Screening of olive germplasm for resistance to Xylella fastidiosa ST53: the state of the art

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Screening of olive germplasm for resistance to Xylella fastidiosa ST53: the state of the art Donato Boscia CNR, Istituto per la Protezione Sostenibile delle Piante, Bari (IT)

GENETIC RESISTANCE TO PATHOGENS AND PESTS Phylloxera Citrus tristeza virus The history of Plant Pathology reports several examples of successes against epidemics and plant diseases Genetic resistances to bacteria, including Xylella, are known in several plant species (i.e. apple, pear, poplar, potato, tomato, alfalfa, corn, cotton, rice, etc.) From susceptibility to immunity: several degrees of resistance/tolerance 2

DIFFERENT SOURCES OF NATURAL RESISTANCE TO XYLELLA FASTIDIOSA (XF) HAVE BEEN DESCRIBED IN GRAPEVINES AND CITRUS LACK OF INFORMATION ON POSSIBLE SOURCES OF RESISTANCE/TOLERANCE IN THE OLIVE GERMPLASM. 3

Biodiversity in olive germplasm: huge number of cultivars in several international collections Variability Inter-cultivar Variability Intra-cultivar (Clones) Need to identify resistant/tolerant cultivars/clones 4

PRELIMINARY FIELD OBSERVATIONS OF A FEW CULTIVARS IN SALENTO, HAVE SHOWN THAT DIFFERENTIAL RESPONSES TO XF INFECTIONS EXIST Ogliarola Leccino 5

Leccino Carolea 6

Leccino Ogliarola 7

Leccino Ogliarola 8

3 main research lines Continuous survey in the infected area for the search of symptomless cultivars and olive seedlings Experimental fields for the evaluation of the susceptibility to different cultivars to Xf infection Overgrafting on infected trees for the faster identification of resistance (Quick Tolerance test) 9

Suitable site in infected area selected for the experimental field 10

First experimental plot, 2015, with the EFSA support Arbequina Arbosana Cellina di Nardò Cima di Melfi Coratina Don Carlo Frantoio FS17 Koroneiki Leccino 11

This plot was extended in April 2016 ASCOLANA TENERA BELLA DI SPAGNA CELLINA DI NARDO CIPRESSINO DOLCE DI CASSANO ITRANA LECCINO MAIATICA NOCELLARA ETNEA NOCELLARA MESSINESE NOCIARA OGLIAROLA OLIASTRO PENDOLINO PERANZANA PICHOLINE SIMONE TERMITE DI BITETTO TOSCANINA 12

Further extended in November 2016 Changlot x Dolce Agogia Changlot x Dolce Agogia Arbequina o.p. Frantoio o.p. Frantoio x Arbosana Frantoio x Arbosana Frantoio x Arbosana Koroneiki o.p. Koroneiki x Empeltre Koroneiki x Empeltre O. cerasiformis O. cerasiformis PicualxJeac9 PicualxJeac9 JA5 - subsp. europaea var. sylvestris MSAC43 - subsp. europaea var. sylvestris OLM28 - subsp. europaea var. sylvestris OLM40 - subsp. europaea var. sylvestris TN10 - subsp. guanchica TN2 - subsp. guanchica 13

2017: 40 NEW SELECTIONS PROVIDED BY IFAPA (CORDOBA) Abou Kanani Arbequina Arbosana Ascolana Tenera Ayvalik Barnea Barri Canino/caninese Changlot Real Chemlal de Kabylie Chetoui Chiquitita Coratina Empeltre Frantoio Galega Gordal Sevillana Grappolo Hojiblanca Kalamon Koroneiki Lastovka Leccino Maarri Manzanilla de Sevilla Mari Mastoidis Megaritiki Memeçik Meski Moraiolo Morrut Picholine Picholine marrocaine Picual Picudo Shengeh/Fishomi Toffahi Uovo di Piccione Uslu Syria Turkey Israel Syria Argelia Tunisia Portugal Greece Greece Croatia Syria Iran Greece Greece Turkey Tunisia France Marocco Iran Egypt 14 Turkey

HALF EXPOSED IN INFECTED AREA BEFORE TO BE TRANSPLANTED NEXT SPRING IN THE FIELD Abou Kanani Arbequina Arbosana Ascolana Tenera Ayvalik Barnea Barri Canino/caninese Changlot Real Chemlal de Kabylie Chetoui Chiquitita Coratina Empeltre Frantoio Galega Gordal Sevillana Grappolo Hojiblanca Kalamon Koroneiki Lastovka Leccino Maarri Manzanilla de Sevilla Mari Mastoidis Megaritiki Memeçik Meski Moraiolo Morrut Picholine Picholine marrocaine Picual Picudo Shengeh/Fishomi Toffahi Uovo di Piccione Uslu Syria Turkey Israel Syria Argelia Tunisia Portugal Greece Greece Croatia Syria Iran Greece Greece Turkey Tunisia France Marocco Iran Egypt 15 Turkey

HALF NEEDLE INOCULATED UNDER CONTROLLED CONDITIONS (IN BARI) Abou Kanani Arbequina Arbosana Ascolana Tenera Ayvalik Barnea Barri Canino/caninese Changlot Real Chemlal de Kabylie Chetoui Chiquitita Coratina Empeltre Frantoio Galega Gordal Sevillana Grappolo Hojiblanca Kalamon Koroneiki Lastovka Leccino Maarri Manzanilla de Sevilla Mari Mastoidis Megaritiki Memeçik Meski Moraiolo Morrut Picholine Picholine marrocaine Picual Picudo Shengeh/Fishomi Toffahi Uovo di Piccione Uslu Syria Turkey Israel Syria Argelia Tunisia Portugal Greece Greece Croatia Syria Iran Greece Greece Turkey Tunisia France Marocco Iran Egypt 16 Turkey

2015: 2 experimental fields in infected area 17

Overgrafting for a «Quick tolerance test» The numbers: 12 hectars 5000 overgrafts 260 cultivars 18

The complete list of the cultivar under screening is now available on the webpage of XF-ACTORS: http://www.xfactorsproject.eu 19

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SOME PRELIMINAR ENCOURAGING RESULTS: Leccino and FS-17 21

Evidences of resistance: Low population of Xylella in Leccino (only 1-2%) and even less (0,5%), in FS-17 vs Ogliarola Comparative analysis of the transcriptome (Leccino vs Ogliarola) show that in infected Leccino there are overexpressed genes involved in mechanisms of resistance previously reported in citrus resistant to xylella 22

Quantitave PCR assay OG LC1 LC2 HC OG= ogliarola salentina; LC= Leccino, HC= healthy control 23

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Identification of genes involved in defense response Genes under study surface receptors expansin (overexpressed in Ogliarola; underexpressed in resistant mandarin (De Souza et al.) Calcium transporting, Calcium protein-kinase, Calcium binding (De La Fuente et al.) Evaluation of expression in susceptible and resistant cultivars in greenhouse on field plants RNA cdna - qpcr 25

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