Workshop - Best of Both Worlds_ Combine KG and Vector search for enhanced R...
28 new traits in advanced breeding populations-a. peil
1. New traits in advanced breeding
populations in apple and peach
WP2 – Establishment of prebreeding material
Results achieved since the beginning
of the project and plans for 2013
2. Reminder of the main objectives of the
WP
• Description of possible donors
• Introduction of new traits in to apple and peach
• Combination of traits
• Pyramiding of resistance traits
• Application and improvement of the fast breeding
approach
Main aim is to develop advanced
material with new traits for the breeders
3. Results achieved since the beginning
of the project
• Task 2.1: Description and evaluation of available plant material regarding
the traits powdery mildew, brown rot –peach, scab resistance (Rvi11,
Rvi12), fire blight resistance (Evereste, Mr5) material with pyramided
powdery mildew resistances (Pl1Pl2Pld, Pl1Pl2Plm)
Powdery Mildew
Brown Rot
4. Selection of parents to increase resistance/tolerance
to brown-rot
BO 92038173 (Contender x Elegant Lady)
Gialla tardiva
Percoca di Romagna 7
MA 25-13-143
(Suncrest X UCD-F8,5-156 )
IFF 638 (Bea x Aline)
New possible parents (also with putative resistance to PPV)
MA 25-10-111
((Romea*Snowqueen) X SD 75)
5. Results achieved since the beginning
of the project
• Task 2.2 Conventional pre-breeding
– crosses were performed to introduce new traits, combine
and or pyramid resistance genes, quality
Number of crosses Pollinated flowers Seeds* Seedlings*
Peach 44 1609
Apple 40 7498 7474 2803
* Not all seeds and seedlings from crosses 2012 are included
6. Traits used in apple and peach
Apple Peach
Introduction of new traits Introduction of new traits
Rvi11 Vbj brown rot resistance
Rvi12 Vb brown rot tolerance
FbE Combining and pyramiding of traits
FbR5 Rm1, Rm2 Green peach aphid
Low chilling Vr1, Vr2 Powdery mildew
Combining and pyramiding of traits maturity time
Rvi2, Rvi4, Rvi6 Vh2, Vh4, Vf Y yellow/white flesh
FbF7 D acid/subacid
Pl1, Pl2, Pld, Plm G peach/nectarine
Sd1 S flat/round
increase quality F melting/non melting
7. Examples for apple
Comparison of Rvi4 symptoms
with presence or absence of
marker CH02C02a
23%
29%20%
28%
Marker for
Rvi11
Marker for Sd1
Marker for
Rvi11 and Sd1
None of the
markers
Quality, Sd1 x Rvi11
8. Results achieved since the beginning
of the project
• Task 2.3:
– Application and improvement of the fast breeding
approach
• Introduction of fire blight resistance in to apple
– Evereste
– Malus xrobusta 5
• Establishment of early flowering lines
– With the transgene on another LG
– Different cultivars with the early flowering gene
– Use of a inducible promoter and a different
flowering gene
9.
10. JKI, Dresden, Germany
Overexpression of the
transcription factor
BpMADS4 from silver
birch (Betula pendula
Roth.) in transgenic line
of Pinova (T1190)
induced precocious
flowering (in vitro, and
also few months after
transfer to greenhouse)
Flachowsky et al., 2007
Plant Breeding
Silver birch Pinova_T1190
BpMADS4
The fast breeding approach
11. The idea behind the approach
Flachowsky H, Le Roux P-M, Peil A, Patocchi A, Richter K, Hanke M-
V. (2011) Applying a high-speed breeding technology to apple
(Malus × domestica) based on transgenic early flowering plants and
marker-assisted selection. New Phytologist 192, 364-377
Le Roux P-M, Flachowsky H, Hanke M-V, Gessler C, Patocchi A
(2011) Use of a transgenic early flowering approach in apple (Malus
× domestica Borkh.) to introgress fire blight resistance from
‘Evereste’. Molecular Breeding, in press.
12. Fire blight
Fotos: http://www.feuerbrand.it/info/infos/krankheitssymtome/bluetenbefall.html; http://www.hortipendium.de/Feuerbrand;
http://www.fohnsdorf.at/cms/umwelt/feuerbrand.asp; http://www.ogv-teisendorf.de/Media/Feuerbrand%20Triebspitze.jpg;
http://www.landwirtschaft.sg.ch/home/landwirtschaftliches/Beratung/Pflanzenbau/Pflanzenschutz/Feuerbrand_/Allgemeine_I
nformationen.html
14. Destroy all infected material
Foto: http://www.nzz.ch/aktuell/schweiz/null-toleranz-bei-feuerbrand-1.17954557
15. Fb_E
Major QTL identified in the
ornamental apple Evereste on
the bottom of Linkage Group
(LG) 12 (R2 = 50-70 %).
Durel et al., 2009 – Genome
Parravicini et al., 2011 – Molecular Plant
Pathology
General objective: selecting apple pre-breeding genotypes carrying major
genes/quantitative trait loci (QTL) for disease resistance from wild apples,
with as less as possible „wild“ genome.
Introgression of a highly efficacious fire blight (FB) resistance locus Fb_E from
Evereste.
Using T1190 at Agroscope ACW
Introduction of fire blight resistance from Evereste
16. X
X
X
X
X
EveresteT1190
F1
3 Fb_E + BpMADS4
Topaz, Diwa® and Maloni Sally ®
Royal Gala
GrannySmith
Braeburn2013
2010
2011
End 2008
beg. 2009
2012
BC‘1
9 Fb_E + BpMADS4
BC‘2
13 Fb_E + BpMADS4
BC‘3
? Fb_E + BpMADS4
BC‘4
? Fb_E without BpMADS4
2014
2018 first flowers of this pre-breeding material
2012/2013
111 seeds were
harvested and
stratified = 25 %
potential BC’3 plants
State of the art
17. We started with one early flowering Pinova line carrying the
transgene on LG 4
To enlarge the applicability of the approach three Pinova lines
carrying the BpMADS4 on different linkage groups (LG) than
LG 4 were created
17
Line Genotype Linkage group Integrated vector backbone
1189 PN 8 None
1236 PN 16 None
1240 PN 14
Ca. 1700 bp after BL (tests
outstanding)
Improvement of the fast breeding approach
18. Other cultivars carrying the early flowering gene
• Successful transformation with early flowering genes under control of 35S
promotor in different apple cultivars
• Generation of transgenic lines of different apple cultivars overexpressing
BpMADS4 under control of 35S
cv. ‘Pinova’:3, ‘Gala’: 5, ‘Santana’: 1, ‘Gala Mitchgla’: 1
• Transgenic lines flowered during in vitro cultivation
18
GA 35S BpMADS4 SAN 35S BpMADS4 PN 35S BpMADS4
vitro cultivation of transgenic apple lines.
Early flowering apple plants during in vitro
cultivation.
Improvement of the fast breeding approach
19. What else?
• Establishment of early
flowering lines using a new
gene (TFL1) under the
control of an inducible
promoter
Pinova with TFL1 under HSP
20. Publications
Flachowsky H, Le Roux P-M, Peil A, Patocchi A, Richter K, Hanke M-V. (2011)
Applying a high-speed breeding technology to apple (Malus × domestica) based
on transgenic early flowering plants and marker-assisted selection. New
Phytologist 192, 364-377
Le Roux P-M, Flachowsky H, Hanke M-V, Gessler C, Patocchi A (2011) Use of a
transgenic early flowering approach in apple (Malus × domestica Borkh.) to
introgress fire blight resistance from ‘Evereste’. Molecular Breeding, in press.
21. Main challenges for 2013
• Identification of new traits to be introduced in
to apple and peach
• Selection of parents for the next crosses
(phenotyping for brown rot)
• Development of early flowering lines with TFL
under the control of a inducible
HeatShockPromoter (HSP)
22. The future
What are you expecting from us?
New traits
New combinations
New genes
Acknowledgements: Melanie Jänsch, Markus Kellerhals, Lucie Leumann, Andrea
Patocchi, Alessandro Liverani, Thierry Pascal, Kathleen Barthel, Henryk
Flachowsky, ...