marker-assisted selection and pyramiding of late blight ... · marker-assisted selection and...
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Marker-assisted selection and pyramiding of late blight resistance
genes in potatoJadwiga Śliwka, Marta Brylińska, Emil Stefańczyk,
Jarosław Plich, Sylwester Sobkowiak
Plant Breeding and Acclimatization Institute –National Research Institute, Młochów, Poland
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Genes for late blight resistance on potato genetic map
Śliwka & Zimnoch-Guzowska 2013
Rpi-mcq1
Rpi-edn2 R9
Rpi-dlc1
R8/Rpi-Smira2
I
VIIIVII
VIVIVIIIII
XIIXIXIX
R1
R3a R3b R5
R6 R7 R10
R11 Rpi-sto2
Rpi-pcs1
Rpi-Smira1
Rpi-vnt1.1
Rpi-vnt1.2
Rpi-vnt1.3
Rpi-nrs1
Rpi-phu1
R2, R2-like
R2-likeBzura
Rpi-blb3
Rpi-abpt
Rpi-bst1
Rpi-mcd1
Rpi-amr3i
Rpi-blb2
Rber,
Rpi-ber1
Rpi-ber2
Rpi-blb1/RB
Rpi-sto1
Rpi-pta1
Rpi-plt1
Rpi-ver1-8
Rpi1
Rpi-rzc1
Rpi-mch1
Rpi-avl1
Rpi-cap1
Rpi-qum1
Rpi-edn3
Rpi-pur1
Rpi-vnt2
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Virulence 2016
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Virulence 2017
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Goals
1. Marker-assisted selection for Rpi-rzc1 on different genetic backgrounds
2. Marker-assisted pyramiding:
Rpi-phu1+Sárpo Mira (Rpi-Smira1+Rpi-Smira2)
Rpi-phu1+Rpi-rzc1
3. Marker-assisted transfer of Rpi-rzc1 from 2x to 4x
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Solanum phureja
Solanum stenotomum
http://www.cgn.wageningen-ur.nl/pgr/collections/crops/potato/species.htm
Rpi-phu1
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• 1970 S. stenotomum x S. phureja comes from CIP, Lima, Peru
Crosses with S. tuberosum 2x
• 1992 first resistant tetraploid (unreduced gametes)
• 2006 mapping the gene (Śliwka et al. 2006, TAG)
• 2009 sequencing, virulent isolate P. infestans EC1; MP1162; = Rpi-vnt1.1
(Foster et al. 2009, MPMI)
• 2010 MAS (Śliwka et al. 2010, JAG)
• 2012 pyramiding with Sárpo Mira
• 2014 GMO field tests in The Sainsbury Laboratory [Jones et al.]
• 2018 Cultivar Gardena by Zamarte Potato Breeding Ltd Group IHAR
Rpi-phu1
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• Originates from:Bolivia, Peru
habitat: inter-Andean valleys, barren, erodedor cultivated soil
2400-3000 m above sea level
• Syn. S. sparsipilum, re-identified as • S. brevicaule
• Selected clones:- resistant to P. infestans
– low tendency to enzymatic blackeningand after-cooking darkening
Solanum ruiz-ceballosii Cárd. 2x (2EBN) VIR8664 (VIR7370)
Ochoa 1990
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rzc flower colourdH Balbina x 99-10/36 rzc
Śliwka et al. 2012 BMC Genet. 13 (1): 11
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New linkage map: 12-4, N=236
Brylińska et al. 2015
Flower colour locus F
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Sárpo MiraGenetic map,
chromosome XI
3 P. infestans isolates,Rpi-Smira1
R2 (%)cM
Tomczyńska et al. 2014
Rpi-Smira2 = R8 on chromosome IX
Vossen et al. 2016
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a
bcc
a a
b b
ab
cc
a
bb
c
– + – +
– – + +
24 21 31 7
resis
tance
MP324
resis
tance
MP324x
MP1498 MP1580
Rpi-Smira1 – + – +
Rpi-phu1 – – + +
n 24 21 31 7
± SD± SEmean
Tukey’s range test for unequal sample sizes with a significance level of 0.05
R8?
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R8 marker
Black’s differential
R1 R8 R9 99
-10
/36
05
-18
/25
Bzu
ra
Sárp
o M
ira
04
-IX
-21
2414 bp (65% of gene length)
05
-18
/3
dH
Bal
bin
a
Ro
bij
n
(–)
2000
1000
2000
1000
Selection for R8/Rpi-Smira2
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R8 marker
2414 bp (65% of gene length)
Alouette Carolus Gardena Kelly
2000
1000
R8 SM (–)
1 2 1 2 1 2 1 2
cultivars
Selection for R8/Rpi-Smira2
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a
bcc
a a
b b
ab
cc
a
bb
c
– + – +
– – + +
10 8 12 2
24 21 31 7
resis
tan
ce
MP324
resis
tance
MP324x
MP1498 MP1580
Rpi-Smira1 – + – +
Rpi-phu1 – – + +
R8 10 8 12 2
n 24 21 31 7
± SD± SEmean
Tukey’s range test for unequal sample sizes with a significance level of 0.05
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population parental lines n
G2P-1 DG 06-508 DG 08-238 109 Rpi-rzc1
G2P-2 DG 97-943 DG 99-10/36 159 Rpi-phu1+Rpi-rzc1
G2P-3 DG 97-2174 DG 99-10/36 120 Rpi-rzc1
Rpi-phu1
Rpi-rzc1
Diploid potato populationsNew more specific marker for Rpi-phu1Flanking markers for Rpi-rzc1 1812 and 1910
R gene
Rpi-phu1+Rpi-rzc1
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detached leaflet tests
pop/isolate MP324 MP324x MP1820
G2P-1 2016, 2018 2017
G2P-2 2016, 2018 2016, 2018 2017
G2P-3 2016, 2018 2017
at least one date (two repeats × three leaflets)
isolate/virulence Black’s differential Rpi-phu1 Rpi-rzc1
MP324 1.2.3.4.5.6.7.8.10.11 avirulent avirulent
MP324x 1.2.3.4.6.7.(10).11 virulent avirulent
MP1820 (13_A2) 1.2.3.4.5.6.7.8.10.11 avirulent avirulent
Rpi-phu1+Rpi-rzc1
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0 1 2 3 4 5 6 7 8 90
10
20
30
40
50
60
MP324
Rpi-rzc1 markern score range
1812 1910
– – 51 1.0 – 5.0
– + 2 9.0
+ + 56 7.0 – 9.0
ind
ivid
uals
resistance
G2P-1
Marker-assisted selection for Rpi-rzc1
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0 1 2 3 4 5 6 7 8 90
10
20
30
40
50
60
MP1820 (13_A2)
Rpi-rzc1 markern score range
1812 1910
– – 51 1.0 – 6.0
– + 2 9.0
+ + 56 6.7 – 9.0
ind
ivid
uals
resistance
G2P-1
Marker-assisted selection for Rpi-rzc1
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0 1 2 3 4 5 6 7 8 90
10
20
30
40
50
60
Rpi-rzc1 markern score range
1812 1910
– – 76 1.0 – 5.0
+ + 44 7.7 – 9.0
MP324in
div
idu
als
resistance
G2P-3
Marker-assisted selection for Rpi-rzc1
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0 1 2 3 4 5 6 7 8 90
5
10
15
20
25
30
35
40
45
Rpi-rzc1 markern score range
1812 1910
– – 76 1.0 – 4.5
+ + 44 6.7 – 9.0
MP1820 (13_A2)in
div
idu
als
resistance
G2P-3
Marker-assisted selection for Rpi-rzc1
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0 1 2 3 4 5 6 7 8 90
20
40
60
80
100
120
MP324
0 1 2 3 4 5 6 7 8 90
10
20
30
40
50
60
70
80
MP324x virulent to Rpi-phu1
0 1 2 3 4 5 6 7 8 90
20
40
60
80
100
MP1820 (13_A2)
ind
ivid
uals
resistance
G2P-2N = 159
Pyramiding: Rpi-phu1+Rpi-rzc1
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MP324
a
b b b
± SD± SEmean
MP324x
a a
b b
MP1820a
b b b
– – + + 1812 Rpi-rzc1 marker– – + + 1910
– + – + Rpi-phu1 marker
51 30 50 28 n
resis
tance
resis
tance
– – + +
– – + +
– + – +
51 30 50 28
Tukey’s range test for unequal sample sizes with a significance level of 0.05
G2P-2
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Summary
1. R8/Rpi-Smira2 is not contributing to resistance in ourdetached leaflet tests
2. R8/Rpi-Smira2 is not present in cv. Gardena3. Markers for Rpi-rzc1 1812 and 1910 work well on
different genetic backgrounds4. Rpi-phu1+Rpi-rzc1 pyramiding done on diploid level5. Transfer of the Rpi-rzc1 gene to tetraploid level: 18
resistant plants obtained in 2x-4x crosses will be crossed further
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Acknowledgements
Dr. Jarosław Plich Laboratory of Breeding MethodsZamarte Potato Breeding Ltd. Group IHAR
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21st Triennial Conference of the European Association for the Potato Research