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Researchrepository.Murdoch.Edu.Au http://tweaket.com/CPGenerator/?id=4872 View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Research Repository MURDOCH RESEARCH REPOSITORY http://researchrepository.murdoch.edu.au This is the author's final version of the work, as accepted for publication following peer review but without the publisher's layout or pagination. Sakalidis, M.L. , Hardy, G.E.St.J. and Burgess, T.I. (2011) Class III endophytes, clandestine movement amongst hosts and habitats and their potential for disease; a focus on Neofusicoccum australe. Australasian Plant Pathology, 40 (5). pp. 510-521. http://researchrepository.murdoch.edu.au/4872 Copyright © Australasian Plant Pathology Society Inc. 2011 It is posted here for your personal use. No further distribution is permitted. 1 of 1 1/09/2011 10:36 AM Author's personal copy Class III endophytes, clandestine movement amongst hosts and habitats and their potential for disease; a focus on Neofusicoccum australe Monique L. Sakalidis & Giles E. StJ. Hardy & Treena I. Burgess Abstract Neofusicoccum australe is a class III endophyte Introduction characterised by a quiescent passive life phase and an active pathogenic life phase as a latent pathogen. The latter life The life strategies of the Botryosphaeriaceae generally fit into stage has been observed worldwide for numerous woody the description of class III endophytes (Rodriguez et al. 2009). horticultural hosts. In this study, we have re-evaluated Many members have a broad host range (Punithalingam GenBank ITSrDNA sequence data to establish the current 1976, 1980), they fruit on senescing tissue (Slippers and host and geographical range of N. australe. Additionally, Wingfield 2007), are found in above ground plant parts we have interrogated the diversity of N. australe in (Johnson et al. 1992) and multiple species are found often Australia using microsatellite markers to ascertain if there within localised infections in the host tissue (Sakalidis et al. are any host or site preference for different genotypes. N. 2011a, b; Spagnolo et al. 2011). However; unlike other Class australe has a widespread distribution across ten countries III fungi they have occasionally been observed in host seeds and colonises 46 hosts from 18 plant families; mainly (Johnson et al. 1998; Bihon et al. 2011) and there is also angiosperms, some coniferous species and one monocot. evidence of infections in horticultural crops occurring via Phylogenetic analysis of the ITSrDNA sequence indicates pruning wounds (Brown and Hendrix 1981; Smith et al. there is a single dominant ITS genotype present in most 1994, 1996; Amponsah et al. 2011; Sakalidis et al. 2011c). locations and there are another 12 rare or moderately rare Pathogenicity trials are successfully used to prove potential genotypes. Populations of N. australe in Australia appear to for disease development (Amponsah et al. 2011; Sakalidis et be highly diverse, and there is no discernable host or habitat al. 2011c) but little is known about possible passive restriction. The dominance of N. australe in native forest (endophytic) or delayed active (latent pathogen) life stages throughout the southwest of Western Australia, and its rarity that may also initiate from the same infections. elsewhere in native vegetation, while being common as a Neofusicoccum australe (Slippers, Crous & M.J. Wingf.) pathogen of horticultural hosts, suggests that this species is Crous, Slippers & A.J.L. Phillips, was originally described endemic to Western Australia. as Botryosphaeria australis from native Acacia spp. in Victoria and exotic Sequoiadendron giganteum in Canberra; Keywords Botryosphaeriaceae . Population . Genetic both in eastern Australia (Slippers et al. 2004). N. australe diversity. Latent pathogens . Distribution had previously been identified as Botryosphaeria ribis from various hosts including Banksia coccinea (Shearer et al. 1995), Banksia caleyi and Eucalyptus marginata (Australia) and a Pistacia sp. (Italy) (Smith and Stanosz 2001) and as Botryosphaeria lutea on Fraxinus augustifolia (Portugal) M. L. Sakalidis (*) : G. E. StJ.S. Hardy Hardy : T· . TI.. I.Bu Burgessrgess (Phillips et al. 2002) and from Protea cynaroides (South State Centre of Excellence in Climate Change, Africa) (Denman et al. 2003). In 2006, an extensive Woodland and Forest Health, School of Biological Sciences, phylogenetic evaluation of the Botryosphaeriaceae identi- Murdoch University, Perth 6150, Australia fied several lineages and Botryosphaeria was only e-mail: [email protected] retained for Botryosphaeria dothidea and Botryosphaeria Author's personal copy M.L. Sakalidis et al. corticis (Crous et al. 2006). Taxa that were characterised variation between isolates of this species. A one bp difference by Fusicoccum-like anamorphs and Dichomera-like syn- between N. australe isolates in the ITS gene region and up to anamorphs were placed into Neofusicoccum; which is three bp differences between isolates in the EF1-a gene where N. australe currently resides (Crous et al. 2006). region has been reported (Lazzizera et al. 2008). There have been many reports describing N. australe as In this study, we have re-evaluated GenBank sequence a significant pathogen of grapevine; it is associated with a data to establish the current host and geographical range variety of symptoms affecting trunk, canes, buds and the of N. australe worldwide. Additionally, we have then fruit. Symptoms include stem cankers, dieback, wedge focussed on the Australian diversity of N. australe to shaped lesions in grapevine stems and fruit rot. N. australe ascertain if there is any host or site preference for different has been associated with symptomatic grapevine in Western genotypes and to compare isolates from WA and eastern Australia (Taylor et al. 2005), southern and eastern Australia. Australia (Cunnington et al. 2007; Pitt et al. 2010), New Zealand (Amponsah et al. 2009, 2011), South Africa (van Niekerk et al. 2004) and USA (California) (Úrbez-Torres et Materials & methods al. 2006; Úrbez-Torres and Gubler 2009). Other fruit crop declines have linked N. australe to olive “drupe rot” in Italy Collection of Australian material (Lazzizera et al. 2008) and dieback symptoms of peach, plum and apricots in South Africa (Damm et al. 2007; Isolates from previous studies in WA were included Slippers et al. 2007). Pavlic et al. (2007) established N. (Burgess et al. 2006; Taylor et al. 2009). They were australe as a potential pathogen of Eucalyptus grandis x E. collected from four locations in the south-west of WA; camaldulensis in which pathogenicity trials of young Yalgorup National Park from E. gomphocephala, S. 2-year-old trees produced stem lesions significantly greater acuminatum, A. rostellifera, B. grandis, A. fraseriana, A. then control trees. In 2008, N. australe was isolated from flexuosa and E. marginata; Woodman’s Point from E. symptomatic wood showing perennial cankers from Eucalyp- gomphocephala, S. acuminatum, A. rostellifera, A. coch- tus globulus and 20-year-old Pistachio vera in northern Spain learis and C. preissii; and two plantations and adjacent (Armengol et al. 2008). Subsequent pathogenicity trials native tree species close to Denmark on the south coast of illustrated a rapid induction of symptoms including leaf WA, with samples taken from E. globulus, E. marginata and wilting and canker formation in 8-month-old Eucalyptus E. diversicolor. Samples, including type cultures, were seedlings, whilst it took 3 months for cankers to development also obtained from VPRI (Victorian Department of in 2-year-old Pistachio seedlings. Primary Industries) and from CMW (collection of the Relatively few reports of N. australe have been recorded Forestry and Agriculture Biotechnology Institute, Uni- in eastern Australian native tree species; the only record of versity of Pretoria, South Africa) in Victoria, New South N. australe was linked to canker development on native Wales and South Australia. Acacia spp. in the original description (Slippers et al. 2004). However, N. australe is the dominant Botryosphaer- World-wide distribution iaceae endophyte found amongst native tree species in the southwest of Western Australia (WA) (Shearer et al. The majority of the N. australe isolates, including those 1995; Burgess et al. 2006; Taylor et al. 2009). Native used in the original descriptions (CMW9073, CMW9072, hosts include Acacia rostellifera, Acacia cochlearis, CMW6837 and CMW6853) can be separated from N. Allocasuarina fraseriana, Agonis flexuosa, B. coccinea, luteum (CMW9076/ICP7818 and CMW10309/CAP002) Banksia grandis, Callitris preissii, Eucalyptus diversi- by five fixed polymorphisms in the ITSrDNA region. A color, E. gomphocephala, E. marginata, Eucalyptus smaller subset of N. australe isolates can only be phylacis, Santulum acuminatum (Shearer et al. 1995; separated from N. luteum by three to four of these Burgess unpublished data; Burgess et al. 2005, 2006; polymorphisms. These polymorphisms were used to Taylor et al. 2009). Slippers et al. (2004) suggested that N. assist in identification of N. australe sequence from australe is endemic to the southern hemisphere, and GenBank (Table 1). current evidence suggests N. australe is endemic to WA (Burgess et al. 2006; Taylor et al. 2009). Molecular identification Interestingly, pathogenicity trials using more than one isolate of N. australe have, in some cases, found significant Molecular identification was used to confirm identification variation in the virulence between isolates (van Niekerk
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