BLE-Projekt
2810OE071 (LfL), 2810OE121 (JKI), 2810OE122 (IPK)
Assessment of genetic hotspots for Phytophthora
resistance and their use as molecular markers
in potato breeding
K. Sieber, G. Forster, A. Berger, A. Schwarzfischer und A. Kellermann
Development of Late Blight (Phytophthora infestans) Resistant
Potato Breeding Material for Organic Farming
Aims and objectives
Field Trial:
150 cultivars and breeding clones have been grown and
examined under organic farming conditions.
Potato Breeding:
We aim at combining late blight resistance with quality traits.
For the selection of breeding material we use a participative
approach on organic farms
Molecular Analyses:
Examination of resistance hotspots and evaluation of genetic
markers for resistance breeding
Fotos: JKI
Dr. Karen Sieber
2
Aims and objectives
Field Trial:
150 cultivars and breeding clones have been grown and
examined under organic farming conditions.
Potato Breeding:
We aim at combining late blight resistance with quality traits.
For the selection of breeding material we use a participative
approach on organic farms
Molecular Analyses:
Examination of resistance hotspots and evaluation of genetic
markers for resistance breeding
Fotos: JKI
Dr. Karen Sieber
3
Field trial
Varieties for field trials 2014
Total: 153 cultivars
•
•
•
•
•
•
Fotos: JKI
JKI (28)
LfL (26)
IPK (10)
German potato breeders (58)
others (26)
reference (5)
Dr. Karen Sieber
4
Field trial
GL
GL: Groß Lüsewitz
UE
UE: Uelzen
artificial infection
natural infection / organic trial field
pest control / maturity test
ND: Neuburg a.d. Donau)
FS: Freising
ND
LL
LL: Landsberg am Lech
Fotos: JKI
Dr. Karen Sieber
FS
5
Field trial
year
number of
varieties
environments
trials per
environment
individuals per plot
2012
117
3
(LL, ND, UE)
2
10
2013
149
1 (FS)+
3 (ND, LL, UE)
2
10
2014
153
1 (FS) +
3 (LL, ND, UE)
2
10
Parameter
Sprouting time
development
growth form
maturity
late blight
yield
tuber shape
tubers per
plant
starch content
tuber size
quality
frying quality
Fotos: JKI
Dr. Karen Sieber
6
Field trial
Trial field in Schrobenhausen 2012
Fotos: JKI
Dr. Karen Sieber
9
Field trial: Phytophthora Monitoring
UE
UE: Uelzen
EU_13_A2
other
ND: Neuburg a.d. Donau)
ND
LL: Landsberg am Lech
Fotos: JKI
LL
Dr. Karen Sieber
10
Field trial
Fotos: JKI
Δ rAUDPC
Δ rAUDPC
Corrected Late Blight Resistance and
Maturity for Black’s Differential Set
days to maturity
days
days to
to maturity
maturity
LL 2014
Walleshausen LK Landsberg
UE 2014
Natendorf LK Uelzen
Dr. Karen Sieber
11
Field trial
Late Blight Resitance and Maturity 2012-2014
0,2
GD 5%
∆rAUDPC
0,1
Hubal
0,0
80
90
100
110
120
130
140
Kuras
-0,1
Saka6
Axona
Toluca
-0,2
SarpoMira
Biogold
Bionica
Vitabella
-0,3
GD 5%
-0,4
JKI
Fotos: JKI
Population
TuebingerCirce
Carolus
maturity [days]
Anuschka
Princess
Lolita
Dr. Karen Sieber
Jelly
Ditta
12
Aims and objectives
Field Trial:
150 cultivars and breeding clones have been grown and
examined under organic farming conditions.
Potato Breeding:
We aim at combining late blight resistance with quality traits.
For the selection of breeding material we use a participative
approach on organic farms
Molecular Analyses:
Examination of resistance hotspots and evaluation of genetic
markers for resistance breeding
Fotos: JKI
Dr. Karen Sieber
13
Potato breeding
Seedling potatoes in Landsberg 2014
Fotos: JKI
Dr. Karen Sieber
14
Potato breeding
Participative selection of breeding clones in Landsberg 2014
Fotos: JKI
Dr. Karen Sieber
15
Potato breeding
Susceptible plants will be sorted out
Fotos: JKI
Dr. Karen Sieber
16
Potato breeding
Harvest of preselected clones in Landsberg n 2014
Fotos: JKI
Dr. Karen Sieber
17
Potato breeding
number of clones
Comparison of Late Blight Resistance Values
Resistance values (1=resistant, 9=susceptible)
breeding clones LL 2014
Fotos: JKI
Dr. Karen Sieber
trial set 2012-2014
18
Potato breeding: resistance vs yield
Fotos: JKI
Dr. Karen Sieber
19
Aims and objectives
Field Trial:
150 cultivars and breeding clones have been grown and
examined under organic farming conditions.
Potato Breeding:
We aim at combining late blight resistance with quality traits.
For the selection of breeding material we use a participative
approach on organic farms
Molecular Analyses:
Examination of resistance hotspots and evaluation of genetic
markers for potato breeding
Fotos: JKI
Dr. Karen Sieber
20
Molecular Analyses: Wild Resistance Donors
S. demissum
S. acaule
S. bulbocastanum
Fotos: JKI
S. pinnatisectum
S. phureja
S. stoloniferum
21
Molecular Analyses: Resistance Loci
STAos2
45/XI
Fotos: JKI
22
Molecular Analyses: Markers for S. demissum
IV
XI
• We applied regions from R1
and R3b as molecular
markers.
• Sequences are identical for
all investigated cultivars.
R1
• R1 appears in ca. 12% of all
trial samples
R3
Fotos: JKI
• R3b appears in ca. 35% of
all trial samples
Dr. Karen Sieber
S. demissum
Products: R1, R3a, R3b
References: Huang et al. 2005,
Rietman 2011, Lokossou et al.
2009; Kim et al. 2012
24
Molecular Analyses: Markers for S. bulbocastanum
VIII
IV
• Markers Blb3 and Abpt
were established for the
blb3 region
• Blb1-820 was used for the
blb1 region
blb3
• 9% of all samples were
positive at the blb3 locus
blb1
Fotos: JKI
• 1 sample was positive at
the blb1 locus.
Dr. Karen Sieber
S. bulbocastanum
Products:
− Blb3, Abpt,
− RB-629, Blb1-820, RB-226,
Sto1
References:
− Lokossou et al. 2009; Kim et
al. 2012;
− Fadina et al. 2009; Zhu et al.
2012
25
Molecular Analyses: Markers for S. bulbocastanum
Late Blight Resistance Values
0,20
0,10
GD 5%
∆rAUDPC
0,00
80
-0,10
90
100
110
120
GD 5%
130
07.1018.05 blb3
94.7082.15 blb3
140
96.7395.03 blb3
04.5214.04 blb3 + blb1
93.6984.07 blb3
04.5214.03 blb3
98.7826.02 abpt
-0,20
05.5049.10 blb3
97.7550.02 blb3
-0,30
Maturity [days]
-0,40
Anuschka
Fotos: JKI
Princess
Lolita
Dr. Karen Sieber
Jelly
Ditta
JKI
26
Molecular Analyses: Wild Resistance Donors
??
S. andigena
S. bulbocastanum
??
S. stoloniferum
Fotos: JKI
27
Molecular Analyses
Late Blight Resistance Values
0,15
n=9
0,10
n=56
n=38
n=9
ΔrAUDPC
0,05
n=14
0,00
ohne Marker
R1
R3b
R1 + R3b
blb3 / abpt
-0,05
-0,10
-0,15
Fotos: JKI
Dr. Karen Sieber
28
Perspectives
IX
• Continue breeding efforts and selection of
breeding clones
• Analyse of SNP results
• Find molecular markers for other Solanum
species that were used as resistance
donors
• Develop markers that allow to distinguish
between resistance genes and their
inactive homologues
• S. venturii
• S. phureja
Fotos: JKI
Dr. Karen Sieber
29
Thank You!
Andi Berger
Georg Forster
Andrea Schwarzfischer
Adi Kellermann
Fotos: JKI
Dr. Karen Sieber
30
Project Team
Fotos: JKI
Dr. Karen Sieber
31