Phytopathologia Mediterranea (2014) 53, 3, 565−592 DOI: 10.14601/Phytopathol_Mediterr-15167 ABSTRACTS Abstracts of oral and poster presentations given at the 9th International Workshop on Grapevine Trunk Diseases, Adelaide, Australia, 18–20 November 2014 The 9th International Workshop on Grapevine Trunk Diseases was held in Adelaide, Australia, 18–20 November, 2014. The meeting was attended by 111 participants and 71 papers were presented either as oral or poster pres- entations in four sessions: Pathogen Identification and Detection, Epidemiology, Host-Pathogen Interactions and Disease Management. A field trip to the Barossa Valley wine region was undertaken to showcase proactive manage- ment of grapevine trunk diseases in Australia. The workshop is the 9th organised by members of the International Council on Grapevine Trunk Diseases (www. icgtd.org), a subject matter committee of the International Society for Plant Pathology (www.isppweb.org). Pathogen Identification and Detection vegetative compatibility barriers. This indicated the likelihood of frequent parasexual recombination. The The identity, distribution and diversity of botry- isolation of genetically similar isolates from single le- osphaeriaceous species in New Zealand vineyards – a sions reinforced this hypothesis. A suite of molecular national perspective. H.J. RIDGWAY1,*, J. BASKAR- tools were developed to aid epidemiology studies. ATHEVAN1,2, N. AMPONSAH1,3, M.V JASPERS1 and Endogenous markers produced for isolates with typi- E.E. JONES1. 1Faculty of Agriculture and Life Sciences, cal pathogenesis showed they could be dispersed at Lincoln University, PO Box 85084, Lincoln University, Lin- least 2 m from the site of conidiation in a single rain/ coln, New Zealand, 7647. 2Ministry for Primary Industries- wind event. The use of a multi-genus PCR-SSCP system Manatū Ahu Matua, PO Box 2095, Auckland 1140, New showed that N. parvum and N. luteum are released year Zealand. 3Plant & Food Research, Old Mill Road, RD 3 round and this probably contributes to their successful Motueka, New Zealand. *E-mail: Hayley.Ridgway@lincoln. invasion of vineyards. Application of these molecular ac.nz tools has provided a comprehensive snapshot of New Zealand vineyards revealing a thriving and diverse In recent years molecular tools have been applied to population of botryosphaeriaceous species that present provide understanding of the population structures a serious concern to the industry. of botryosphaeriaceous species in New Zealand vine- yards. A national survey of symptomatic material from 43 vineyards showed that 88% had infection by Grapevine trunk diseases studies in British Colum- botryosphaeriaceous species. Vine age had the strong- bia: the status of long known diseases in a young and est correlation with incidence, with the least infection emerging grape-growing region. J.R. ÚRBEZ-TOR- in grapevines 1–5 years old (30%). Sequencing of taxo- RES*, P. HAAG, P. BOWEN and D.T. O’GORMAN. Ag- nomic genes identified nine species. In contrast to other riculture and Agri-Food Canada, Pacific Agri-Food Research countries, N. luteum and N. parvum were predominant Centre, 4200 Highway 97, Box 5000, Summerland, BC V0H species with Lasiodiplodia theobromae notably absent. 1Z0, Canada. *E-mail: [email protected] As with other countries, research showed that dis- tribution is likely to be related to climate. Analysis of Grapevine trunk diseases (GTD) are caused by a wide populations demonstrated that, despite predominantly range of taxonomically unrelated fungi and are current- asexual reproduction, the genetic diversity of isolates ly recognized as one of the major threats to the indus- within species was high. Frequent hyphal anastomoses try’s future economic sustainability. Previously, there and fusions were observed in dual culture with weak was no information available regarding the status of www.fupress.com/pm ISSN (print): 0031-9465 565 © Firenze University Press ISSN (online): 1593-2095 Abstracts GTD in British Columbia (BC), the second largest grape- Pathogenic fungi were isolated from 207 symptomatic growing region in Canada. Therefore, following some grape trunks. DNA isolation from fungal colonies was preliminary investigations studies to determine the in- conducted with the NucleoSpin Plant II kit. The ITS4 cidence, spread, and significance of GTD in BC started and ITS5 primers were used for PCR amplification of in 2010. Field surveys were conducted in 191 vineyards the ITS1-5.8S-ITS2 region. Purified amplicons were se- that included assesment of foliar symptomatology from quenced by Mycrosinth, Austria. Consensus sequences over 60,000 vines and isolations from 463 symptomatic were aligned with reference sequences using Clustal X vines. These studies revealed the presence of all GTD and manually corrected with GeneDoc. Phylogenetic in BC, including Botryosphaeria dieback, esca, Eutypa analyses were performed with MEGA 5.05. To date, dieback, young vine decline (black foot and Petri dis- 366 pure fungal cultures were isolated and identified. ease), and Phomopsis dieback. Phenotypic character- The majority of the isolates (56%) were determined as istics, including colony growth and micromorphology Diplodia seriata (Botryosphaeriaceae). Other fungi, like of reproductive structures along with DNA analyses Fusarium sp. (13.9%), Alternaria sp. (9.3%), Aspergillus of four genes (ACTIN, β-tubulin, TEF-1α, ITS1-5.8S- sp. (7.9%), Mucor sp. (7.1%), Diaporthe sp. (3.3%), Tricho- ITS2) allowed us to identify i) five different black foot derma sp. (1.4%), Epicoccum sp. (0.5%), Penicillium (0.3%) fungi in the genera Cylindrocarpon and Ilyonectria, ii) and Xanthomendoza sp. (0.3%) were also identified from seven species associated with esca and Petri disease, grapevine trunk samples. including the novel species Phaeoacremonium canadensis and Phaeoacremonium roseum, iii) several botryospha- eriaceous and diatrypaceous taxa, and iv) species in the Fungal pathogens involved in Grapevine Trunk Dis- Diaporthe/Phomopsis genera. Pathogenicity tests for all eases in China. J.Y. YAN1, W. ZHANG1, M. LIU1, X.H. identified fungi were completed and the characteristic LI1,* and K.D. HYDE1, 2. 1 Institute of Plant and Environ- ‘tiger-stripe’ symptoms commonly observed on esca- ment Protection, Beijing Academy of Agriculture and For- infected vines were reproduced when ‘Baco noir’ on estry and Sciences, Beijing 100097, China. 2 Institute of Ex- 3309C potted vines were artificially inoculated with cellence in Fungal Research, Mae FahLuang University, Chi- Phaeomoniella chlamydospora and different Togninia/ ang Rai. 57100, Thailand. *E-mail: lixinghong1962@163. Phaeoacremonium species under greenhouse conditions. com This study represents the first attempt to demystify the status of GTD in BC, a young and emerging grape- The grapevine cultivated area in China (both table- and growing region with unique climatic conditions. wine-grapes) has significantly increased from 125,000 hectares to 625,000 hectares between 2002 and 2013. As the cultivation area has increased, diseases became a Endophytic and pathogenic fungi associated with major problem for the grape industry in China. Apart Grapevine Trunk Diseases in the Tokaj wine region, from grape downy and powdery mildews, grapevine Hungary. CS. KOVÁCS1,*, E. SÁNDOR1, Z. BIHARI2 trunk diseases are the main factors limiting both vine- and F. PELES1. 1University of Debrecen, Faculty of Agricul- yard longevity and productivity. Woody parts of the tural and Food Sciences and Environmental Management, vine are infected by fungi, which lead to serious decline Food Science Institute, Böszörményi Street 138., H-4032 of the vineyard. In the past 10 years, grapevine trunk Debrecen, Hungary; 2Research Institute for Viticulture and diseases have become serious in some regions. Botry- Enology, Könyves Kálmán Street 54., H-3915 Tarcal, Hun- osphaeria dieback, being one of the most devastating gary. *E-mail: [email protected] diseases in China has been studied previously. In this study, another two trunk diseases were studied. Dis- Grapevine trunk diseases (GTD) are amongst the most eased sample survey conducted throughout the coun- important diseases in vineyards worldwide. GTD are try has shown Diaporthe spp. and Pestalotiopsis spp. are caused by several pathogenic fungi including, Phaeo- involved in the grape trunk diseases in China. Morpho- acremonium aleophilum, Phaeomoniella chlamydospora, Eu- logical and molecular phylogeny analysis confirmed typa lata, Fomitiporia mediterranea, Diplodia seriata among four Pestalotiopsis species and five Diaporthespecies as- many others. However, there is currently very limited sociated with grape trunk diseases. information about the status of GTD pathogens in the Tokaj wine region in Hungary. In this study, sympto- matic vine samples showing GTD foliar symptoms Detection of trunk pathogen inoculum in young vine- were collected from vineyards in four areas in the Tokaj yards to encourage adoption of preventative practices. region in 2013: Szemere (Degenfeld), Dorgó (Disznókő), D.P. LAWRENCE1,*, P.T. FUJIYOSHI1, R. TRAVADON1, Szarvas (Kereskedőház) and Várhegy (Patrícius). The D.A. CANTU2, A. MORALES-CRUZ2, P.E. ROLSHAUS- woody tissue samples were cut into small pieces, and EN3 and K. BAUMGARTNER1. 1United States Depart- were disinfected before plating on malt extract agar ment of Agriculture, Agricultural Research Service, Crops and incubated at room temperature for 3–8 days. Pathology and Genetics Research Unit, Davis, CA, USA. 566 Phytopathologia Mediterranea 9th International Workshop on Grapevine Trunk Diseases 2Department of Viticulture and Enology, University of Cali- and are designed for the identification of one or a small fornia, Davis, CA, USA. 3Department of Botany and Plant number of species at a time. Hence, considering that a Sciences, University of California, Riverside, CA, USA. *E- wide range of taxonomically unrelated fungal species mail: [email protected] are involved in the YVD complex and that many of these can be found in the nursery propagated material Because there are no efficient eradicative practices for prior to planting in commercial sites, there is a critical the wood infections caused by trunk diseases, routine need for a diagnostic approach capable to simultane- use of preventative practices is critical to maintain vine- ously identify and rapidly differentiate a much larger yard productivity. To help convince California growers number of YVD fungi at once. Accordingly, we have that early adoption of such practices is necessary in designed a DNA-based macroarray that contained over young vineyards, even in the absence of canopy symp- 100 species-specific oligonucleotide probes targeting toms, we evaluated spore traps (microscope slides and unique polymorphisms of the β-tubulin gene region. Rotorod® air sampler, coated in petroleum jelly) as a Probe specificity was determined by hybridizing over detection tool. Spores were trapped in three young (3–5 140 fungal isolates representing 73 different species year-old, asymptomatic) and three mature (13–15 year- from 8 genera including ex-type specimens from dif- old, symptomatic) vineyards, 12 sites total, in each of ferent geographical grape-growing regions. The DNA two regions (Napa and San Joaquin Counties) from De- macroarray precisely identified and discriminated over cember 2013 to March 2014. Traps were collected one 60 different fungal species including those associated day after each of eight rain events. For culture-based with YVD. The array was validated by testing artifi- detection, serial dilutions of spore suspensions were cially inoculated grapevine cuttings and soil as well as plated on PDA. For DNA-based detection, PCR primers commercial field samples. The high level of specific- were designed for the most aggressive and prevalent ity demonstrated by this DNA array showed it to be a trunk pathogens (Eutypa lata, Neofusicoccum parvum, promising detection system for accurate identification Phaeomoniella chlamydospora). Culture-based detection of YVD pathogens in a single test. revealed E. lata in 11 sites, Botryosphaeriaceous fungi (e.g. N. parvum) in nine sites, and P. chlamydospora in no sites. DNA-based detection revealed N. parvum in eight Two clonal lineages of Cadophora luteo-olivacea in sites, P. chlamydospora in two sites, and E. lata in no sites. Spain and South Africa revealed by multilocus ISSR Fewer pathogens were detected in culture and none markers. D. GRAMAJE1,*, M. LEON2, P.W. CROUS3 and by PCR from Rotorod® traps compared to microscope J. ARMENGOL2. 1Department of Crop Protection, Institute slides. Additional years of trapping and more specific of Sustainable Agriculture (IAS), Spanish National Research primers based on genome sequences of the pathogens Council (CSIC), Alameda del Obispo s/n, P.O. Box 4084, may resolve these conflicting results. Regardless, our 14080 Córdoba, Spain. 2Instituto Agroforestal Mediterráneo, finding of similar pathogens cultured from microscope Universidad Politécnica de Valencia, Camino de Vera s/n, slides in both young and mature vineyards suggests 46022 Valencia, Spain. 3CBS-KNAW Fungal Biodiversity that preventative practices should be adopted in young Centre, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands. vineyards. *E-mail: [email protected] Cadophora luteo-olivacea is a fungal trunk pathogen of Design, development and implementation of an oli- grapevine which has been recently isolated from vines gonucleotide array for identification and detection of showing decline symptoms in several grape growing fungal pathogens responsible for causing young vine regions worldwide. In this study, 80 C. luteo-olivacea decline. J.R. ÚRBEZ-TORRES*, P. HAAG, J. DICK, P. isolates (65 from Spain and 15 from South Africa) were BOWEN and D.T. O’GORMAN. Agriculture and Agri- studied. They were collected in the 1999–2010 period Food Canada, Pacific Agri-Food Research Centre, 4200 from vines in young vineyards, from planting material Highway 97, Box 5000, Summerland, BC V0H 1Z0, Cana- showing black vascular streaking and decline symp- da. *E-mail: [email protected] toms characteristic of Petri disease or from nursery samples and planting material at different stages of the Black foot and Petri disease are principle causes of propagation process. These isolates were studied by young vine decline (YVD) and mortality in young vine- means of Inter Simple Sequence Repeat (ISSR) analysis. yards. To date, over 45 fungi in the genera Cadophora, A total of 55 polymorphic ISSR markers were obtained Campylocarpon, Cylindrocadiella, Ilyonectria/Cylindro- with the four ISSR primers used. The ISSR markers re- carpon, Phaeomoniella, and Togninia/Phaeoacremonium vealed 40 multilocus genotypes (MLGs) in the global have been associated with YVD. Though significant population, with MLG13, MLG14, MLG29 and MLG31 advances have been made on molecular identification being the most frequent. Minimum spanning network and diagnostics of YVD fungal pathogens during the analysis showed that the MLGs from South Africa clus- last decade, many have been PCR-based techniques tered around the most frequent genotype, while the Vol. 53, No. 3, December, 2014 567 Abstracts genotypes from Spain were distributed all across the tion in the greenhouse, indicating the ability of these network. Bayesian and principal component analyses Cadophora species to degrade grapevine wood. This identified two highly differentiated genetic clusters in study showed that Cadophora species infecting grape- the Spanish and South African C. luteo-olivacea popu- vine in North America are diverse and should help fur- lations, with no intermediate genotypes among these ther morphological and molecular identification of this clusters. This suggests several events of introduction, obscure group of fungi. although movement within the Spanish provinces may have occurred repeatedly given the frequent retrieval of the same genotype in distant locations. Our findings Identification of Cylindrocladiella species isolated provide new insights into the population genetic struc- from rootstock wood cankers in California. L.J. BET- ture of C. luteo-olivacea and highlight the need to pro- TIGA1,*, S.T. KOIKE1, W.D. GUBLER2 and T. NGUYEN duce healthy and quality planting material in grape- 2. 1 University of California Cooperative Extension, 1432 Ab- vine nurseries to avoid its spread throughout different bott Street, Salinas, California 93901, USA. 2 Department of grape growing regions. Plant Pathology, University of California, Davis, California 95616, USA. *E-mail: [email protected] The role of Cadophora species as trunk pathogens in In late summer 2012, 2-year old grapevines (cv. Char- North American grape-growing regions. D.P. LAW- donnay on 101-14 Mgt) in a vineyard located in the RENCE1, R. TRAVADON1,*, S. ROONEY-LATHAM2, Salinas Valley, California displayed black-foot disease. W.D. GUBLER3, P.E. ROLSHAUSEN4 and K. BAUM- Internal tissues of roots and rootstock exhibited brown GARTNER1. 1United States Department of Agriculture to black streaking and sections. Approximately 2% of - Agricultural Research Service, Crops Pathology and Ge- vines were affected. Surface disinfested pieces of symp- netics Research Unit, Davis, CA 95616, USA. 2Plant Pest tomatic wood, placed on PDA, consistently yielded Diagnostics Branch, California Department of Food & Ag- two types of fungal colonies, one primarily yellow in riculture, 3294 Meadowview Road, Sacramento, CA 95832- color, and the other reddish. However, for all isolates 1448, USA. 3Department of Plant Pathology, University of the general morphological features were similar and California, Davis, CA 95616, USA. 4Department of Botany consisted of penicillate conidiophores, cylindrical hya- and Plant Sciences, University of California, Riverside, CA line conidia, and hastate vesicles. Single-spore cultures 92521, USA. *E-mail: [email protected] of ten yellow and eight red isolates were used for mo- lecular characterization. Based on the morphological Cadophora species are recently acknowledged as puta- and molecular data, yellow isolates were confirmed as tive trunk pathogens in independent investigations Cylindrocladiella lageniformis and red isolates were iden- from California, South Africa, Spain, Uruguay, and tified as C. peruviana. To demonstrate Koch’s postulates, Canada. In particular, Cadophora luteo-olivacea has been 14-day-old colonies of three C. lageniformis and four C. associated with vines affected by Petri disease and peruviana isolates were inoculated onto potted plants of Esca, and indeed has been shown to cause the char- cv. Riesling by inserting mycelial agar plugs beneath acteristic black streaking in controlled inoculations. In cuts into the epidermis of shoots. The two Cylindroclad- North America, little is known regarding the diversity iella species were recovered from discolored tissue in all and relative importance of Cadophora species, their geo- canes of their respective inoculated plants. Analysis of graphic distribution, and their ability to degrade grape- the recovered isolates demonstrated that these matched vine wood. Accordingly, we characterized 41 Cadophora the morphological and molecular characteristics of the isolates recovered from decayed grapevine wood from original isolates. Control canes, inoculated with sterile a broad geographic area in North America, using mor- agar plugs, did not develop symptoms and no patho- phological and molecular analyses, and pathogenic- gen was isolated. To our knowledge this is the first doc- ity assays. Morphological characterization (sizes of umentation of Cylindrocladiella lageniformis and C. peru- conidia, hyphae, conidiophores, and conidiogenous viana associated with a grapevine disease in California. cells) identified five groups of isolates, which corre- These pathogens were also isolated from grapevines in sponded to five species as revealed by phylogenetic 2014, indicating that the disease may be more broadly analyses of ITS, translation elongation factor 1-alpha, distributed. and beta-tubulin: C. luteo-olivacea, C. melinii/fastigiata, C. malorum, and two Cadophora sp. nov. Restricted to the northeastern US were C. malorum and the two Cadopho- First molecular characterization of Phomopsis viticola ra sp. nov., whereas C. luteo-olivacea was only recovered and Diplodia seriata isolated from Esca-BDA diseased from California. C. melinii/fastigiata was present both in grapevines in Northern Tunisia. A. BEN GHNAYA- California and Ontario, Canada. Pathogenicity assays CHAKROUN1,2,3, A. REZGUI1,2,3, J. VALLANCE2,3, I. produced lesions significantly longer than those of the KHAROUBI1, M. DRIDI4, M.R. HAJLAOUI4, P. REY2,3,* non-inoculated control plants after 24 months incuba- and N. SADFI-ZOUAOUI1. 1 Laboratoire des Microorgan- 568 Phytopathologia Mediterranea 9th International Workshop on Grapevine Trunk Diseases ismes et Biomolécules Actives, Département de Biologie, ‘Baco blanc’ (a hybrid grape used in Armagnac produc- Faculté des Sciences de Tunis. Campus Universitaire, 2092 tion). During the spring season, 42- and 58-year-old Tunis, Tunisie. 2 Université de Bordeaux, Bordeaux Sciences vines were sampled; they showed no foliar symptoms Agro, ISVV, UMR1065 SAVE, F-33140 Villenave d’Ornon, of Esca or Eutypa dieback. The methods used for stud- France. 3 INRA, UMR1065 Santé et Agroécologie du Vigno- ying the microflora were isolation and subsequent se- ble (SAVE), ISVV, F-33140 Villenave d’Ornon, France. 4 quencing the rDNA internal transcribed spacer region Laboratoire de Protection des végétaux, Institut National de (ITS), and a molecular fingerprinting method, Single- la recherche Agronomique de Tunisie (INRAT) 2049, Ariana strand conformation polymorphism analysis (SSCP). Tunisie. *E-mail: [email protected] Among the 421 cultured strains used for ITS sequenc- ing, 7 plant-pathogenic fungal species, particularly Esca and Black Dead Arm (BDA) are two major those involved in Esca (42-year-old vines) and Eutypa grapevine trunk diseases (GTDs)diseases, which oc- dieback (58-year-old vines), were identified, including cur worldwide. Although Esca and BDA have started Phaeomoniella chlamydospora, Fomitiporia mediterranea to cause considerable damage in Tunisian vineyards, and Eutypa lata. Fungal communities differed accord- knowledge of the symptoms and microflora associated ing to vine age, based on both cultured-based identi- with these GTDs is still incomplete in Tunisia, The ob- fication and SSCP profiles. However, communities did jective of this research was to study and characterize not differ significantly among the cordon or the upper, the fungal microflora that colonize the wood tissues central, or basal parts of the trunk. Regardless of vine of Esca-BDA symptomatic and asymptomatic vines. age, numerous mycoparasites, such as Trichoderma spp. To do this, apparently healthy and necrotic wood tis- and Bionectria sp., and saprobes were also isolated. The sues were collected from diseased and control plants in absence of foliar symptoms may be associated with an eight vineyards in the north of Tunisia, and analyzed equilibrium resulting from competition between patho- using microbiological and molecular approaches. Our genic and mycoparasitic fungi in the woody tissues. results showed that diverse fungal assemblages made up of potentially plant pathogenic and numerous sap- rophytic fungal species (mainly Aspergillus spp. and Al- Molecular and phenotypic characterisation of Phaeo- ternaria alternata) colonize the wood tissues (apparently moniella chlamydospora isolates from the demarcated healthy wood and necrotic) of the sampled vines. The wine region of Dão (Portugal). J. SOFIA1,*, A. PORTU- ITS sequencing of the different isolates confirmed the GAL1, H. PAIVA de CARVALHO1, N. MESQUITA1, T. presence of Phomopsis viticola and Diplodia seriata, two NASCIMENTO2, M.T. GONÇALVES1 and C. REGO2. fungi described in the literature as involved in GTDs. 1Centre for Functional Ecology, Department of Life Sciences, The molecular analysis also highlighted that Alternaria University of Coimbra. Calçada Martim de Freitas, 3000- alternata is the main saprophytic strain colonizing both 446. Coimbra. Portugal. 2Instituto Superior de Agronomia, healthy and diseased wood. A halophilic bacterium Technical University of Lisbon, Tapada da Ajuda, 1349-017 (strain J9) was tested as an antagonistic agent against Lisboa, Portugal. *E-mail: [email protected] the isolated fungi using in vitro confrontations. The re- sults, especially those concerning D. seriata, were prom- Phaeomoniella chlamydospora is considered one of the ising. main causal agents of Esca and Petri disease, which are responsible for worldwide grapevine decay, as in the Portuguese demarcated wine region of Dão. In this old Pathogenic fungi and mycoparasites colonizing the and well-known wine region, regional grapevine cul- functional wood of old vines with no Eutypa die- tivars are mainly used with a low penetration of alien back or Esca foliar symptoms. E. BRUEZ1,2, K. BAUM- varieties, and most of the propagation material is pro- GARTNER3, S. BASTIEN1,2, R. TRAVADON3, L. GUER- duced in local nurseries. In the present work, we used IN-DUBRANA1,2 and P. REY1,2,*. 1Université de Bordeaux, 69 isolates of Pa. chlamydospora, obtained from symp- Bordeaux Sciences Agro, UMR1065 SAVE, F-33140 Ville- tomatic Esca and Petri disease grapevines, from differ- nave d’Ornon, France. 2INRA, UMR1065 Santé et Agroé- ent locations across the Dão region. We compared the cologie du Vignoble (SAVE), ISVV, F-33140 Villenave morphological characteristics, geographic origin and d’Ornon, France. 3United States Department of Agriculture, ITS (Internal Transcribed Spacers) sequence variations Agricultural Research Service, University of California, One to determine intraspecific variability and population Shields Avenue, Davis, CA 95616, USA. *E-mail: prey@ structure. Furthermore, results will likely allow viru- bordeaux.inra.fr lence assessment and prediction. In France, grapevine trunk diseases Esca and Eutypa dieback tend to be less severe in vineyards older than Current situation of the fungal trunk pathogens of 25 years. We compared the fungal microflora colonizing grapevines in Chile. B.A. LATORRE* and G.A. DIAZ. the functional (i.e. non-necrotic) wood of the cultivar Departamento de Fruticultura y Enología, Facultad de Vol. 53, No. 3, December, 2014 569 Abstracts Agronomía e Ingeniería Forestal, Pontificia Universidad sponsible for the most harmful trunk diseases of grape- Católica de Chile, Vicuna Mackenna 4860, Macul, Santiago, vines. The MYCORRAY system consists of a disposable Chile. *E-mail: [email protected] Array Tube containing a DNA microarray chip and a cost-effective standalone array reader suitable for use Trunk diseases in grapevine (Vitis vinifera) have been by non-specialists, composed of an optical system and identified as a major problem in the wine and table an image processing system. Additionally, a simple grapes worldwide, reducing the productivity, quality and inexpensive methodology for non-lethal sampling and longevity of vineyards. The present study exam- and DNA extraction has been developed for optimum ined 694 wood samples from the cordons and trunks of detection of GTDs in vine material. Seven DNA extrac- vines with trunk disease symptoms in 67 Chilean vine- tion methods were compared to ensure maximum fun- yards located between Copiapó (27°18’ S) and Los An- gal DNA extraction and purification coupled with the geles (37°42’ S) from 2009 to 2012. A total of 1,363 fun- highest sensitivity and minimum testing costs. Sam- gal isolates were obtained from diseased cordons and pling using a drill from four points in the scion and two trunks with dark brown streaking, yellowish spongy points on the rootstock that were pooled together gave cankers and brown hard V-shaped cankers. On the ba- the best results. Using traditional isolation techniques sis of morphology and molecular analysis, a total of 11 in combination with a nested PCR approach for patho- fungal genera were identified at varying frequencies: gen detection, this sampling minimized the number of Phaeomoniella chlamydospora (85%); Botryosphaeriaceae DNA extractions required. The MYCORRAY system in (56%) including Diplodia mutila, D. seriata, Neofusicoc- combination with this sampling protocol is therefore cum parvum and Spencermartinsia viticola; Inocutis sp. likely to provide a robust, efficient tool to help manage (47%); Diatrypaceae (Cryptovalsa ampelina and Eutypella GTDs worldwide. leprosa) (4.8%); Seimatosporium botan (1.7%); Phomopsis viticola (0.4%); Cylindrocarpon sp. (0.4%); and Phaeo- Epidemiology acremonium aleophilum (0.2%). All species were patho- genic, inducing dark brown streaking on various aged Botryosphaeriaceae conidial dissemination in two grapevine wood tissue. In conclusion, several fungal different locations with Mediterranean semiarid cli- species are associated with grapevine trunk diseases mate in the Valparaíso region of Chile. D. VALENCIA, in the Chilean vineyards being Pa. chlamydospora, D. se- C. TORRES and X. BESOAIN*. Facultad de Agronomía, riata and Inocutis sp. the most frequent isolated species. Pontificia Universidad Católica de Valparaíso, Av. Brasil These are pathogens that can be found alone or they can 2950, Valparaíso, Chile. *E-mail: [email protected] coexist in the same plant. There were no evidences of Eutypa lata affecting grapevines in Chile. The dissemination of Botryosphaeriaceae causing die- back in vineyards has recently received increased atten- tion. Rain and high relative humidity trigger conidial MYCORRAY: a DNA microarray tool for the early, release into the environment which can infect prun- reliable, simultaneous identification of the grape- ing wounds during this time. However, the seasonal vine trunk diseases fungal agents. J. WOODHALL2, abundance of conidia varies among production areas G. MARCHI1, T. CINELLI1, B. GINETTI1, D. BOSSIO1, and different latitudes. The goal of this study was to J. TOMLINSON2, C. HARRISON2, N. BOONHAM2 determine the effects of rainfall, relative humidity and and L. MUGNAI1,*. 1Dipartimento di Scienze delle Pro- temperature on the moment of maximal dispersion duzioni Agroalimentari e dell’Ambiente (DISPAA) sez. of Botryosphaeriaceae conidia in two Mediterranean Patologia Vegetale ed Entomologia, P.le delle Cascine, 28, semiarid temperate climate locations in Valparaíso Re- 50144 Firenze, Italy. 2The Food and Environment Research gion of Chile. To achieve this, five microscope slides Agency, Sand Hutton, York, YO41 1LZ, UK. *E-mail: laura. covered with liquid Vaseline were placed in four vine- [email protected] yards of two locations: Casablanca (with high marine influence) and Panquehue (with low marine influence) Grapevine trunk diseases (GTD) including the Esca both displaying symptoms of Botryosphaeria die- complex, Eutypa dieback, Botryosphaeria canker, and back. The slides were replaced every week and sent to Black foot, often involve more than one causal agent PUCV Phytopathology Laboratory in order to observe and several causal agents may coexist in the same vine. the presence of conidia presence over a one-year time Diagnoses with currently available diagnostic methods period. In addition, isolations were carried out from only detect one organism per test, making the detec- plants with Botryosphaeria dieback at each location. tion of all GTD causal agents involved impractical and Species were identified using morphological and mo- prohibitively expensive. With this purpose, a microar- lecular techniques. In Casablanca, a spore “peak” was ray tool (named MYCORRAY) is under development detected during August, coinciding with the last rain- which aims for early, reliable, simultaneous identifica- fall recorded. Release started again at the beginning of tion of the 11 most relevant grapevine trunk fungi re- autumn 2014 and has continued to the current time and 570 Phytopathologia Mediterranea 9th International Workshop on Grapevine Trunk Diseases according to winter rain patterns. In Panquehue loca- Protection Division, ARC Infruitec-Nietvoorbji, Private Bag tion, spore release was observed during spring, while X5026, Stellenbosch, 7599, South Africa. 2Department of in fall-winter release was null or very low. The Botry- Plant Pathology, University of Stellenbosch, Private Bag X1, osphaeriaceae species identified were Diplodia seriata, Matieland, 7602, South Africa. *E-mail: [email protected] Dothiorella sp. and Neofusicoccum sp. Phaeomoniella chlamydospora and Phaeoacremonium spe- cies have been associated with Petri disease in grape Genetic analysis of Neofusicoccum parvum and N. growing regions around the world. Spores of these path- luteum isolates from nurseries and vineyards indi- ogens are aerially dispersed and infect vines through cates different infection sources. R. BILLONES-BAA- susceptible wounds. However, no information is avail- IJENS1,2,*, J. BASKARATHEVAN1,3, M.V. JASPERS1, E.E. able about the time when these spores are released in JONES1, R.H. CRUICKSHANK1 and H.J. RIDGWAY1. South African vineyards. A study was undertaken in six 1Faculty of Agriculture and Life Sciences, Lincoln Uni- vineyards and two rootstock mother blocks within the versity, PO Box 85084, Lincoln University, Lincoln, New Western Cape Province, over two seasons, from May– Zealand, 7647. 2National Wine and Grape Industry Centre November 2012 and March–November 2013. Micro- (NWGIC), Charles Sturt University, Mckeown Drive, Wag- scope slides coated with petroleum jelly were affixed to ga Wagga, NSW 2678 Australia. 3Plant Health and Envi- five vines (or rootstock mother plants) in each vineyard. ronment Laboratory, Ministry for Primary Industries, PO The slides were changed weekly, washed in 5ml sterile Box 2095, Auckland 1140, New Zealand. *E-mail: rbaaijens water and passed through 5- and 0.45-μm filters. The @csu.edu.au filters were backwashed and the suspensions plated onto PDA. Colonies resembling Petri disease pathogens Surveys in 2007–08 showed that Neofusicoccum parvum were recorded, sub-cultured and identified on the basis and N. luteum are the two most prevalent and virulent of cultural characteristics (Pa. chlamydospora), and with species found in New Zealand vineyards and nurser- species-specific primers or sequencing of the partial ies. However, N. parvum is more common in vineyards betatubulin and actin genes (Phaeoacremonium species). while N. luteum dominates in nurseries. Pathogenicity Spores of various species were detected within each of studies also showed that N. parvum was more aggres- the vineyards and inoculum was available whenever sive and produced more severe, darker external le- pruning or sucker wounds were made in winter and sions on canes than N. luteum. This study used genetic spring. Over the two seasons 13 Phaeoacremonium spe- data to elucidate population origins of N. parvum and cies were detected, namely Pm. aleophilum, Pm. alvesii, N. luteum from vineyards and nurseries. Vineyard and Pm. australiense, Pm. viticola, Pm. parasiticum, Pm. sco- nursery isolates of N. parvum (n=79) and N. luteum lyti, Pm. sicilianum, Pm. subulatum, Pm. iranianum, Pm (n=64) were genotyped using five universally-primed inflatipes, Pm. venezuelense, Pm. prunicola and Pm. griseo- polymerase chain reaction (UP-PCR) primers. The five olivacea, the latter two for the first time in South Afri- primers were able to amplify a total of 51 loci for N. par- can vineyards. Pa. chlamydospora and Pm. aleophilum vum (66% polymorphic) and 54 loci for N. luteum (44% were the only species detected in all the vineyards. This polymorphic). Phylogenetic analysis using parsimony study confirms the need for effective wound protection (PAUP) showed that 92% and 78% of the N. parvum strategies. and N. luteum populations, respectively, were of unique genotypes. The neighbour joining trees showed that N. parvum from nurseries clustered separately from the Studying and modelling the summer development vineyard isolates indicating the two populations were of esca foliar symptoms. P. LECOMTE1, E. BRUEZ1,4, genetically distinct. This supported the initial hypoth- J. GERBORE1,2, P. PIERI3, L. GUERIN-DUBRANA1,4, esis that the nursery infections by this aggressive spe- D. BARKA1, Y. MEZIANI1, C. BENETREAU1, M. FER- cies were intercepted during the grading process and, MAUD1 and P. REY1,4. 1INRA, Université de Bordeaux, therefore, there was less movement of this species to ISVV, UMR1065 SAVE, CS 20032, F-33882, Villenave the vineyards. In contrast, the N. luteum nursery and d’Ornon cedex, France. 2BIOVITIS, F-15400 Saint Etienne de vineyard populations showed high genetic similarities. Chomeil. 3INRA, Université de Bordeaux, ISVV, UMR1287 This indicated that the less distinct symptoms caused EGFV, F-33140 Villenave d’Ornon, France. 4Université de by this species are not graded out and, therefore, these Bordeaux, Bordeaux Sciences Agro, ISVV, UMR1065 SAVE, infections can easily migrate from the nurseries to the CS 20032, F-33882, Villenave d’Ornon cedex, France. *E-mail: vineyards. [email protected] A survey carried out in Aquitaine vineyards (France) Spore release patterns of Petri disease fungi in South from 2004 to 2006 showed that the occurrence of esca African vineyards and rootstock mother blocks. M.A. foliar symptoms had a similar progressive pattern in BALOYI1, L. MOSTERT2 and F. HALLEEN1,2,*. 1Plant all the plots surveyed. This pattern corresponded to a Vol. 53, No. 3, December, 2014 571 Abstracts sigmoidal increase in the incidence of vines exhibit- etation Index (NDVI) acquired by the UAV and GLSD ing typical symptoms. In general, regardless of the site symptoms, and the ability to discriminate symptomatic and year, the leaf-symptomatic vines increased uni- from asymptomatic plants. High-resolution multispec- formly over time, reaching a maximum incidence by tral images were acquired over 2012 and 2013 in June the end of July. To further study this phenomenon, an- and July in an experimental vineyard, located in Tusca- other survey was carried out from 2012 to 2014 in the ny (Italy), where vines had been surveyed and mapped Bordeaux region. It was based on regular monitoring since 2003. Each vine was mapped with Leica Differ- of vines twice a week. The data confirmed previous re- ential-GPS, and classified for foliar symptoms appear- sults and were used to develop a logistic model aimed ance and disease severity weekly since the beginning at characterizing the period of symptom appearance of each season. In 2013, single leaves from the vineyard over time. This period corresponded to the increase were analyzed for physiological parameters using a in average temperatures and to a putative establish- Portable Photosynthesis System (Li-Cor 6400). Remote ment of water restriction. This model could be used sensing and ground observation data were analyzed to to study the effect of environmental factors, i.e. soil, promptly identify the early stages of symptoms, even climate and the susceptibility of cultivars. Esca symp- before visual detection. The objective of this research toms were mostly associated with longitudinal xylem was to develop an innovative method aimed primarily discolorations located just under the bark on cor- at conducting a quantitative and qualitative analysis of dons or trunks. The origin of this peculiar symptom the symptom spread, and then to realize a predictive is still unclear even though it has been attributed to tool of the GLSD symptoms onset. infections by Botryosphaeriaceae. In the same survey, vines with recent symptoms were identified and in- vestigated. Diverse fungal communities were isolated Seasonal susceptibility of grapevine pruning wounds from discoloured or “apparently healthy wood”. The to artificial infection with Diplodia seriata and Phae- percentages of Botryosphaeriaceae isolated from these omoniella chlamydospora in Catalonia, NE Spain. G. samples were not significantly different. Furthermore, ELENA* and J. LUQUE. IRTA Cabrils, Ctra. de Cabrils km their absence in some samples indicated that these 2, 08348 Cabrils, Spain. *E-mail: [email protected] fungi cannot be responsible (at least alone) for the xy- lem discolorations. Dispersal of grapevine trunk diseases in the vineyard relies on the infection of pruning wounds by airborne pathogen spores. The aim of this study was to evaluate Unmanned Aerial Vehicle based remote sensing the period in which pruning wounds remain suscep- as a predictive tool for the onset of grapevine leaf tible to infection by the pathogenic fungi Diplodia se- stripe disease symptoms. S.F. DI GENNARO1,2,*, M. riata and Phaeomoniella chlamydospora. An experimental BENANCHI3, A. MATESE1, J. PRIMICERIO1, L. GEN- ‘Tempranillo’ vineyard located in our institution was ESIO1, A. PALLIOTTI2, O. FACINI4, S. DI MARCO4 and pruned at two different seasons: early in autumn, and L. MUGNAI3. 1Istituto di Biometeorologia (IBIMET), CNR, late in winter. In both seasons, pruning wounds (N=20 Via G. Caproni 8, 50145 Florence, Italy. 2Dipartimento di per treatment) were artificially inoculated with a co- Scienze Agrarie e Ambientali, Università degli Studi di nidial suspension of each pathogen separately at dif- Perugia, Borgo XX Giugno 74, 06128 Perugia, Italy. 3Di- ferent periods after pruning (1 day, and after 1, 2, 4, 8 partimento di Scienze delle Produzioni Agroalimentari e and 12 weeks). For the control treatment, pruned canes dell’Ambiente (DiSPAA), Sez. Patologia vegetale ed Ento- were treated with sterile distilled water. Four months mologia, Università degli Studi di Firenze, Piazzale delle after the inoculations, the percentage of fungal recov- Cascine 28, 50144 Florence, Italy. 4 Istituto di Biometeorolo- ery from canes was determined. The experiment was gia (IBIMET), CNR, Via Gobetti 101, 40129 Bologna, Italy. repeated twice in the seasons 201213 and 20132014. The *E-mail: [email protected] reisolation percentage of D. seriata fell below 35% af- ter 2 weeks in the early pruning, while a comparable Grapevine Leaf Stripe Disease (GLSD) foliar symptoms decrease in the late pruning was not observed until 8 were proved by several authors to be linked to a dras- weeks. Phaeomoniella chlamydospora showed a simi- tic alteration of photosynthetic function, as well as an lar pattern since the reisolation percentage decreased activation of defence responses in affected grapevines, dramatically after 4 weeks in the early pruning while several days before the appearance of the first visible susceptibility in late pruning only fell below 25% after symptoms on leaves. This study suggests a method 8 weeks. The results indicated that pruning wounds to investigate the correlation between high-resolution remain less susceptible to pathogen infection after an multispectral images acquired by Unmanned Aerial early pruning as compared to a late pruning, thus sug- Vehicles (UAV) and GLSD foliar symptoms monitored gesting that a shift of the traditional late pruning in the by ground surveys. This approach previously showed area to an early pruning would reduce the risk of path- a high correlation between Normalized Difference Veg- ogen infection in Catalonia. 572 Phytopathologia Mediterranea 9th International Workshop on Grapevine Trunk Diseases Grapevine sucker wounds as infection ports for trunk olivacea, Phaeoacremonium spp. and Phaeomoniella chla- disease pathogens. G. MAKATINI1, C. MUTAWILA1, F. mydospora. All trunk pathogens, with the exception of HALLEEN1,2 and L. MOSTERT1,*. 1Department of Plant E. lata, were detected in samples from the four vine- Pathology, University of Stellenbosch, Private Bag X1, Mat- yards evaluated by nested-PCR, being Ca. luteo-olivacea ieland, 7602, South Africa. 2Plant Protection Division, ARC and Phaeoacremonium spp. the most frequent. We found Infruitec-Nietvoorbji, Private Bag X5026, Stellenbosch, the co-occurrence of 2, 3 or 4 different fungal trunk 7599, South Africa. *E-mail: [email protected] pathogens in the same sample. Cadophora luteo-olivacea and Phaeoacremonium spp. were the combination most The susceptibility of sucker wounds to different trunk frequently detected. In addition, fungal isolation from disease pathogens was assessed from natural and ar- liquid samples in semi-selective culture medium for tificial infections. Sucker wounds were sampled from Ca. luteo-olivacea, Phaeoacremonium spp. and Pa. chla- three wine and two table grape vineyards during 2011 mydospora was also performed, but only Ca. luteo-oli- and 2012 in the Western Cape province of South Africa. vacea was recovered from samples collected in 3 out of Isolations were made from 161 sucker wounds and the 4 vineyards evaluated. Shears artificially infested with fungal cultures were identified. Sixtytwo percent of suspensions of conidia or mycelial fragments of Diplo- wounds were naturally infected by at least one of the dia seriata, Eutypa lata, Ca. luteo-olivacea, Pm. aleophilum trunk pathogens. The most predominant trunk disease and Pa. chlamydospora, were used to prune 1-year-old pathogens isolated from sucker wounds of field wine grapevine cuttings of 110 Richter rootstock. Fungal re- and table grape cultivars, respectively were Phomopsis isolation from the cuttings four months after pruning (Po.) viticola (46%; 18%), Diplodia (D.) seriata (30%; 9%) confirmed that infested shears were able to infect them and Phaeomoniella (Ph.) chlamydospora (27%; 5%). Lower through pruning wounds. Our results demonstrate incidences of Phaeoacremonium aleophilum (18%), Eu- the potential of inoculum present on shears to infect typella sp. (3%), Cryptovalsa ampelina (2%), Diplodia sp. grapevines. (1%) and Neofusicoccum australe (1%) were obtained, however, only from wine grapes. Sucker wounds on 1-year-old potted grapevine plants of Chardonnay cul- Co-infection by Neofusicoccum luteum and N. parvum tivar were inoculated with spore suspensions of Eutypa influences direction of lesion expansion but not total lata, N. parvum, Pa. aleophilum, Ph. chlamydospora and lesion size. J. BASKARATHEVAN1,2, E.E. JONES2, M.V. Po. viticola in the glasshouse. After 4 months all the in- JASPERS2 and H.J. RIDGWAY2,*. 1Ministry for Primary oculated pathogens could be re-isolated. Sucker wound Industries-Manatū Ahu Matua, PO Box 2095, Auckland susceptibility was further ascertained under field con- 1140, New Zealand. 2Faculty of Agriculture and Life Scienc- ditions on Cabernet Sauvignon vines by artificial inoc- es, Lincoln University, PO Box 85084, Lincoln University, ulation of the same pathogen species. After 5 months, Lincoln 7647, New Zealand. *E-mail: Hayley.Ridgway@ only Po. viticola, N. parvum and Ph. chlamydospora could lincoln.ac.nz be re-isolated. The duration of susceptibility of field sucker wounds to Ph. chlamydospora was assessed for 4 In a national survey of symptomatic vines a total of 238 weeks. The wounds remained susceptible for 4 weeks samples were collected yielding 336 isolates of botry- with a decline in susceptibility after one week. This osphaeriaceous species. From this collection 18 lesions study showed that sucker wounds are susceptible to were systematically sampled to determine if multiple the major trunk disease pathogens. species were present. Twelve lesions contained multi- ple species. Diplodia seriata was found most frequently in combination with other species (40%) but Neofusico- Potential role of shears on the infection of grapevine cum luteum and N. parvum were found most frequently by fungal trunk pathogens. C. AGUSTÍ-BRISACH, M. together (20%). To determine whether co-infection by LEÓN, J. GARCÍA-JIMÉNEZ and J. ARMENGOL*. In- these two Neofusicoccum species was synergistic, trunks stituto Agroforestal Mediterráneo, Universidad Politécnica of 1-year-old Sauvignon blanc vines were co-inoculated de Valencia, Camino de Vera s/n, 46022-Valencia, Spain. *E- with two isolates each of N. luteum and N. parvum, ei- mail: [email protected] ther together or alone. Each pair of isolates for each spe- cies consisted of a weakly virulent and highly virulent Four Spanish vineyards were surveyed during prun- isolate, with virulence established in previous experi- ing in 2012. In each vineyard, shears were washed ments. Following inoculation the presence of both spe- with a sterile solution of 0.2% Tween-20 after prun- cies in one lesion was confirmed by PCR. The results ing ten grapevines. Twenty liquid samples were col- showed that in co-inoculated lesions mean total lengths lected from a total of 200 plants. Molecular detection were not larger than the lesion length produced by the of fungal trunk pathogens in these samples was per- most virulent of the isolate pair (P≤0.05), irrespective formed by nested-PCR using specific primers to detect of the virulence of the second isolate. However, when Botryosphaeriaceae spp., Eutypa lata, Cadophora luteo- the distances from the inoculation points to lesion Vol. 53, No. 3, December, 2014 573 Abstracts edges were analysed, results showed that there were Aranha Avenue, 540, Zip Code 95700-000 Bento Gonçalves, significantly greater lengths below than above inocula- Rio Grande do Sul, Brazil. 2 Embrapa Grape & Wine, Livra- tion points for all combinations (P≤0.05). In addition, mento Street, 515, Bento Gonçalves, Rio Grande do Sul, Bra- co-inoculation of two weakly virulent isolates reduced zil. 3 Plant Protection Division, ARC Infruitec-Nietvoorbji, upward movement (P≤0.05). A decrease in the endo- Private Bag X5026, Stellenbosch, 7599, South Africa. *E- phytic movement of N. luteum beyond the lesion was mail: [email protected] also observed in all co-inoculations. Overall, the results demonstrated that there were no synergistic effects of Several factors affect the quantity and quality of grape- co-inoculation with these two common species. vine production in Brazil. This includes the grapevine decline phenomenon and the various disease complex- es associated with it. For example, in Brazil, there is The genetic structure of Botryosphaeriaceae isolated very little knowledge about Esca and Petri disease and from vineyards in south eastern Australia as revealed the distribution of the pathogens associated with it. The by molecular markers. Y. QIU, C.C. STEEL, G.J. ASH objective of this research was to study Phaeomoniella and S. SAVOCCHIA*. National Wine and Grape Industry chlamydospora (Pchl) distribution in different cultivars Centre, Charles Sturt University, School of Agricultural and and plant parts from vineyards in the Serra Gaúcha re- Wine Sciences, Locked Bag 588, Wagga Wagga NSW 2678 gion of the Rio Grande do Sul state in southern Brazil. Australia. *E-mail: [email protected] Grapevines with decline symptoms were collected in 21 vineyards and taken to the laboratory for destructive Genetic variation within four species of Botryopha- sampling and analysis. Vitis vinífera cultivars included eriaceae isolated from south eastern Australian vine- Chardonnay (14 plants), Merlot (1) and Cabernet Sau- yards (Lower and Upper Hunter Valley and Mudgee) vignon (7). Vitis labrusca cultivars included Isabel (1 was investigated using a hierarchical sampling and a plant), Bordô (10) and Niágara (4). Isolations were made range of PCR primers. A set of 100 primers (Microsatel- from symptoms in spurs, arms, upper part of the trunk, lite Primer Set #9; Biotechnology laboratory, University graft union and basal end of the rootstock. Pchl inci- of British Columbia, Canada), (ACC), R1 (consensus dences ranged from 2.5 to 27.4% in V. labrusca and 4.8 5 region from a plant intron splice junction) and M13 to 8.8% in V. vinifera. From the different plant parts Pchl (core sequence of the M13 minisatellite region) prim- incidences ranged from 7.3 to 13.1%. From the different ers were screened with a selection of isolates from all internal symptoms, Pchl was most frequently isolated four species. Of these, five of the 100 primer set, (ACC), from black spotting (17.1 %) and soft and spongy rot- 5 M13 and R1 were selected and resulted in 280 highly ted wood (15.4%), followed by brown-red wood (7.8%), reproducible polymorphic bands across 127 Diplodia black line (7%) and wedge shaped necrosis (6.3%). seriata, 23 Neofusicoccum parvum, 18 Botryosphaeria doth- idea and three Lasiodiplodia theobromae isolates. Cluster Host Pathogen Interaction analysis with the integrated locus matrix separated the isolates into four distinguishable groups according to Induction of grapevine defence systems using the species (cophenetic correlation = 0.97). Within D. seriata oomycete Pythium oligandrum against a pathogenic differentiation by region was evident. Furthermore, for fungus involved in Esca. A. YACOUB1,2,3, J. GERBORE4, this species there was variation in genotype between N. MAGNIN1,2, J. VALLANCE2,1, D. GRIZARD4, R. vineyards in several instances. Isolates of N. parvum GUYONEAUD3 and P. REY2,1,*. 1INRA, UMR1065 Santé showed greater homogeneity between vineyards and et Agroécologie du Vignoble (SAVE), ISVV, F-33140 Ville- regions, while B. dothidea showed homogeneity be- nave d’Ornon, France. 2Université de Bordeaux, Bordeaux tween regions from which they were isolated (Lower Sciences Agro, ISVV, UMR1065 SAVE, F-33140 Villenave and Upper Hunter Valley). The results obtained in this d’Ornon, France. 3UMR CNRS 5254/IPREM-EEM, IBEAS, study were effective in determining the genetic varia- Université de Pau et des Pays de l’Adour, 64013Pau, France. tion within the Botryosphaeriaceae species examined 4BIOVITIS, 15400 Saint Etienne de Chomeil, France. *E- and could be used to guide sampling in future surveys mail: [email protected] and population studies. The oomycete, Pythium oligandrum, has been reported to control several diseases and is able to induce plant Ocurrence of Phaeomoniella chlamydospora in Vitis defense systems. In order to control grapevine diseases, vinifera and V. labrusca vineyards of the Serra Gaú- we isolated P. oligandrum strains from the rhizosphere cha region of the Rio Grande do Sul state in southern of vines cultivated in 12 vineyards of the Bordeaux re- Brazil. M.A.K. ALMANÇA1,*, C. RUSIN1, G. GIOTTO1, gion. Pythium spp. with echinulated-oospores were fre- G.T. MARTINELI1, M.R. DE OLIVEIRA1 , F.R. CAVAL- quently isolated from the root system of the sampled CANTI2 and F. HALLEEN3. 1 Rio Grande do Sul Educa- vines, with P. oligandrum being the most frequently tion, Science and Technology Federal Institute, Osvaldo identified species (91% of strains, analyses based on ITS 574 Phytopathologia Mediterranea