fr.) Keissl. isolate obtained from black spot disease of pomegranate in Israel (Accession No. JN247826.1, ITS and Accession No. JN247836.1, beta tubulin) (2). As all the sequences obtained showed 100% homology, ITS and beta tubulin sequences of a representative isolate (1516B) were submitted to GenBank (KF199871 and KF199872, respectively). In addition, a PCR reaction with specific primers (C_for/C_rev) designed to recognize highly virulent isolates of A. alternata causing black spot of pomegranate was used with all isolates (2). A characteristic fragment of ~950 bp was amplified in two isolates: 1552B and 1707B. Pathogenicity was assessed on plants and detached fruit of pomegranate cv. Mollar (1). Two-year-old pomegranate trees were inoculated with isolates 1552B and 1707B by spraying a conidial suspension (106 conidia/ml) onto the upper and lower leaf surfaces. Five plants per fungal isolate were used and five control plants were sprayed with sterile water. Plants were covered with transparent plastic bags and incubated in a growth chamber for 1 month at 25°C, with a 12-h photoperiod. One-month-old fruits were surface sterilized in 1.5% sodium hypochlorite solution for 1 min and rinsed twice in water. Two filter paper squares (5 × 5 mm) were dipped in the conidial suspensions and placed on the fruit surface. Inoculated fruit were incubated in a humid chamber in the dark at 25°C. Ten fruit per fungal isolate were used and 10 control fruit were inoculated with sterile water. Black spots were visible on inoculated leaves and fruit, 10 and 3 days after inoculation, respectively. Symptoms were not observed on controls. The fungus was re-isolated from leaf and fruit lesions, confirming Koch's postulates. Leaf black spot of pomegranate caused by A. alternata was first described in India in 1988, and later in Israel in 2010 affecting both fruit and leaves (1). To our knowledge, this is the first report of the disease in Spain, where it could represent a threat for pomegranate cultivation due to the increasing amount of area dedicated to this crop. References: (1) D. Ezra et al. Australas. Plant Dis. Notes 5:1, 2010. (2) T. Gat et al. Plant Dis. 96:1513, 2012. (3) N. L. Glass and G. C. Donaldson. Appl. Environ. Microbiol. 61:1323, 1995. (4) E. G. Simmons. Alternaria: An identification manual. CBS Fungal Biodiversity Center, Utrecht, Netherlands, 2007.

First Report of Olpidium bornovanus and O. virulentus on Melon in Italy.

A survey for the presence of Olpidium spp. on melon (Cucumis melo L.) was conducted during the beginning of 2013 in central Italy in an unheated greenhouse, located in the melon-producing coastal area of north Latium (central Italy, Viterbo Province) (42°23'09.31″N, 11°30'46.10″E) with a history of monosporascus root rot and vine decline (MRRVD). For this aim, 10 soil samples were collected adjacent to the roots of plants symptomatic of MRRVD, represented by root lesions and rots and loss of smaller feeder roots. Olpidium was baited from collected infested soil by growing melon (cv. Dinero) plants for 45 days. Bait plants grown in sterilized soil were used as negative controls. All the baited melon roots were analyzed by morphological and molecular methods. For the morphological analysis, feeder roots were clarified in a 1.5% KOH solution for 24 h (2) and observed under a light microscope to record the presence or absence of sporangia and resting spores of Olpidium spp., which were observed in baited melon plants grown in infested soil and not in control roots. In particular, stellate resting spores were referred to as O. virulentus because this species cannot be distinguished from O. brassicae, which does not colonize melon. O. bornovanus had smooth-walled resting spores with a honeycomb-like pattern (2). For molecular analysis, DNA was extracted from 21 melon roots and tested by multiplex PCR to confirm Olpidium spp. identification (2). Based on molecular identification, O. virulentus was identified in 40% of samples, and O. bornovanus was identified in 10%. There were no mixed infections in the same sample. Two amplified PCR products, corresponding to O. bornovanus and O. virulentus expected fragment sizes of 977 and 579 bp respectively, were sequenced (GenBank Accession Nos. KF661295 and KF661296). BLAST analysis of the sequences showed 99% nucleotide identity with O. bornovanus isolate CH from Japan collected in melon roots (AB205215) and O. virulentus isolate HY-1 from Japan collected in lettuce roots as reported by Sasaya and Koganezawa (3) (AB205204, formerly O. brassicae). At the end of the experiment, the root systems of all inoculated plants appeared brown, whereas neither symptoms nor sporangia and resting spores were observed in roots of control plants. Olpidium spp. are root-infecting plant pathogens of melon (4), acting as vectors of Melon necrotic spot virus (MNSV) and other destructive plant viruses (1). Moreover, they are directly involved in the induction of germination of ascospores of Monosporascus cannonballus, the causal agent of MRRVD of cucurbits (4). To our knowledge, this is the first report of O. virulentus and O. bornovanus on melon in Italy. References: (1) A. Alfaro-Fernández et al. J. Phytopathol. 91:1250, 2009. (2) J. A. Herrera-Vásquez et al. Mycol. Res. 113:602, 2009. (3) T. Sasaya and H. Koganezawa. J. Gen. Plant Pathol. 72:20, 2006. (4) M. E. Stanghellini and I. J. Misaghi. Phytopathology 101:794, 2011.

Fungal Planet description sheets: 214-280.

Novel species of microfungi described in the present study include the following from South Africa: Cercosporella dolichandrae from Dolichandra unguiscati, Seiridium podocarpi from Podocarpus latifolius, Pseudocercospora parapseudarthriae from Pseudarthria hookeri, Neodevriesia coryneliae from Corynelia uberata on leaves of Afrocarpus falcatus, Ramichloridium eucleae from Euclea undulata and Stachybotrys aloeticola from Aloe sp. (South Africa), as novel member of the Stachybotriaceae fam. nov. Several species were also described from Zambia, and these include Chaetomella zambiensis on unknown Fabaceae, Schizoparme pseudogranati from Terminalia stuhlmannii, Diaporthe isoberliniae from Isoberlinia angolensis, Peyronellaea combreti from Combretum mossambiciensis, Zasmidium rothmanniae and Phaeococcomyces rothmanniae from Rothmannia engleriana, Diaporthe vangueriae from Vangueria infausta and Diaporthe parapterocarpi from Pterocarpus brenanii. Novel species from the Netherlands include: Stagonospora trichophoricola, Keissleriella trichophoricola and Dinemasporium trichophoricola from Trichophorum cespitosum, Phaeosphaeria poae, Keissleriella poagena, Phaeosphaeria poagena, Parastagonospora poagena and Pyrenochaetopsis poae from Poa sp., Septoriella oudemansii from Phragmites australis and Dendryphion europaeum from Hedera helix (Germany) and Heracleum sphondylium (the Netherlands). Novel species from Australia include: Anungitea eucalyptorum from Eucalyptus leaf litter, Beltraniopsis neolitseae and Acrodontium neolitseae from Neolitsea australiensis, Beltraniella endiandrae from Endiandra introrsa, Phaeophleospora parsoniae from Parsonia straminea, Penicillifer martinii from Cynodon dactylon, Ochroconis macrozamiae from Macrozamia leaf litter, Triposporium cycadicola, Circinotrichum cycadis, Cladosporium cycadicola and Acrocalymma cycadis from Cycas spp. Furthermore, Vermiculariopsiella dichapetali is described from Dichapetalum rhodesicum (Botswana), Ophiognomonia acadiensis from Picea rubens (Canada), Setophoma vernoniae from Vernonia polyanthes and Penicillium restingae from soil (Brazil), Pseudolachnella guaviyunis from Myrcianthes pungens (Uruguay) and Pseudocercospora neriicola from Nerium oleander (Italy). Novelties from Spain include: Dendryphiella eucalyptorum from Eucalyptus globulus, Conioscypha minutispora from dead wood, Diplogelasinospora moalensis and Pseudoneurospora canariensis from soil and Inocybe lanatopurpurea from reforested woodland of Pinus spp. Novelties from France include: Kellermania triseptata from Agave angustifolia, Zetiasplozna acaciae from Acacia melanoxylon, Pyrenochaeta pinicola from Pinus sp. and Pseudonectria rusci from Ruscus aculeatus. New species from China include: Dematiocladium celtidicola from Celtis bungeana, Beltrania pseudorhombica, Chaetopsina beijingensis and Toxicocladosporium pini from Pinus spp. and Setophaeosphaeria badalingensis from Hemerocallis fulva. Novel genera of Ascomycetes include Alfaria from Cyperus esculentus (Spain), Rinaldiella from a contaminated human lesion (Georgia), Hyalocladosporiella from Tectona grandis (Brazil), Pseudoacremonium from Saccharum spontaneum and Melnikomyces from leaf litter (Vietnam), Annellosympodiella from Juniperus procera (Ethiopia), Neoceratosperma from Eucalyptus leaves (Thailand), Ramopenidiella from Cycas calcicola (Australia), Cephalotrichiella from air in the Netherlands, Neocamarosporium from Mesembryanthemum sp. and Acervuloseptoria from Ziziphus mucronata (South Africa) and Setophaeosphaeria from Hemerocallis fulva (China). Several novel combinations are also introduced, namely for Phaeosphaeria setosa as Setophaeosphaeria setosa, Phoma heteroderae as Peyronellaea heteroderae and Phyllosticta maydis as Peyronellaea maydis. Morphological and culture characteristics along with ITS DNA barcodes are provided for all taxa.

Evidence for multiple introductions and clonality in Spanish populations of Fusarium circinatum.

Fusarium circinatum is thought to have been moved around the world with pine planting stock consisting, most probably, of infected seed. In this effort, we investigate the genetic structure of F. circinatum in Spain and globally. In total, 223 isolates were studied from five regions in northern Spain and eight countries. Eight microsatellite markers revealed 66 multilocus genotypes (MLGs). Minimum spanning network analysis of MLGs by region within Spain as well as globally, discriminant analysis of principal components, and analysis of molecular variance revealed that Spanish populations are significantly differentiated and structured into two distinct groups, each one including one of the dominant genotypes observed. This result suggests that two independent introductions occurred into Spain that subsequently underwent clonal divergence and admixture. This result is further supported by the linkage disequilibrium and clonality observed for F. circinatum populations in northern Spain. The maintenance of differentiation between the clusters could result from the lack of or rare sexual reproduction in Spain. Possible introduction pathways from other countries and subsequent routes of dispersion of F. circinatum in Spain are discussed.

Effect of Environmental Factors on Mycelial Growth and Conidial Germination of Fusicladium eriobotryae, and the Infection of Loquat Leaves.

In Spain, loquat scab, caused by Fusicladium eriobotryae, is usually controlled by fungicides when there are favorable conditions for infection. Lacking specific data on the effect of weather conditions on infection by F. eriobotryae, infection periods are predicted based on the Mills table for apple scab. Experiments were conducted to determine the influence of temperature, wetness duration, relative humidity (RH), and dry periods on mycelial growth, conidial germination, and infection of loquat leaves by F. eriobotryae. F. eriobotryae was able to grow and the conidia to germinate in a wide range of temperatures, whereas no germination occurred at RH < 100%. Maximum conidial germination occurred with 24 h of wetness and germination was strongly reduced by >10 h of dryness interrupting the wetness period. Loquat infection occurred between 10 and 20°C, and disease incidence and severity increased as the duration of wetness period increased. The combined effect of temperature and wetness duration on conidial germination of F. eriobotryae and infection of leaves was described by combining ß and Gompertz equations, while the effect of dry periods on reducing the conidial germination was described by a logarithmic equation. The equations developed in this work provided a reasonable fit of the biological processes investigated and could be used for better disease control; they could be further integrated in a disease prediction system for scheduling fungicide sprays against loquat scab.

Monosporascus eutypoides, a Cause of Root Rot and Vine Decline in Tunisia, and Evidence that M. cannonballus and M. eutypoides Are Distinct Species.

Three Monosporascus eutypoides-like isolates recovered from cucurbit plants with symptoms of Monosporascus root rot and vine decline in Tunisia were compared to 28 isolates of M. cannonballus from 12 countries for phenotypic, genomic, and pathogenicity characteristics. Morphologically, M. cannonballus and M. eutypoides-like cultures were similar, each producing fertile perithecia in culture containing globose, smooth, dark brown to black ascospores. Nevertheless, all M. cannonballus isolates had one ascospore per ascus, while M. eutypoides-like isolates had mainly two to three ascospores per ascus (rarely one). The employment of the internal transcribed spacer (ITS) of nuclear ribosomal DNA, the elongation factor 1-α (EF-1α), and the ß-tubulin (ß-tub) gene sequence diversity analyses and the resulting phylogenies identified a level of polymorphism that enabled separation of M. cannonballus and M. eutypoides-like isolates. All isolates of M. cannonballus had identical EF-1α and ß-tub sequences irrespective of very diverse geographic origins, which were different from the EF-1α and ß-tub sequences of the M. eutypoides-like isolates (96 and 97% similarity, respectively). Similar results were obtained for the ITS region of rDNA. In addition, of three M. eutypoides-like isolates tested for pathogenicity, all three were pathogenic on watermelon, two were pathogenic on muskmelon, but only one was pathogenic on cucumber. The results demonstrate that the M. eutypoides-like isolates belong to the species M. eutypoides, and that M. cannonballus and M. eutypoides are distinct species.

First Report of Sirosporium celtidis Causing a Foliar Disease of European Hackberry in Spain.

Hackberry (Celtis australis L.) is widely used for reforestation and as shade tree in parks and roadside plantings in southern Europe (4). In autumn 2011, a foliar disease was observed affecting several trees planted in a garden area located in Alzira (Valencia province, eastern Spain). Symptoms appeared on lower leaf surfaces as reddish to dark brown velvety irregular spots, later becoming grayish brown on the upper surface. Most of the infected trees were prematurely defoliated. Spots on lower leaf surfaces were covered by mycelium, conidiophores, and conidia. Fungal isolates were recovered directly from the structures present on the lesions and by surface-disinfecting small fragments of symptomatic leaf tissue in 0.5% NaOCl, double-rinsing the sections in sterile water, and plating the sections onto potato dextrose agar (PDA) amended with 0.5 g of streptomycin sulfate per liter. Single conidium cultures made onto PDA were maintained for 2 months at 25°C in darkness for morphological examination. Conidia were thick walled, dark reddish brown, often markedly curved or coiled, cylindrical to obclavate, smooth, wrinkled, or verrucose, typically multicellular, 2 to 40 transversely septate and occasionally with 1 to 3 longitudinal or oblique septa that were often constricted, 20 to 96 (44.9) × 6 to 9 (7.1) µm, with an inconspicuous scar at the base. Morphological characters corresponded to the description of Sirosporium celtidis (Biv. ex Spreng) M. B. Ellis published in 1963 (3). The internal transcribed spacer (ITS) region of the rDNA was amplified with the primers ITS1 and ITS4 from DNA extracted from the isolate AL1, and sequenced (GenBank Accession No. JX397963). The sequence was identical to that obtained from an isolate of S. celtidis from the Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands (CBS 289.50). Pathogenicity tests were conducted on five 2-year-old hackberry trees by spraying onto the upper and lower leaf surfaces a conidial suspension of S. celtidis (approximately 50 ml/plant, 106 conidia/ml of water). Five control plants were sprayed with sterile water. Plants were covered with clear plastic bags and incubated in a growth chamber for 72 h at 25°C with a 12-h photoperiod. First leaf spots were visible on inoculated plants after 7 days, but symptoms were not observed on control plants. The fungus was reisolated from leaf lesions on inoculated plants, confirming Koch's postulates. S. celtidis was first described in Sicily in 1815 (3) and has been recorded on various hackberry species in Mediterranean countries and the USA (1,2). To our knowledge, this is the first report of the disease in Spain. The economic and ecological significance of the pathogen in natural ecosystems in Spain remains to be determined but it could certainly become a serious problem for nurseries and urban plantings. References: (1) S.O. Cacciola. 2000. Plant Dis. 84, 492. (2) D. H. Linder. 1931. Ann. Mo. Bot. Garden 18, 31. (3) M. B. Ellis. 1963. Mycological Papers, No. 87. Commonw. Mycol. Inst. Kew, England. (4) S. Pauleit et al., Urban For. Urban Green. 1:83, 2002.

First Report of Cylindrocladiella parva and C. peruviana Associated with Black-foot Disease of Grapevine in Spain.

From 2007 to 2009, Cylindrocladiella-like isolates were recovered from grapevine (Vitis vinifera L.) roots with symptoms of black-foot disease in Spain, where the causal agents of this disease have been previously reported as Campylocarpon and Cylindrocarpon species (1,2). Three representative isolates were selected to confirm their identity: CPa1 and CPa2 from Asturias (northern Spain), and CPe523 from Cuenca (central Spain). Isolates were incubated on malt extract agar (MEA) and Spezieller Nährstoffarmer Agar (SNA) with carnation leaves (4) at 25°C for 10 days in darkness. On MEA, colonies developed light brown, cottony mycelium. On SNA, all three isolates produced chlamydospores in chains, and conidia were zero-to one-septate, but CPa1 and CPa2 produced longer conidia (10.4 to 18.9 [15.3] × 1.7 to 3.1 [2.4] µm) than CPe523 (6.4 to 12.3 [9.7] × 1.6 to 3.3 [2.4] µm). A fragment of the beta-tubulin gene from all isolates was sequenced with primers T1 and Bt2b (1) and deposited in GenBank (Accession Nos. JQ693133, JQ693134, and JQ693135). CPa1 and CPa2 showed high similarity (99%) to Cylindrocladiella parva (AY793486) and CPe523 showed high similarity (99%) to C. peruviana (AY793500), which is in agreement with the corresponding morphological features of these species (4). Pathogenicity tests were conducted with inoculum produced on wheat (Triticum aestivum L.) seed soaked for 12 h in 300 ml of distilled water and autoclaved three times. Inoculum was prepared by inoculating two fungal disks (8 mm in diameter) of a 2-week-old culture of each isolate grown on potato dextrose agar to wheat seed and incubation at 25°C for 4 weeks. One-month-old grapevine seedlings were planted individually in 220-cc pots filled with a potting medium of sterilized peat moss and 10 g of inoculum, and grown in the greenhouse at 25°C in a completely randomized design. Controls were inoculated with sterile, noninoculated wheat seed. There were six replicate plants per isolate, with an equal number of controls, and the experiment was repeated once. Symptoms developed in all plants by 20 days post-inoculation and consisted of reduced vigor, necrotic root lesions, and occasionally mortality, all of which resembled the symptoms from grapevines in the field from which the isolates were originally recovered. Mean shoot dry weights of inoculated plants (0.25, 0.16, and 0.28 g for CPa1, Cpa2, and CPa523, respectively) were significantly lower (P < 0.05) than that of the controls (0.74 g). Mean root dry weights of inoculated plants (0.28, 0.16, and 0.29 g for CPa1, Cpa2, and CPa523, respectively) were also significantly lower (P < 0.05) than that of the controls (0.68 g). Isolates recovered from the roots of inoculated plants were identical morphologically and molecularly to C. parva and C. peruviana, thereby satisfying Koch's postulates. No symptoms were observed on the control plants. These Cylindrocladiella spp. have been reported from nurseries or vineyards in South Africa and New Zealand (3). To our knowledge, this is the first report of C. parva and C. peruviana associated with black-foot disease of grapevine in Spain, and in Europe. References: (1) S. Alaniz et al. Plant Dis. 91:1187, 2007. (2) S. Alaniz et al. Plant Dis. 95:1028, 2011. (3) E. E. Jones et al. Plant Dis. 96:144, 2012. (4) L. Lombard et al. Mycol. Progress DOI 10.1007/s11557-011-0799-1, 2012.