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Morphological and Molecular Characterisation of Periconia Pseudobyssoides Sp

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Morphological and Molecular Characterisation of Periconia Pseudobyssoides Sp Mycol Progress DOI 10.1007/s11557-013-0914-6 ORIGINAL ARTICLE Morphological and molecular characterisation of Periconia pseudobyssoides sp. nov. and closely related P. byssoides Svetlana Markovskaja & Audrius Kačergius Received: 23 April 2013 /Revised: 26 June 2013 /Accepted: 9 July 2013 # German Mycological Society and Springer-Verlag Berlin Heidelberg 2013 Abstract Anamorphic ascomycetes of the genus Periconia, and in other European countries, 34 species of anamorphic occurring on invasive Heracleum sosnowskyi and on other fungi was established, including Periconia spp. which frequent- native Apiaceae plants were examined during this study. On ly occurred. Part of Periconia specimens were identified as P. the basis of morphological, cultural characteristics and ITS byssoides Pers., which is widely distributed on Apiaceae and sequences a new species of Periconia closely related to other herbaceous plants, but several specimens differed from P. Periconia byssoides, is described and illustrated. The new byssoides and other known Periconia species by morphological species Periconia pseudobyssoides, collected on dead stalks and cultural characters. These specimens represented a separate of Heracleum sosnowskyi, is characterized by producing taxonomic entity which is proposed here as a new species. brownish verruculose mycelium on malt-extract agar, and Most Periconia species are widely distributed terrestrial differs from P. byssoides and other known Periconia species saprobes and endophytes colonizing herbaceous and woody by producing reddish-brown, macronematous conidiophores plants in various geographical regions and habitats (Ellis with numerous percurrent proliferations, often verruculose at 1971, 1976;Matsushima1971, 1975, 1980, 1989, 1996;Rao the apex immediately below the conidial head, verrucose and Rao 1964;Subramanian1955; Subrahmanyam 1980; ovoid conidiogenous cells arising directly from the swollen Lunghini 1978; Saikia and Sarbhoy 1982; Muntañola- apical cell cut off by a septum from the stipe apex, and Cvetković et al. 1997, 1998, 1999;Mel’nik 2000; Minter spherical, reddish-brown, characteristically ornamented ver- et al. 2001;CarmaránandNovas2003), but several species of rucose conidia. Detailed description and illustration of mor- this genus are economically important and well-studied plant phological characters, cultural characteristics and ITS se- pathogenic fungi that cause various diseases of roots and leaves, quences barcoding are also provided for P. byssoides. like Periconia circinata (L. Mangin) Sacc. and P. macrospinosa Lefebvre & Aar. G. Johnson (Odvody et al. 1977;Romeroetal. Keywords Anamorphic . Ascomycota . Pleosporales . 2001). Aquatic saprobes are represented only by three species: Taxonomy . ITS . rDNA P. prolifica Anastasiou, P. abyssa Kohlm. and P. variicolor S.A. Cantrell, Hanlin & E. Silva known from mangrove, marine and hypersaline environments (Tubaki and Ito 1973; Kohlmeyer Introduction 1977; Alias and Jones 2000; Prasannarai and Sridhar 2001; Cantrell et al. 2007). During a study of microfungi associated with Heracleum Periconia Tode is a polyphyletic anamorphic ascomycete sosnowskyi Manden. (Apiaceae), an invasive plant in Lithuania genus belonging to the Pleosporales with a rather compli- cated taxonomy and unclear phylogenetic affinity (Seifert et al. 2011). Following the main generic characters, * S. Markovskaja ( ) Periconia is characterized by producing pale to dark Laboratory of Mycology, Institute of Botany, Nature Research Centre, Žaliųjų ežerų str. 49, LT-08406 Vilnius, Lithuania brown, smooth or rarely verruculose macronematous (mostly e-mail: [email protected] with a stipe and apical conidial head, branched or un- branched), and micronematous (undifferentiated) conidio- č A. Ka ergius phores. Conidiogenous cells are monoblastic or polyblastic, Vokė Branch of Lithuanian Research Centre for Agriculture and Forestry, Žalioji sq. 2, Trakų Vokė, LT-02232 Vilnius, Lithuania discrete, ellipsoidal to spherical directly formed on a stipe or e-mail: [email protected] on branches. Sometimes the apex is sterile and conidiogenous Mycol Progress cells with conidia arise in the lower and basal part of the stipe. study was to identify, analyze and characterize Periconia The conidiogenous cells give rise to acropetal conidial chains isolates obtained from Heracleum sosnowskyi and other by a holoblastic process (except the basipetal production of Apiaceae plants in Lithuania based on morphology and cul- the conidial chains in P. prolifica), but conidial maturation is tural features as well as rDNA ITS sequence data, including basipetal (Cole and Samson 1979; Bunning and Griffiths re-examination and barcoding of P. byssoides isolates and 1982, 1984). Conidia are catenate, non-septate, spherical, discussion of the taxonomic position of the species concerned. ellipsoidal to oblong, smooth or ornamented, pale to dark brown. According to online databases such as Index Fungorum (CABI 2011) and MycoBank (Robert et al. 2005) Materials and methods the genus includes ca. 183 published names with more than 20 transferred to other genera, but currently only ca. 40 species Cultural and morphological studies are recognized as genuinely belonging to Periconia (Carmarán and Novas 2003;Kirketal.2008). Specimens of dead stalks of Heracleum sosnowskyi and other The genus was erected by Tode (1791)withPericonia Apiaceae plants were collected from various localities of Lithu- lichenoides Tode as type species. Afterwards, the type speci- ania in 2003 and repeatedly in 2011. All specimens were studied men was lost and only Tode’s drawings were left available for by light microscopy (LM); specimens of the new species were taxonomical comparison. From Tode’s expanded account of P. also examined by scanning electron microscopy (SEM). Prior to lichenoides it is known that the fungus was found on stems of examination, specimens were incubated in moist chambers at herbaceous plants belonging to various genera somewhere in room temperature and then studied under a dissecting micro- Germany. Persoon (1801)describedP. byssoides Pers. also scope Nikon SMZ 800; to obtain axenic cultures, conidia were collected in Germany on stems of unidentified herbaceous transferred to malt extract agar (MEA) media. Colonies on MEA plants and very similar to P. lichenoides. In the middle of the developed only from specimens collected in 2011. Colony twentieth century, Mason and Ellis (1953) examined six of characters and growth characteristic were studied on MEA Persoon’s collections labeled Periconia byssoides and noted plates incubated for 2–4weeksinthedarkat25±1°Candlater that P. lichenoides is probably conspecific with the latter after 1 month’s growth under light at room temperature to species or with Periconia cookei E.W. Mason & M.B. Ellis promote sporulation. Growth rates were determined as the av- and considered Sporocybe byssoides Fr. a synonym of P. erage increase in radius in mm per day of three replicate plates. byssoides. Some years later Hughes (1958)revisedthegenus Axenic cultures were subjected to molecular analysis. Periconia, and after a detailed comparison of all species he Descriptions and illustrations were made from fresh prep- designated P. byssoides as lectotype on the basis of the arations in distilled water and in 25 % lactic acid, LM photo- protologue of Sporocybe byssoides published by Fries micrographs were made with a digital camera Nikon DS-Fi1 (1932), and this lectotypification was legitimate according to mounted on a Nikon SMZ 800 binocular and with digital Art. 7.8 as associated with the sanctioning protologue of Fries camera Pentax *istDS mounted on a Olympus CX41 light (Seifert et al. 2011). Modern morphological and cultural char- microscope at magnifications up to×100. SEM images were acterizations of P. byssoides are available in various publica- obtained using a Hitachi TM1000 electron microscope. Mea- tions (Mason and Ellis 1953;Ellis1971; Matsushima 1975) surements of 50 mature conidia are indicated in the formal and also in Mycobank (Robert et al. 2005), but corresponding (extreme–)A–B–C(−extreme) where A and C are limits of the DNA sequence data are still absent. The ribosomal internal 95 % interval and B the arithmetic mean. Dimensions (only transcribed spacer (ITS) gene region is generally applied as the extremes) of other structures are given as a range of at least universal DNA barcode marker for fungi (Begerow et al. 30 measurements. Colors were determined according to 2010; Schoch et al. 2012). Molecular data of the complete Kornerup and Wanscher (1978). The holotype and other dried ribosomal ITS gene region of P. byssoides based on type or specimens, axenic cultures and permanent slides are deposited lectotype material is not available in National Center for in the Herbarium of Institute of Botany, Nature Research Biotechnology Information (NCBI) GenBank or elsewhere. Centre, Vilnius, Lithuania (BILAS). The nomenclatural nov- Only two ITS data of specimens identified as P. byssoides, but elty is deposited in MycoBank. both of them not collected in Europe (one from North America (Canada), the other from Australia) are available in Global DNA extraction, amplification, sequencing and analysis Mirror System of DNA Barcode Data (GMS-DBD 2010), but the data in this database are unfortunately incomplete, without Genomic DNA was extracted from fresh axenic cultures any ecological and morphological details. The specimens grown on Malt Extract Agar (MEA) solid medium
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