First Report of Phaeoacremonium inflatipes, P. iranianum, and P. sicilianum Causing Petri Disease of Grapevine in Spain

Abstract

In 2008, four isolates of Phaeoacremonium, morphologically and genetically different from known Phaeoacremonium spp. in Spain, were isolated from rootstocks of young grapevine (Vitis vinifera) plants showing Petri disease symptoms including low vigor, reduced foliage, and dark streaking of the xylem in Badajoz Province (western Spain; cv. Syrah on SO4 rootstock), Tarragona Province (eastern Spain; cv. Garnacha on 161 49 C rootstock), and Balearic Islands (eastern Spain; cv. Tempranillo on Rupestris de Lot rootstock). Single-conidial isolates were obtained and grown on potato dextrose agar (PDA) and malt extract agar (MEA) at 25°C for 2 to 3 weeks in the dark until colonies sporulated (3). Identification was based on morphological characteristics (1–3). Phaeoacremonium inflatipes W. Gams, Crous & M. J. Wingf. and P. iranianum L. Mostert, Gräf., W. Gams & Crous were detected in Badajoz Province and P. sicilianum Essakhi, Mugnai, Surico & Crous in Tarragona Province and Balearic Islands. Colonies of P. inflatipes were gray on PDA and gray-brown on MEA. Conidiophores were branched, 15 to 37 (mean 25) μm long. Conidia were hyaline, oblong-ellipsoidal or obovoid, 3 to 5.5 (mean 4) μm long, and 1.2 to 1.9 (mean 1.6) μm wide. Colonies of P. iranianum were brownish gray on PDA and pale brown on MEA. Conidiophores were unbranched and 18 to 47.5 (mean 29) μm long. Conidia were hyaline, oblong-ellipsoidal, 3 to 5 (mean 4) μm long, and 1 to 1.8 (mean 1.5) μm wide. Colonies of P. sicilianum were pale brown on PDA and brown to pale orange on MEA. Conidiophores were branched and 13 to 55 (mean 32.5) μm long. Conidia were hyaline, allantoid, 3 to 8.5 (mean 6) μm long, and 1.5 to 2 (mean 1.8) μm wide. Identity of isolates Pin-2, Pir-4, Psi-1, and Psi-2 was confirmed by sequencing a fragment of the beta-tubulin gene with primers T1 and Bt2b (P. inflatipes, isolate Pin-2: GenBank Accession No. FJ872407, 100% similarity to Accession No. AY579323; P. iranianum, isolate Pir-4: GenBank Accession No. FJ872406, 99% similarity to Accession No. EU128077; P. sicilianum isolates Psi-1 and Psi-2: GenBank Accession Nos. FJ872408 and No. FJ872409, 100% similarity to Accession No. EU863489). Pathogenicity tests were conducted using Pin-2, Pir-4, and Psi-1 isolates. One-year-old callused and rooted cuttings of 110 R rootstock cultivated in sterile peat were wounded at the uppermost internode with an 8-mm cork borer. An 8-mm mycelium plug from a 2-week-old culture was placed into the wound. Wounds were wrapped with Parafilm. Ten cuttings per fungal isolate were used. Ten control plants were inoculated with 8-mm noncolonized PDA plugs. Plants were maintained in a greenhouse at 25°C. Within 2 months, all Phaeoacremonium-inoculated cuttings exhibited shoots with poor growth, small leaves, short internodes, and black streaks in the xylem. The mean shoot weight per plant was 1.8 g in P. inflatipes-inoculated plants, 1.9 g in P. iranianum-inoculated plants, and 1.6 g in P. sicilianum-inoculated plants, all lower than the control treatment (6.8 g). Control plants did not show any symptoms. All fungal species were reisolated from wood of all inoculated cuttings, completing Koch's postulates. Their identity was confirmed with the methods described above. To our knowledge, this is the first report of P. inflatipes, P. iranianum, and P. sicilianum causing Petri disease in Spain. References: (1) P. W. Crous et al. Mycologia 88:786, 1996. (2) S. Essakhi et al. Persoonia 21:119, 2008. (3) L. Mostert et al. Stud. Mycol. 54:1, 2006.

Phaeoacremonium species diversity on woody hosts in the Western Cape Province of South Africa

“… Costa Rica ( Groenewald et al 2001 ) Prunus armeniaca South Africa (this study) Prunus persica South Africa (this study) Psidium guajava South Africa (this study) Quercus robur South Africa (this study) Quercus virginiana USA ( Groenewald et al 2001 ) Salix sp. South Africa (this study) Soil USA ( Rooney et al 2001 ) Vitis vinifera Chile ( Mostert et al 2006 ); Iran ( Mohammadi et al 2013 ); Spain ( Gramaje et al 2009a ) P. iranianum Actinidia chinensis Italy ( Mostert et al 2006 ) Alnus glutinosa Iran ( Kazemzadeh Chakusary et al 2017 ) Cinnamomum camphora South Africa (this study) Crataegus rhipidophylla Iran ( Sami et al 2014 ) Cydonia oblonga Iran ( …”

“…Phaeoacremonium has a global distribution, with species being reported from South, Central and North America, Europe, Scandinavia, Ukraine, the Middle East, Far East, Oceania and Africa ( Gramaje et al 2015 ). Most Phaeoacremonium species are associated with wood diseases of plants, e.g., Petri and esca diseases of grapevines ( Crous et al 1996 , Essakhi et al 2008 , Gramaje et al 2009a , b , White et al 2011 , Raimondo et al 2014 ) and dieback and cankers on other fruit trees and woody hosts ( Damm et al 2008b , Gramaje et al 2012 , Mohammadi et al 2014 , Sami et al 2014 , Carlucci et al 2015 , Hashemi & Mohammadi 2016 , Kazemzadeh Chakusary et al 2017 ). Eleven Phaeoacremonium species are also known from human infections ( Mostert et al 2005 , Gramaje et al 2015 ).…”

Phaeoacremonium: From esca disease to phaeohyphomycosis

“…on grapevines has been extensively investigated with different inoculation methods, such as soaking bases of grapevine cuttings or seedlings in spore suspensions (Larignon & Dubos 1997;Scheck et al 1998;Gramaje et al 2007Gramaje et al , 2008Aroca et al 2008;Aroca & Raposo 2009), insertion of mycelial plugs/conidial suspensions into side wounds or cut ends of the grapevine stems (Mugnai et al 1999;Sparapano et al 2001;Feliciano et al 2004;Santos et al 2006;Halleen et al 2007;Zanzotto et al 2008;Gramaje et al 2009aGramaje et al , 2013Romero-Rivas et al 2009) Various pathogenicity studies have investigated symptom expression of P. minimum. Inoculation studies have shown that P. minimum can cause brown wood streaking (Khan et al 2000;Sparapano et al 2000b;Feliciano et al 2004;Halleen et al 2007;Gramaje et al 2010), reduced shoot growth Zanzotto et al 2008;Gramaje et al 2010) and esca symptoms on leaves and berries (Sparapano et al 2001;Feliciano et al 2004).…”

“…(Damm et al 2008;Gramaje et al 2012;Olmo et al 2014). (Essakhi et al 2008); Hungary (Essakhi et al 2008); Iran (Mohammadi 2011); South Africa (White et al 2011); Spain (Gramaje et al 2007); USA (Groenewald et al 2001 ; Italy (Essakhi et al 2008); South Africa (White et al 2011); Spain (Gramaje et al 2009a (Auger et al 2005); Greece (Essakhi et al 2008); Hungary (Essakhi et al 2008); Iran , Israel (Essakhi et al 2008); Italy (Crous et al 1996); France (Larignon & Dubos 1997); Germany (Fisher & Kassemeyer 2003); Mexico (Hern andez-Mart ınez et al 2008); Portugal (Rego et al 2000); South Africa (Crous et al 1996), Spain (Armengol et al 2001); Turkey (Ari 2000); Uruguay (Abreo et al 2011); USA (Crous et al 1996) Yugoslavia (Crous et al 1996 Petri disease is associated with stunted growth and dieback of young grapevines, and can cause losses of up to 50 % in newly planted vineyards (Pascoe & Cottral 2000). Young vines infected with Phaeoacremonium species and Phaeomoniella chlamydospora can later develop esca symptoms following further colonisation by several basidiomycetous species (Mugnai et al 1999).…”

Studies on the epidemiology of black-foot disease of grapevine in Spain

“…In Spain, the main fungal agents associated to this disease are Pa. (Gramaje et al, 2007;Aroca et al, 2008a;Gramaje et al, 2008Gramaje et al, , 2009aGramaje et al, , b, 2011a. In addition, Ca.…”