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

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

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

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)

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

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

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

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

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

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.

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_Informationen.html

Fire blight solutions

Foto: http://www.stadt-lienz.at/system/web/zusatzseite.aspx detailonr=218784046

Foto: http://tirol.orf.at/news/stories/2543474/

Foto: http://www.nzz.ch/aktuell/schweiz/null-toleranz-bei-feuerbrand-1.17954557

Foto: http://www.gartentipps.com/feuerbrand-melden-es-besteht-meldepflicht.html

Destroy all infected material

Foto: http://www.nzz.ch/aktuell/schweiz/null-toleranz-bei-feuerbrand-1.17954557

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

X

X

X

X

X

Evereste T1190

F1 3 Fb_E + BpMADS4

Topaz, Diwa® and Maloni Sally ®

Royal Gala

GrannySmith

Braeburn 2013

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

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

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

What else?

• Establishment of early flowering lines using a new gene (TFL1) under the control of an inducible promoter

Pinova with TFL1 under HSP

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.

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)

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, ...