1. 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

2. 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

3. 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

4. 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

5. 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)

6. Methods: Phenotyping
 artificial scab test on common scab races in
greenhouse
 growth character in container field
 powdery mildew resistance in container field

7. 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

8. 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

9. 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

10. New possible traits for crosses 2014
• Self-thinning
• Low carb (low fructose, low sorbitol)

11. Apple genotypes described by Agroscope
Traits need to be
confirmed with new
know-how on the
markers and false
positive signals

12. 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

13. 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.

14. 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, …

15. 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)

16. 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

17. 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

18. 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

19. 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

20. 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

21. 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

22. 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.