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Characteristics of the pre-breeding parent lines developedApple: Fire blight, apple scab and powdery mildew
Markus Kellerhals, Lucie Leumann, Simone Schütz, Isabelle Baumgartner, Agroscope WädenswilAndreas Peil, Julius Kühn Institute, Dresden-Pillnitz
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
Combination
Female parent
Traits Male parent
Traits seeds seedlings (labelled)
selected for container field
selection incontainer field
1112 ACW 16924
Rvi6, Rvi4*, Pl2, PlD
ACW 16102
Rvi6, Rvi2, Rvi4* Pl1
258 251 133 57
1113 ACW 16956
Rvi6, Rvi2, Pl2 ACW 16924
Rvi6, Rvi4*, Pl2, PlD
185 171 68 33
1114 ACW 17053
Rvi6, Pl2, FBF7 ACW 16102
Rvi6, Rvi2, Rvi4*, Pl1
330 303 89 55
total 773 725 290 1451207 ACW
18933Rvi12b? (Vb2?) Anna low chill 2 2 1 1
1208 ACW 18921
Rvi12b? (Vb2?) Cv good fruit size
fruit quality, size, FBF7
80 58 41 4
1209 ACW 18913
Rvi12b? (Vb2?) Cv good fruit size
fruit quality, size, FBF7
224 159 83 11
total 306 219 125 16
* = pin point pits in progenies observed
Agroscope Crosses 2012 (ctnd.)
Combination
Female parent
Traits Male parent Traits seeds seedlings (labelled)
selected for container field
selection incontainer field
1210 ACW 18933
Rvi12b?(Vb2?)
Cv good fruit size
Quality, size, FBF7
221 109 30 10
- ACW 18941
Rvi12b?(Vb2?)
Cv. Good fruit size
Quality, size, FBF7
398 (frozen) - - -
1211 ACW 18954
Rvi12b?(Vb2?)
Anna low chill 104 19 10 0
1212 ACW 18971
Rvi12b?(Vb2?)
Anna low chill 44 41 20 6
total 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 parentsTrait Resistance Molecular marker(s)Scab resistance Rvi6 (Vf) CHVf1 (SSR)Scab resistance Rvi2 (Vh2) CH02B10 (SSR), OPL19 (SCAR)Scab resistance Rvi4 (Vh4) CH02C02a (SSR)Mildew resistance Pl1 AT20 (SCAR)Mildew resistance Pl2 CH04H02 (SSR)Mildew resistance PlD CH03C02 (SCAR)Fire blight resistance QTL FBF7 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 seeds
1311 Cv low allergenic
low allergenic Berlepsch low allergenic 258
1312 Cv low allergenic
low allergenic Goldparmäne(King of the Pippins)
low allergenic 185
1313 Ladina litchi flavor ACW 20728 mango or passionfruit flavor
330
total 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/article-2298958/Geneticists-search-perfect-apple-hypoallergenic-variety-doesn-t-cause-allergies.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 hypo-allergenic 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 resistanceLinkage group Origin
Major genes
Pl1 12 Malus ×robusta
Pl2 11 Malus zumi
Plbj 10 Malus baccata jackiiPld 12 D12
Plm 11 Mildew immune selection (MIS)
Plw 8 White angel
QTLs 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 plants
without PM
Pl1 Pl2 Plm Pl1Pl2 Pl1Plm Pl2Plm Pl1Pl2Plm
GD 06005/9 50 17 2 7 4 4
GD 06005/21 50 23 6 6 8 3
GD 06005/26 50 17 1 1 14 1
GD 06005/55 50 20 1 2 14 3
GD 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 1 5 9 1 3
Results from genotyping (GD x Pl1Pl2Pld)
Mother Father Seeds planted
No. of plants
without PM
Pl1 Pl2 Pld Pl1Pl2 Pl1Pld Pl2Pld Pl1Pl2Pld
GD AA03 2,20 50 13 1 12
GD AA03 2,33 50 29 1 1 7 5 15
GD AA03 2,37 50 21 2 3 10 1 5
GD AA03 2,45 50 21 1 3 2 6 2 7
GD AA03 2,58 50 32 1 3 3 4 11 8
GD AA03 2,60 50 18 2 8 8
Additional crosses for pyramiding of powdery mildew resistances
Mother Father Resistances Resistancesseedlings
No.
Mother Father
Ariwa ACW8259 Rvi6, Pl1 Pl2 No seeds
Ariwa ACW13597 Rvi6, Pl1 Pld No seeds
Ariwa ACW8099 Rvi6, Pl1 Rvi6, Pl2, FBF7 Rvi6, Pl1, Pl2, FBF7Rvi6, Pl1, Pl2
87
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, PlmRvi2, Rvi4, FbF7, Sd1, Pl1, Pl2Rvi2, Rvi4, FbF7, Pl1, Pl2Rvi2, Rvi4, Sd1, Pl1, Pl2Rvi2, Rvi4, Pl1, Pl2Rvi2, Rvi4, FbF7, Sd1, Pl1, PlmRvi2, Rvi4, FbF7, Pl1, PlmRvi2, Rvi4, Sd1, Pl1, PlmRvi2, Rvi4, Pl1, Plm
112822134
Resistance to fire blight from Mr5 (ETH and JKI)
• Resistance gene of Mr5 has been isolated and function proved in transgenic GalaBroggini 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 Mr5using the fast breeding approach (JKI)
PinovaT11902011
2012 (F2) 12120
13103 2013 (F3) Fuji
Golden Del.
(Idared x Mr5)x
x
x
14100 (16 seedlings) 2014 (F4)
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.