Published on February 28, 2014
Characteristics of the pre-breeding parent lines developed Apple: Fire blight, apple scab and powdery mildew Markus Kellerhals, Lucie Leumann, Simone Schütz, Isabelle Baumgartner, Agroscope Wädenswil Andreas Peil, Julius Kühn Institute, Dresden-Pillnitz YOUR LOGO
Objective: innovation Establishment of pre-breeding material Vb (Rvi12) scab resistance (from Hansen’s baccata 2): two different genes involved: Rvi12a and Rvi12b Pyramiding of resistances against scab, powdery mildew and fire blight Low chilling requirement Low allergenic Rich flavour
Agroscope Crosses 2011 and 2012 Combina tion 1112 1113 1114 total 1207 1208 1209 Female parent ACW 16924 ACW 16956 ACW 17053 ACW 18933 ACW 18921 ACW 18913 Traits Male parent Rvi6, Rvi4*, ACW Pl2, PlD 16102 Rvi6, Rvi2, Pl2 ACW 16924 Rvi6, Pl2, FBF7 ACW 16102 Traits seeds Rvi6, Rvi2, Rvi4* Pl1 Rvi6, Rvi4*, Pl2, PlD Rvi6, Rvi2, Rvi4*, Pl1 seedlings (labelled) selected for container field selection in container field low chill Rvi12b? (Vb2?) Cv good fruit size Rvi12b? (Vb2?) Cv good fruit size fruit quality, size, FBF7 fruit quality, size, FBF7 total * = pin point pits in progenies observed 251 133 57 185 171 68 33 330 303 89 55 773 2 725 2 290 1 145 1 80 58 41 4 224 159 83 11 306 Rvi12b? (Vb2?) Anna 258 219 125 16
Agroscope Crosses 2012 (ctnd.) Combina tion 1210 1211 1212 total Female parent ACW 18933 ACW 18941 ACW 18954 ACW 18971 Traits Male parent Traits Rvi12b? (Vb2?) Rvi12b? (Vb2?) Rvi12b? (Vb2?) Rvi12b? (Vb2?) Cv good fruit size Cv. Good fruit size Anna Quality, size, FBF7 Quality, size, FBF7 low chill Anna low chill seeds seedlings (labelled) selected for container field selection in container field 221 109 30 10 398 (frozen) 104 - - - 19 10 0 44 41 20 6 369 169 60 16
Methods: Genotyping Seedlings of crosses 1112, 1113 and 1114 were analysed by Agroscope (company www.ecogenics.ch) with molecular markers for scab, mildew and fire blight resistance present in their parents Trait Scab resistance Scab resistance Scab resistance Mildew resistance Mildew resistance Mildew resistance Fire blight resistance QTL Resistance Rvi6 (Vf) Rvi2 (Vh2) Rvi4 (Vh4) Pl1 Pl2 PlD FBF7 Molecular marker(s) CHVf1 (SSR) CH02B10 (SSR), OPL19 (SCAR) CH02C02a (SSR) AT20 (SCAR) CH04H02 (SSR) CH03C02 (SCAR) AE10_375 (SCAR), GE-8019 (SCAR)
Methods: Phenotyping artificial scab test on common scab races in greenhouse growth character in container field powdery mildew resistance in container field
Results: crosses 1112, 1113 and 1114 145 plants (of 725 labelled seedlings) with at least two scab resistance markers (Rvi6 and/or Rvi2 and/or Rvi4) and at least one for powdery mildew resistance (Pl1 or Pl2 and/or PlD) 55 of them carry also FBF7 QTL for enhanced fire blight resistance phenotypic scab resistance in artificial scab test powdery mildew resistance and good growth character evaluated in container field
Results: crosses 1207, 1208, 1209, 1210, 1211 and 1212 Mother trees: derived from a cross between Gala x Hansens baccata 2 displays no scab symptoms in the field were selected negative with the Markers Hi02d05 and CH02h11b for Rvi12 (Vb) are expected to carry an other scab resistance (here called Vb2) → so far no molecular markers available 32 plants of 388 labelled seedlings selected phenotypic scab resistance in artificial scab test powdery mildew resistance and good growth character selected in container field
Crosses 2013 Combination Female parent Traits Male parent Traits 1311 Cv low allergenic Cv low allergenic Ladina low allergenic Berlepsch low allergenic 258 low allergenic Goldparmäne low allergenic (King of the Pippins) ACW 20728 mango or passionfruit flavor 185 1312 1313 total litchi flavor seeds 330 773
New possible traits for crosses 2014 • Self-thinning • Low carb (low fructose, low sorbitol)
Apple genotypes described by Agroscope Traits need to be confirmed with new know-how on the markers and false positive signals
Low allergenic apples ‘Some people who are allergic may simply say they don’t like apples, since they’ve a very mild reaction after eating them,’ Eric van de Weg, plant scientist at Wageningen University in the Netherlands told Fresh Plaza. Read more: http://www.dailymail.co.uk/sciencetech/article2298958/Geneticists-search-perfect-applehypoallergenic-variety-doesn-t-causeallergies.html#ixzz2rxZTbvCn
Different approaches towards low allergenic apples Dr. Alessandro Botton, a plant physiologist at the Department of Agronomy, Food, Natural Resources, Animals and Environment in the University of Padua worked as a plant physiologist on the ISAFRUIT project to unravel the apple's genetic markers. Botton is currently working on the AGER Project to identify naturally occurring hypoallergenic proteins in order to find a way to breed different apple varieties into a hypo-allergenic variety of apples.
Resistance to powdery mildew Overview on powdery mildew resistance Linkage group Origin Major genes Pl1 Malus ×robusta Pl2 11 Malus zumi Plbj 10 Malus baccata jackii Pld 12 D12 Plm 11 Mildew immune selection (MIS) Plw QTLs 12 8 White angel 1, 2, 3, 5, 7, 8, 9, 10, 13, 14, 16, 17 Malus hupehensis, Malus sargentii, Dülmener Rosenapfel, Discovery, TN10-8, …
Pyramiding of powdery mildew resistances • available material, lines with – Pl1Pl2Plm (Pl2 and Plm both on LG11, recombination required) – Pl1Pl2Pld (Pl1 and Pld both on LG12, recombination required) • very small fruit size, bad quality, described in D2.1 • improvement of size quality (crosses with GD)
Selection of seedlings with pyramided powdery mildew resistances Phenotyping of seedlings in the glasshouse Genotyping of resistant seedlings marker Pl1 AT20SCAR Pl2 CH04h02, N18, U2 Pld CH03c02 Plm N18, AC20, U2
Results from genotyping (GD x Pl1Pl2Plm) Mother Father Seeds planted No. of Pl1 Pl2 Plm Pl1Pl2 Pl1Plm Pl2Plm Pl1Pl2Plm plants without PM GD 06005/9 50 17 GD 06005/21 50 23 GD 06005/26 50 GD 06005/55 GD 2 7 4 4 6 6 8 3 17 1 1 14 1 50 20 1 2 14 3 06006/7 50 28 1 1 4 16 6 GD 06006/8 50 21 3 1 2 14 1 GD 06006/32 50 20 8 2 3 7 GD 06006/57 50 21 1 1 5 9 1 1 3
Results from genotyping (GD x Pl1Pl2Pld) Mother Father Seeds planted No. of Pl1 Pl2 Pld Pl1Pl2 Pl1Pld plants without PM GD AA03 2,20 50 13 GD AA03 2,33 50 29 GD AA03 2,37 50 21 GD AA03 2,45 50 21 GD AA03 2,58 50 32 GD AA03 2,60 50 18 1 12 5 7 Pl1Pl2Pld 1 1 Pl2Pld 15 2 3 10 1 5 1 3 2 6 2 7 1 3 3 4 11 8 8 8 2
Additional crosses for pyramiding of powdery mildew resistances Mother Father Resistances Mother Resistances seedlings Father No. Ariwa ACW8259 Rvi6, Pl1 Pl2 No seeds Ariwa ACW13597 Rvi6, Pl1 Pld No seeds Ariwa ACW8099 Rvi6, Pl1 Rvi6, Pl2, FBF7 Rvi6, Pl1, Pl2, FBF7 Rvi6, Pl1, Pl2 8 7 Ariwa ACW8259 Rvi6, Pl1 Pl2 Rvi6, Pl1, Pl2 8 06006/7 E6 33,124 Rvi2, Rvi4, FbF7, Sd1 Pl1, Pl2, Plm Rvi2, Rvi4, Pl1, Pl2, Plm Rvi2, Rvi4, FbF7, Sd1, Pl1, Pl2 Rvi2, Rvi4, FbF7, Pl1, Pl2 Rvi2, Rvi4, Sd1, Pl1, Pl2 Rvi2, Rvi4, Pl1, Pl2 Rvi2, Rvi4, FbF7, Sd1, Pl1, Plm Rvi2, Rvi4, FbF7, Pl1, Plm Rvi2, Rvi4, Sd1, Pl1, Plm Rvi2, Rvi4, Pl1, Plm 1 1 2 8 2 2 1 3 4
Resistance to fire blight from Mr5 (ETH and JKI) • Resistance gene of Mr5 has been isolated and function proved in transgenic Gala Broggini et al. (2014) Engineering fire blight resistance into the apple cultivar 'Gala' using the FB_MR5 CC-NBS-LRR resistance gene of Malus × robusta 5. Plant Biotechnology Journal accepted Transgenic Gala lines
Introduction of fire blight resistance from Mr5 using the fast breeding approach (JKI) 2011 PinovaT1190 x 2012 (F2) 2013 (F3) 2014 (F4) 12120 (Idared x Mr5) x 13103 Golden Del. x Fuji 14100 (16 seedlings) MAS: with markers surrounding the fire blight resistance gene FB_MR5
Conclusions Pyramiding a broad range of disease resistance genes and factors is supposed to be promising in respect of durability of the resistance. To integrate scab, mildew and fire blight resistances in the same genotype is an added value provided a high fruit quality and productivity can be achieved. Further breeding objectives such as low chilling, low allergenic, high flavor and eventually self-thinning and health/nutritional aspects were or will be considered.
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