Isolation and pathogenicity of Colletotrichum spp. causing anthracnose of strawberry in south west Spain

Abstract

Anthracnose, caused by Colletotrichum spp., is a major disease of cultivated strawberry, Fragaria× ananassa. This study identifies the Colletotrichum spp. which causes strawberry anthracnose in the southwest of Spain. A survey of the region was carried out, and the strains isolated were identified as C. acutatum by using the polymerase chain reaction (PCR) with genus and species-specific primers, demonstrating that this species is currently the causal agent of strawberry anthracnose in the studied region. The pathogenicity of C. acutatum and C. gloeosporioides strains was evaluated on two principal strawberry cultivars (cvs Camarosa and Ventana) under field conditions, the latter being more pathogenic than the former.

Phylogenetic relationships and genome organisation of Colletotrichum acutatum causing anthracnose in strawberry

“…Colletotrichum acutatum isolates used in this study were obtained from: (i) a previous survey carried out in Spain from strawberry anthracnose fruit rot (Garrido et al 2007); (ii) the International Mycological Institute (IMI), and (iii) the Central Science Laboratory (CSL) collection, which includes isolates from Australia, Canada, France, Germany, Japan, The Netherlands, New Zealand, Norway, Portugal, Switzerland, USA and UK, isolated from strawberry; isolates from lupin, rose and phlox were also included (Table 1). Isolates from this collection were kindly supplied by Dr. Neil Boonham from CSL.…”

“…Genus-specific primers Col1 (5´-AAC CCT TTG TGA ACR TAC CTA-3´) and Col2 (5´-TTA CTA CGC AAA GGA GGC T-3´) were used for identification of the genus Colletotrichum (Garrido et al 2007;Martínez-Culebras et al 2003); species-specific primers CaInt2 (5´-GGG GAA GCC TCT CGC GG-3´) for C. acutatum (Garrido et al 2007;Sreenivasaprasad et al 1996) and CgInt (5´-GGC CTC CCG CCT CCG GGC GG-3´) for C. gloeosporioides (Mills et al 1992) were each coupled in PCR reactions with the universal primer ITS4 (White et al 1990). Two independent experiments of PCR amplification were performed with each pair of primers.…”

“…The nucleotide sequences of the ITS1, 5.8S RNA gene, and ITS2 were studied for each isolate. Thirty-four isolates were sequenced in this work, ten isolates had been previously sequenced (Garrido et al 2007), and the sequences of the eight remaining isolates had been previously reported by Martínez-Culebras et al (2003) (GenBank database accession numbers are given in Table 1). A neighbour-joining phylogenetic tree, with the 5.8S-ITS sequences, was estimated using the Kimura two-parameter model and a bootstrap test of 5000 runs.…”

“…have been focused on just one or two aspects of the disease: traditionally, with cultural or morphological studies (Afanador-Kafuri et al 2003;Denoyes-Rothan and Baudry 1995;Smith and Black 1990), and nowadays, using molecular techniques including isoenzyme comparisons, Restriction Fragment Length Polymorphisms (RFLP) analyses of mitochondrial DNA, Amplified Fragment Length Polymorphism (AFLP), AT rich analyses, Random Amplified Polymorphic DNA (RAPD), and ITS sequence analyses for specific PCR identification (Buddie et al 1999;Garrido et al 2007Garrido et al , 2009Freeman et al 1993;Sreenivasaprasad et al 1996;Talhinhas et al 2005). These techniques have improved the accuracy and reduced the time necessary for the identification and classification of Colletotrichum spp.…”

Olive – Colletotrichum acutatum: An Example of Fruit-Fungal Interaction

“…1). The Colletotrichum acutatum can affect up to 100% of the fruit on a olive tree during humid (or rain) autumns where susceptible olive cultivars are grown (Casado-Díaz et al, 2006;Freeman et al, 2002;Garrido et al, 2008;Peres et al, 2005;Talhinhas et al, 2011;Trapero & Blanco, 2008). Moreover, a poor quality and low stability olive oil is obtained from olives harvested in areas affected by anthracnose, presenting alterations in oil color (red), high acidity and other typical organoleptic characteristics (Carvalho et al, 2006;Moral et al, 2008;Talhinhas et al, 2009Talhinhas et al, , 2011 (Fig.1).…”