ARTICLE 317 , Z. Wil- 2 rubra, Jan. Quercus Key words: #" entomogenous fungi Ips Populus tree diseases vascular wilt j\" 1 2 \ /U+U et al. 2013, van der Nest et al. and IMA · 7(2): 317–323 (2016) 317–323 · 7(2): FUNGUS IMA causes !\ /U+U \OJ ';9;U; jX , Andi Wilson 1 is a fungus et al. /='+O/='+" 5 K" 1 S. Seegmueller Iowa: fagacearum Huntiella bhutanensis Ips schmutzenhorferi that infests "## SEQUENCED STRAIN USA: ACCESSION NUCLEOTIDE SEQUENCE NUMBER The AND METHODS MATERIALS Ceratocystis fagacearum \ /U+U # G" !\$ # O" M# _ !MGO_$ J G# \OJL{DG M" "# Ceratocystidaceae (Wilken /='?" comparisons et al. 2016), particularly in order to draw 1991, 61535 - dried culture). ] J" 6 ]O {]K====== "GD 2

) previously known as C. populicola, C. {D 1 is a major causal agent of vascular wilt of oaks is a major causal agent X et al. 1944, Ceratocystidaceae spp. and Pinus G& 1 2 Ceratocystis fagacearum Huntiella bhutanensis and Huntiella C. fagacearum falls outside of ; Quercus spp.) and other Fagaceae ( 7G5" 1 are presented.

species in the USA and Canada. Grosmannia penicillata species in the USA Ceratocystis fagacearum X7J O7#"###" 2 . The data are thus intended to compliment the . 5#"##" C. harringtonii, Grosmannia penicillata, Ceratocystis fagacearum, !""#$ Ceratocystis fagacearum causes a damaging and The aim of this study was to sequence the genome of Ceratocystidaceae 5#""MG"O"_!MGO_$#J 5#]MG"O"_!MGO_$#JG#Z" disease in Populus Article info:JP'J/='UZGPUD/='UZP/*D/='U sapwood on various conifers, including Picea in Picea spinulosa Abstract: Africa LPO"¨‡ Huntiella bhutanensis and other trees in the family Fagaceae. Ceratocystis harringtonii, © 2016 International Mycological Association © 2016 International Mycological conditions: distribute and transmit the work, under the following are free to share - to copy, You Attribution: 1 2 O5" Ceratocystis fagacearum, C. harringtonii, Gros- C. harringtonii, fagacearum, for Ceratocystis sequences Draft genome penicillata, mannia IMA Genome-F 7 IMA helm de Beer Non-commercial:No derivative works:if you get Any of the above conditions can be waived 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. # # Nothing in this license impairs or restricts the author’s moral rights. permission from the copyright holder. in the eastern and north-central US (Henry in the eastern and north-central US Billings 2000, Juzwik et al. 2004). Based on ecological and phylogenetic data, however, Ceratocystis fagacearum Draft genome sequence for the oak Draft genome sequence pathogen is a wilt pathogen of oak ( IMA Genome-F 7A Genome-F IMA P'=++*9Q#"/='U=>=/'' VOLUME 7 · NO. 2 fagacearum the genus Ceratocystis5O et al. (2014). Its taxonomic position as a discrete genus is "!5O$ important vascular wilt disease known as (Appel 1995). Infection occurs in spring through wounds commonly made die rapidly and the pathogen Trees during pruning operations. can pass from one tree to another via root grafts resulting in As the trees die, pressure pads rows or patches of dying trees. EL" C. fagacearum produces a fruity odour attractive to insects These include (Lin & Phelan 1992, Cease & Juzwik 2001). L#" # " # infections (Cease & Juzwik 2001, Juzwik et al. 2004). " et al.

Biodiversity Centre (CBS), Utrecht, The Netherlands. The IMA Genome-F 7B #""/+ \/@E"!XGP /= @ Q O J G#$ Draft genome sequence of the poplar '== )"Qq 7 " 5DG " L# jE pathogen Ceratocystis harringtonii

ARTICLE et al. (2004) and sequenced on the Genomics Analyzer IIx platform (Illumina) at the Genome Centre (University 7 " # !Populus trichocarpa) has #\#5\G$7!;+=L †# U==L L $ " "# according to standard Illumina protocols and produced (Tuskan et al. 2006). This tree species is well known for its †"#E'== resistance to fungal infection, and to date only a few pathogens j#==+† # !j < '**+$ M" " \q\ ] >+' currently causing severe disease on plantation-grown poplar (CLCBio, Aarhus). This software was also used to produce #Melampsora (e. g. # " " de novo D et al. /===$ # Marssonia # # " 7 !" X et al. 2004). In order to gain increased †##"JJG\X/=!O‡et understanding of the adaptations allowing pathogens to al./=''$""]M//'!O‡ #"#X#" Pirovano 2012). The online version of the de novo prediction Melampsora larici-populina, was sequenced. This provided a software AUGUSTUS was employed to predict the putative powerful tool for studies in molecular plant-pathogen interactions "#! " " +== cankers on infected trunks and rooted cuttings (Wood & D+=#?/;=+" M '*U; K et al. /==+$ M ]\ # ?U* @ G]J7J U>=; ##"### >$ 7 " "" D 5OJ\<?>M"OJ\< American poplar species P. trichocarpa, P. balsamifera, and "OJ\<"#'?;9 P. tremuloides, X P. nigra BUSCO groups searched (Simao et al. 2015). almost entirely resistant to infection (Johnson et al. 2005). One Comparison of these genome statistics revealed no C. harringtonii elicits host defences is "##"#C. fagacearum # L "L and those determined in previous studies (Wilken et al. molecular pattern protein. This toxin is similar to the well- 2013, van der Nest et al./='?"et al./='+ characterized ceratoplatanin and cerato-ulmin, which are /='UO/='+$7†" known in C. platani and Ophiostoma novo-ulmi, respectively taxonomic position (de Beer et al. /='?$ M "L (Comparini et al./==*qet al./=';et al. " 2013). of the molecular and evolutionary processes underlying the In order to extend our knowledge on plant-pathogen "#" # resistance against fungal stem infection in poplar, the genome Authors: M.A. van der Nest, P.M. Wilken, E.T. Steenkamp, of C. harringtonii was sequenced. This genome sequence \Z\”\+•\ #" ' which plant pathogens infect woody stems and cause canker *ContactP"D¨#‡ disease in a highly resistant tree species.

SEQUENCED STRAIN

Poland: E Populus maximowiczii x P. laurifolia x P. nigra ‘Italica’ (P. xberolinensis$ '*>> J.

318 IMA FUNGUS IMA Genome – F7

Gremmen!\OJ''=>9‚\'?>9*!EL$Z Ceratocystidaceae!/9M;/Z"et al. 2016, Wilken ARTICLE jXU'+;;L$ et al. 2013, van der Nest et al. /='?$ 5 genome size, the C. harringtonii genome contains similar #9*" /@E"!XGP/@QOJ Authors: A. Hammerbacher*, P.M. Wilken, M.A. van der G#$'==3"Qq#'=" Z\\Z\+•\ the method of Barnes et al. (2001). The Genomics analyzer ' Ilx platform (Illumina) at the Genome Centre (University *Contact: ¨#‡ # \# 5 \G$ # †" "7;+=LU==LL were made using standard Illumina protocols. Sequence IMA Genome-F 7C #† the software package CLC Genomics (CLCBio, Aarhus, 5$ " # " † ! Draft genome sequence of # ==+$ Q 7 Grosmannia penicillata "##"JJG\X/= (Boetzer et al./=''$""]M The asexual morph of Grosmannia penicillata //'!O‡ /='/$7" Leptographium penicillatum (Grosmann 1931), "O" " # # " J"L\<"!OJ\<$#!J " # Ips typographus. et al./='+$\"+==# The sexual morph was discovered soon afterwards and Ceratostomella penicillata (Grosmann 1932), ] !'*;U$ from other Ceratostomella spp. and introduced a new generic RESULTS AND DISCUSSION name, Grosmannia, with G. penicillata as type species. J† Grosmannia was treated as a synonym J†"#C. harringtonii5DG of Ophiostoma !J‡ '*;* GE '*+/ K #;'=+>=;?L""#'=' "/=='$Ceratocystis (Bakshi 1951, Hunt 1956, G#";=*>/>9/ '*9'$ ´# et al. (2006) re-instated the " " # *U 7 ‡ # name to accommodate the sexual morphs of Leptographium #"/U"#''=E spp. After the implementation of the one fungus one name 7"]\#?99@D+= principles (Hawksworth 2011), Leptographium, as the older "‡#UUG#*/=" generic name, currently has preference over Grosmannia (de # 9'; " +== !E" O " /=';$ & # mitochondrial genome sequence data). BUSCO analysis two genera, L. lundbergii and G. penicillata, group in distinct #'?;9OJ\<" lineages of which the generic status needs reconsideration ';/>"LU>E !O "/=';$M" fragmented or missing. Analysis using AUGUSTUS (Stanke includes G. penicillata '> Leptographium and et al./==U$UU/>"# Grosmannia species, are referred to as the G. penicillata Genome sizes differ widely among , species complex in Leptographium s. lat. ( Six et al., 2011, for example the Fusarium graminearum and Trichoderma Linnakoski et al/='/O "/=';$ reesii " E ‡ # ;U ;? Grosmannia penicillata occurs on various conifers !‡ et al. /==9 {" et al. 2015), including Picea and Pinus X!K " while the genome of Ceratocystiopsis minuta is only 21 /=='$_ ‡ !" et al. 2016). The estimated genome (Coleoptera; Curculionidae; Scolytinae$ size of C. harringtonii !/U $ " "" L" Ips typographus "## !K "/=='q et al. 2012). Inoculation

VOLUME 7 · NO. 2 319 et al.

studies have indicated that G. penicillata is not pathogenic to ]\#+9>;@ its hosts (Jankowiak et al./==*j et al. 2015). This is with other species of Leptographium s. lat. for which whole unlike polonica,Oet " G. penicillata has a similar al. (2014), that is a common associate of I. typographus and genome size to that of Leptographium lundbergii !/UU$ !&et al.'*9; !" et al. 2015), and slightly smaller than those of G.

ARTICLE jet al. 2015). clavigera!/*9$!5] et al. 2011) and L. procerum In this study, we determined the genome sequence of !/9U$!D et al. 2014). G. penicillata 7 7 *?U @ ?/ @ "#E fragmented, and 1.2 % missing, BUSCOs. Augustus predicted evolutionary relationships of this species and other genera #9>';""#U>'9L and species in the Ophiostomatales. exon and 1995 are single-exon genes. As the type species of Grosmannia, the genome sequence of G. penicillata will E SEQUENCED STRAIN "7# "Ophiostomatales where Norway: Akershus: Ås, isol. Picea abiesK'*9=H. †""" Solheim!\/U??‚\OJ''U==9ZjXU'+;UM are of particular interest. dried culture) Authors: "'\+•\–\ NUCLEOTIDE SEQUENCE ACCESSION _^\' NUMBER *Contact: 75"¨#‡

The genomic sequence of Grosmannia penicillata !\ /U?? \OJ ''U==9$ 55OKQXOqQ IMA Genome-F 7D ]OqK‰======7 qK‰='====== Draft genome sequence for the bark beetle-associated fungus Huntiella MATERIALS AND METHODS bhutanensis

Grosmannia penicillata\/U??# Huntiella bhutanensis (; Microascales) is a !\$#MG" #"Ceratocystidaceae. The species was O" _ !MGO_$ # previously accommodated in Ceratocystis as C. bhutanensis JG#]5DGE##‡L (van Wyk et al. 2004). However, a recent taxonomic review of "5" et al. this and related genera led to the re-circumscription of these !/=';$7L!;+=+;=" #"""" insert size) were prepared and sequenced using the Illumina and molecular characteristics (de Beer et al. 2014). Thus, H. &J† /=== # '== " < bhutanensis, " 6†"" as Huntiella omanensis and H. moniliformis, were assigned Trimmomatic v. 0.36 (Bolger et al. 2014). The genome was to the genus Huntiella (de Beer et al. 2014). ""JG;*!O et al. Huntiella bhutanensis## /='/$7###JG6 Ips schmutzenhoferi (Coleoptora; Curculionidae; ###"JJG\XL;=!O‡ Scolytinae) or their galleries found on Picea spinulosa in et al. /=''$G " ]M ‰'L Bhutan (van Wyk et al. 2004). This was only the second report 10 (Boetzer & Pirovano 2012). The completeness of the of any ophiostomatoid fungi from this country and represented O" #Ceratocystidaceae J"L\<"!OJ\<'''$"" locality (van Wyk et al./==?$5 #" !J et al. /='+$ 7 # L coding genes was determined using Augustus v 3.2.2 (Stanke associates in the Ceratocystidaceae, H. bhutanensis is not et al. 2006). considered a primary pathogen and in inoculation studies, it gave rise to only small necrotic lesions on P. spinulosa trees !{ et al. 2012). RESULTS AND DISCUSSION The non-pathogenic nature of H. bhutanensis is consistent with other Huntiella species that are weak Over 11 million read pairs and 1.4 million single reads were "!O et al. 2014). It also shares #†"De-novo" many morphological characteristics with other Huntiella SPAdes resulted in 293 scaffolds which were larger than 500 " E 7###'**# ascomatal necks, and hat-shaped ascospores (van Wyk et ##"JJG\X""]M7 al. 2004). It is unusual amongst species in the genus in that it D+=#;'U{‡#/U;; "? \! et al. 2004) and it gives off a

320 IMA FUNGUS IMA Genome – F7

putrid odour unlike the sweet smelling odours typical of other RESULTS AND DISCUSSION ARTICLE Huntiella species (van Wyk et al. 2004). The aim of this study was to produce a good quality Sequencing of the Huntiella bhutanensis #"#H. bhutanensis for use in future ;+99U/*9""#'=' " 7"†#;+9///*; genera in Ceratocystidaceae. Genome sequences are ""#*U7" #H. moniliformis (van der Nest of H. bhutanensis ‡#/U>>" et al. 2014a) and H. omanensis (van der Nest et al./='?$ " # '/U¬7 ??9 ## " # " +== D+= # /=' 9=9 in the elucidation and comparison of ecological strategies, E ]\ # ?>* @ G]J7J E#L# > /U' *\OJ''?/9*jX+>9=>L$ The genome of H. bhutanensis+ that of its phylogenetically-close relative H. omanensis and possesses 1 134 fewer genes (van der Nest et al. /='?$ NUCLEOTIDE SEQUENCE ACCESSION &"#H. bhutanensis and H. NUMBER moniliformis /+? /U> U9;/>/U'] The genome sequence of H. bhutanensis will serve as an J"655OKQXDGQ]O # " KJ====== 7 species of this genus and will aid in the characterization of their KJ='====== #E#"

Authors: A. Wilson*, M.A. van der Nest, P.M. Wilken, MATERIALS AND METHODS Z\+•\' *Contact: G¨#‡ Huntiella bhutanensis \ 9/'> !\$ # M G"O"_!MGO_$ ACKNOWLEDGEMENTS #JG#_†# \OJL{DG M" O \ !\OJ$ 7 L# ] j _ 7 D ] 5DG " ! # $ # j method of Barnes et al. (2001) and sequenced using the 5"5J7QDjM\#XE7 ]G‡__E_#\5 &O"!MGO_#$D ] \ # \# 5 !\G$ j M !DjM$ !] 9>;;/$ J†" L#;+=LU==L‡ the genomes of Ceratocystis fagacearum, C. harringtonii, Huntiella were prepared and sequenced following Illumina protocol. bhutanensis"_"G ¡ " # ¡!_G¡$"#DjM " \q\ ] >+' !\q\O G$7"# genome sequence using the de novo# REFERENCES #"7#JJG\X/=!O‡ et al./=''$##" G5D!'**+$7"P#7E ""]M epidemic. Annual Review of Phytopathology 33P'=;M''9 //'!O‡ /='/$7Lde novo G 5D { 7q j !'**=$ D # gene prediction software AUGUSTUS was used to predict oak wilt fungus and other Ceratocystis spp. in Texas. European #"#! ""#Fusarium graminearum GEKG!'*+/$G""Ceratostomella (Stanke et al. 2004). Genome completeness was assessed Sacc., Ophiostoma Syd. und Rostrella Zimmermann. Antonie with the Benchmarking Universal Single-Copy Orthologs van Leeuwenhoek 18P/='M/'; !OJ\<'//$!J et al. 2015) using the fungal O O{ !'*+'$ J # # Ceratocystis, with a dataset. discussion on fungi causing sap-stain in Britain. Mycological Papers 35P'M'U O G D J G 5 ] GG 5 , et al. !/='/$ JGP " "

VOLUME 7 · NO. 2 321 et al.

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mycangia. Fungal Biology 119P'=>+M'=*/ Ceratocystis moniliformis. IMA Fungus 5P';+M'?= ARTICLE D ] J" O 5 J O &5 !/===$ jE K O _ " O5 \ 5O et al. Melampsora xcolumbiana#M. medusae and (2004) Ceratocystis bhutanensis sp. nov., associated with the M. occidentalis. Mycological Research 104P/U'M/>? Ips schmutzenhoferi on Picea spinulosa in Bhutan. jGO64JK!/='+$"# Studies in Mycology 50P;U+M;>* fungi on Picea abies in Slovenia. Forest Pathology. 45P/*=M/*> J X7" K O ´ " jE K &" & " K !/==?$ Ceratocystis O5!/=';$_G]LM'P&[: draft nuclear species infecting stem wounds on Eucalyptus grandis in South genome sequence for the plant pathogen, &[. Africa. Plant Pathology 53P?'?M?/' IMA Fungus 4P;+>M;+9 j5K<!'**+$5" " O5 G { GLD MG O qG O et crops poplar and willow. Biomass and Bioenergy 9PU*M>* al. !/='+$ _G ]L M ?P 5# " † # J‡ !'*;*$ ´ "‡ ‡‡= Chrysoporthe austroafricana, Diplodia scrobiculata, Fusarium polskim. Planta Polonica 7P'M+? nygamai, Leptographium lundbergii, Limonomyces culmigenus, J MGj_{X‰´ Stagonosporopsis tanaceti, and punctulata. IMA X!/='+$OJ\/;IMA Fungus 30P/'>M//> & !'*9'$ A monograph of Ceratocystis and MGM5!'*U;$&[, the cause of a Ceratocystiopsis. Athens, GA: University of Georgia Press. stem canker of quaking aspen. Forest Science 9P/;/M/;+ DGOqG\KG5KXO´ ´#j5O´K{"O5"K!/==U$ et al. !/='?$ _G ]LM ;P 5# " # Amanita L""Ceratocystiopsis and Grosmannia jacksonii, Ceratocystis albifundus, Fusarium circinatum, Huntiella distinct from Ophiostoma. Studies in Mycology 55P>+M*> omanensis, Leptographium procerum, Rutstroemia sydowiana, and Sclerotinia echinophila. IMA Fungus 5P?>;M?9U D G O 5 ‰ q D { j 5 et al. !/='?$ _G ]LM /P Ceratocystis manginecans, Ceratocystis moniliformis, Diplodia sapineaP 5# " sequences of Diplodia sapinea, Ceratocystis manginecans, and

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