Mycosphere Two new species of Diatrypaceae from coastal wattle in Coorong National Park, South Australia Trouillas FP 1, Sosnowski MR2 and Gubler WD1* 1Department of Plant Pathology, University of California, Davis, California 95616, USA 2South Australian Research and Development Institute, Adelaide, SA 5001, Australia Trouillas FP, Sosnowski MR, Gubler WD 2010 – Two new species of Diatrypaceae from coastal wattle in Coorong National Park, South Australia. Mycosphere 1(2), 183–188. In the present study, two species of Diatrypaceae were isolated from the wood of Acacia longifolia subsp. sophorae shrubs in the Coorong National Park, South Australia. Based on habitat, host, morphological observations and literature review, the isolates are described as the new species Diatrype brunneospora and Eutypella australiensis. These new taxa are fully described and illustrated and sequences of the internal transcribed spacer region of the nuclear ribosomal DNA are also provided. Key words – Australia – Diatrypaceae – New species – Taxonomy Article Information Received 16 June 2010 Accepted 13 July 2010 Published online 3 August 2010 *Corresponding author – Walter D. Gubler – e-mail – [email protected] Introduction been made (Rappaz 1987). Among them, eight Species of Diatrypaceae (Xylariales) are genera have been recognized (Rappaz 1987); commonly found on stems of various woody these include Cryptosphaeria Ces. & De Not. plants around the world. They are generally (four species), Diatrype Fr. (56 species), Dothi- considered to be saprotrophs, although some deovalsa Speg. (three species), Echinomyces F. species seem to be especially well established Rappaz (two species), Eutypa Tul. & C. Tul. in wood of recently dead host plants (Tiffany & (26 species), Eutypella (Nitschke) Sacc. (76 Gilman 1965). A few species in this family species), Leptoperidia F. Rappaz (four species) constitute serious plant pathogens, among them and Rostronitschkia Fitzp. (one species). Eutypa lata (Pers.) Tul. & C. Tul. (syn.: E. In North America, the works of Glawe armeniacae Hansf. & Carter) has been parti- and Rogers (1984), and Tiffany and Gilman cularly well investigated as the causal agent of (1965), constitute important additional taxono- Eutypa dieback of several woody crops world- mic studies of Diatrypaceae. Recently, Vasil- wide (Carter 1991). Cryptosphaeria lignyota yeva and Stephenson (2004, 2005, 2006, 2009) (Fr.) Auersw. (syn.: C. populina (Pers.) Sacc.) described various species of Diatrypaceae from and Eutypella parasitica R.W. Davidson & the Great Smoky Mountains National Park in R.C. Lorenz in this family represent severe the eastern United States, as well as Arkansas pathogens of forest trees in the United States and Texas. In California, 11 diatrypaceous and Europe (Davidson and Lorenz 1938, Hinds species were newly reported and described 1981, Jurc et al. 2006). from Vitis vinifera and various other woody The taxonomy of octosporous Diatry- host plants (Trouillas & Gubler 2004, Trouil- paceae has been revised and descriptions of las et al. 2010). The diversity of Diatrypaceae approximately 120 morphological species have from Argentina was also recently investigated 183 and new species as well as new records were Coorong is a wetland ecosystem and estuary, proposed (Carmaran et al. 2009). particularly valuable for its unique ecology and The current generic delineation and clas- cultural significance. Acacia longifolia subsp. sification in the Diatrypaceae as suggested by sophorae is a common shrub indigenous to Rappaz (1987) is based primarily on characters south eastern Australia and restricted mainly to of the teleomorphic states, including stroma coastal dunes. The taxa isolated for this study morphology and organization of perithecia. differed from known species in various aspects However, much overlap of taxonomic charac- of ascospores and ascomata morphology, in ters exists between the current diatrypaceous habitat and host, and deserve description as genera. Vasilyeva and Stephenson (2004) new species. acknowledged that the concept of Diatrype as delimited by Rappaz (1987) is somewhat Methods problematic, as there is, in some instances, no Pieces of dead stems of Acacia longifolia clear separation between this genus and either subsp. sophorae with black and effuse stromata Eutypa or Eutypella. Moreover, phylogenetic typical of Diatrypaceae were collected in classifications of Diatrypaceae were attempted December 2008 from the Coorong National using molecular data as well as morphological Park in South Australia. Microscopic analyses characters but evolutionary relationships in the and photography were conducted using stan- family remained uncertain (Acero et al. 2004, dard light microscopy with a Leica DMLB Carmaran et al. 2006, Trouillas et al. 2010). microscope mounted with a Leica DFC480 The diatrypaceous mycota of Australia is digital camera. Stroma and perithecial charac- remained largely unexplored. A few species teristics were examined using a Leica MZ95 have been studied, mainly because of their stereo microscope. Examinations and measure- occurrence on Vitis vinifera grapevines (Carter ments of microscopic characters were carried 1991, Mostert et al. 2004, Pitt et al. 2009). The out in water. Melzer’s reagent also was used to occurrence of Diatrypaceae on native assess amyloid reaction of the apical ascus ring. Australian host plants has been given little Thirty ascospores, ten asci (spore bearing consideration. In his account of Australian portion only) and ten perithecia of each species fungi, Cooke (1892) reported nine putative isolate were measured. Pure fungal cultures species of Diatrypaceae. An additional three were obtained from hyphal tips and grown on species were described from intertidal host potato dextrose agar (PDA) (Difco Labora- plants in north Queensland: Cryptovalsa tories, Detroit) dishes incubated under 24-hours halosarceicola K.D. Hyde on Halosarcia fluorescent lighting. Thirty conidia per isolate halocnemoides in a mangrove at Cairns Airport were measured in water as conidial masses (Hyde 1993a), Eutypa bathurstensis K.D. Hyde became visible from colonies. Sequences of the & Rappaz (Hyde 1993b) and Eutypella naqsii nuclear internal transcribed spacer (ITS) region K.D. Hyde (Hyde 1995) on Avicennia sp. at of the ribosomal DNA (rDNA) were obtained Bathurst Heads. In a study of fungi associated following standard DNA extraction procedure with Acacia and Eucalyptus plants in Melville and amplification by polymerase chain reaction Island in the Northern Territory of Australia, (PCR) using primers ITS1 and ITS4 (White et Yuan (1996) reported Cryptovalsa protracta al. 1990). Sequences have been deposited into (Pers.) De Not., Diatrype stigma (Hoffm.) Fr. GenBank. Holotypes and ex-type cultures were and Eutypella scoparia (Schwein.) Ellis & deposited at the Australian Scientific Collec- Everh. To our knowledge, the above references tions Unit, NSW Industry & Investment, constitute the only studies that illustrate the Orange, NSW, Australia (DAR). diatrypaceous biota in Australia. During the course of surveys for Taxonomy Diatrypaceae associated with Vitis vinifera in South Australia, two original diatrypaceous Diatrype brunneospora Trouillas, Sosnowski species were collected on native Acacia & Gubler, sp. nov. Figs 1–6 longifolia subsp. sophorae (coastal wattle) MycoBank 518611 Fabaceae) from Coorong National Park. The 184 Mycosphere Figs – 1–6 – Morphology of Diatrype brunneospora. 1. Asci and paraphyses. 2. Mature (brown) and immature (light brown) ascospores showing typical oil drops. 3. Cylindrical to clavate ascus, with flat apex. 4. Filiform conidia. 5. Colony after 6 days on PDA dishes (85 mm diam) incubated under 24-hours fluorescent lighting. 6. Perithecial cavities and emerging perithecial ostioles in the wood of Acacia longifolia subsp. sophorae. Bars = 10 µm in 1–4; 1 mm in 6. Etymology – referring to the brown Stromata effuse, embedded in decor- ascospores. ticated wood and incorporating host tissues, Stromata lata effusa, in ligno decorticato. rather eutypoid, blackening the host surface. Perithecia immersa, monosticha, globosa vel Perithecia in one layer, globose to ovoid, ovoidea, 0.4−0.5 mm diam, ostiolis quadri- irregularly scattered, 0.4−0.5 mm diam, sulcatis, 0.3−0.4 mm diam. Asci octospori, ostioles commonly 4-sulcate, occasionally 5- clavulato-cylindracei, parte sporifera 65−85 × sulcate, generally emerging separately, 0.3−0.4 6−7.5 µm, annulis apicalibus haud amyloideis mm diam. Asci cylindrical to clavate with flat ornati. Ascoporae oblongae, brunneae, oleosae, apex and easily breaking stipes, without (6.5−)8−10(−12.5) × 3.5−4.5 µm. Conidia amyloid reaction in Melzer’s reagent, 8-spored, filiforma, subhyalina, (20−)22−32(−34) × p. sp. 65−85 × 6−7.5 µm. Ascospores oblong, 1−1.5 µm. Hab. in ramis emortuis Acacia dark brown at maturity, usually with two oil longifolia subsp. sophorae, in South Australia. droplets, (6.5−)8−10(−12.5) × 3.5−4.5 µm. 185 Figs – 7–12 – Morphology of Eutypella australiensis. 7. Allantoid subolivaceous ascospores. 8. Filiform conidia. 9. Knobbed apex of an ascus. 10. Clavate and long-stipitate ascus. 11. Colony after 6 days on PDA dishes (85 mm diam) incubated under 24-hours fluorescent lighting. 12. Emerging perithecial ostioles in the wood of Acacia longifolia subsp. sophorae. Bars = 10 µm in 7– 10; 1 mm in 12. Cultural characteristic – Colonies white Trouillas CNP01, herb. and ex-type culture = on PDA, growing slowly and covering half DAR 80711, GenBank