Piramydizing resistance genes in grape: a breeding program for the selection of elite cultivars

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Piramydizing resistance genes in grape: a breeding program for the selection of elite cultivars Foria S., Monte C., Testolin R., Di Gaspero G., Cipriani G. Dipartimento di Scienze AgroAlimentari, Ambientali e Animali University of Udine _Italy

Azerbaijan 7,000 years BP

10.000 vine varieties known in the world 13 represents more than one third of the world vine area 33 varieties represents 50% «international varieties» Cabernet-Sauvignon one of the most cultivated wine grape in the world (5% world vine area)...

The most important grape cultivars of the world (OIV, 2015) Cultivar Destination Area (1000 ha) Countries 1 Kyoho Table 365.000 China 2 Cabernet-Suavignon Wine 340.000 Many 3 Sultanina Table, dried and wine 300.000 Many 4 Merlot Wine 266.000 France 5 Tempranillo Wine 231.000 Spain 6 Airen Wine aand Brandy 218.000 Spain 7 Chardonnay Wine 211.000 Many 13 Trebbiano toscano/ugni blanc Wine and Brandy 111.000 Italy, France

Distribution of the genus Vitis

Grape has 800 resistant genes, but it is not resistant to the pathogens evolved overseas

Powdery Mildew - Erysiphe necator (syb Uncinula necator) - Leaves, shoots, rachis, stems, inflorescences and maturing berries are all susceptible

Downy Mildew - Plasmopora viticola - Leaves, inflorescences and young bunches are all susceptible

Sources of resistance The genetic basis has been evaluated and mapped in several genetic backgrounds: - Hybrids from North American Vitis species; - Two M. rotundifolia accessions; - Two Near Eastern V. vinifera cultivars (Kishmish vatkana and Dza. kara); - Several Chinese Vitis species. NBS LRR genes TIR NB LRR genes CC NB LRR genes

Resistant Genes R locus Chromosome R gene type Rpv1 12 TIR NB - LRR Rpv2 18 NBS - LRR Rpv3 18 TIR NB - LRR Rpv8 14 NBS - LRR Rpv10 18 NBS - LRR Rpv12 14 CC NB - LRR Run1 12 TIR NB - LRR Ren1 13 CC NB - LRR Ren4 18 unknown 21 Rpv genes 12 Ren/Run genes http://www.vivc.de/ Update: October 04, 2017

1999 2018 Vc_001 Vc_571 Merlot, Cabernet Sauvignon, Sauvignon blanc, Chardonnay, Traminer, Pinot, Aglianico, Sangiovese, Tocai F Regent, Bianca, 20/3, V. aestivalis Cynthiana, Pannonia, 99-1-48, Kishmish vatkana, SK-00-1/7, Solaris, Vc34_113 (Soreli)

Crosses - Emasculation - Collection of pollen - Pollination - Seeds harvesting - Germination in glasshouse

First year: sampling for genotyping and phenotyping

Phenotypic evaluation

Selection Name Cross 34.111 34.113 76.026 55.098 55.100 Fleurtai Soreli Sauvignon Kretos Sauvignon Nepis Sauvignon Rytos Tocai x 20/3 Tocai x 20/3 Sauvignon x 20/3 Sauvignon x Bianca Sauvignon x Bianca Fleurtai Soreli Sauvignon Kretos Sauvigno n Nepis Sauvignon Rytos

Selection Name Cross 58.083 32.078 31.122 31.125 36.030 Cabernet Eidos Cabernet Volos Merlot Kanthus Merlot Chorus Julius Cab Sauvignon x Bianca Cab Sauvignon x 20/3 Merlot x 20/3 Merlot x 20/3 Regent x 20/3 Cabernet Cabernet Merlot Merlot Julius Eidos Volos Kanthus Chorus

Resistance gene pooling - Pyramidization Resistant Elite 1 X Resistant Elite 2 = Rpv3 Rpv12 Run1 Ren1

Vc_XXX 2004 2013 SO4 Gujot (2.80 m X 1 m) 70 cross combinations 1101 progenies 1 174 progenies per cross MAS on seedlings Genes Rpv3 Rpv12 Run1 Ren1 Markers UDV305 UDV737 Sc36_7 Sc81_9.1 Sc34_8 Sc35_2 Sc47_6 Sc47_20 Phenothipic selection on seedlings

Number of crosses including at least one of the resistance genes Gene pooling 59 49 Combination with 4 R 19% Combination with 1 R 11% 26 Combination with 3 R 28% Combination with 2 R 42% 2 9 6 10 13 1

Phenotype evaluation berry color R 1% berry color ND 11% bunch evaluation 3 0% bunch evaluation ND 12% berry color B 28% berry color W 60% berry color W berry color B berry color R berry color ND bunch evaluation 2 4% bunch evaluation 1 84% bunch evaluation 1 bunch evaluation 2 bunch evaluation 3 bunch evaluation ND productivity low 1 11% productivity high 2 2% productivity very high 3 1% habitus week 1 1% habitus vigorous 2 0% productivity medium 86% productivity low 1 productivity high 2 productivity very high 3 productivity medium habitus normal 99% habitus week 1 habitus vigorous 2 habitus normal

Resistance to other diseases Botrytis 1 1% Botrytis 2 0% sour rot 1 [PERCENTUALE] sour rot 2 [PERCENTUALE] Botrytis 1 sour rot 1 Botrytis - no infections 99% Botrytis 2 Botrytis - no infections sour rot - no infections [PERCENTUALE] sour rot 2 sour rot - no infections

Vc_156 (99-1-48 x Pinot noir) Progenies 174 Cross Combinations 1 berry color W 85 berry color B 81 berry color R 0 berry color - 8 bunch evaluation 1 157 bunch evaluation 2 8 bunch evaluation 3 1 bunch evaluation - 2 dead plants 6 productivity low 1 8 productivity high 2 0 productivity very high 3 0 productivity medium 166 habitus week 1 0 habitus vigorous 2 0 habitus normal 174 Botrytis 1 0 Botrytis 2 0 Botrytis - no infections 174 Acid rot 1 34 Acid rot 2 1 Acid rot - no infections 139 berry color B 49% bunch evaluation 2 5% bunch evaluation 3 1% bunch evaluation 1 94% berry color W 51% berry color W berry color B bunch evaluation 1 bunch evaluation 2 bunch evaluation 3

New selections Pinot Progenies Downy mildew genotyping Powdery mildew genotyping Field resistance to downy mildew Filed resistance to powdery mildew Vc_109-033 W Rpv 1, Rpv 12 Run 1, Ren 3 +++ +++ Vc_109-052 W Rpv 12 Ren 3 ++ + Vc_156-1017 W Rpv 1, Rpv 12 Run 1 +++ +++ Vc_156-869 W Rpv 12 none +++ - Vc_156-680 B Rpv 1, Rpv 12 Run 1 +++ +++ Vc_156-537 B Rpv 1, Rpv 12 Run 1 ++ +++ Vc_156-312 B Rpv 12 none +++ -

Vc156_1017 99-1-48 X Pinot noir Vigour: high Productivity: high Bud burst: early Maturation: early High resistance to powdery and downy mildew Leaf similar to Croatina, easy berry detachment, short brush Harvest Cluster weight (g) Production per plant (kg) Production per ha (t) o Brix Total acidity (g/l) 2/09/2016 220 3.4 12.1 22.0 5.5 3.6 21/08/2017 212 4.0 14.3 21.4 5.4 3.7 Location Centro Sperimentale VCR, Guyot, 2.8 x 1 ph

Wine test

Vc156_1017 99-1-48 X Pinot noir Fruity with spicy aromas, full body and persistant, buona struttura Year Total acidity (g/l) ph Tartaric acid (g/l) Malic acid (g/l) Sugar free extract (g/l) Alcohol, % 2016 5.1 3.5 1.9 1.8 20.8 13.4 2017 6.2 3.1 4.5 1.7 21.5 13.0 Location Centro Sperimentale VCR, Guyot, 2.8 x 1

Conclusions or open questions How many resistance genes Genetic drag concerns Quality demanding products Wine versus table grape cultivars

Thanks to