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Phylogeny and Morphology of Four New Species of <I Persoonia 25, 2010: 1–10 www.persoonia.org RESEARCH ARTICLE doi:10.3767/003158510X524150 Phylogeny and morphology of four new species of Lasiodiplodia from Iran J. Abdollahzadeh 1,3, A. Javadi 2, E. Mohammadi Goltapeh3, R. Zare 2, A.J.L. Phillips 4 Key words Abstract Four new species of Lasiodiplodia; L. citricola, L. gilanensis, L. hormozganensis and L. iraniensis from various tree species in Iran are described and illustrated. The ITS and partial translation elongation factor-1 se- Botryosphaeriaceae α quence data were analysed to investigate their phylogenetic relationships with other closely related species and EF-1α genera. The four new species formed well-supported clades within Lasiodiplodia and were morphologically distinct ITS from all other known species. Lasiodiplodia phylogeny Article info Received: 11 March 2010; Accepted: 29 June 2010; Published: 27 July 2010. taxonomy INTRODUCTION as synonyms of L. theobromae since he could not separate them on morphological characters. However, on account of its Members of the Botryosphaeriaceae (Botryosphaeriales, morphological variability and wide host range it seems likely that Dothideomycetes, Ascomycota) are cosmopolitan and occur L. theobromae is a species complex. Recent studies based on on a wide range of monocotyledonous, dicotyledonous and sequence data have confirmed this and eight new species have gymnosperm hosts (von Arx & Müller 1954, Barr 1987). They been described since 2004 (Pavlic et al. 2004, 2008, Burgess are associated with various symptoms such as shoot blights, et al. 2006, Damm et al. 2007, Alves et al. 2008). stem cankers, fruit rots, dieback and gummosis (von Arx 1987) There have been no studies on the Lasiodiplodia species in and are also known as endophytes (Slippers & Wingfield Iran apart from a few reports of L. theobromae. In a survey of 2007). Based on 28S rDNA sequence data Crous et al. (2006) Botryosphaeriaceae in Iran some Lasiodiplodia isolates that showed that Botryosphaeria is polyphyletic and they divided differed from L. theobromae in terms of morphology and ISSR it into several genera distinguishable by conidial morphology fingerprinting profile were found. The aim of this study was to and phylogenetic data. Botryosphaeria was thus restricted characterise these isolates in terms of anamorph morphology to species with Fusicoccum anamorphs. However, the clade and phylogenetic analysis. containing Diplodia/Lasiodiplodia could not be fully resolved. In a multigene genealogy Phillips et al. (2008) resolved and separated this clade into six genera including Diplodia, Lasio­ MATERIALS AND METHODS diplodia, Neodeightonia, Barriopsis, Phaeobotryon and Phaeo­ botryosphaeria. Morphological characters of the anamorphic Fungal isolation and teleomorphic states also supported the separation of During a survey of Botryosphaeriaceae in different regions of these genera. Iran in 2005–2007 some 30 Lasiodiplodia-like isolates were col- Lasiodiplodia species are common, especially in tropical and lected from various tree species showing symptoms of branch subtropical regions where they cause a variety of diseases dieback, cankers and fruit rot. Isolations were made from single (Punithalingam 1980). According to Sutton (1980) the genus conidia or by directly plating out pieces of diseased tissue after is based on Lasiodiplodia theobromae. The main features that surface sterilization (1–4 min in 70 % ethanol). Representative distinguish this genus from other closely related genera are the isolates were deposited in the culture collection of the Iranian presence of pycnidial paraphyses and longitudinal striations on Research Institute of Plant Protection (IRAN, Tehran, Iran) mature conidia. Thus far 20 species have been described and and the Centraalbureau voor Schimmelcultures (CBS, Utrecht, they are differentiated on the basis of conidial and paraphyses The Netherlands). Isolates included in the morphological and morphology. The more recently described species (described phylogenetic analyses are listed in Table 1. since 2004) have been separated not only on morphology, but also on the basis of ITS and EF-1α sequence data. Punithalin- Morphology and culture characteristics gam (1976) included several of the species known at that time To induce sporulation, isolates were transferred to 2 % water agar with sterilised pine needles on the agar surface and incu- 1 Present address: Plant Protection Department, Agriculture Faculty, Uni- bated under mixed near-UV and cool-white fluorescent light in versity of Kurdistan, P.O. Box 416, Sanandaj, Iran; corresponding author e-mail: [email protected]. a 12 h light-dark regime for 2–6 wk at 25 °C. Vertical sections 2 Department of Botany, Iranian Research Institute of Plant Protection, P.O. through conidiomata were made for some isolates with a Leica Box 1454, Tehran 19395, Iran. CM1100 cryostat microtome. Structures were mounted in 100 % 3 Plant Pathology Department, Agriculture Faculty, Tarbiat Modares Univer- lactic acid and digital images were recorded with a Leica DFC sity, P.O. Box 14115-336, Tehran, Iran. 320 camera on a Leica DMR HC microscope. Measurements 4 Centro de Recursos Microbiológicos, Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829- were made with the Leica IM500 measurement module. From 516 Caparica, Portugal. measurements of 50 conidia the mean, standard deviation © 2010 Nationaal Herbarium Nederland & Centraalbureau voor Schimmelcultures You are free to share - to copy, distribute and transmit the work, under the following conditions: Attribution: You must attribute the work in the manner specified by the author or licensor (but not in any way that suggests that they endorse you or your use of the work). Non-commercial: You may not use this work for commercial purposes. No derivative works: You may not alter, transform, or build upon this work. For any reuse or distribution, you must make clear to others the license terms of this work, which can be found at http://creativecommons.org/licenses/by-nc-nd/3.0/legalcode. Any of the above conditions can be waived if you get permission from the copyright holder. Nothing in this license impairs or restricts the author’s moral rights. 2 Persoonia – Volume 25, 2010 α AY640258 GU945339 GU945340 DQ103559 GU973866 GU973867 EF622059 AY573227 EU673310 DQ458877 GU945343 GU945345 GU945334 GU945335 GU945337 GU973861 GU973865 DQ103568 DQ103569 AY905562 GU945341 GU945342 GU973863 EF-1 DQ458874 AY573223 DQ458879 AY573219 DQ458869 DQ458880 DQ458882 DQ458881 DQ458883 EU430267 EU430268 DQ458885 DQ458884 AY573220 DQ103557 DQ103567 GU945344 GU945338 GU945336 EU144065 EU144066 EF622064 EF622063 EF445396 EF445395 EF622057 AY622054 GU973860 GU973862 GU973864 DQ458878 DQ458873 GenBank AY640255 DQ458894 AY259093 ITS GU945353 GU945354 DQ103552 GU973874 GU973875 EF622080 AY259100 AY259090 DQ458892 GU945355 GU945357 GU945346 GU945347 GU945349 GU973869 GU973873 DQ103547 DQ103548 AY905555 GU945352 GU945351 GU973871 DQ458889 AY573206 DQ458886 DQ458895 DQ458897 DQ458896 DQ458898 AY210344 AY210345 DQ458900 DQ458899 AY25909 DQ103550 AY639594 GU945356 GU945350 GU945348 EU144050 EU144051 EF622084 EF622083 AY343482 EF445362 EF622077 AY622074 GU973868 GU973870 GU973872 DQ458893 DQ458888 A.Aptroot A. Shekari Abdollahzadeh/A.J. Javadi S. Mohali Abdollahzadeh/A.J. Javadi Abdollahzadeh/A.J. Javadi Unknown A. Alves A. M. E. Sánchez/A.Trapero D. Pavlic Abdollahzadeh/A.J. Javadi Abdollahzadeh/A.J. Javadi N. Khezrinejad A. Javadi Abdollahzadeh/A.J. Javadi Abdollahzadeh/A.J. Javadi Abdollahzadeh J. S. Mohali S. Mohali J. Luque & S. Martos Z. Solel A.J.L. Phillips J. Abdollahzadeh/A.J. Javadi Abdollahzadeh/A.J. Javadi Unknown L. Mugnai A.J.L. Phillips Collector L.L. Huillier Aa H.A. van der Swart W.J. M.J. Wingfield M.J. Wingfield A. Gure A. Gure M.J. Wingfield M.J. Wingfield A.J.L. Phillips Burgess/B.T.I. Dell D. Pavlic Abdollahzadeh/A.J. Javadi Abdollahzadeh/A.J. Javadi Abdollahzadeh/A.J. Javadi Burgess T.I. Burgess T.I. O. Rangel O. Rangel Halleen F. U. Damm J. Carranza-Velásquez Unknown Unknown Abdollahzadeh/A.J. Javad Unknown Z. Solel A. Funk New Guinea Iran Iran Venezuela Iran Iran Zaire Bet Dagan, Israel Portugal Portugal Spain Africa South Iran Iran Iran Iran Iran Iran Unknown Venezuela Venezuela Spain Locality Iran Iran Iran Netherlands Italy USA Netherlands Africa South Indonesia Indonesia Ethiopia Ethiopia Mexico Mexico Portugal Australia Africa South Iran Iran Iran Australia Western Australia Western Colombia Colombia Africa South Africa South Costa Rica Unknown Iran Iran Iran Bet Dagan, Israel Canada . field soil field soil ­ ­ sp. sp. sp sp. sp. sp. sp. sp. sp. sp. Vitis vinifera Vitis Eucalyptus urophylla Fruit on coral reef coast Quercus suber Quercus ilex Vitis Citrus Citrus Syzygium cordatum Olea Mangifera indica Mangifera indica Juglans Citrus Coffea Citrus Citrus Mangifera indica Coccos Acacia mangium Acacia mangium vinifera Vitis Malus pumila Cupresuss sempervirens Phoenix dactylifera Pinus nigra Pinus radiata Pinus patula Pinus patula Pinus africana Pinus africana Pinus greggii Pinus greggii vinifera Vitis Santalum album Unknown Syzygium cordatum Mangifera indica Mangifera indica Salvadora persica Adansonia gibbosa Adansonia gibbosa Cassava Cassava vinifera Vitis Prunus salicina Gmelina arborea Unknown Unknown Mangifera indica Unknown Unknown Unknown Substrate Cupresuss sempervirens Tsuga heterophylla Tsuga CMW 13488 CBS 164.96 CBS 112546 CJA57 IRAN 921C IRAN 1502C 12539 WAC CBS 115812 CBS 115038 IRAN 1521C CBS 112549
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