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
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
Virulence 2016
Virulence 2017
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
Solanum phureja
Solanum stenotomum
http://www.cgn.wageningen-ur.nl/pgr/collections/crops/potato/species.htm
Rpi-phu1
• 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
• 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
rzc flower colourdH Balbina x 99-10/36 rzc
Śliwka et al. 2012 BMC Genet. 13 (1): 11
New linkage map: 12-4, N=236
Brylińska et al. 2015
Flower colour locus F
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
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?
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
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
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
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
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
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
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
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
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
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
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
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
Acknowledgements
Dr. Jarosław Plich Laboratory of Breeding MethodsZamarte Potato Breeding Ltd. Group IHAR
21st Triennial Conference of the European Association for the Potato Research