Studies on cherry leaf spot at Dresden- Pillnitz
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1 Studies on cherry leaf spot at Dresden- Pillnitz Mirko Schuster Institute for Breeding Research on Fruit Crops
2 Cherry breeding Disease resistance Cherry leaf spot (Blumeriella jaapii) Leaf spot on sweet cherry Leaf spot on sour cherry
3 Leaf spot fungus, Blumeriella jaapii - Life cycle Pathogen: - Ascomycet, Blumeriella jaapii (Rehm) Arx (syn. Coccomyces hiemalis Higgins) Vegetative cycle (conidia) April Sept. H I C J A A - Myceli between mesophyll cells B Acervulus at the lower site of a leaf C - Macro-conidia Generative cycle (ascospores) April-June B D Premature apothecia G F F - Micro-conidiophore and trichogyne G - Trichogyne with micro-conidia H - Apothecia containing asci Nach Jakobsen at al., 1986) D I - Asci J - Ascospores
4 Objectives: Development of a artificial leaf spot test Description of rating scales Evaluation of cherry genotypes and cherry wild species resistant to leaf spot Studies about the inheritance of leaf spot resistance Use of resistant donors in cherry breeding
5 Leaf spot fungus, Blumeriella jaapii - Isolation Infected cherry leaf Acervuli with conidia (Hochapfel, 1952) Acervuli with conidia on the lower leaf surface
6 Leaf spot fungus, Blumeriella jaapii Isolation/Propagation Leaf spot fungus-isolates on potato dextrose agar Mycelia of B. jaapii
7 Leaf spot fungus Propagation / 1 st inoculation test Propagation of the fungus / conidia Inoculation method
8 Results of the 1 st inoculation test = Successful infection, but low infection rate on the leaves Use of conidia suspension from naturally infected leaves
9 Germination test of conidia - Dry stored infected leaves Conidia germinated after 3, 5, 7, 11 months and more
10 Inoculation test Inoculation suspension Collection of infected leaves in the orchard Drying of the leaves at room temperature Storage of the dry leaves in an exsiccator
11 Inoculation test Inoculation suspension Leaving of the dry infected leaves in a wet chamber for one day
12 Inoculation test Inoculation suspension Collection of the conidia from the lower surface of leaf
13 Inoculation test Inoculation suspension Counting of the concentration of conidia and diluting of the inoculation suspension
14 Inoculation test Preparation of Petri dishes for inoculation test
15 Inoculation test Inoculation Leaving of the leaves in the Petri dishes Moistening of the leaves with water Inoculation of the leaves with the suspension
16 Inoculation test Evaluation Evaluation of the reaction to leaf spot after days susceptible resistant genotypes
17 Inoculation test Test material Leaf spot infection on old leaves of sour cherry cv. Schattenmorelle
18 Inoculation test Test material Seedling 82 ( Almaz x o.p.) 1. leaf 2. leaf 3. leaf 4. leaf x 1,0 x 1,0 x 1,0 x 1,0 x 2,5 x 2,5 x 2,5 x 2,5 x 4,0 x 4,0 x 4,0 x 4,0 3. or 4. leaf for the inoculation test
19 Inoculation test Evaluation Clone 'DK1,7,3' Clone 'DK1,7,13' Resistant and susceptible genotypes
20 Inoculation test Evaluation sheet + scale
21 Inoculation test Rating scale Reaction type 1 no symptoms, green leaf 2 chlorotic or necrotic points or small pigmented lesions 3 larger lesions, partly with aerial mycelium and stunted sporulating lesions 4 2 to 10 sporulating lesions, acervuli 5 11 or more sporulating lesions, acervuli (modified after Wharton et al. 2003)
22 Cherry leaf spot Field evaluation F2 seedling populations: F2 population: 'Namati' x clone 18,167 'Sunburst' Prunus canescens clone 3,105 (P. cerasus x P. maackii) x 'Fanal' clone 3,106 (P. cerasus x P. maackii) x 'Fanal' 'Schattenmorelle' x clone 3,105 'Schattenmorelle' x clone 3,106 'Schattenmorelle' 'Gerema' 'Ujfehertoi fürtös' 'Coralin'
23 Cherry leaf spot Rating scale Evaluation (two rating scales): Reaction type scale 1 5 Distribution of infection scale 1 9 Reaction type on the leaf (R-type): 1 - no symptoms, green leaf 2 - chlorotic or necrotic points 3 - larger lesions, partly with aeial mycelium and stunted sporulating acervuli 4 - sporulating acervuli with chlorotic and necrotic lesions 5 - heavily sporulating acervuli Distribution of infection on the tree (R-distribution): 1 - no symptoms, green leaf 3 - chlorotic or necrotic points, scattered infected leaves 5 - infected leaves >50 % of the tree 7 - infected leaves, 50 % of the tree defoliated 9 - heavily sporulating acervuli on all leaves, >90 % of the tree defoliated
24 Cherry leaf spot Rating scale (R-distribution) Distribution of infection on the tree (R-distribution): 1 no symptoms, green leaf, no leaves drop down 3 chlorotic or necrotic points, scattered infected leaves, first leaves drop down 5 infected leaves % of the tree 7 infected leaves, 50 % of the tree defoliated 9 heavily sporulating acervuli on all leaves, >90 % of the tree defoliated
25 Cherry leaf spot Rating scale Evaluation (rating scale): Reaction type scale 1 5 Distribution of infection scale 1 9 Reaction type on the leaf (R-type): 1 - no symptoms, green leaf 2 - chlorotic or necrotic points 3 - larger lesions, partly with aeial mycelium and stunted sporulating acervuli 4 - sporulating acervuli with chlorotic and necrotic lesions 5 - heavily sporulating acervuli Distribution of infection on the tree (R-distribution): 1 - no symptoms, green leaf 3 - chlorotic or necrotic points, scattered infected leaves 5 - infected leaves % of the tree 7 - infected leaves, 50 % of the tree defoliated 9 - heavily sporulating acervuli on all leaves, >90 % of the tree defoliated Resistant (R) R-type 1-3 / R-distribution 1-5 Susceptible (S) R-type 4-5 / R-distribution 6-9
26 Problem? - Hypersensitive resistance reaction type Hypersensitive reaction / defoliation DK1,5,45 (clone 3,105 x Fanal) LS r-type - 2N! LS r-distrib. - 7
27 Evaluation of cherry genotypes and Prunus species as to resistance to cherry leaf spot Prunus species R-type P. serrulata 1 P. maackii 1 P. canescens 1 P. kurilensis 1 P. nipponica 1 P. insisa 1 P. subhirtella 1 P. sargentii 1 P. dawyckensis 4 Sour cherry R-type Almaz* 0 Ciganus 59 2 Csengödi 2 Hartai 1 Karneol 1 Korai Pipacsmeggy 1 Köröser Gierstädt 2 Morina 2 Paljus 2 Wishnja* 1 *interspecific hybrid Sweet cherry R-type Krupnoplodnaja 4 Lapins 3 Namati 4 Napoleon 3 Naprumi 4 Sunburst 4 Vinka 4 40 sour cherry - susceptible
28 Inheritance of cherry leaf spot resistance P. canescens Segregation analysis of leaf spot resistance : Combination resistant susceptible total χ 2 P % Namati' x clone '18,167' 41 39* 80* 0, * 7 susceptible seedlings dead from : 1 Segregation: resistant : susceptible 1 : 1 (41 : 39) one dominant resistance gene Detection of a QTL on LG 4 for leaf spot resistance from P. canescens (Cooperation with MSU East Lansing, A. Iezzoni) Travis Stegmeir, Mirko Schuster, Audrey Sebolt, Umesh Rosyara, George W. Sundin and Amy Iezzoni Cherry Leaf Spot Resistance in Cherry (Prunus) is Associated with a Quantitative Trait Locus on Linkage Group 4 Inherited from P. canescens. Molecular Breeding (in process)
29 Inheritance of cherry leaf spot resistance P. maackii Segregation analysis of leaf spot resistance : Cross combination resistant susceptible total χ 2 1 : 1 P % clone 3,105 x Fanal , clone 3,106 x Fanal , Schattenmorelle x clone 3, , Schattenmorelle x clone 3, , Segregation: resistant : susceptible 1 : 1 one dominant resistance gene
30 Use of resistant donors in cherry breeding Diploid (2x): P. canescens F1 x P. avium F2 x P.avium F3 x P. avium Tetraploid (4x): P. maackii P. padus P. serotiona P. spinosa F1 x P. cerasus F2 x P. cerasus F3 x P. cerasus
31 Thank you for your attention
32
33 Cherry leaf spot fungus, Blumeriella jaapii - symptoms At first on the upper surface of leaves small, circular, purple spots appear which eventually turn brown Infected leaves turn yellow, the area around lesions may remain green During wet weather, white to light pink spore masses appear on the lower surface in the centre of the spots Serious defoliation of trees can cause uneven ripening and poor fruit quality The tree vigour and winter-hardiness of the flower buds and the wood is reduced
34 Inoculation test Evaluation scale DK1,7,5 scale 1 no symptoms, green leaf Korall scale 1-2 chlorotic or necrotic points, small pigmented lesions DK1,7,16 scale 3 larger lesions, partly with aerial mycelium and stunted sporulating lesions DK1,7,10 DK1,7,13 scale 5 11 or more sporulating lesions
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