Fungal Planet description sheets: 281-319.

Novel species of fungi described in the present study include the following from South Africa: Alanphillipsia aloeicola from Aloe sp., Arxiella dolichandrae from Dolichandra unguiscati, Ganoderma austroafricanum from Jacaranda mimosifolia, Phacidiella podocarpi and Phaeosphaeria podocarpi from Podocarpus latifolius, Phyllosticta mimusopisicola from Mimusops zeyheri and Sphaerulina pelargonii from Pelargonium sp. Furthermore, Barssia maroccana is described from Cedrus atlantica (Morocco), Codinaea pini from Pinus patula (Uganda), Crucellisporiopsis marquesiae from Marquesia acuminata (Zambia), Dinemasporium ipomoeae from Ipomoea pes-caprae (Vietnam), Diaporthe phragmitis from Phragmites australis (China), Marasmius vladimirii from leaf litter (India), Melanconium hedericola from Hedera helix (Spain), Pluteus albotomentosus and Pluteus extremiorientalis from a mixed forest (Russia), Rachicladosporium eucalypti from Eucalyptus globulus (Ethiopia), Sistotrema epiphyllum from dead leaves of Fagus sylvatica in a forest (The Netherlands), Stagonospora chrysopyla from Scirpus microcarpus (USA) and Trichomerium dioscoreae from Dioscorea sp. (Japan). Novel species from Australia include: Corynespora endiandrae from Endiandra introrsa, Gonatophragmium triuniae from Triunia youngiana, Penicillium coccotrypicola from Archontophoenix cunninghamiana and Phytophthora moyootj from soil. Novelties from Iran include Neocamarosporium chichastianum from soil and Seimatosporium pistaciae from Pistacia vera. Xenosonderhenia eucalypti and Zasmidium eucalyptigenum are newly described from Eucalyptus urophylla in Indonesia. Diaporthe acaciarum and Roussoella acacia are newly described from Acacia tortilis in Tanzania. New species from Italy include Comoclathris spartii from Spartium junceum and Phoma tamaricicola from Tamarix gallica. Novel genera include (Ascomycetes): Acremoniopsis from forest soil and Collarina from water sediments (Spain), Phellinocrescentia from a Phellinus sp. (French Guiana), Neobambusicola from Strelitzia nicolai (South Africa), Neocladophialophora from Quercus robur (Germany), Neophysalospora from Corymbia henryi (Mozambique) and Xenophaeosphaeria from Grewia sp. (Tanzania). Morphological and culture characteristics along with ITS DNA barcodes are provided for all taxa.

Leaf Blight of Buxus sempervirens in Northern Forests of Iran Caused by Calonectria pseudonaviculata.

Buxus sempervirens subsp. hyrcana (Pojark.) Takht. (boxwood) is an evergreen shrub/tree in Caspian hyrcanian forests covering the Alborz mountain range of northern Iran. During the summer of 2012, a sudden leaf and twig blight disease of boxwood was observed throughout the northern forests of Iran. Disease symptoms included circular dark spots on leaves leading to defoliation, and longitudinal brown-black streaks on the shoots. Diseased plant material was collected from the Guilan and Mazandaran areas, placed in moist chambers, and incubated at 20°C to induce sporulation. Single conidia were plated onto half-strength potato dextrose agar supplemented with 250 mg/L streptomycin and incubated at 25°C under near-ultraviolet light. Isolates were transferred to carnation leaf agar and incubated at 25°C under near-ultraviolet for morphological characterization, and representative isolates were deposited into the culture collection of the CBS-KNAW Fungal Biodiversity Centre under accession numbers CBS 134431 and CBS 134432. Gross morphological characters were determined by mounting fungal structures in lactic acid and 50 measurements at 1,000× magnification were made for all taxonomically informative characters. The observed macroconidiophores consisted of a stipe bearing a penicillate suite of reproductive branches and a stipe extension, terminating in a naviculate vesicle. The stipe extensions were septate, hyaline (85 to 160 × 2 to 4 µm), terminating in a naviculate vesicle, 6 to 11 µm in diam. Conidia were cylindrical, rounded at both ends, straight, with one septum (55 to 68 × 4 to 6 µm). These morphological observations agreed to those provided for C. pseudonaviculata Lombard, M. J. Wingf. & Crous (1,2). To confirm morphological identification, DNA sequence data were generated for the ITS1-5.8S-ITS2 region of the rDNA, and a fragment of the translation elongation factor 1-alpha gene region (3). These sequences were compared to other sequences of C. pseudonaviculata in GenBank (100% similarity for both loci), which confirmed the morphological observations. Sequences were submitted to GenBank under the accession numbers KC736850 and KC736851 for ITS, and KC736852 and KC736853 for TEF. Koch's postulates were proven by spraying a 3 × 106 conidia/ml conidial suspension of isolate CBS 134431 onto 1-year-old B. sempervirens subsp. hyrcana plants until run-off, and covering them for 24 h with a plastic bag to maintain high humidity. Control plants were sprayed with sterile water. Ten plants were used for each treatment and maintained in a greenhouse at 20 to 22°C with 95% relative humidity. Symptoms similar to those observed in nature developed within 4 days of inoculation and the test fungus was successfully reisolated from the inoculated plants. No symptoms were observed on the control plants. Boxwood blight caused by C. pseudonaviculata, was first reported in the United Kingdom in the mid-1990s and has since become widespread, causing epidemics globally (1,2,4). To our knowledge, this study represents the first report of boxwood blight in its native environment and in Iran. References: (1) P. W. Crous et al. Sydowia 54:23, 2002. (2) B. Henricot and A. Culham. Mycologia 94:980, 2002. (3) L. Lombard et al. Stud. Mycol. 66:31, 2010. (4) M. R. Saracchi et al. J. Plant Pathol. 90:581, 2009.

First Report of Pilidiella granati Causing Dieback and Fruit Rot of Pomegranate (Punica granatum) in Iran.

Iran is the largest producer of pomegranate (Punica granatum) in the world, with more than 60,000 ha currently in production. In the spring of 2011, a decline and dieback of young pomegranate trees (7 to 10 years old) were observed in the Kheir area of Fars Province. Dieback and twig blight developed toward the lower part of the stem, resulting in death of aerial tree parts and growing suckers from roots. Surface-disinfected tissues of diseased plants were plated on potato dextrose agar (PDA) and malt extract agar media. Isolates were separated into two groups that had either pale green or white aerial mycelia and sporulated after 5 to 7 days at 25°C. Pycnidia were globose and black with thin, membranous, pseudoparenchymatic walls, 80 to 140 µm in diameter. Conidia were hyaline, one-celled, elongate to fusiform, straight, and 11 to 17 × 4 to 6 µm (average 14 × 4.7 µm). Cardinal minimum growth temperatures were 8 to 10°C, optimum at 27 to 30°C, and maximum at 35°C. Radial growth rate at 30°C was 8 to 9 mm per day. Representative isolates were deposited in the CBS-KNAW Fungal Biodiversity Centre, the Netherlands (CPC 19625 = CBS 130974 and CPC 19626 = CBS 130975; GenBank JN815312 and JN815313, respectively). Genomic DNA was extracted with the UltraClean Microbial DNA Isolation Kit (MoBio Laboratories, Inc., Solana Beach, CA) and the internal transcribed spacer (ITS) region of the nrDNA operon of two isolates were sequenced as described previously (1). On the basis of morphology (3), the causal organism was identified as Pilidiella granati Sacc. This identification was corroborated by the ITS sequence data, which was identical for both colony types to GenBank HQ166057 (identities = 614 of 614 [100%]). Pathogenicity tests were conducted using two representative isolates from each group on 5-month-old P. granatum trees with 10 replicates under greenhouse conditions; 5-mm mycelial plugs from the edge of 7-day-old colonies on PDA were placed under the bark of twig wounds. Uncolonized PDA plugs were used as noninoculated controls. Pathogenicity was also tested on nonwounded fruit by placing colonized 5-mm-diameter mycelial plugs on surface-disinfected pomegranate fruits; noncolonized PDA plugs were used as controls. All treated fruit were placed in plastic bags and maintained at 25°C for 10 days. Isolates were found to be pathogenic on twigs after 2 months, giving rise to brown lesions that were 2 to 5 cm long. No lesions were observed on the controls. Furthermore, the fungus was reisolated from all infected tissues, satisfying Koch's postulates. On pomegranate fruit, the fungus colonized the fruit after 5 to 8 days, followed by the appearance of fruit rot symptoms leading to the formation of abundant pycnidia covering the skin after 10 days. No decay was observed in control inoculations. Pilidiella granati has previously been reported as a pathogen of P. granatum fruit from Europe, Asia, and the United States (2). To our knowledge, this is the first report of this pathogen causing dieback and fruit rot of pomegranate in Iran. References: (1) J. Frank et al. Persoonia 24:93, 2010. (2) L. Palou et al. Online publication. doi:10.5197/j.2044.0588.2010.022.021. New Dis. Rep. 22:21, 2010. (3) J. M. Van Niekerk et al. Mycol. Res. 108:283, 2004.

The Occurrence of Charcoal Disease Caused by Biscogniauxia mediterranea on Chestnut-Leaved Oak (Quercus castaneifolia) in the Golestan Forests of Iran.

The chestnut-leaved oak (Quercus castaneifolia) is native to the Alborz Mountains, including the Golestan Forests, in northern Iran. Trees grow up to 35 (-50) m tall with a trunk up to 2.5 (-3.5) m in diameter. During 2010, we received reports of a decline of oak trees in the Ghorogh Region of the Golestan Forests. The decline began with discolorations and browning of the leaves, resulting in drying of the foliage. Viscous liquid exudates were observed on the trunks, resulting in a brown-black discoloration of phloem and bark. In January 2011, all infected trees were dead and exhibited symptoms of charcoal disease with carbonaceous, perithecial stromata erupting from the bark on stems. Perithecia were obovoid, containing short-stipitate, amyloid asci with dark brown, ellipsoid ascospores, 14 to 19 × 7 to 9 µm, with straight germ slits along the spore length. On the basis of these morphological characteristics, the fungus was identified as Biscogniauxia mediterranea. Blast searches of the NCBI GenBank nucleotide database were done using ITS sequences derived from three cultures (CBS 129072 to 129074). GenBank Accession Nos. JF295127 to JF295129 of the isolated fungus differed by one nucleotide from B. mediterranea (GenBank Accession No. AF280624) (1,3). Pathogenicity tests were conducted using an isolate of B. mediterranea under greenhouse conditions. Six-month-old Q. castaneifolia seedlings were inoculated by means of stem wounds with a mycelial plug of colonized potato dextrose agar. After 6 months, typical decline disease symptoms associated with charcoal disease were observed and the same fungus was reisolated. Perithecia were observed on the surface of black carbonaceous stromata, which usually developed on stems of inoculated plants. The decline is known as charcoal disease because fungal growth induces a typical charcoal-black surface on diseased branches and trunks. The pathogen can easily spread through large cavity vessels, colonize bark and woody tissues, and is able to kill the host in a single, growing season. B. mediterranea causes necrosis on stems and branches of Quercus spp., including Q. suber, Q. cerris, Q. frainetto, Q. pubescens (4), Q. alba, Q. ilex, Q. imbricaria, Q. lusitanica, Q. palustris, and Q. pyrenaica (2) in Europe, North America, Africa, New Zealand, and Asia (China and India). On the basis of our current knowledge, Q. castaneifolia represents a new host of B. mediterranea, and this is the first report of this fungal pathogen from Iran causing charcoal disease on Q. castaneifolia trees in the Golestan Forest. Given its new introduction into Iran, it is highly likely that it will spread to species of Fagus, Zelkova, and other woody hosts in the area. References: (1) J. Collado et al. Mycologia 93:875, 2001. (2) D. L. Hawksworth. No. 359. Description of Pathogenic Fungi and Bacteria. CMI, Kew, UK, 1972. (3) A. Mazzaglia et al. Mycol. Res. 105:952, 2001. (4) A. Vannini and G. Scarascia Mugnozza. Eur. J. For. Pathol. 21:193, 1991.

Root and Crown Rot of Anthurium Caused by Calonectria ilicicola in Iran.

In the autumn of 2008, a severe disease of Anthurium andraeanum with wilting and root and crown rot symptoms was observed in a greenhouse in the Varamin area of Tehran. A species of Calonectria was isolated consistently from symptomatic tissues on 2% potato dextrose agar (PDA). The fungus produced perithecia and a Cylindrocladium anamorph when incubated on carnation leaf agar under near-ultraviolet light at 25°C. Perithechia were reddish brown, subglobose to ovoid, and 300 to 400 µm in diameter. Asci were clavate, hyaline, 90 to 140 × 12 to 19 µm, and tapering to a long thin stalk. Ascospores were fusoid, straight to slightly curved, 1- (-3) septate, and (30-) 37 to 50 (-65) × (4-) 5 to 6.5 (-7) µm (mean = 45 × 6 µm; n = 30). Penicillate conidiophores gave rise to stipe extensions that terminated in sphaeropedunculate vesicles (6-) 7 to 10 (-12) µm in diameter. Conidia were hyaline, cylindrical, rounded at both ends, straight, (45-) 70 to 82 (-90) × (4-) 5 to 6.5(-7) µm (mean = 62 × 6 µm; n = 30), and (1-) 3-septate. On the basis of morphology, the fungus was identified as Calonectria ilicicola Boedijin & Reitsma. Koch's postulates were fulfilled by spray inoculating 1-month-old seedlings with a conidial and mycelial suspension (105 particles per ml) of the fungus obtained from 14-day-old single-spore colonies grown on PDA at 25°C. Following inoculation, all plants were maintained in plastic bags in a glasshouse at 25 ± 1°C. After 15 to 25 days, symptoms resembling those seen in the diseased glasshouse were detected on inoculated plants. C. ilicicola was reisolated from the artificially infected tissues. No symptoms were detected on the control plants. Nucleotide sequences of the internal transcribed spacer (ITS) regions of the nrDNA operon and the partial histone H3 gene were determined for derived strain CPC 16334 as described previously (1,3). The ITS sequence (GenBank Accession No. GU057378) matched 100% (644/644 bp) with the sequence of C. ilicicola strain CBS 463.76 (GenBank AF493963) and the histone H3 sequence (GenBank GU057379) matched 99% (456/458 bp; due to two versus three AC repeats in the sequence) with that of C. ilicicola strain CBS 112217 (GenBank AY725686). To our knowledge, this is the first report of Calonectria and Cylindrocladium genera and the disease caused by C. ilicicola from Iran. References: (1) R. Cheewangkoon et al. Persoonia 23:55, 2009. (2) P. W. Crous and M. J. Wingfield. Mycotaxon 51:341, 1994. (3) P. W. Crous et al. Stud. Mycol. 50:415, 2004.