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In vitro assessment of the antagonism of Trichoderma spp. isolates for the control of Ciborinia camelliae The European and Mediterranean Plant Protection Organization (EPPO) includes Ciborinia camelliae in the A2 list as a quarantine organism (EPPO, 2011). At the moment, there is no fully effective treatment to cure camellias of the disease. In recent years the use of microbial antagonists for control of plant fungal diseases has received much attention. Fungi of the genus Trichoderma have been shown to be effective as biocontrol agents of several soil borne phytopathogens (Benítez et al., 2004), and could be useful agents to control fungal diseases affecting aerial parts of plants. In this work, we evaluated the in vitro antagonistic activity of two species of Trichoderma (T. harzianum and T. asperellum), obtained in two camellia nurseries in Japan (Genkai and Hisatoni), against Ciborinia camelliae. INTRODUCTION MATERIAL AND METHODS RESULTS AND DISCUSSION When grown in dual culture, overgrowth and sporulation of Trichoderma isolates on all C. camelliae isolates were observed (Figure 1). A reduction of the colony diameter of C. camelliae was also noted (table 3), thus indicating an antagonistic effect of Trichoderma isolates on the pathogen. The most effective antagonist was Trichoderma harzianum, which causes the highest growth inhibition for the three C. camelliae isolates (table 4). However, T. asperellum Genkai hardly inhibited the growth of the pathogen isolate EFA 866.2 and was less effective than the other two Trichoderma isolates to antagonize the isolate EFA 909.1. In contrast, Trichoderma asperellum Hisatoni was as effective as T. harzianum to inhibit growth of those two isolates of C. camelliae (EFA 866.2 and EFA 909.1). Our results suggest that T. harzianum Hisatoni and T. asperellum Genkai could be effective antagonists of C. camelliae, given its rapid colony formation and its ability to inhibit growth of C. camelliae in vitro in the short term. The suitability of applying inocula of these Trichoderma isolates as a control method of camellia flower blight must now be tested in greenhouse and field conditions. Some authors have found that fungi showing antagonism under controlled growth conditions are not effective in the field due to soil characteristics and/or competence with indigenous microbial populations (Garveba et al., 2004). REFERENCES Benítez T., Rincón A.M., Limón M.C, Codón A.C. 2004. Biocontrol mechanisms of Trichoderma strains. International Microbiology 7: 249-260. EPPO. 2011. Ciborinia camelliae. http://www.eppo.org/QUARANTINE/listA2.htm. Last access: November 2011. Garveba P., van Veen J.A., van Elsas J.D. 2004. Microbial diversity in soil: Selection of microbial populations by plant and soil type and implications for disease suppressiveness. Annual Review of Phytopathology 42: 243-270. Two isolates of Trichoderma asperellum and one of Trichoderma harzianum were obtained from the greenhouse soil of two camellia nurseries in Japan (Genkai and Hisatoni) (table 1). Three isolates of Ciborinia camelliae were obtained from flowers of Camellia japonica plants growing in Pontevedra (NW Spain) (table 2). The competition between Ciborinia camelliae and the three antagonists (T. asperellum Genkai, T. asperellum Hisatoni and T. harzianum Hisatoni) were evaluated in dual cultures. For each Trichoderma and C. camelliae isolates combination, mycelial discs of the potential antagonist and the pathogen were placed simultaneously 3 cm apart on a Petri dish. Colony growth of the pathogen and the antagonists was recorded after 4 and 6 days. Growth of C. camelliae cultivated alone was used to estimate the percent growth inhibition of the pathogen by the antagonists. a Estación Fitopatolóxica do Areeiro, Deputación de Pontevedra, Subida a la Robleda s/n, 36153 Pontevedra, Spain b Departamento de Producción Vegetal, Universidad de Santiago de Compostela, Campus Universitario s/n, 27002 Lugo, Spain Aguín O. 1a, Dalama J. a, Sainz M.J. b, Salinero M.C. a, Mansilla J.P. a T C Distribution map of Ciborinia camelliae (EPPO, 2011). Isolate of Trichoderma asperellum in PDA medium. Isolate of Ciborinia camelliae in PDA medium. Table 3. Colony diameter of three isolates of Trichoderma spp. and three isolates of C. camelliae in control cultures after 4 and 6 days of growth at 20 ºC. Symptoms of Ciborinia camelliae on flowers of Camellia japonica. Table 4. Percent growth inhibition of three isolates of Ciborinia camelliae caused by Trichoderma spp. after 6 days of growth in dual cultures. For each growth parameter, values followed by a different letter are significantly different for P < 0.05. Figure 1. Dual culture of Trichoderma asperellum Genkai (T) and C. camelliae EFA 866.5 (C) after 4 (a) and 6 (b) days of growth, showing overgrowth of the antagonist. ab Table 1. Isolates of Trichoderma spp. obtained from greenhouse soil in two camellia nurseries in Japan. Table 2. Isolates of Ciborinia camelliae obtained from flowers of Camellia japonica plants growing in Pontevedra (NW Spain). ISOLATION OF ANTAGONISTIC FUNGI ISOLATION OF CIBORINIA CAMELLIAE IN VITRO ANTAGONISM TESTS Ciborinia camelliae Khon is a highly specialized pathogen that attacks the flowers of many species of Camellia, causing the disease known as camellia flower blight. It is considered the main phytosanitary problem for cultivars of Camellia spp. with ornamental value, since it causes the premature fall of the flower. Legend Present (national record) + Present (subnational record)