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 Leaf Spot Caused by Myrothecium roridum on Coffea canephora in Brazil.

Coffea canephora (conilon coffee) represents approximately 30% of the coffee marketed worldwide. The state of Espírito Santo is the largest conilon coffee-producing state in Brazil. In 2013 and 2014, leaves with a leaf spot were observed on most of the conilon coffee seedlings in a commercial nursery in Laranja da Terra, Espírito Santo, Brazil. The infected leaves were deposited in the VIC Herbarium (VIC 42482) and a pure single-spore culture of the pathogen was deposited in the culture collection of the Universidade Federal de Viçosa (Accession No. COAD 1729). The initial symptoms were circular, brown to dark brown lesions with yellow margins occurring on both leaf surfaces. In high humidity, concentric rings formed and the lesions expanded rapidly to reach up to 30 mm in diameter, and later became dark brown with a grayish center. Black sporodochia with white, and marginal mycelial tuffs bearing black spore masses were observed in the older lesions. These symptoms were consistent with those of Myrothecium leaf spot reported on Coffea spp. (3). Microscopic observation revealed aseptate, hyaline, and cylindrical conidia, rounded at both ends, greenish to black in mass, and 5 to 6 µm long and 1 to 2 µm wide. The symptoms and morphological characteristics described above matched the description of Myrothecium roridum Tode (4). To confirm this identification, DNA was extracted using a Wizard Genomic DNA Purification Kit and the sequence of an internal transcribed spacer (ITS) region was obtained and deposited in GenBank (Accession No. KJ815095). The sequence of the ITS region exhibited 100% identity over 561 bp with another M. roridum sequence in GenBank (JF343832). To verify the pathogenicity of the fungus, healthy leaves of the C. canephora clones 12v and 14 (four seedlings each) were wounded superficially with a sterilized needle and inoculated by spraying them with a suspension of M. roridum conidia (106 conidia ml-1). The seedlings were covered with plastic bags and incubated in a growth chamber at 25°C under a photoperiod of 12 h light/12 h dark for 5 days. The control seedlings were sprayed with distilled water and incubated similarly. Fifteen days after inoculation, symptoms in all inoculated seedlings were consistent with those initially observed on the naturally infected seedlings, whereas the controls remained healthy. Re-isolation and identification confirmed Koch's postulates. M. roridum has a wide host range, and symptoms were similar to those reported in other hosts of the pathogen in Brazil (2,3). There is only one report of M. roridum on C. canephora in Colombia (1); however, this pathogen was previously reported on C. arabica in Brazil, Colombia, Costa Rica, Guatemala, India, Indonesia, Puerto Rico, and the Virgin Islands (1,3). To our knowledge, this is the first report of a leaf spot caused by M. roridum on conilon coffee in Brazil. The cultivation of conilon coffee is increasing and the reported leaf spot disease affects the quality of the seedlings in nurseries. It is therefore important to conduct a thorough study of management strategies for this disease. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab. ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases , 27 May 2014. (2) A. M. Quezado Duval et al. Braz. J. Microbiol. 41:246, 2010. (3) S. F. Silveira et al. Fitopatol. Bras. 32:440, 2007. (4) M. Tulloch. Mycol. Pap. No. 130. CMI, Wallingford, UK, 1972.

Colletotrichum boninense Causing Anthracnose on Coffee Trees in Brazil.

In Brazil, dieback and necrosis of leaves and berries of coffee trees (Coffea arabica and C. canephora) are common symptoms of anthracnose disease caused by Colletotrichum gloeosporioides (Penz.) Sacc. In April 2010, these symptoms were observed in 100% of the plants from different coffee plantations in the Brazilian states of Espírito Santo and Bahia. Ten isolates were obtained from symptomatic leaves and berries from these areas. Of the 10 isolates, one had distinct conidial morphology with hyaline and ellipsoid conidia measuring 10 to 16 × 5.0 to 7.5 µm and melanized irregular or spatulated-shaped appressoria measuring 7.5 to 11.0 × 5.5 to 8.5 µm, formed either solitary or concatenated, which concurred with the conidia description of Colletotrichum boninense. In order to confirm the identity of this isolate, the internal transcribed spacer (ITS) rRNA region and the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene were sequenced (GenBank Accession Nos. JF683320 and JF331654, respectively) and compared to sequences from a database of C. boninense, confirming that the isolate was definitely C. boninense sensu lato, since it was exactly identical to other sequences in a large clade of isolates. To verify the pathogenicity of C. boninense in coffee and to compare the symptoms with those caused by C. gloeosporioides, leaves and berries were inoculated with the isolate of C. boninense and one representative isolate of C. gloeosporioides, both expressing the GFP (green fluorescent protein) gene. The isolates were grown for 7 days on potato dextrose agar and a conidial suspension (106 conidia × ml-1) was used to inoculate the organs, wounded and non-wounded, at different stages of development. In non-wounded organs, the conidial suspension was inoculated on the surface, and in leaves and berries used as control, the suspensions were substituted for sterile water. Leaves and berries were wounded with a sterilized needle and inoculated with 20 and 10 µl of the conidial suspension, respectively. Inoculated materials were incubated at 25°C and 100% relative humidity. The experiment was performed twice and evaluated daily for a week. No symptoms were observed on the control and non-wounded organs, while wounded organs exhibited typical anthracnose symptoms for both species. In berries, C. gloeosporioides consistently caused more severe symptoms at a faster rate than C. boninense. Both fungi caused necrosis in young but not old leaves. Typical acervuli were observed on the lesions and the fungus was successfully recovered from the inoculated tissues, which was confirmed by fluorescence microscopy, fulfilling Koch's Postulates. C. boninense has been identified as a pathogen causing anthracnose in a range of hosts worldwide. However, in Brazil, it has only been reported in pepper (Capsicum annuum) (3), passion fruit (Passiflora) (4), Hippeastrum (1) and in the medicinal plant Maytenus ilicifolia (2). To our knowledge, this is the first report of C. boninense associated with anthracnose of coffee trees in Brazil. Since the symptoms are similar to those caused by C. gloeosporioides, it can be stated that both species are associated with this disease in commercial coffee plantations in Brazil. Therefore, control strategies should consider the occurrence of C. boninense. References: (1) D. F. Farr et al. Mycol. Res. 110:1395, 2006. (2) S. A. Pileggi et al. Can. J. Microbiol. 55:1076, 2009. (3) H. J. Tozze et al. Plant Dis. 93:106, 2009. (4) H. J. Tozze et al. Australas. Plant Dis. Notes 5:70, 2010.

First Report of Leaf Blight on Rubus brasiliensis Caused by Colletotrichum acutatum in Brazil.

There are more than 300 blackberry (Rubus) species worldwide. Rubus brasiliensis Mart. is a native Brazilian species found in tropical forests. In January 2009, samples of R. brasiliensis with severe leaf blight were collected from an area of rain forest in the city of São Miguel do Anta, State of Minas Gerais, Brazil. Dark spots began developing in the young leaves and progressed to necrotic spots with occasional twig dieback. From the spots, a fungus was isolated with the following morphology: acervuli that were 20 to 50.0 × 50 to 125.0 µm and hyaline amerospores that were ellipsoid and fusiform and 7.5 to 23.75 × 2.5 to 5.0 µm. On the basis of these morphological characteristics, the fungus was identified as Colletotrichum acutatum. In Brazil, C. acutatum is reported in apple, citrus, strawberry, peach, plum, nectarine, olive, medlar, and yerba-mate, but it was not reported as the causal agent of leaf blight in R. brasiliensis. A sample was deposited in the herbarium at the Universidade Federal de Viçosa, Minas Gerais, Brazil (VIC 31210). One representative isolate, OLP 571, was used for pathogenicity testing and molecular studies. Identity was confirmed by amplifying the internal transcribed spacer (ITS) regions of the ribosomal RNA with primers ITS4 (3), CaInt2 (a specific primer for C. acutatum [2]) and CgInt (a specific primer for C. gloeosporioides [1]). Isolates of C. acutatum (DAR78874 and DAR78876) and C. gloeosporioides (DAR78875) obtained from Australian olive trees were used as positive controls. The primers ITS4 and CaInt2 amplified a single DNA product of 500 bp expected for C. acutatum. OLP 571 was grown for 7 days on potato dextrose agar. Young leaves of R. brasiliensis were inoculated with a conidial suspension (106 conidia/ml) on young leaves. Inoculated plants were maintained in a moist chamber for 2 days and subsequently in a greenhouse at 25°C. Necrotic spots similar to those described were detected on young leaves 3 days after the inoculation. Control leaves, on which only water was sprayed, remained healthy. The same fungus was reisolated from the inoculated symptomatic tissues. To our knowledge, this is the first report of C. acutatum causing leaf blight in the native species of R. brasiliensis in Brazil. References: (1) P. R. Mills et al. FEMS Microbiol. Lett. 98:137, 1999. (2) S. Sreenivasaprasad et al. Plant Pathol. 45:650, 1996. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.