WG1&2 COST Action FA 1303 Vienna, Austria, February 25-26, 2016 Influence of grapevine pruning systems on the diversity of wood-colonizing fungi Casdar 2013-2016 Ministry (Public) + Profession (Private) Project SCRI Proposal F (UMRSAVE-INRA PECH Rouge) + USA (USDA) K. Baumgartner, C. Chevrier, B. Diarra, D. Lawrence, P. Lecomte, H. Ojeda, D. Renault, P. Rey, R. Travadon, J. Vallance
COST Action FA 1303, Influence of grapevine pruning, Introduction Démarrage en Août 2014 Fin en 2016
COST Action FA 1303, Influence of grapevine pruning, Introduction English literature: Eutypa Dieback: Smith and Wishart 1955; Carter 1991; Gu et al 2005 Esca, Bot dieback: Mugnai 1999; Urbes-Torres 2011; Bertsch et al 2013 Many French reports: References Trunk disease Training system Pruning Rozier, 1781 (Abbot) Esca, X Dezeimeris, 1891 Esca, X Lafon, 1921 (Dubos, 2002) Arnaud et Arnaud, 1931 Esca X X Geoffrion, 1977 Esca X Bolay, 1979 Eutypa and Phomopsis Cordeau et al., 1984 Eutypa X X Boubals et Mur, 1990 Eutypa X Geoffrion et Renaudin, 2002 Esca X X Dumot, 2003; et al., 2012 Eutypa X Lecomte et al., 2007 2012 Esca X X Dal et al., 2008 Esca X X
COST Action FA 1303, Influence of grapevine pruning, Introduction COST Action FA 1303WG4, 2 nd meeting -October 8, 2015 Bordeaux Science Agro Management of GTDs in the vineyard -Cultural practices S. Di Marco 14:45 Zanutto Alessandro (or Marco Simonit), Preparatori d uva. Grapevine pruning respecting the sap flow and vine surgery: two agronomictechniques to reduce the damage caused by the trunk diseases 15:15 Francois Dal. Pruning, regreffage and curettage to reduce the risk or the effects of GTDs 15:30 Riccardo Castaldi: Possible relationship between vineyardsmechanization and Esca disease 16:00-16:30 Discussion 16:30-16:50 Coffee break We need more data to know more or to more convince!
COST Action FA 1303, Influence of grapevine pruning, Introduction Grapevine pruning systems influence the diversity of wood-colonizing fungi Invited Conference Bordeaux July, 2015 Kendra Baumgartner, Renaud Travadon, Daniel Lawrence USDA Agricultural Research Service, Davis, California Pascal Lecomte, Barka Diarra INRA, UMR 1065 SAVE, Université de Bordeaux, ISVV, CS 20032, 33882 Villenave d'ornon cedex, France Patrice Rey, David Renault, Jessica Vallance Université de Bordeaux, ISVV, UMR 1065 SAVE, Bordeaux Sciences Agro, CS 20032, 33882 Villenave d Ornon cedex, France Hernán Ojeda INRA, Unité expérimentale de Pech Rouge, 11430 Gruissan, France
COST Action FA 1303, Influence of grapevine pruning, Introduction Minimal pruning Spur pruning Fewer pruning wounds: Less wood necrosis? Fewer trunk pathogens? More pruning wounds: More wood necrosis? More trunk pathogens?
COST Action FA 1303, Influence of grapevine pruning, Material & Methods Location: Pech-Rouge (South East of France) Syrah (MS) et Mourvèdre (R) /140R ; 14 years old Minimal pruning since the year 4 and spur pruning since the year of planting Esca symptoms (wood and foliage) were recorded in 2013 Only non-leaf and non wood symptomatic vines were collected in December 2013 8 vines per treatment ; vines were cut lengthwise Mid-trunks were photographed for image analysis (proportion of wood necrosis) 4 wood samples were collected per vine for isolation and ITS-DNA sequencing, of for wood DNA extraction allowing SSCP profiles Results of isolation (number of wood chips that yielded a taxon) were analysed using different tools : some measures of relative abundance, and Canonical Correspondence Analysis (CCA) to identify specific pathogens and to examine the effects of cultivar, pruning system, trunk location, wood type
COST Action FA 1303, Influence of grapevine pruning, Material & Methods Necrosis was quantified by converting the image to black and white, and then calculating the percentage of trunk area occupied by black pixels as % necrosis.
COST Action FA 1303, Influence of grapevine pruning, Material & Methods Trunk location Top Close to canopy & more pruning wounds. Base Close to roots & soilborne spores of Esca pathogens, root pathogens. Close to graft union. Heartwood Wood type Sapwood Young xylem Typically not discolored Old xylem Always discolored Sometimes necrotic
COST Action FA 1303, Influence of grapevine pruning, Material & Methods 2 pruning systems 2 cultivars 2 trunk locations 2 wood types 8 replicate vines 128 wood samples 2.048 wood chips
COST Action FA 1303, Influence of grapevine pruning, Material & Methods We recorded fungi isolated from each wood chip and thus were able to use this as a quantitative measure of colonization on a per sample basis. Subcultured fungi were initially grouped according to similar colony and spore morphology. Representative cultures of each group were sequenced and those representing the unique operational taxonomic units were referred to as taxa.
COST Action FA 1303, Influence of grapevine pruning, Results ANOVA. Each column is the mean of six vines. Error bars are 95% confidence limits. Means with overlapping error bars are not significantly different (Tukey s test, α=0.05, P 0.05).
COST Action FA 1303, Influence of grapevine pruning, Results Esca incidence (% vines with wood and/or foliar symptoms) 46% 16% 32% 12%
COST Action FA 1303, Influence of grapevine pruning, Results Community diversity : 88 taxa - 85 Ascomycetes, 3 Basidiomycetes Ascomycete taxa per order
COST Action FA 1303, Influence of grapevine pruning, Results 15 Most Frequently Isolated Taxa Trunk pathogens Wood-colonizing endophytes Botryosphaeria dieback Alternaria tenuissima Diplodia seriata Aureobasidium pullulans Neofusicoccum parvum Bionectria ochroleuca Esca Paraconiothyrium brasiliense Phaeomoniella chlamydospora Paraconiothyrium variabile Togninia minima Pestalotiopsis microspora Phomopsis dieback Diaporthe ampelina Unknown ecology Diaporthe foeniculina Phaeosphaeria sp. 1 Phaeosphaeria sp. 2 Plant pathogen Fusarium brachygibbosum
q D COST Action FA 1303, Influence of grapevine pruning, Results Community diversity Measures of abundance-based species diversity (e.g., species richness) converted to effective number of species (Hill numbers) q reflects sensitivity to rare species 0 D species abundance not accounted for equivalent to species richness 1 D all species weighted by their frequencies equivalent to exponential of Shannon entropy 2 D abundant species favored equivalent to inverse of Simpson concentration Chao et al., 2014. Unifying Species Diversity, Phylogenetic Diversity, Functional Diversity, and Related Similarity and Differentiation Measures Through Hill Numbers. Annual Review of Ecology, Evolution, and Systematics 45: 297-324.
COST Action FA 1303, Influence of grapevine pruning, Results Comparing diversity between cultivars. Mourvedre - 63 taxa Syrah - 62 taxa 37 shared More taxa estimated from Mourvedre more singletons Similar diversity measures when taxa weighted by frequencies (q = 1) and when abundant taxa favored (q = 2). Singletons mean taxa recovered from only one wood piece
COST Action FA 1303, Influence of grapevine pruning, Results Comparing diversity between pruning systems Spur pruning - 66 taxa Minimal pruning - 58 taxa 36 shared Diversity measures higher in spur-pruned vines than in min-pruned vines
COST Action FA 1303, Influence of grapevine pruning, Results Canonical correspondence analysis
COST Action FA 1303, Influence of grapevine pruning, Results SSCP Profiles showed: Interaction cultivar/training system Syrah spur-pruned vines Interaction wood type/training system Heartwood of spur-pruned vines
Spur pruned vines Trunk pathogens Wood-colonizing endophytes Botryosphaeria dieback Alternaria tenuissima Diplodia seriata Aureobasidium pullulans Neofusicoccum parvum Bionectria ochroleuca Esca Paraconiothyrium brasiliense Phaeomoniella chlamydospora Paraconiothyrium variabile Togninia minima Pestalotiopsis microspora Phomopsis dieback Diaporthe ampelina Unknown ecology Diaporthe foeniculina Phaeosphaeria sp. 1 Phaeosphaeria sp. 2 Plant pathogen Fusarium brachygibbosum
Minimal pruned vines Trunk pathogens Wood-colonizing endophytes Botryosphaeria dieback Alternaria tenuissima Diplodia seriata Aureobasidium pullulans Neofusicoccum parvum Bionectria ochroleuca Esca Paraconiothyrium brasiliense Phaeomoniella chlamydospora Paraconiothyrium variabile Togninia minima Pestalotiopsis microspora Phomopsis dieback Diaporthe ampelina Unknown ecology Diaporthe foeniculina Phaeosphaeria sp. 1 Phaeosphaeria sp. 2 Plant pathogen Fusarium brachygibbosum
COST Action FA 1303, Influence of grapevine pruning, Conclusions Spur-pruned vines had more necrosis and higher disease incidence. Pruning system was more important than cultivar, in terms of fungal diversity. Spur-pruned vines had greater fungal diversity. Pruning system, cultivar and wood type were significant, in terms of composition of the fungal community. Spur-pruned vines had different, but not much more, trunk pathogens. A paper was submitted in Fungal Ecology (Travadon et al., )
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