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

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.