12 apple characteristics of prebreeding material kellerhals markus

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Characteristics of the pre-breeding parent lines developedApple: Fire blight, apple scab and powdery mildew

Markus Kellerhals, Lucie Leumann, Simone Schütz, Isabelle Baumgartner, Agroscope WädenswilAndreas Peil, Julius Kühn Institute, Dresden-Pillnitz

Objective: innovation

Establishment of pre-breeding material Vb (Rvi12) scab resistance (from Hansen’s

baccata 2): two different genes involved: Rvi12a and Rvi12b

Pyramiding of resistances against scab, powdery mildew and fire blight

Low chilling requirement Low allergenic Rich flavour

Agroscope Crosses 2011 and 2012

Combination

Female parent

Traits Male parent

Traits seeds seedlings (labelled)

selected for container field

selection incontainer field

1112 ACW 16924

Rvi6, Rvi4*, Pl2, PlD

ACW 16102

Rvi6, Rvi2, Rvi4* Pl1

258 251 133 57

1113 ACW 16956

Rvi6, Rvi2, Pl2 ACW 16924

Rvi6, Rvi4*, Pl2, PlD

185 171 68 33

1114 ACW 17053

Rvi6, Pl2, FBF7 ACW 16102

Rvi6, Rvi2, Rvi4*, Pl1

330 303 89 55

total 773 725 290 1451207 ACW

18933Rvi12b? (Vb2?) Anna low chill 2 2 1 1

1208 ACW 18921

Rvi12b? (Vb2?) Cv good fruit size

fruit quality, size, FBF7

80 58 41 4

1209 ACW 18913

Rvi12b? (Vb2?) Cv good fruit size

fruit quality, size, FBF7

224 159 83 11

total 306 219 125 16

* = pin point pits in progenies observed

Agroscope Crosses 2012 (ctnd.)

Combination

Female parent

Traits Male parent Traits seeds seedlings (labelled)

selected for container field

selection incontainer field

1210 ACW 18933

Rvi12b?(Vb2?)

Cv good fruit size

Quality, size, FBF7

221 109 30 10

- ACW 18941

Rvi12b?(Vb2?)

Cv. Good fruit size

Quality, size, FBF7

398 (frozen) - - -

1211 ACW 18954

Rvi12b?(Vb2?)

Anna low chill 104 19 10 0

1212 ACW 18971

Rvi12b?(Vb2?)

Anna low chill 44 41 20 6

total 369 169 60 16

Methods: Genotyping

Seedlings of crosses 1112, 1113 and 1114 were analysed by Agroscope (company www.ecogenics.ch) with molecular markers for scab, mildew and fire blight resistance present in their parentsTrait Resistance Molecular marker(s)Scab resistance Rvi6 (Vf) CHVf1 (SSR)Scab resistance Rvi2 (Vh2) CH02B10 (SSR), OPL19 (SCAR)Scab resistance Rvi4 (Vh4) CH02C02a (SSR)Mildew resistance Pl1 AT20 (SCAR)Mildew resistance Pl2 CH04H02 (SSR)Mildew resistance PlD CH03C02 (SCAR)Fire blight resistance QTL FBF7 AE10_375 (SCAR), GE-8019 (SCAR)

Methods: Phenotyping

artificial scab test on common scab races in greenhouse

growth character in container field powdery mildew resistance in container field

Results: crosses 1112, 1113 and 1114

145 plants (of 725 labelled seedlings) with at least two scab resistance markers (Rvi6 and/or Rvi2 and/or Rvi4) and at least one for powdery mildew resistance (Pl1 or Pl2 and/or PlD)

55 of them carry also FBF7 QTL for enhanced fire blight resistance

phenotypic scab resistance in artificial scab test powdery mildew resistance and good growth

character evaluated in container field

Results: crosses 1207, 1208, 1209, 1210, 1211 and 1212

Mother trees: derived from a cross between Gala x Hansens baccata 2 displays no scab symptoms in the field were selected negative with the Markers Hi02d05 and

CH02h11b for Rvi12 (Vb) are expected to carry an other scab resistance (here

called Vb2) → so far no molecular markers available 32 plants of 388 labelled seedlings selected phenotypic scab resistance in artificial scab test powdery mildew resistance and good growth character

selected in container field

Crosses 2013

Combination Female parent

Traits Male parent Traits seeds

1311 Cv low allergenic

low allergenic Berlepsch low allergenic 258

1312 Cv low allergenic

low allergenic Goldparmäne(King of the Pippins)

low allergenic 185

1313 Ladina litchi flavor ACW 20728 mango or passionfruit flavor

330

total       773

New possible traits for crosses 2014

• Self-thinning• Low carb (low fructose, low sorbitol)

Apple genotypes described by Agroscope

Traits need to be confirmed with new

know-how on the markers and false

positive signals

Low allergenic apples

‘Some people who are allergic may simply say they don’t like apples, since they’ve a very mild reaction after eating them,’ Eric van de Weg, plant scientist at Wageningen University in the Netherlands told Fresh Plaza.

Read more: http://www.dailymail.co.uk/sciencetech/article-2298958/Geneticists-search-perfect-apple-hypoallergenic-variety-doesn-t-cause-allergies.html#ixzz2rxZTbvCn

Different approaches towards low allergenic apples

Dr. Alessandro Botton, a plant physiologist at the Department of Agronomy, Food, Natural Resources, Animals and Environment in the University of Padua worked as a plant physiologist on the ISAFRUIT project to unravel the apple's genetic markers. Botton is currently working on the AGER Project to identify naturally occurring hypo-allergenic proteins in order to find a way to breed different apple varieties into a hypo-allergenic variety of apples.

Resistance to powdery mildew

Overview on powdery mildew resistanceLinkage group Origin

Major genes

Pl1 12 Malus ×robusta

Pl2 11 Malus zumi

Plbj 10 Malus baccata jackiiPld 12 D12

Plm 11 Mildew immune selection (MIS)

Plw 8 White angel

QTLs 1, 2, 3, 5, 7, 8, 9, 10, 13, 14, 16, 17

Malus hupehensis, Malus sargentii, Dülmener Rosenapfel, Discovery, TN10-8, …

Pyramiding of powdery mildew resistances

• available material, lines with– Pl1Pl2Plm (Pl2 and Plm both on LG11, recombination required)– Pl1Pl2Pld (Pl1 and Pld both on LG12, recombination required)

• very small fruit size, bad quality, described in D2.1

• improvement of size quality (crosses with GD)

Selection of seedlings with pyramided powdery mildew resistances

Phenotyping of seedlings in the glasshouse Genotyping of resistant seedlings

marker

Pl1 AT20SCAR

Pl2 CH04h02, N18, U2

Pld CH03c02

Plm N18, AC20, U2

Results from genotyping (GD x Pl1Pl2Plm)

Mother Father Seeds planted

No. of plants

without PM

Pl1 Pl2 Plm Pl1Pl2 Pl1Plm Pl2Plm Pl1Pl2Plm

GD 06005/9 50 17 2 7 4 4

GD 06005/21 50 23 6 6 8 3

GD 06005/26 50 17 1 1 14 1

GD 06005/55 50 20 1 2 14 3

GD 06006/7 50 28 1 1 4 16 6

GD 06006/8 50 21 3 1 2 14 1

GD 06006/32 50 20 8 2 3 7

GD 06006/57 50 21 1 1 1 5 9 1 3

Results from genotyping (GD x Pl1Pl2Pld)

Mother Father Seeds planted

No. of plants

without PM

Pl1 Pl2 Pld Pl1Pl2 Pl1Pld Pl2Pld Pl1Pl2Pld

GD AA03 2,20 50 13 1 12

GD AA03 2,33 50 29 1 1 7 5 15

GD AA03 2,37 50 21 2 3 10 1 5

GD AA03 2,45 50 21 1 3 2 6 2 7

GD AA03 2,58 50 32 1 3 3 4 11 8

GD AA03 2,60 50 18 2 8 8

Additional crosses for pyramiding of powdery mildew resistances

Mother Father Resistances Resistancesseedlings

No.

Mother Father

Ariwa ACW8259 Rvi6, Pl1 Pl2 No seeds

Ariwa ACW13597 Rvi6, Pl1 Pld No seeds

Ariwa ACW8099 Rvi6, Pl1 Rvi6, Pl2, FBF7 Rvi6, Pl1, Pl2, FBF7Rvi6, Pl1, Pl2

87

Ariwa ACW8259 Rvi6, Pl1 Pl2 Rvi6, Pl1, Pl2 8

06006/7 E6 33,124 Rvi2, Rvi4, FbF7, Sd1 Pl1, Pl2, Plm Rvi2, Rvi4, Pl1, Pl2, PlmRvi2, Rvi4, FbF7, Sd1, Pl1, Pl2Rvi2, Rvi4, FbF7, Pl1, Pl2Rvi2, Rvi4, Sd1, Pl1, Pl2Rvi2, Rvi4, Pl1, Pl2Rvi2, Rvi4, FbF7, Sd1, Pl1, PlmRvi2, Rvi4, FbF7, Pl1, PlmRvi2, Rvi4, Sd1, Pl1, PlmRvi2, Rvi4, Pl1, Plm

112822134

Resistance to fire blight from Mr5 (ETH and JKI)

• Resistance gene of Mr5 has been isolated and function proved in transgenic GalaBroggini et al. (2014) Engineering fire blight resistance into the apple cultivar 'Gala' using the FB_MR5 CC-NBS-LRR resistance gene of Malus × robusta 5. Plant Biotechnology Journal accepted

Transgenic Gala lines

Introduction of fire blight resistance from Mr5using the fast breeding approach (JKI)

PinovaT11902011

2012 (F2) 12120

13103 2013 (F3) Fuji

Golden Del.

(Idared x Mr5)x

x

x

14100 (16 seedlings) 2014 (F4)

MAS: with markers surrounding the fire blight resistance gene FB_MR5

Conclusions

Pyramiding a broad range of disease resistance genes and factors is supposed to be promising in respect of durability of the resistance.

To integrate scab, mildew and fire blight resistances in the same genotype is an added value provided a high fruit quality and productivity can be achieved.

Further breeding objectives such as low chilling, low allergenic, high flavor and eventually self-thinning and health/nutritional aspects were or will be considered.