Olive tree session outputs Partners involved in olive

Olive tree session outputs

Partners involved in olive research Institut National de la Recherche Agronomique (MOROCCO ) Institut de l’Olivier (TUNISIA) Ecole Nationale Supérieure Agronomique ( ALGERIA) Hellenic Agriculture Organization-Institute of Olive Tree, Subtropical Crops & Viticulture (GREECE)

Olive - WPs and tasks WP 1. Phenotyping for traits relevant to resilient fruit systems production WP 2. Genetic dissection and methods for selection of relevant traits for resistance to biotic and abiotic stresses T 1. 1 Evaluation of plant response to biotic and abiotic stresses in germplasm collections √ T 1. 2 Phenotyping segregating progenies to decipher genetic determinism for traits relevant to predefined ideotypes √ T 1. 3 Environment and management effect on adaptive traits √ T 1. 4 Omics analysis of environmental adaptation mechanisms √ T 2. 1 Genome-wide genotyping and genetic characterization of germplasm resources √ T 2. 2 Linkage and association mapping √ T 2. 3 Development of GS models for resilience traits WP 3. Exploitation of local biodiversity and development of pre-breeding materials WP 4. Communication, dissemination, training and demonstration T 2. 4 Development of markers for resistance/tolerance QTLs √ T 3. 1 Development of pre-breeding materials to pyramid resilience traits √ T 3. 2 Setting-up of reference collections in multiple environments √ T 3. 3 Progeny evaluation in MB √ T 4. 5 FREECLIMB data collection and management √

WP 1. Phenotyping for traits relevant to resilient fruit systems production Task 1. 1 Evaluation of plant response to biotic and abiotic stresses in germplasm collections Targeted traits and Number of cultivars for each partner ENSA Abiotic stress Biotic stress IO INRA-MO IOSV √ (5) √ (10) Heat on flowers √ (10) Flowering √ (36) √ (4) √(100) √(40) Drought √ (10) √ (4) √ (5) √ (40) Salinity √ (10) √ (4) Olive fruit fly Verticillium wilt. √ (10) √ (100) √ (40) √ (10)

WP 1. Phenotyping for traits relevant to resilient fruit systems production Task 1. 1 Evaluation of plant response to biotic and abiotic stresses in germplasm collections Targeted trait: Early flowering to avoid heat stress Years 1, 2 and 3 Participating partners: ALL Measurements: Record once every week phenological stages according to Sanz Cortes et al. , 2002. Start as soon as possible for as more cultivars as possible. Simple: date, cultivar, stage of flowering (number of stage) Example: 10/04/2019, Frantoio, 50 Koroneiki, 48 Arbequina, 52 etc. In case of available human resources, more measurements that are useful are: 1. Flowering load of the tree. Scale from 0% (no flowers at all) to 100% (tree full of flowers) 2. Number of inflorescences (we measure the length of flowering shoot and the number of inflorescence, so that the result is number of inflor. per meter of shoot) in 4 -10 shoots (3 -4 trees) per cultivar. 3. Percentage of hermaphrodite flowers (we collect 20 inflorescence from 3 -4 trees per cultivar, for each inflorescence we count total number of flowers and number of hermaphrodite flowers.

WP 1. Phenotyping for traits relevant to resilient fruit systems production Task 1. 1 Evaluation of plant response to biotic and abiotic stresses in germplasm collections Targeted trait: Tolerance to high temperature stress Years 1, 2 and 3 Participating partners: ENSA, INRA-MO, IOSV Growing of potted plants 1) outdoors, under natural conditions 2) inside a greenhouse / growth chamber with elevated temperature Use uniform plants of the same age and overall condition. Apply the same care, fertilizer, pest control, etc. Use 5 -10 cultivars, preferably local, (for example, 4 local and 1 international, or 8 local and 2 international) Suggested international: Koroneiki, Arbequina. All plants are initially grown outdoors. One week before flower opening (in estimation) one group of plants is transferred to the greenhouse for higher temperature and the other remains outside. Measurements: Number of inflorescence / plant for all plants one week before flower opening. Pollen germination in vitro. Number of fruits 2 weeks after full bloom. Add measurements according to your facilities and expertise e. g. gas exchange, gene expression, enzyme activity, spectral signatures, etc. Use temperature recorder inside and outside the greenhouse (preferably hourly T).

WP 1. Phenotyping for traits relevant to resilient fruit systems production Task 1. 1 Evaluation of plant response to biotic and abiotic stresses in germplasm collections Targeted trait: Tolerance to Drought Participating partners: ALL Open-field in adult trees and/or potted young plants Plants under different water regimes (0%; 50% and 100%). measurement of different parameters : Photosynthetic rate, Stomatal conductance, Leaf temperature, Spectral signature Xylem vulnerability to embolism (Morocco); Leaves and fruit size ; Oil content and components; Reproductive phenology …. Years 1, 2 and 3

WP 1. Phenotyping for traits relevant to resilient fruit systems production Task 1. 1 Evaluation of plant response to biotic and abiotic stresses in germplasm collections Targeted traits : Resistance to Salinity Participating partners: ENSA, IOSV open-field in adult trees or Potted young plants Plants under different salinity regimes. Mineral elements determination measurement of physiological parameters Years 1, 2 and 3

WP 1. Phenotyping for traits relevant to resilient fruit systems production Task 1. 1 Evaluation of plant response to biotic and abiotic stresses in germplasm collections Targeted traits : Resistance to Olive fruit fly Participating partners: INRA-MO, IOSV Field monitoring of olive fruit fly population by traps Fruit sampling to determine infestation parameters according to Garantonakis et al. , 2016 Years 1, 2 and 3

WP 1. Phenotyping for traits relevant to resilient fruit systems production Task 1. 1 Evaluation of plant response to biotic and abiotic stresses in germplasm collections Targeted traits : Resistance to Verticillium wilt Participating partners: IOSV - Artificial inoculation of non-defoliating V. dahliae isolate - visual symptom assessment - Destructive sampling of plant - pathogen re-isolation and RT-PCR quantification

WP 1. Phenotyping for traits relevant to resilient fruit systems production T 1. 2. Phenotyping segregating progenies to decipher genetic determinism for traits relevant to predefined ideotypes. Targeted traits : Resistance to Olive fruit fly Participating partners: IO; INRA-MO Segregating progeny from cross : Coratina x Chemlali Picholine x marocaine x Arbequina Picholine marocaine x Picholine de languedoc Field monitoring of olive fruit fly population by traps Fruit sampling to determine infestation parameters according to Garantonakis et al. , 2016 Targeted trait: Early flowering to avoid heat stress Participating partners: IO Segregating progeny from cross : Coratina x Chemlali As described in T 1. 1 based on the paper by Sanz Cortes et al. , 2002

WP 1. Phenotyping for traits relevant to resilient fruit systems production T 1. 3 Environment and management effect on adaptive traits. Years 2 and 3 Firstly SSR characterization for the common olive cultivars between partners Two main traits : - Olive fruit fly - Flowering Algeria (18 cultivars) Akerma, Azerdji, Tabelout , Aîmel, Picual, Arbequine. . . Sets of Common cultivars in different areas Frantoio; Leccino; Manzanille de sevilla (4 partners) Koroneiki; Mastoidis; P. languedoc; P. marocaine (INRA; IOSV; IO) 34 cultivars (2 places) Greece (16 cultivars) Koroneiki, Mavreya, Kolybada, leccino, Frantoio… Olive Core collection Marrakech (100 cultivars) Tunisia (13 cultivars) Chetoui, Dhokar, Gerboui, Picholine marocaine …

WP 1. Phenotyping for traits relevant to resilient fruit systems production T 1. 4. Omics analysis of environmental adaptation mechanisms Years 2 and 3 Participating partners: INRA-MO High throughput RNA sequencing q. RT-PCR using candidate genes c. DNA librairies prepared and delivered to genotyping laboratories Cultivars with contrasted behavior in field Different level of susceptibility/resistance to fruit fly Different levels of flowering Experiemental trials (heat in green house/irrigation) Different water stress Plants under heat during flowering Identification of Differencially Expressed genes (which genes involved and how much are activated)

WP 1. Phenotyping for traits relevant to resilient fruit systems production Deliverables • D 1. 1: Validation of existing protocols for abiotic stress assessment (M 24) • D 1. 2: List of accessions combining resistance/tolerance to biotic and abiotic stresses (M 24; update M 36) • D 1. 3: Phenotypic data for genetic analyses provided (M 24) • D 1. 4: New parents of interest to be used in crossing, pre-breeding or to be tested in orchards (M 24; update M 36) • D 1. 5: Characterization of adaptive mechanisms signatures in model fruit species (M 36)

WP 2. Genetic dissection and methods for selection of relevant traits for resistance to biotic and abiotic stresses T 2. 1 Genome-wide genotyping and genetic characterization of germplasm resources Participating partners: ALL SSR characterization and authentication of plant material in common between different partners DNA will be extracted and delivered to IOSV for genotyping

WP 2. Genetic dissection and methods for selection of relevant traits for resistance to biotic and abiotic stresses T 2. 2 Linkage and association mapping Participating partners: IO; INRA-MO GBS data core collection 3 segregation populations (Be. FORE Project output) Genetic maps (QTL mapping) Variability within cultivars (SNP) (GWAS) - Resistance/susceptibility to olive fruit fly - maybe flowering ? Assistance for GBS for IO (population hybrid) ? ?

WP 2. Genetic dissection and methods for selection of relevant traits for resistance to biotic and abiotic stresses T 2. 4 Development of markers for resistance/tolerance QTLs. Participating partners: IO is mis-stated in the proposal instead of INRA-MO QTL regions If detected Resequencing allelic variants and markers dvp Validation with other germplsm and progenies

WP 2. Genetic dissection and methods for selection of relevant traits for resistance to biotic and abiotic stresses Deliverables • D 2. 1: High-quality and high-throughput genotyping data for olive genetic materials (M 18) • D 2. 2: Genomic regions controlling variation for resistance to biotic and abiotic stresses (M 24; update M 36) • D 2. 3: Genomic tools to improve selection/introgression of resilience traits (GS and GWAS models accounting for environmental and management interactions) (M 24; update M 36)

WP 3. Exploitation of local biodiversity and development of pre-breeding materials T 3. 1 Development of pre-breeding materials to pyramid resilience traits. High performant cultivars regarding traits of interest Germplasm Bank Type of materials (e. g. selection, seedling, accession, progenies etc. ) Greece accession Cultivar name Cross parents or origins (if available) Kalamon Greece accession Lefkolia Serron Koroneiki Arbequina Greece Spain Morocco Tunisia Tunisia Algeria accession accession accession Lechin de Sevilla Azeradji Picholine marocaine Chemlali Chétoui Oueslati Coratina Koroneiki SIGOISE LIMLI TABELOUT Spain Algeria Morocco Tunisia Italy Greece Algeria Trait / Resistance to Ve rticilliumwilt, salinity Verticillium wilt, early flowering, high temperature olive fruit fly Olive fruit fly; drought; cold; salinity; Early flowering; Drought; salinity; Drought, salinity Drought, fusarium, Pseudomonas savastanoi Fusicladium oleagineum Salinity Early flowering, salinity; moderately tolerant of cold and drought Moderately to cold and very resistant to drought moisture resistant, early flowering IOSV make some crosses, probably other partners will procced this years M 0012. More crosses next years

WP 3. Exploitation of local biodiversity and development of pre-breeding materials T 3. 1 Development of pre-breeding materials to pyramid resilience traits. Seeds issued from crosses will be shared between different partners to be planted and evaluated in different climatic conditions

WP 3. Exploitation of local biodiversity and development of pre-breeding materials Deliverables D 3. 1: Pre-breeding material combining a sound horticultural value, fruit quality and resilience traits relevant for MB production (M 36) D 3. 2: Lists of genotypes to include in replicated multi-site collections (M 36) D 3. 3: Setting up of progeny evaluation network under Mediterranean conditions (M 36)

Personnel exchange Communication between partners to decide on topics e. g. multispectral imaging measures, etc. Facilities needed Laboratories for genotyping activities (SSR for all partners; GBS for IO)

Thank you