NOTAS CIENTÍFICAS Leaf blight and defoliation of Eugenia spp. caused by Cylindrocladium candelabrum and C. spathiphylli in Brazil Luiz Sebastião Poltronieri1, Rafael Ferreira Alfenas2, Jaqueline Rosemeire Verzignassi3, Acelino Couto Alfenas2, Ruth Linda Benchimol1, Tathianne Pastana de Sousa Poltronieri1 1 Embrapa Amazônia Oriental, Tv. Enéas Pinheiro, S/N, CEP 66095-100, Belém, PA. 2Departamento de Fitopatologia, Universidade Federal de Viçosa, CEP 36570-000, Viçosa, MG. 3Embrapa Gado de Corte, Rodovia BR 262, km 4, Caixa Postal 154, CEP 79002-970, Campo Grande, MS. Autor para correnpondência: Jaqueline Rosemeire Verzignassi (jaqueline@cnpgc.embrapa.br) Data de chegada: 21/12/2009. Aceito para publicação em: 06/11/2010. 1679 ABSTRACT Poltronieri, L.S.; Alfenas, R.F.; Verzignassi, J.R.; Alfenas, A.C; Benchimol, R.L.; Poltronieri, T.P.S. Leaf blight and defoliation of Eugenia spp. caused by Cylindrocladium spathiphylli and C. candelabrum in Brazil. Summa Phytopathologica, v.37, n.2, p.147-149, 2011. Leaf blight and defoliation of Eugenia stipitata Mc Vaugh and Eugenia patrisii Vahl, caused respectively by Cylindrocladium candelabrum (Calonectria scoparia) and C. spathiphylli (Calonectria spathiphylli) are reported in the state of Pará, Brazil. On both host species, the disease is characterized by dark brown lesions of different sizes and shapes. A whitish bright sporulation, resembling Cylindrocladium is observed on the necrotic lesions by using a stereomycroscope or a pocket lense (10-20 X). Under fa vora ble condit ion s a nd d ependi ng on t he level of i nfec tion, intense premature tree defoliation may also be found.Although the conidial germination and mycelial growth were higher at 25ºC for b ot h s p e c i es , C . c a n d e l a b r u m wa s m ore s en s i t i ve t o t h e variation of temperature (10, 20, 30 and 40 ºC) than C. spathiphylli. This is the first report of C. candelabrum and C. spathiphylli on Eugenia stipitata (araçá-boi) and on Eugenia patrisii (ubaia-daamazônia), respectively in Brazil. Keywords: foliar lesions, Myrtaceae. RESUMO Poltronieri, L.S.; Alfenas, R.F.; Verzignassi, J.R.; Alfenas, A.C; Benchimol, R.L.; Poltronieri, T.P.S. Lesões foliares e queda de folhas em Eugenia spp. causadas por Cylindrocladium spathiphylli e C. candelabrum no Brasil. Summa Phytopathologica, v.37, n.2, p.147-149, 2011. Relata-se a incidência de mancha, queima foliar e desfolha, causadas respectivamente por Cylindrocladium candelabrum (Calonectria scoparia) em Eugenia stipitata Mc Vaugh (araçá-boi) e C. spathiphylli (Calonectria spathiphylli) em Eugenia patrisii Vahl (ubaia-da-amazônia), no Estado do Pará, Brasil. Em ambos os hospedeiros, a doença caracterizase por lesões necróticas de diferentes dimensões e formas, coloração marrom escura, contendo uma massa branca brilhante de esporulação típica de Cylindrocladium, facilmente observada sob microscópio estereoscópico ou com o auxílio de uma lente de bolso de 10 – 20 x de aumento. Sob condições favoráveis à infecção, as plantas podem sofrer intensa queda prematura de folhas. Embora, o maior número de conídios germinados e o maior crescimento micelial foram obtidos a 25ºC para ambas as espécies, C. candelabrum foi mais sensível às variações de temperatura (10, 20, 30 e 40 ºC) que C. spathiphylli. Este é o primeiro relato de C. candelabrum em Eugenia stipitata e C. spathiphylli em Eugenia patrisii no Brasil. Palavras-chave adicionais: lesões foliares, Myrtaceae. Plants of Eugenia stipitata Mc Vangh (araçá-boi) and Eugenia patrisii Vahl showing leaf blight and defoliation were found in the Experimental field of Emb 0 x 4,5-5,0 µm and globose vesicle typical of Cylindrocladium spathiphylli Schoult., El-Gholl & Alfieri (Calonectria spathiphylli El-Gholl, J.Y. Uchida, Alfenas, T.S. Schub., Alfieri & A. R. Chase) (Figure 2), sensu Crous (3). Inoculations to confirm the pathogenicity of both fungi were made by placing inoculum plugs (5 mm diameter) on the adaxial portion of the leaf limb of the respective plant species. After inoculation the plants were maintained in a moist chamber for 48 h and then transferred to a green house. Three days later typical symptoms of the disease were observed and re-isolations of the fungi to PDA confirmed their pathogenicity on the respective host species. This is the first report of C. candelabrum on E. stipitata and C. spathiphylli on Eugenia patrisii Summa Phytopathol., Botucatu, v. 37, n. 2, p. 147-149, 2011 in Brazil, although C. scoparium Morgan and C. gracile (Bugn.) Boesew. were already found causing fruit and root decay in araçá-boi in Pará State and in Federal District, respectively (4, 5). The two fungal species reported here have an ample geographic distribution and a wide host range (3). Conidial germination and the mycelial growth of both fungi were evaluated at 5, 15, 20, 25, 30 and 35ºC in the dark. For germination, 25 µl of a 1x104 conidia.mL-1 suspension were added into three cavities of a sterile glass slide. Each treatment contained three replicates, in a completely randomized design. After 4h of incubation in the dark, germination was interrupted by addition lacto phenol-cotton blue and the percentage of germination was estimated by scoring 600 randomly chosen conidia per treatment, under a optical mycroscope (200 X). Conidia presenting differentiated germ tubes, independently on their 147 A y = -0.2575x2 + 11.62x - 53.781 R2 = 0.7188 Germination (%) 100 80 60 40 20 0 A 0 10 20 Temperature (ºC) 30 Germination (%) 100 40 B 80 60 40 y = -0.1416x2 + 7.9277x - 25.839 R2 = 0.9207 20 0 0 10 20 30 40 Temperature (ºC) B Figure 1. Leaf blight caused by Cylindrocladium spp. in the field. A - C. candelabrum in Eugenia stipitata; B – C. spathiphylli in Eugenia patrisii. Figure 3. Conidial germination at different temperatures. A – C. Candelabrum; B - C. spathiphylli. Mycelial growth (g) 3.5 y = -0.0094x2 + 0.3815x - 1.6365 R2 = 0.8743 A 2.5 1.5 0.5 -0.5 0 10 20 30 40 A B Figure 2. Conidiophores containing vesicle and conidia typical of Cylindrocladium: A – C. spathiphylli; B - C. candelabrum. Mycelial growth (g) Temperature (ºC) y = -0.0101x2 + 0.4366x - 1.7306 R2 = 0.8769 3.5 B 2.5 1.5 0.5 -0.5 0 10 20 30 40 Temperature (ºC) Figure 4. Mycelial growth at different temperatures. A – C. candelabrum; B - C. spathiphylli. 148 Summa Phytopathol., Botucatu, v. 37, n. 2, p. 147-149, 2011 size were considered germinated. Germination of C. candelabrum and C. spathiphylli conidia varied significantly with the temperature of incubation, but it was higher at 25°C for both species (Figure 3). However, C. candelabrum was more sensitive to temperature variation than C. spathiphylli (Figure 3). On the other hand, based on the regression equation, the estimated optimum temperature for conidial germination was of 23°C for C. candelabrum and 28°C for C. spathiphylli. For the mycelium growth one culture disk (4 mm diameter) of each fungus was transferred separately to each 125 mL Erlenmeyer flask, containing 50 mL of a semi-synthetic liquid growth media (1), following incubation at the same temperatures tested for spore germination. Five replicates in a completely randomized experimental design were employed. At 10 days of incubation the mycelial mat was filtered in a Buchner funil containing a Whatman no. 1 filter paper. After removal of the water excess by pressing the mycelial mat in germ test filter paper, the fresh weight was determined. The highest mycelial growth of both species was at 25°C. Based on the regression equation (Figure 4), the estimated optimum temperature was 20°C for C. candelabrum and 23°C for C. spathiphylli. As for conidial germination, C. candelabrum was also more sensitive to the temperature variation than C. spathiphylli. Generally, the effect of temperature on mycelial growth for both fungi studied in the present Summa Phytopathol., Botucatu, v. 37, n. 2, p. 147-149, 2011 work was similar to that found for C. ilicicola isolated from eucalyptus (2). This findings may have important implications on the occurrence of these fungi according to the temperature oscillation around the year and may explain the wide geographical distribution of C. candelabrum in less warm southeaster regions in Brazil. LITERATURE CITED 01. Alfenas, A.C. Extração de proteínas para eletroforese. In: ALFENAS, A.C. (Ed.). Eletroforese e marcadores bioquímicos em plantas e microorganismos. 2. ed. Viçosa: Ed. UFV, 2006. p. 85-114. 02. Alfenas, A.C.; Matsuoka. K.; Ferreira. F.A.; Hodges. C.S. Identificação, características culturais e patogenicidade de três espécies de Cylindrocladium, isoladas de manchas de folha de Eucalyptus spp. Fitopatologia Brasileira, Brasília, v.4, p.445-449, 1979. 03. Crous, P.W. Taxonomy and pathology of Cylindrocladium (Calonectria) and allied genera. St. Paul: American Phytopathological Society, 2002. 278p. 04. Junqueira, N.T.V.; Fialho, J.F.; Ramos, V.H.V.; Leão, A.J.P. Doenças e potencial de produção do araçá-boi (Eugenia stipitata) nos cerrados. Fitopatopatologia Brasileira, Brasília, v.22, p.272, 1997. 05. Nunes, A.M.L.; Stein, R.L.B.; Albuquerque, F.C. Araçá-boi (Eugenia stipitata): um novo hospedeiro de Cylindrocladium scoparium. Fitopatologia Brasileira, Brasília, v.20, p.488-490, 1995. 149