presentation

Prebreeding for resistance – examples
from barley and wheat
Frank Ordon
www.jki.bund.de
Prebreeding
Effective
Screening methods
Identification of
donors of resistance
Genetics and marker
development
H. vulgare
Cultivars
Landraces
H. vulgare ssp.
spontaneum
H. bulbosum
Hordeum sp.
n=30
Institute for Resistance Research and Stress Tolerance
Marker based exploitation
Marker assisted selection
Marker assisted backcrossing
Pyramiding
etc.
Barley yellow mosaic virus disease
First discovery in Europe: 1978 Germany
1980 UK
Cultivar
BaYMV/BaMMVBaYMV/BaMMV- Yield
Reaction
t/ha
relative
Asorbia (6(6-rowed)
Corona (6(6-rowed)
Romanze (2(2-rowed)
Marinka (2(2-rowed)
resistant
susceptible
resistant
susceptible
5.33
3.53
4.20
2.38
100
65
100
57
Yuka (6(6-rowed)
Grete (6(6-rowed)
Duet (2(2-rowed)
Angora (2(2-rowed)
resistant
susceptible
resistant
suceptible
7.66
4.10
6.30
4.24
100
54
100
67
W. Huth
10 m2 plots, 3 replications; LSD (5%) = 0.25 t/ha and 0.35 t/ha; yield
yield in ATM
BaMMV, BAMMV-SIL, BaMMV-EIK, BaYMV, BaYMV-2
Institute for Resistance Research and Stress Tolerance
Economic losses caused by BaMMV/BaYMV
Acreage (2008): 1418200 ha
Yield: 6.61 t = 9374302 t
W. Huth
Barley price: 140 € pro t
Economic value: 1312402280 Euro
50% of barley acreage potentially infested (Huth 1988 ): 709100 ha
Moderate yield loss of 25%: 1171787 t
Economic loss: 164050180 €
Institute for Resistance Research and Stress Tolerance
Screening for resistance and genetic analysis
BaMMV BaYMV BaYMV-2 No.
Genotypes
Hordeum vulgare ssp. vulgare
201
+
86
+
53
+
+
43
Hordeum vulgare ssp. spontaneum
+
+
3
2
- = negative ELISA (resistant), + = positive ELISA ( susceptible)
Ordon et al. 1993, Habekuss et al. 2000
F2
Segregation
r : s
χ2**
7r:9s
H. vulgare ssp vulgare
Chikurin Ibaraki 1 x Ogra
Russia 57 x Diana
Bulgarian 347 x Diana
49 : 69
68 : 81
61 : 89
0.233
0.214
0.574
98
95
95
Bulgarian 347 x Russia 57
Taihoku A x Chikurin Ibaraki 1
Bulgarian 347 x Chikurin Ibaraki
42 : 73
50 : 68
53 : 63
2.442
0.091
0.0114
100
100
100
Cross
Institute for Resistance Research and Stress Tolerance
Infect.rate (%)
Götz & Friedt 1993, Ordon & Friedt 1993
Kartierung und Entwicklung molekularer Marker
Rym14HB
rym7
rym11
rym1
rym13
rym9
rym8
rym4
rym5
rym6
rym10
Rym16HB
rym15
rym3
Chikurin
Ibaraki 1
100cM
7H(1)
2H
3H
4H
Distanz
Marker
cM
name
BaYMV/BaYMV-2
Resistenz
Bmac0306
Bmac0163
0.0
Bmac0113
3.2
Ebmac0970
1.1
Bmag0394
5H
Institute for Resistance Research and Stress Tolerance
1H(5)
6H
5H(7)
mod. Graner et al. 2000, Ordon et al. 2005
Pyramiding of BaMMV/BaYMV Resistance Genes
BaYMV BaYMV-2 BaMMV BaMMV-SIL BaMMV-EIK
rym4
r
s
r
r
r
rym5
r
r
r
s
s
rym9
s
s
r
r
r
rym11
r
r
r
r
r
rym4 x rym9 x rym11
rym4 x rym9
rym4
rym4 x rym11 rym9 x rym11
rym5
Institute for Resistance Research and Stress Tolerance
rym5 x rym9 x rym11
rym5 x rym9
rym9
rym5 x rym11
rym11
Pyramiding of BaMMV/BaYMV Resistance Genes
1 Locus homozygous
Kreuzung
P
rym4
RYM9
rym4
RYM9
F1
F1´
X
RYM4
rym9
rym4
RYM11
RYM4
rym9
rym4
RYM11
rym4
rym9
RYM11
RYM4
RYM9
RYM11
rym4
RYM9
RYM4
RYM11
rym4
RYM11
RYM9
RYM4
rym11
RYM9
RYM4
RYM9
rym4
rym11
RYM9
rym9
rym11
RYM4
rym11
rym4
rym11
RYM9
RYM4
RYM11
RYM9
RYM9
RYM4
RYM11
RYM4
RYM11
RYM9
RYM11
RYM4
rym9
RYM11
RYM4
RYM9
RYM11
rym4
rym9
RYM11
rym4
RYM9
RYM11
rym4
RYM9
RYM11
rym4
RYM9
RYM11
rym4
RYM9
RYM11
rym4
RYM9
RYM9
RYM11
rym11
RYM4
RYM4
rym9
RYM9
RYM11
rym11
RYM4
RYM9
RYM11
rym4
RYM4
RYM9
rym11
RYM4
rym9
RYM9
RYM11
RYM9
RYM11
RYM4
rym9
RYM11
RYM4
RYM9
RYM11
rym4
rym9
RYM11
RYM4
RYM9
RYM11
RYM4
RYM9
RYM11
RYM4
RYM9
RYM11
RYM4
RYM9
RYM11
rym4
RYM9
RYM11
RYM4
rym9
RYM11
RYM4
RYM9
RYM11
rym4
rym9
RYM11
rym4
RYM9
RYM9
rym11
RYM9
rym11
rym4
Bmac 29 (rym4)
rym4
rym4
HVM 67 (rym9)
rym9 x rym4
X
rym9 x rym11
11
rym11 x rym4
X
rym11 x rym9
3
rym5 x rym9
X
rym5 x rym11
3
rym9 x rym5
X
rym9 x rym11
3
rym11 x rym5
X
rym11 x rym9
8
-
RYM9
HVM 03 (rym11)
14
6
rym11
rym4
rym11
5
1
rym9
rym4
RYM11
rym4 x rym9
X
rym4 x rym11
rym4
RYM4
RYM11
X
x
X
x
2 Loci heterozygous
3 Loci
heterozygous
8
-
rym11
DH Line
Production
A1
rym4
rym9
rym1
1
rym4
rym9
RYM
11
rym4
RYM
9
rym1
1
rym4
RYM
9
RYM
11
rym4
rym9
rym1
1
rym4
rym9
RYM
11
rym4
RYM
9
rym1
1
rym4
RYM
9
RYM
11
Marker Selection
Werner K, W Friedt, F Ordon, 2005. Mol. Breeding 16: 45-55
Institute for Resistance Research and Stress Tolerance
rym4, rym9 and
rym11 fixed
rym4 and
rym9 fixed
rym4 and
rym11 fixed
Pyramiding of BaMMV/BaYMV Resistance Genes
4r
4r
4r
9r
4r
9r
9r
5r
5r
11r
11r
9r
11r
11r
rym4 x rym9 x rym11
rym5 x rym9 x rym11
20 von 107 DHs, Chi2 = 0,072
27 von 187 DHs, Chi2 = 2,186
9r
9r
11
11
4r
4r
9
9
11r
11r
4/5
4/5
9r
9r
11r
11r
5r
5r
9
9
11r
11r
5r
5r
9r
9r
11
11
rym4 x rym9
rym4 x rym11
rym9 x rym11
rym5 x rym11
rym5 x rym9
31 von 107 DHs
17 von 107 DHs
42 von 294 DHs
21 von 187 DHs
31 von 187 DHs
Chi2 =7,489
Chi2 =0,086
Chi2 =0,033
Chi2 =1,642
Chi2 =0,629
Werner K, W Friedt, F Ordon, 2005. Mol. Breeding 16: 45-55
Institute for Resistance Research and Stress Tolerance
Gen Identification
Rym4/Rym5
High resolution mapping
A
P11M36 0.10
P21M68
B
MWG838
0.10
0.05
0.10
0.97
0.05
rym4
MWG838+
Y57C10+
E37*M36
E37*M63
E35*M81
E39*M39
STSE31*M41+
rym5
0.034
E62*M67
0.82
P20M46 0.05
B368N12
OP-Z04
0.34
0.88
OP-AF18H971
E45*M69
MWG010
BMac029
0.34
0.66
figure taken from:
Buchanan, Gruissem, Jones (2000)
Biochemistry & Molecular Biology of plants
OP-AF18H971+
E45*M69
MWG010+
BMac029*
Pellio et al. 2005
BAC-contig
Co-segregation
proximal
100 kb
Institute for Resistance Research and Stress Tolerance
eIF4E allele mining
1000 accessions selected
year1
resequencing
known haplotypes
40 novel exon haplotypes
resistance tests
13 susceptible
27 resistant haplotypes
year2/3
test crosses,
resistance tests
year3
Institute for Resistance Research and Stress Tolerance
non allelic genes
identification of
8 novel eIF4E alleles
A. Graner
Leaf rust (Puccinia hordei) in wheat
Use of Triticum monococcum
Lr37
2007
2005
Institute for Resistance Research and Stress Tolerance
Accession
7038
7041
36594
37351
37353
BGRC 42007
BGRC 042013
BGRC 042017
EK 2
PI 167526
PI 272560
PI 352473
PI 352475
PI 355538
PI 428174
TRI 00620
TRI 17633
TRI 17668
TRI 577/81
TRI 645/76
TRI 1988/76
TRI 1989/76
TRI 1996/75
TRI 2125/91
TRI 4304/74
TRI 00620
PTR
r
r
r
r
r
r
r
r
r
r
r
mr
r
r
mr
r
r
Pow dery Mildew
s
r
r
r
r
r
s
s
s
s
r
r
r
r
r
r
r
r
r
r
r
r
Leaf rust
r
r
s
s
s
r
r
r
r
r
s
r
mr
r
r
r
r
r
r
r
r
r
T.monococcum
(resistant)
T. monococcum (medium
susceptibility)
Wheat (susceptible)
Prehaustorial resistance
48 h post inoculation
96 h post inoculation
168 h post inoculation
10 d pst inoculation
Balken = 20 µm
A. Serfling
Institute for Resistance Research and Stress Tolerance
Preliminary results of QTL-analyses
Institute for Resistance Research and Stress Tolerance
Stem rust (Puccinia graminis)
IRAN 2008
http://image.space.rakuten.co.jp/lg01/88/0000061588/25/imgb9
01d9fazikbzj.jpeg
Institute for Resistance Research and Stress Tolerance
Analyses of Pgt races
Shewa 13 races (DPBTR, KHHTT, MLBDC, PTHSR, QLDGH, QMQTR,
RKHSR, RRTTR, TCHTT, THHST, TTHSR, TTKSR, TTTTR)
Arsi
8 races (KCCST, PTHSR, RMTTM, RRTTR, TTHSH, TTHSR, TTHTR,
TTKSR)
Bale
7 races (KCCST, MRHLR, PTHSR, RRTTR, TTHSR, TTKSR)
NWE
8 races (HRTSH, KCCST, KRHST, QQQCM, RRTTR, TMHSR, TTHSR,
TTKSR)
Total
22 races
. 2009
Admassu et al
Institute for Resistance Research and Stress Tolerance
Effectiveness of resistance genes
Differential host
Resistance
gene
Shewa
Arsi
NWEa
Bale
Total
No.
%
No.
%
No.
%
No.
%
No.
%
CnSSr9g
Sr9g
51
94.4
39
100
33
100
25
96.2
148
97.4
W2691Sr10
Sr10
53
98.1
39
100
31
93.9
25
96.2
148
97.4
ISr11-Ra
Sr11
39
72.2
37
94.9
32
97
22
84.6
130
85.5
W2691Sr13
Sr13
10
18.5
2
5.1
1
3
9
34.6
22
14.5
Combination VII
Sr17
48
88.9
39
100
33
100
25
96.2
145
95.4
CS_T_mono_deriv
Sr21
43
79.6
34
87.2
27
81.8
26
100
130
85.5
Bt Sr30 Wst
Sr30
21
38.9
19
48.7
17
51.5
9
34.6
66
43.4
W2691SrTt-2
Sr36
12
22.2
7
17.9
2
6.1
7
26.9
28
18.4
Triumph
SrTmp
18
33.3
9
23.1
5
15.2
4
15.4
36
23.7
McNair 701
SrMcN
54
100
39
100
33
100
26
100
152
100
Admassu et al.2009
Institute for Resistance Research and Stress Tolerance
Genetic Mapping of stem rust resistance gene Sr13
Sr13 – effective against Ug99 and other races
Crossing between the susceptible cultivar (Morocco) and resistant line
(Khapstein/9*LMPG )
158 F2 population developed and phenotyped using well characterized
Pgt isolate
10 – 15 progenies of each F2 plant re-tested
A
0
XSSR1
11
XSSR2
15
XSSR3
18
XSr13
24
XSSR4
37
XSSR5
Admassu et al.(in prep.)
Institute for Resistance Research and Stress Tolerance
Pre-breeding in the future
Puccinia triticina
Einkorn (schwach
anfällig)
Weizen (anfällig)
-Sequence information
-omics Technologies
Einkorn (resistent)
Identification of genes
and networks
G. Hammer et al. 2006: Trends in Plant
Science 11, 587-593
48 h nach Inokulation
96 h nach Inokulation
Detection of allelic diversity
168 h nach Inokulation 10 d nach Inokulation
mit Braunrost (Puccinia triticina)
Balken = 20 µm
Pre- breeding and breeding on
the allele level
Institute for Resistance Research and Stress Tolerance
Prof. Dr. Andreas Graner
Dr. Nils Stein
Prof. Dr. Dr. h.c. W. Friedt
Dr. Jens Weyen
Dr. Andrea Schiemann
Dr. Konstanze Scheurer
Dr. Bettina Pellio
Dr. Kay Werner
Dr. Dragan Perovic
Dr. Antje Habekuß
Dr. Albrecht Serfling
Dr. Volker Lind
Belayneh Admassu
DFG
GFP
BMBF
BMELV
KAAD
EU
Institute for Resistance Research and Stress Tolerance