07/12/2017 1 NORDAPP meeting on fruit and berry breeding September 4-5, 2017, Dobele and Jelgava, Latvia The Danish Apple Collection characterization and utilization by Carsten Pedersen, with contribution from Bjarne Larsen, Maren Korsgaard, Marian Ørgaard, Mikael Agerlin Petersen and Torben Toldam-Andersen. University of Copenhagen, Denmark, Sean Myles and Kyle Gardner, Dalhausie University, Canada, The collection and some resistance characters The Apple-oasis project Genetic Diversity and Genome-Wide Association Mapping of Fruit Quality Traits a PhD study
07/12/2017 2 The Danish Apple Collection: Part of NordGen https://www.nordgen.org/nak/index.php?act=startsearch&letter=a About 300 cultivars of mainly Danish origin Characters recorded for: The tree: Shape Flowers: 6 Leaves and shoots: 6 Harvest and use: 4 Outer qualities of the fruit: Inner qualities of the fruit: Keeping qualities: 2
07/12/2017 3 Replica of the Danish Apple Collection at Gl. Estrup grown without spraying 212 of these have been evaluated from 2006 to 2009 for: Apple scab on fruits and leaves (Venturia inaequalis) Fruit tree canker (Neonectria ditissima) Powdery mildew (Podosphaera leucotricha) Visual field-evaluation on a scale from 1-9 on tree level Rosy apple aphid (Dysaphis plantaginea) Sooty blotch and flyspeck (SBFS) or apple summer disease
1 8 1 8 15 22 29 36 43 50 57 64 71 78 85 92 99 106 113 120 127 134 141 148 155 162 169 176 183 190 197 204 15 22 29 36 43 50 57 64 71 78 85 92 99 106 113 120 127 134 141 148 155 162 169 176 183 190 197 204 Disease Disease severity severity 07 07 06 06 05 05 04 04 03 03 02 02 01 00 Scab on fruits, Scab on average fruits 2006-9 and leaves Cultivars ordered according to to disease severity Augustæble Augustæble, gult Bedstefars Æble Blankholm Bodil Neergård Bodils Æble Bredæble Brostrøm Guldreinette Brændekilde Brøndæble Bøghs Citronæble C. J. Hansen 603 C.J.H. 6-18 Dansk Rosenhæger Dauglykkeæble Degneæble Diana Herschendsgave 07/12/2017 4 Hugonæble Hyltofte Æble Højbjerg Sommeræble Hørningsholm Ildrød Eilstrup Kronprins Frederik Kundbyæble Kyholm Reinette Ladbyæble/Fynsk udv I Langeland/Fynsk udv IV Lise Legind æble Lunds Doucin Lyngby Æble Maren Nis Marselisborg Sommeræb Mikkel Pedersæble Nonnetit Bastard Ondrup Moseæble Ormslevæble Oudrupgaards Høstæble Scab on fruits Drejæble Scab on leaves Edle Wendelborg Ejby æble Else Lindstow Pigeon fra Juellinge Ferskenrødt SommeræblePigeon fra Maribo Filippa 'Anka' Rifbjerg Gylling Flaskehalser Ringkloster Kammerjunk Flintinge Rødby Æble Fynsk udvalg V Sildig Sandholt Æble Fynsk udvalg VI Skarridsø Æble Glostrup Guldreinet Skovfoged Gravenfin Sorøæble Grænseæble Spiseæble Vejle Guldborg Søbjergæble Guldæble Sønderskov Hans Mathiesen Yduns æble Henrik Jensen Æble Aabyæble Herschendsgave Hans Wassard
Disease severity 07/12/2017 5 08 Mildew 07 06 05 04 2006-8 2006-9 03 02 01 00 Cultivars
07/12/2017 6 00 01 02 03 04 05 06 07 1 6 11 16 21 26 31 36 41 46 51 56 61 66 71 76 81 86 91 96 101 106 111 116 121 126 131 136 141 146 151 156 161 166 171 176 181 186 191 196 201 206 211 Disease severity Cultivars Canker 2006-8 2006-9
07/12/2017 7 General disease index: 2006-9 average of: Scab on leaves Scab on fruits Mildew Canker Hans Wassard 1.0 Bedstefars Æble 1.1 Brøndæble 1.1 Langeland/Fynsk udv IV 1.1 Lise Legind æble 1.1 Sildig Sandholt Æble 1.1 Antonius 1.1 Hans Mathiesen 1.1 Oudrupgaards Høstæble 1.1 Filippa 1.2 Dansk Rosenhæger 1.2 Flintinge 1.2 Guldæble 1.2 Hugonæble 1.2 Maren Nis 1.2 Skenkelsø Æble 1.2 Conclusion: A number of the cultivars are showing high resistances towards scab, mildew and canker. Augustæble 1.3 Brostrøm Guldreinette 1.3 Farum Æble 1.3 Fynsk udvalg VI 1.3 Kundbyæble 1.3 Ludvigsens Æble 1.3 Ringkloster Kammerjunker 1.3 Sukkeræble 1.3 Søde Æbler fra Aalsrode 1.3 Augustæble, gult 1.3 Aunhusæble 1.3 Bodils Æble 1.3 C. J. Hansen 603 1.3 Edle Wendelborg 1.3 Fynsk udvalg V 1.3 Lundbytorp 1.3 Lyngby Æble 1.3 Ormslevæble 1.3 Pigeon Rød Vinter 1.3 Sorøæble 1.3 Søbjergæble 1.3 Aabyæble 1.3 Kathrineæble 1.3 Rødhætte Æble 1.3
07/12/2017 8 The Apple-oasis project: Crowd breeding Seeds from ca. 50 selected cultivars distributed to apple-enthusiastic people in autumn 2013 The cultivars were selected to be robust and tasty The progenies are from random pollination (more than 1000 possible fathers) Totally about 10000 apple seeds were distributed and seedlings from many of these are now growing up. Indicate cultivars found to have very good disease resistance under unsprayed conditions at Gl. Estrup. Many of the others may have as well.
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07/12/2017 10 Breeding Potential in Danish Apple Cultivars: Genetic Diversity and Genome-Wide Association Mapping of Fruit Quality Traits A PHD STUDY BY BJARNE LARSEN SUPERVISIORS: TORBEN TOLDAM-ANDERSEN (POMETUM) MARIAN ØRGAARD (BOTANY) CARSTEN PEDERSEN (MOLECULAR PLANT BREEDING) SUPPORT: DEPARTMENT OF PLANT AND ENVIRONMENTAL SCIENCES OTHER PEOPLE INVOLVED: MIKAEL AGERLIN PETERSEN (FOOD SCIENCE) SEAN MYLES AND KYLE GARDNER (DALHAUSIE UNIVERSITY, CANADA)
07/12/2017 11 Materials: The Pometum collection: 287 Danish cultivars A set of reference cultivars primarily from other European countries: 86 The Assens collection: 76 A selection of M. sieversii, M. sylvestris and small-fruited, ornamental cultivars: 36 Methods: Extended with ca. 400 more cultivars! Genotyping with 15 SSR-markers (485 acc.) : Diversity, parentages, duplicates Genotyping S-RNase alleles Determine ploidy levels by flow cytometry Genotyping by sequencing (GBS) (380 acc.): ca. 30.000 SNP-markers : Diversity, parentages, duplicates Phenotypic traits for GWAS: - Apple-key traits - Sugars and acids - Apple juice aroma volatiles (YDUN-project) GWAS using TASSEL
Parent offspring relationships: 07/12/2017 12
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Duplicates: 07/12/2017 15
07/12/2017 16 Genotyping by sequencing (GBS): Relative cheap (380 cultivars for 90.000 dkr) Ca. 6 dkr/marker or 237 dkr/genotype (apple cultivar) Produces many SNP-markers Mostly random locations
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07/12/2017 18 Conclusions: High diversity in The Pometum Apple collection Parent-offspring relationships and duplicates can be determined by both SSR-markers and SNP-markers Ploidy-levels can also be determined by GBS (based on level of heterozygosity) High-throughput S-RNase genotyping protocol established. GWAS: major genes can easily and precisely be mapped GWAS: minor genes much more challenging requires many accessions and accurate phenotyping SNP-markers provided for MAS of important traits High-quality phenotyping is important for future GWAS Verifications of candidate genes is another future goal
07/12/2017 19 Suggestions for topics for Nordic collaborations: 1. Study alleles of some of the candidate genes in Nordic cultivars/breeding material: Harvest time would be interesting. The aim would be to validate the significance of the gene and develop a marker for MAS 2. Phenotype the SNP-genotyped 380 cultivars for canker and storage rot susceptibility in order to map involved genes and develop markers for MAS 3. Collect trees (grafting) from the Apple-oasis project consisting of half-sip families from (50) selected cultivars for pre-breeding e.g. with markers from 1 and 2.