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First report of curvularia trifolii causing curvularia blight in agrostis stolonifera in South of Portugal
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Abstract(s)
Algarve region (Portugal) has nearly 40 golf courses with a significant economic impact. Summer surveys on 10 golf courses detected an unknown disease on one course in 2009 and on another course in 2012 and 2013 at 29 to 30°C daily average. The second course had symptoms on about 25% of the turf of two putting greens. Diseased bentgrass (Agrostis stolonifera L.) had a green dappled pattern with irregular patches of turfgrass on yellowed leaves. Prior to decaying, affected leaves turned brown and then gray. Crown and leaf sheath infections resulted in dark brown dry rot. No lesions were observed on the roots. Leaves were surface disinfected with 5% commercial bleach (0.225% sodium hypochlorite) and cultured on potato dextrose agar (PDA). Ten fungal colonies grew from the leaf tissue, and brown mycelia, conidiophores, and conidia were observed under a microscope. Conidia were ventricose pyriform, mostly abruptly curved, 20 to 36 µm (30 µm, SD = 4) × 7 to 12 µm (10.5 µm, SD = 1.3) (n = 50), predominantly three-septate, with a prominent hilum and enlarged and darkened central cells. Colonies grown on PDA were black-brown with a black reverse side. Conidia differed in size, 15.4 to 24.6 µm (19.99 µm, SD = 3.00) × 6 to 11 μm (8.68 µm, SD = 1.54) (n = 50) and morphology (cylindrical or slightly curved). These characteristics were consistent with Curvularia trifolii (Kauffm.) Boedijn. (Ellis 1971; Falloon 1976; Khadka 2016). Species identification of the representative isolate A2 1.12 was confirmed by analysis of nucleotide sequences of the ITS1-5.8S-ITS2 region using primers ITS1 and ITS4 (White et al. 1990) and GPDH gene region with primer set gpd (Koike et al. 2013). BLAST searches of GenBank showed a high similarity of the isolate ITS sequence (MG029439) to the reference sequence JN712458 of C. trifolii (99% identity) and GPDH sequence (MK570108) with LT715803.1 (97.88% identity). The maximum likelihood phylogenetic tree showed that our isolate clustered with C. trifolii. The pathogenicity assay of this isolate was conducted in greenhouse on A. stolonifera ‘Penncross’. The isolate was grown on PDA (25°C, 10 days). Five pots (100 ml) were filled with a sand and peat mix (9:1 v/v) with 0.06 g of seeds per pot, covered with a fine sand layer. Turfgrass was cut once a week beginning 2 weeks after seeding and was fertigated with 0.5 g/liter of Peter’s foliar feed (27 + 15 + 12; N + P2O5 + K2O; and micronutrients; Scotts, Heerlen, The Netherlands). To obtain a conidial suspension for inoculation, cultured plates were scraped with a sterilized spreader and water. The suspension was filtered through a sterile gauze. Conidia were counted under a microscope (400×) with a hemocytometer. The suspension was adjusted to 8 × 103 conidia/ml, and 10 ml was sprayed per pot. Pots maintained humidity for 2 days under microtunnels. The first disease symptoms appeared 3 days after inoculation. Bentgrass from the five pots developed Curvularia blight and rotted crown symptoms. Control plants (five pots treated with water) did not display symptoms. This trial was repeated once. On PDA, C. trifolii was reisolated from leaf lesions and morphologically identified, confirming Koch’s postulates. Ellis (1971) referred to the presence of C. trifolii in Portugal, but no region, symptom description, or grass species was detailed. Sivanesan (1987) reported C. trifolii in Portugal only on Lolium multiflorum. Therefore, this is the first report of C. trifolii in Algarve, affecting A. stolonifera. This disease can increase maintenance costs in greens in this area.
Description
Keywords
Fungi Turfgrass Pathogen detection
Citation
Publisher
American Phytopathological Society