Plant Pathology (2010) 59, 1170 Doi: 10.1111/j.1365-3059.2010.02297.x First report of anthracnose and fruit mummification of olive fruit (Olea europaea) caused by Colletotrichum acutatum in Brazil H. S. S. Duarte a , P. G. C. Cabral a , O. L. Pereira a *, L. Zambolim a , E. D. Gonc ¸alves b , J. Vieira Neto b , E. M. Zambolim a and V. Sergeeva c a Departamento de Fitopatologia, Universidade Federal de Vic ¸osa, Vic ¸ osa, Minas Gerais, 36570-00; b Empresa de Pesquisa Agropecua ´ria de Minas Gerais (EPAMIG), Maria da Fe ´ , Minas Gerais, 37517-000, Brazil; and c Centre for Plants and the Environments, University of Western Sydney, Locked Bag 1797, South Penrith, DC, NSW 1797, Australia The olive tree is an arboreal species belonging to the family Oleaceae with recognized importance in the production of olive oils and olives. In December 2008, typical lesions of anthracnose, with mature fruit mum- mification were observed in olive tree fields in Maria da Fe ´ , in the state of Minas Gerais, Brazil. A fungus was isolated directly on potato dextrose agar (PDA) from conidia collected from pink to orange masses on infected fruit. A typical fruit sample was deposited in the local herbarium (VIC 31209). The isolate showed a pink colony on PDA, producing sporodo- chia with a mass of hyaline amerospores with pointed ends. Based on these morphological characteristics the fungus was identified as Colletotrichum acutatum, which has been reported to cause anthracnose on olives trees in other countries and most recently in Australia (Sergeeva et al., 2008). In Brazil, C. acutatum is reported to cause disease on fruit of apple, citrus, strawberry, peach, plum, nectarine, medlar, and on yerba-mate (Kimati et al., 2005). Identity was confirmed by extracting the DNA of a monoconidial iso- late, OLP 570, and amplifying the ITS region of the rRNA by polymerase chain reaction with primer ITS4 and specific primers for C. acutatum (CaInt2; Sreenivasaprasad et al., 1996) and C. gloeosporioides (CgInt; Mills et al., 1992). Isolates of C. acutatum (DAR78874 and DAR78876) and C. gloeosporioides (DAR78875) obtained from Australian olives trees were used as positive controls. The primers ITS4 and CaInt2 ampli- fied a single DNA product of 490 bp, as expected for C. acutatum. Patho- genicity was confirmed by placing a 40-mm disk of PDA colonized with OLP 570 on 40 olive fruits that were either intact or slightly wounded. Non-colonized PDA disks were used as negative controls. The inoculated fruits were transferred to Gerbox-type boxes with high humidity and kept in a growth chamber at 25°C. Typical anthracnose symptoms were observed only on the slightly wounded inoculated fruit 4 days post-inocu- lation with subsequent fruit mummification after a further 3 days. This is the first report of C. acutatum causing anthracnose and mummification of olive fruit in Brazil. Acknowledgements Authors Duarte, Cabral, Pereira and Zambolim thank the CNPQ and Gonc ¸ alves and Vieira Neto thank FAPEMIG for financial support. References Kimati H, Amorim L, Rezende JAM, Bergamin Filho A, Camargo LEA, 2005. Manual de Fitopatologia-Doenc ¸as das Plantas Cultivadas. Sa ˜o Paulo, Brazil: Agrono ˆ mica Ceres Ltda. Mills PR, Sreenivasaprasad S, Brown AE, 1992. Detection and differentiation of Colletotrichum gloeosporioides isolates using PCR. FEMS Microbiology Letters 98, 137–44. Sergeeva V, Spooner-Hart R, Nair NG, 2008. First report of Colletotrichum acutatum and C. gloeosporioides causing leaf spots of olives (Olea europaea) in Australia. Australasian Plant Disease Notes 3, 143–4. Sreenivasaprasad S, Sharada K, Brown AE, Mills PR, 1996. PCR-based detection of Colletotrichum acutatum on strawberry. Plant Pathology 45, 650–5. *E-mail: oliparini@ufv.br. Accepted 26 January 2010 at http://www.bspp.org.uk/ndr where figures relating to this paper can be viewed. Plant Pathology (2010) 59, 1170 Doi: 10.1111/j.1365-3059.2010.02356.x Botryosphaeria dothidea causing canker of grapevine newly reported in China X. Li a , J. Yan a , F. Kong ab , G. Qiao a , Z. Zhang c and Z. Wang b * a Institute of Plant and Environment Protection, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097; b Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193; and c College of Enology, Northwest A & F University, Yangling, Shanxi 712100, China In the summer of 2009 in Zhejiang, Jiangsu, Guangxi, Shanxi, Beijing, Tianjin and Hebei Provinces, a decline of grapevines (Vitis vinifera) was identified with symptoms of dieback on one or more shoots, accompanied by elongated oval-shaped cankers in the wood, shrivelling and rot of fruit clusters. A total of 121 diseased samples were collected from affected areas and from these samples more than 1000 infected tissue pieces were used for pathogen isolation. Tissue from the lesion margins was surface- sterilized by placing in 75% ethanol for one min, then rinsing with steril- ized water (three times) before placing on potato dextrose agar plates incubated at 28°C for 5 days (Niekerk et al., 2004). Hyphae from the col- ony margins were transferred to fresh agar plates to obtain pure isolates. Botryosphaeria dothidea was isolated from 44 of the diseased samples. Colonies of B. dothidea were white, becoming grey or dark brown with age. Pycnidia started to develop after 10 days. Conidia measured 17–32 · 4–9 lm. Some isolates were used for ITS sequence analysis to confirm the morphological identification. The primers ITS1 and ITS4 were used to amplify part of the nuclear rDNA operon by PCR (White et al., 1990). Ten ITS sequences were obtained (GenBank Accession Nos. GU226844 to GU226850, GU226852, GU226854 and GU226855). All sequences showed 99–100% homology with B. dothidea (Urbez-Torres et al., 2006). This is the first report of grapevine canker associated with B. dothidea in China. Acknowledgement The research was supported by the Earmarked Fund for Modern Agro- industry Technology Research System (Ref. No. nycytx-30). References Niekerk JMvan, Crous PW, Groenewald JZ, Fourie PH, Halleen F, 2004. DNA phylogeny, morphology and pathogenicity of Botryosphaeria species on grapevines. Mycologia 96, 781–98. Urbez-Torres JR, Leavitt GM, Voegel TM, Gubler WD, 2006. Identification and distribution of Botryosphaeria spp. associated with grapevine cankers in California. Plant Disease 90, 1490–503. White TJ, Bruns T, Lee S, Taylor JW, 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ, eds. PCR Protocols: A Guide to Methods and Applications. New York, USA: Academic Press, 315–22. *E-mail: wangzhy0301@sina.com. Accepted 29 June 2010 at http://www.bspp.org.uk/ndr where figures relating to this paper can be viewed. 1170 ª 2010 The Authors Plant Pathology ª 2010 BSPP