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Phenotyping for Resilient Olive Tree Traits

This project aims to evaluate olive tree responses to biotic and abiotic stresses and identify traits relevant to resilient fruit systems production. It involves partners from Morocco, Tunisia, Algeria, and Greece, and focuses on traits such as early flowering, tolerance to high temperature, drought, salinity, olive fruit fly, and Verticillium wilt. Various measurements and assessments will be conducted over a span of three years.

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Phenotyping for Resilient Olive Tree Traits

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  1. Olive tree session outputs

  2. Partnersinvolved 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)

  3. Olive - WPs and tasks

  4. WP1. 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 eachpartner

  5. WP1. 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: Earlyflowering to avoidheat 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.

  6. WP1. 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).

  7. WP1. 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 Years 1, 2 and 3 • Participating partners: ALL • Open-fieldin adult trees and/or potted young plants • Plants under different water regimes (0%; 50% and 100%). • measurement of different parameters : Photosyntheticrate, Stomatalconductance, Leaftemperature, Spectral signature • Xylemvulnerability to embolism (Morocco); • Leaves and fruit size ; • Oil content and components; • Reproductive phenology ….

  8. WP1. 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 Years 1, 2 and 3 Participating partners: ENSA, IO, IOSV open-fieldin adult trees or Potted young plants Plants under different salinity regimes. Mineral elements determination measurement of physiological parameters

  9. WP1. 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 Years 1, 2 and 3 • Participating partners: INRA-MO, IOSV • Field monitoring of olive fruit fly population by traps • Fruit sampling to determine infestation parametersaccording to Garantonakis et al., 2016

  10. WP1. 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 Verticilliumwilt 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

  11. WP1. Phenotyping for traits relevant to resilient fruit systems production T1.2 . 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 : Coratinax Chemlali Picholine x marocaine x Arbequina Picholinemarocaine x Picholine de languedoc Field monitoring of olive fruit fly population by traps Fruit sampling to determine infestation parametersaccording to Garantonakis et al., 2016 Targeted trait: Earlyflowering to avoidheat stress Participating partners: IO Segregating progeny from cross : Coratinax Chemlali As described in T1.1 based on the paper by Sanz Cortes et al., 2002

  12. WP1. Phenotyping for traits relevant to resilient fruit systems production T1.3 Environment and management effect on adaptive traits. Years 2 and 3 Firstly SSR characterization for the common olive cultivars between partners Greece (16 cultivars) Koroneiki, Mavreya, Kolybada, leccino, Frantoio… Olive Core collection - Marrakech (100 cultivars) Algeria (18 cultivars) Akerma, Azerdji, Tabelout , Aîmel, Picual, Arbequine... Two main traits : - Olive fruit fly - Flowering Sets of Common cultivars in different areas Tunisia (13 cultivars) Chetoui, Dhokar, Gerboui, Picholine marocaine … Frantoio; Leccino; Manzanille de sevilla (4 partners) Koroneiki; Mastoidis; P. languedoc; P. marocaine (INRA; IOSV; IO) 34 cultivars (2 places)

  13. WP1. Phenotyping for traits relevant to resilient fruit systems production T1.4. Omicsanalysis of environmental adaptation mechanisms Years 2 and 3 • Participating partners: INRA-MO High throughput RNA sequencing qRT-PCR using candidate genes cDNA librairies prepared and delivered to genotypinglaboratories Cultivars withcontrastedbehavior in field Experiemental trials (heat in green house/irrigation) Differentlevel of susceptibility/resistance to fruit fly Different water stress Plants underheatduringflowering Differentlevels of flowering Identification of DifferenciallyExpressedgenes (whichgenesinvolved and how much are activated)

  14. WP1. Phenotyping for traits relevant to resilient fruit systems production Deliverables • D 1.1: Validation of existing protocols for abiotic stress assessment (M24) • D 1.2: List of accessions combining resistance/tolerance to biotic and abiotic stresses (M24; update M36) • D 1.3: Phenotypic data for genetic analyses provided (M24) • D 1.4: New parents of interest to be used in crossing, pre-breeding or to be tested in orchards (M24; update M36) • D 1.5: Characterization of adaptive mechanisms signatures in model fruit species (M36)

  15. WP2. Genetic dissection and methods for selection of relevant traits for resistance to biotic and abiotic stresses T2.1 Genome-wide genotyping and genetic characterization of germplasm resources • Participating partners: ALL SSR characterization and authentication of plant material in commonbetweendifferentpartners DNA willbeextracted and delivered to IOSV for genotyping

  16. WP2. Genetic dissection and methods for selection of relevant traits for resistance to biotic and abiotic stresses T2.2 Linkage and association mapping • Participating partners: IO; INRA-MO GBS data core collection 3 segregation populations (BeFORE Project output) Geneticmaps Variabilitywithin cultivars (SNP) (QTL mapping) (GWAS) • Resistance/susceptibility to olive fruit fly • maybeflowering ? Assistance for GBS for IO (population hybrid) ??

  17. WP2. Genetic dissection and methods for selection of relevant traits for resistance to biotic and abiotic stresses T2.4 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 withothergermplsm and progenies

  18. WP2. 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 (M18) • D 2.2: Genomic regions controlling variation for resistance to biotic and abiotic stresses (M24; update M36) • D 2.3: Genomic tools to improve selection/introgression of resilience traits (GS and GWAS models accounting for environmental and management interactions) (M24; update M36)

  19. WP3. Exploitation of local biodiversity and development of pre-breeding materials T3.1 Development of pre-breeding materials to pyramid resilience traits. High performant cultivars regarding traits of interest IOSV makesome crosses, probablyotherpartnerswillproccedthisyearsM0-012. More crosses nextyears

  20. WP3. Exploitation of local biodiversity and development of pre-breeding materials T3.1 Development of pre-breeding materials to pyramid resilience traits. Seedsissuedfrom crosses willbesharedbetweendifferentpartners to beplanted and evaluated in differentclimatic conditions

  21. WP3. 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 (M36) D 3.2: Lists of genotypes to include in replicated multi-site collections (M36) D 3.3: Setting up of progeny evaluation network under Mediterranean conditions (M36)

  22. Personnel exchange Communication betweenpartners to decide on topicse.g. multispectralimagingmeasures, etc. Facilitiesneeded Laboratories for genotypingactivities (SSR for all partners; GBS for IO)

  23. Thankyou

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