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V Symposium on Ecological Network, Palma, 10-12 de noviembre de 2021., Understanding microbial associations through co-occurrence network analysis is a very effective tool to detect patterns of copresence and mutual exclusion among members of the plant-associated microbiome. By using 16S and ITS rRNA metagenomics equencing, we studied the associations of bacterial and fungal communities present in xylem sap of plantlets and adult olive trees of 'Picual', 'Arbequina', 'Arbosana', 'Frantoio' and 'Hojiblanca' olive varieties. Results based on the analysis of amplicon sequence variant and network assembly analysis showed distinct microbial association behaviors according to olive plant age and genotype. Thus, Pseudomonas, Bosea and Meiothermus showed the highest positive associations in the xylem of adult trees whereas Azospirillium, Rhizobium and Brevundimonas showed the greatest positive interactions in olive plantlets. Furthermore, MCODE method identified clusters of highly interconnected regions for each olive genotype, where Alternaria and Phaeococcomyces genera were unique to ‘Arbequina’; Rhizobium, Bradyrhizobium and Brevibacillus were exclusive to ‘Arbosana’; Gardnerella and Rathayibacter to ‘Frantoio’; Acinetobacter and Micrococcus to ‘Hojiblanca’ and Arthrobacter, Lawsonella, Massilia, Sphingopyxis, Thermaerobacter and Variovorax to ‘Picual’. Interestingly, Pseudomonas and Anoxybacillus were positioned at the network cores obtained when data were analyzed by plant age and plant genotype, respectively. Network association analysis has revealed new knowledge regarding specific microbial associations occurring among olive genotypes at different plant age, offering the identification of keystone xylem-inhabiting microbial species that may result determinant for olive growth and health.

Additional file 6. Table of groups of olive cultivars according to Verticillium wilt resistant response.

En España existe una gran riqueza varietal de olivo (Olea europaea subsp. europaea var europaea) que ha sido ampliamente caracterizada y evaluada. Aun así, varios estudios recientes han evidenciado la presencia de variedades locales de olivo desconocidas, poniendo de manifiesto la necesidad de profundizar en trabajos de prospección, conservación y caracterización varietal. Por ello, en los últimos años, se están llevando a cabo trabajos de prospección en diferentes puntos del país (Andalucía, Aragón, Cataluña y La Rioja). Dichos trabajos han permitido la localización de 290 olivos cultivados de interés y recogida de material vegetal para su posterior identificación. Dicho material ha sido discriminado mediante un conjunto de 96 EST-SNPs recientemente desarrollado en el Banco Mundial de Germoplasma de Olivo del IFAPA (BGMO, Córdoba). El empleo de los marcadores EST-SNPs permitió la identificación de un total de 154 nuevas variedades en las cuatro regiones prospectadas y la detección de casos de sinonimias y homonimias. Además, los marcadores empleados pusieron en evidencia la presencia de una variabilidad relativamente amplia con unos rangos de heterocigosidad esperada y observada de 0,31-0,5 y 0,29-0,81, y valores medios de 0,46 y 0,53, respectivamente. El estudio de las relaciones genéticas entre las variedades prospectadas y el conjunto de variedades nacionales conservadas en el BGMO reveló una clara agrupación de variedades andaluzas y variedades del norte de España, indicando de esta manera una posible selección local de variedades de olivo en nuestro país. En concordancia con estudios anteriores en olivo, estos resultados ponen de manifiesto la eficiencia del conjunto de 96 EST-SNPs para la identificación varietal en olivo y su utilidad para mejorar el manejo de colecciones de recursos genéticos del cultivo.

Trabajo presentado en el XVI Congreso Nacional de Ciencias Hortícolas, celebrado en Córdoba del 17 al 22 de octubre de 2021., Los microorganismos endófitos representan una estrategia innovadora para el manejo de las enfermedades vasculares en el olivo actuando como una defensa natural innata para hacer frente a la infección por patógenos que colonizan el xilema como son Verticillium dahliae y Xylella fastidiosa. Sin embargo, pocos estudios han caracterizado el efecto de la variedad y la edad de la planta en la estructura y diversidad microbiana del xilema del olivo. Por lo tanto, este trabajo se ha centrado en la caracterización mediante un enfoque metagenómico de las comunidades bacterianas y fúngicas presentes en el fluido xilemático de plantones jóvenes y olivos adultos de las variedades de olivo ‘Picual’, ‘Arbequina’, ‘Arbosana’, ‘Frantoio’ y ‘Hojiblanca’. Se han identificado un total de 440 ASVs bacterianos y 96 ASVs fúngicos que variaron en función de la edad de la planta alcanzando una diversidad (alfa) microbiana mayor en plantas adultas que en los plantones jóvenes y presentando una estructura microbiana (beta-diversidad) diferente en función de la edad, pero no de la variedad. Por otra parte, las variedades ‘Arbosana’ y ‘Hojiblanca’ mostraron una mayor riqueza bacteriana y fúngica, respectivamente. Globalmente, Meiothermus (24.61%), Anoxybacillus (18.21%) y Paenibacillus (13.15%) fueron los géneros bacterianos más representativos mientras que Aureobasidium fue el hongo más abundante (50.51%), seguido de Plectosphaerella (6.94%) y Malassezia (6.38%). Estos resultados contribuyen a mejorar la caracterización y el conocimiento de la variabilidad del microbioma xilemático de olivo, generando una información muy útil para optimizar el manejo de las enfermedades vasculares del olivo mediante el uso de sus propios microrganismos endofíticos como agentes control biológico., Estudio financiado por los proyectos XF-ACTORS 727987 (EU-H2020) y AGL2016-75606-R (MICINN y FEDER-EU).

Trabajo presentado en el Congress of Microbiology and Biotechnology (Microbiotec 19), celebrado en Coimbra del 5 al 7 de diciembre de 2019., Up to date, the bacterial and fungal microbial communities from the olive (Olea europaeaL.) root systems have not been simultaneously studied. In this work, we show that microbial communities from the olive root endosphere are less diverse than those from the rhizosphere. But more relevant was to unveil that olive belowground communities are mainly shaped by the genotype of the cultivar when growing under the same environmental, pedological and agronomic conditions. Furthermore, Actinophytocola, Streptomyces and Pseudonocardia are the most abundant bacterial genera in the olive root endosphere, Actinophytocola being the most prevalent genus by far. In contrast, Gp6, Gp4, Rhizobium and Sphingomonas are the main genera in the olive rhizosphere. Canalisporium, Aspergillus, Minimelanolocus and Macrophominaare the main fungal genera present in the olive root system. Interestingly enough, a high proportion of so far unclassified fungal sequences at class level were detected in the rhizosphere. From the belowground microbial profiles here reported, it can be concluded that the genus Actinophytocolamay play an important role in olive adaptation to environmental stresses. Moreover, the huge unknown fungal diversity suggests that there are still some fungi with important ecological and biotechnological implications that have yet to be discovered.

Resumen del trabajo presentado en la 6th International Conference on the Olive Tree and Olive Products, celebrada en Sevilla (España) del 15 al 19 de octubre de 2018., The wide olive genetic patrimony has revealed high variability for most of the agronomic and oil quality traits of interest in olive growing. Few studies, however, have addressed the interaction of this variability with the environment, a subject of particular interest considering the natural high instability of the Mediterranean climate and the challenge of the predicted climate change. The current work presents results on the interaction between genotype and environment from multi-environment trials of olive cultivars and breeding selections, planted in different edaphoclimatic conditions of Andalusia, Southern Peninsular Spain and Canary Islands. For most of the agronomic and oil quality characters evaluated (flowering phenology, flower quality, pattern of oil accumulation, fatty acid composition and phenol content and composition), significant genotype and environment effects have been observed. For example, olive cultivars grown in Tenerife under much milder winter temperatures than in the Iberian Peninsula showed substantially earlierflowering and oil accumulation. Only in the case of flowering phenology was no significant genotype effect found. Furthermore, a strong genotype x environment effect was highly consistent in all characters considered. Regarding resistance to disease, such as Verticillium wilt, the variability of results from both natural and artificial inoculations also tends to indicate a considerable environmental effect and the need for careful testing of disease evolution. All this information strongly suggests the necessity of comparative trials of olive cultivars for both adequate choice of cultivar and final selection in breeding programs.

6 Páginas.-- 2 Figuras, There is an increasing need for diversification of olive (Olea europaea L.) cultivars, mainly because of the new intensive industry and the current trend of establishing olive productions in new areas outside the Mediterranean basin. Olive breeding has traditionally been focused on the improvement of agronomic traits. However, the nutritional quality of virgin olive oil (VOO) is lately being considered as a breeding target because of the growing weight of scientific evidence supporting the positive impact of VOO on human health. Increasing VOO consumption, through improvement of its nutritional and sensory properties, will in turn have a positive effect on both consumer health and olive-producing areas. VOO metabolites responsible for its nutritional and sensory quality include components such as phenolics and volatile compounds, which are synthesized during the oil extraction process by endogenous enzymes. Here, we aimed to investigate olive diversity regarding the key VOO quality metabolites of olive accessions cultivated around the world. For this purpose, oils from 97 olive cultivars representative of the diversity found at the World Olive Germplasm Collection (CAP-UCO-IFAPA), located at IFAPA Centre “Alameda del Obispo” Cordoba, Spain, were obtained and the phenolic and volatile fractions were further analyzed. The component profiles were compared with those of a similar subset of seedlings arisen from the cross of two widely cultivated olive cultivars having contrasting oil compositions ('Arbequina' and 'Picual'). Results showed that a single cross provides enough variability to produce new olive cultivars with improved quality. The set of cultivars from the collection displayed, in general, higher levels of variability than the progeny cross for the main groups of volatile compounds. However, the latter displayed a wider variability than the set of cultivars under study for most of the main classes of phenolic compounds in the oil, except the lignans., Funding for this research came from the OLEAGEN project of Genoma España and AGL2011-24442 project from the Programa Nacional de Recursos y Tecnologías Agroalimentarias, both financed by the Spanish Government.

The irrigation management for olive orchards under future weather conditions requires the development of advanced tools for considering specific physiological and phenological components affected by the foreseen changes in climate and atmospheric [CO2].

6 Figuras.-- 1 Tabla, Different scientific evidences suggest that the long term dietary consumption of virgin olive oil (VOO) seems to be related to an attenuation of the inflammatory response and reduction of the associated risk of chronic inflammatory disease states. VOO phenolic compounds are claimed to be the main responsible for these positive health benefits. They are mainly synthetized from phenolic glucosides present in the olive fruit by the action of glucosidases occurring when they come together once the olive fruit is crushed during olive oil extraction. The genetic variability of the major phenolic compounds was studied in a representative sample of olive cultivars (Olea europaea L.) from the World Olive Germplasm Collection established at IFAPA Centre “ Alameda del Obispo in Cordoba, Spain. The most abundant phenolic components found in VOO are the secoiridoid derivatives resulting from the enzymatic hydrolysis of oleuropein, ligstroside and demethyloleuropein present in the fruit, which showed to be on average their main phenolic glucosides. The mean content of phenolic compounds in the oils was 494.51 µg/g oil, displaying a variability range of 63.74-1432.04 µg/g oil. The mean content of phenolic compounds in the fruits was 12384.23 µg/g fruit with a range of 3754.13-30696.39 µg/g fruit. Total phenolic compounds in the fruits and the oils were significantly correlated (r = 0.66). Thus, the composition and biochemical status of the olive fruit seems to be the most important variables determining the synthesis of the VOO phenolic compounds during the oil extraction process. On the other hand, the content of oleuropein and derivatives in the oils and fruits showed a correlation coefficient (r = 0.64) lower than that observed for ligstroside and derivatives (r = 0.73). These findings might be related to the higher oxidation rates of the former due to the action of oxidative enzymes during the oil extraction process as a consequence of the orthodiphenolic structure they possess. Data on phenolic composition would be of interest for the selection of optimal parents in olive breeding programs with the aim of obtaining new cultivars with improved nutritional quality

8th European Symposium on Plant Lipids. Malmö, Sweden, 02-05/07/2017.

The use of dwarfing rootstocks represents an optimum mechanism to control the vigour of fruit tree species. Exceptionally, olive trees are grown on their own roots and due to excessive vigour, only a very limited number of cultivars have been successfully adapted to high-density orchards. Through prospection and phenotyping of different wild subspecies, a wide range of genetic variability of the species Olea europaea is being explored to identify genotypes that can be used as rootstocks to optimize intensive olive cultivation (e.g. low vigour, low susceptibility to Verticillium dahliae and high tolerance to abiotic stress). Here, early vigour parameters were studied in genotypes belonging to the subsp. europaea (var. sylvestris), guanchica, cuspidata and crosses of cuspidata x sylvestris. Homogeneous plants were obtained from in-vitro germinated seedlings and further micropropagation. After ex-vitro acclimatization and soil planting, plant height, diameter of the main axis at different heights and length of the longest shoot were measured eight months later. Significant correlations were obtained among the different vigour parameters and high values of broad sense heritability were obtained for all of them. A wide variability in vigour parameters was observed with some clear differences regarding the origin of the plant materials. Thus, the lowest vigour was obtained in sylvestris genotypes from Jaen (Spain) and Marrakech (Morocco) provenances, while the highest vigour was obtained in sylvestris genotypes from Cadiz (Spain) and genotypes from crosses cuspidata x sylvestris. However, a wide variability was obtained in all cases, particularly in genotypes from guanchica origin. Future works will be carried out in order to determine the relationship among vigour parameters measured at different plant age and to define the most efficient strategies for selection at early stages of breeding programs. Funding source: RECUPERA-2020 and contract agreement between IFAPA and Viveros Sevilla (CAICEM15-02).

Virgin olive oil (VOO) represents the primary dietary lipid source in the Mediterranean diet and has been linked to its positive health benefits. VOO metabolites responsible for its nutritional and organoleptic quality include certain components that are transferred almost unchanged from fruit to oil (fatty acids and tocopherols) and other compounds that are synthesized during the oil extraction process by endogenous enzymes from precursors already present in the fruit (volatile and phenolic compounds). The health - promoting characteristics of VOO seems to be related to the high level of oleic acid, an adequate balance of saturated/unsaturated fatty acids, the presence of tocopherols and, specially, the contents of phenolic compounds. Meanwhile, volatile compounds are responsible for its aroma, whereas phenolic compounds are also responsible for the bitter and pungent taste notes of this oil. The final content of these metabolites related to VOO quality is controlled by the levels of expression/activity of the corresponding genes/enzymes involved in their biosynthesis which are cultivar dependent. Given that oil quality is one of the main objectives of the breeding programs aiming at obtaining new olive cultivars, knowledge of the content of these key components for the VOO quality might be very useful for the selection. Moreover, the combination of biochemical and molecular studies would allow to identify the most important genes/enzymes in relation to the oil quality and to know their regulatory mechanisms, so that this knowledge may be used in the molecular marker assisted selection of olive clones with improved oil quality features. For this purpose, the natural variability of the olive species was assessed in a collection of cultivars from the World Olive Germplasm Bank (WOGB) located at Córdoba, and the progeny of a cross of Picual x Arbequina olive cultivars. Analytical data showed that the cultivar collection displayed, in general, a higher level of variability than the progeny cross for most of the metabolites assessed, and that a single cross of olive cultivars might apparently provide enough degree of variability for most of the oil quality traits sought in a breeding program.

The cultivation of olive is widely spread in Croatian coastal area and islands while wild olive (Olea europaea subsp. europaea var. sylvestris) populations can be found only in some specific sites, mainly on islands, where olive cultivation is limited or absent. Ninety trees from four wild olive populations sampled in Brijuni, Pag, Hvar and Lastovo islands and 20 most important Croatian olive cultivars were analyzed by eight microsatellite markers in order to test their diversity and genetic relationships. The SSR markers revealed high level of diversity with a total of 124 alleles and mean values of Ho 0, 811 and He 0, 818. Factorial correspondence analysis (FCA) revealed the separation of the samples into two groups. The first group consisted of cultivated olives and wild olive trees belonging to populations from the islands of Brijuni and Hvar while the second was comprised of wild olives from the islands of Pag and Lastovo. These results suggest that the wild olives from the islands of Pag and Lastovo could be considered as genuine wild olives or true oleasters while the populations from the islands of Brijuni and Hvar probably represent feral forms (olivasters), which may be seedlings of the cultivated olives or products of hybridization between the true oleasters and cultivars. The high diversity found in wild olive trees and the presence of true oleasters may be of great interest for future genetic improvement programs in Croatia?

El conocimiento detallado del germoplasma silvestre de olivo, de su variabilidad genética y distribución, representa una vía alternativa en la mejora de esta especie ofreciendo así nuevos materiales de interés como posibles genitores y/o patrones. Estudios recientes de diversidad genética (realizados con marcadores SSRs, DArTs y cpDNA) y evaluación morfo-agronómica in situ y ex situ de poblaciones silvestres de olivo prospectadas en España indican que dentro del germoplasma silvestre de olivo existe una gran variabilidad que es necesario conservar. En este sentido, como resultado de los trabajos de prospección realizados en los últimos años, se está creando una amplia colección de acebuches y especies afines procedentes de condiciones medio- ambientales diferentes. Dicha colección se está evaluando para varios caracteres morfo-agronómicos de interés, entre ellos para encontrar fuentes de resistencia a la Verticilosis del olivo causada por el hongo de suelo Verticillium dahliae Kleb. Además, para estudiar la conveniencia de los acebuches en el programa de mejora se han comparado dos progenies de polinización libre, una procedente de un acebuche de la provincia de Cádiz y la otra de la variedad 'Picual'. Como era de esperar, en la progenie del acebuche se encontraron frutos de menor tamaño y de bajo contenido en aceite, aunque se observaron amplios rangos de variación para ambos caracteres. No obstante, las plantas de semilla de esta progenie mostraron un periodo juvenil significativamente más corto que la progenie de 'Picual'. Por otro lado, se evaluaron dos progenies de cruzamientos dirigidos entre 'Picual' y dos acebuches que mostraron un periodo juvenil mas corto que cruzamientos entre variedades. Fruto de varios proyectos (Oleagen, RF-2006-0017, RF 2009-00005, RTA 2010-36, PP.PEI.PEI2011.1) nuestros resultados sugieren la potencial utilidad del germoplasma silvestre de olivo en programas de mejora y la necesidad de profundizar en su conservación e investigación.

Maslina je jedana od najduže uzgajanih voćnih kultura. Danas je poznato nekoliko tisuća sorta od kojih se mnoge još uvijek uzgajaju, a veliki ih je broj sačuvan u nacionalnim, regionalnim i internacionalnim kolekcijama. Problem sinonima i homonima je prisutan u svim kolekcijama, a rješenje istih je u pravilnim, točnim i pouzdanim metodama identifikacije. Tradicionalno najkorištenije su kvalitativne morfološke metode koje imaju niz prednosti i nedostataka. Između ostalog, navedene metode su subjektivne i nedovoljno standardizirane. Jedan o mogućih načina riješavanja navedenih nedostataka je u primjeni novih informatičkih dostignuća kao što je analiza fotografije. U dosadašnjim morfološkim istraživanjima, bilo onim kvalitativnim ili kvantitativnim, ne navodi se optimalan broj repeticija, odnosno optimalna veličina uzorka. Cilj ovog istraživanja je statistički odrediti oprimalnu veličinu uzorka, što je preduvjet sigurne i učinkovite karakterizacije. Analizom fotografije i statističkim izračunom upotrebom indeksa pouzdanosti (IP) utvrđen je optimalan broj repeticija. Vrijednost indeksa pouzdanosti se kontinuirano samnjuje s povečanjem broja repeticija. Broj repeticija nakon kojeg više ne dolazi do značajnog smanjenja IP, određen je kao optimalni. Kvantitativne metode karakterizacije imaju veliku perspektivu u morfološkim istraživanjima masline, a optimalan broj repeticija po uzorku je 30.

Most of the breeding works carried out in olive up to date has been based on the current ancient cultivars that have been produced by empiric selection of the growers through centuries. However, the high level of variability found in wild olive populations indicates that they could have genes of interest for obtaining olive cultivars adapted to the new trends of olive growing. In order to test the suitability of wild olives for breeding, we compared the segregation of an open pollination progeny of a wild olive located in a forest in Cadiz, Southern Spain, with a progeny obtained from free pollinization of a ‘Picual’ tree located in the World Olive Germplasm Bank of Cordoba, Spain. As expected, the wild progeny beared fruits of low weight and oil content. However, surprisingly, wild seedlings showed a significant lower juvenile period than the ones coming from ‘Picual’ open pollination progeny. This trait is very important for breeders and, quite likely, has not been a criteria of selection of the current cultivars. Additionally, seedlings from two progenies coming from the cross of ‘Picual’ with two wild olive trees have been evaluated. Fruit characteristics were intermediate between ‘Picual’ seedlings and wild seedlings. However, a great heterosis effect has been observed as those seedlings had much higher height and trunk size than seedlings coming from the cross of two cultivars. Accordingly, the average juvenile period of ‘Picual’ x wild progenies were much lower than the one observed in the progenies of the cross of two cultivars. All these results suggest the utility of wild germplasm in olive breeding that needs to be further investigated.

Wild olive populations are a distinctive element of Mediterranean flora and an important genetic source for resistance to biotic and abiotic stresses in olive selection and breeding. The present study represent the first attempt of in situ agro-morphological evaluation of wild olive trees in Spain. Two provinces of Andalusia (Cadiz and Jaen), Southern Spain, were selected based on previous results on wild olive populations (Belaj et al. 2007). SSR markers have also been used for the study of the genetic diversity and preliminary results are discussed.

Wild olives (Olea europaea subsp. europaea var. sylvestris) represent a distinctive element of Mediterranean flora and an important genetic source for resistance to biotic and abiotic stresses in olive selection and breeding. In this study, genetic diversity among 48 wild genotypes sampled from four different sites from two provinces of Andalusia (Cadiz and Jaen), Southern Spain, was evaluated by means of SSR markers and agro-morphological traits. SSR markers revealed high genetic variability among the genotypes. The cluster analysis based on their polymorphism indicated a certain grouping of wild olives according to their sampling sites. The 20 agro-morphological traits evaluated showed a high variation between genotypes and significant correlation coefficients were obtained among the values recorded in two consecutive harvest seasons, 2007/08 and 2008/09 (r: 0.59-0.78). Traits such as pit shape, fruit symmetry (position A) and oil content on dry basis were very useful in genotype discriminating. The average values obtained in wild olives for fruit size and oil content were lower than the previously reported in cultivated material. However, it is worth mentioning that individuals with fruit weights (1.3 g) and olive oil percentage in dry matter (33.8%) comparable to the values found in some olive cultivars, were also found. With both DNA based and agro morphological descriptors, higher levels of variability were found within each site than between sites. Genetic variation observed among the wild olive germplasm at the DNA level was higher than the agro-morphological traits, indicating the efficiency of SSR markers for detecting genetic diversity among wild olive genotypes and their relationships. Molecular data obtained by SSR markers together with in situ morphological and agronomical characterization of wild olive trees confirmed the high diversity found within the wild populations evaluated. These results indicate the richness of wild olive genetic resources in Andalusia.

En el presente trabajo, se muestran los resultados de la caracterización morfológica y agronómica de 48 individuos procedentes de las provincias de Jaén y Cádiz realizada en las campañas 2007/08 y 2008/09. Se han muestreado 4 localidades en cada una de las dos provincias. Todos los caracteres evaluados presentaron un alto nivel de variabilidad, evidenciándose las diferencias con el olivo cultivado. En cuanto a la caracterización agronómica, cabe destacar una alta correlación (r2 entre 0.78 y 0.52) entre los valores obtenidos en ambas campañas, lo que refuerza el interés de la evaluación in situ de germoplasma de olivo silvestre. Para todos los caracteres estudiados se ha encontrado mayor variabilidad dentro de localidades que entre localidades, con coeficientes de variación entre 4 y 27 . Esto está de acuerdo con los resultados obtenidos con marcadores moleculares (SSR) donde también se ha encontrado mayor variabilidad genética dentro de cada población que entre poblaciones. Los valores medios de las medidas morfológicas llevadas a cabo en, hojas, fruto y endocarpo concuerdan con estudios previos en acebuche. En general, como cabía esperar, los frutos encontrados han sido, de media, más pequeños y con menos rendimiento graso que el olivo cultivado. No obstante hay que indicar que se han encontrado individuos con peso de fruto de 1, 3 g y otros con rendimiento en seco del 33, 8%, comparables a los valores de algunas variedades. Estos resultados indican que dentro del germoplasma de acebuche de Andalucía existe una gran variabilidad que es necesario conservar.

Poznavanje genetske raznolikosti divljih populacija maslina kao i genetskog odnosa između divljih populacija i lokalnih kultivara maslina od velikog su zanimanja za rekonstrukciju povijesti uzgoja maslina. U svrhu analize genetske raznolikosti unutar i između sedam populacija divljih maslina i šest skupina lokalnih kultivara iz dvije važne uzgojne regije (Andaluzija i Levant) u Španjolskoj korišteno je osam mikrosatelitnih biljega. Kultivari su pokazali znatno veću razinu zapažene heterozigotnosti dok su vrijednosti očekivane heterozigotnosti bile veće kod divljih populacija. U dendrogramu po Fitchu i Margoliashu upotrebom matrice genetskih udaljenosti na temelju udjela zajedničkih alela uočene su tri glavne skupine koje uključuju uglavnom divlje masline iz Andaluzije (Skupina I), divlje i kultivirane masline s Levanta (Skupina II), te kultivare iz Andaluzije (Skupina III). Rezultati analize molekularne varijance (AMOVA) su pokazali da je postojala značajna razlika između divljih i kultiviranih maslina iz Andaluzije, a između divljih i kultiviranih s Levanta razlika nije postojala. U skladu s ovim rezultatima, Bayesovska metoda razvrstavanja u skupine na temelju modela (STRUCTURE) omogućila je otkrivanje triju genskih skupova: andalužanski divlji, levantinski i andalužanski kultivirani. Navedeni rezultati mogli bi ukazivati na činjenicu da divlje masline iz Andaluzije predstavljaju oleastre, dok divlje s Levanta uključuju uglavnom feralne oblike nastale iz sjemena kultiviranih maslina ili hibridizacijom između oleastera i kultivara.

Se analizaron 158 individuos, 107 olivos silvestres localizados en las comunidades de Andalucía, Valencia y Catalu&ntilde ; a y 51 variedades locales originarias de estas comunidades aportadas por el Banco Mundial de Germoplasma de Olivo del IFAPA Alameda del Obispo de Córdoba. Se emplearon 8 locus SSR, dando lugar a la amplificación de 132 alelos. El número medio de alelos por locus fue de 15, variando entre 6 y 32. En todo el material se obtuvieron elevados valores de heterocigosidad observada (Ho), diversidad genética (He) y contenido de información polimórfica (PIC). La mayor parte de la diversidad genética (84, 07%), se debió a diferencias entre individuos. Curiosamente las diferencias entre grupos (cultivado vs. silvestre), no fueron significativas, sugiriendo que la diferenciación genética entre ambos no es tan clara.

Wild olives (Olea europaea subsp. europaea var. sylvestris) include true oleasters (corresponding to the wild forms present in natural areas) and feral forms, which may be seedlings of the cultivated clones or result from hybridisation between the true oleasters and olive cultivars. The knowledge of the genetic variability of wild olive populations and the study of their relationships may be of great interest for their possible use as a new source of variability in breeding programmes. For that purpose, eight microsatellite markers have been used to study the genetic variation within and between 11 wild olive populations from Southern and North-Eastern Spain and mainland and insular Italy. Our study uncovered abundant allelic variation over eight loci with an average of 18.5 per locus alleles and high values of observed and expected heterozygosity (mean values of 0.717 and 0.843 respectively). Although most of the genetic diversity was attributable to differences among individuals within populations (91.8%), significant phi-values among populations suggested the existence of genotypic differentiation. The analysis of STRUCTURE allowed the detection of four different gene pools: OS, OI, F1 and F2 despite the overall low interpopulation differentiation of this tree species. The gene pools OS and OI predominantly found in southern Spain and Italian islands, respectively may correspond to genuine wild olive germplasm. The gene pools F1 and F2 detected in North-Eastern regions of Spain and in continental Italy likely represent feral gene pools. The existence of four gene pools and the evidence of differences between the genuinely wild olive germplasm and the feral forms were in overall concordance with the Fitch-Margoliash tree based on Nei's genetic distance.

Genotipizacija kultivara molekularnim tehnikama na razini DNA omogućava preciznu identifikaciju, klasifikaciju i kvantitativnu procjenu genetske raznolikosti. Točna procjena postaje sve bitnija jer sužavanjem genetske raznolikosti može doći i do smanjenja plastičnosti kultura u prilagodbi budućim klimatskim promjenama kao i promjenama u populacijama patogena i u agrikulturnoj praksi. Mikrosateliti ili Pojavljajuće jednostavne sekvence (Single Sequence Repats - SSRs) su kratki segmenti DNA u kojim se specifični motiv sastavljen od jedne do šest nukleotidnih baza ponavlja. Ove regije su česte, javljaju se nasumično po biljnom genomu i obično pokazuju veliku varijabilnost između jedinki, populacija i vrsta. Da bi se koristili kao molekularni biljezi, mikrosatelitni lokusi se pojedinačno amplificiraju pomoću lančane reakcije polimerazom (PCR) koristeći oligonukleotidne početnice (primers) specifičnima za sekvence DNA koje omeđuju sekvencu SSR. Polimorfizam u dužini amplificiranih fragmenata uzrokovan je razlikama u broju ponavljanja motiva. Imajući na umu velik broj dostupnih molekularnih sustava biljega, mikrosateliti su vrlo dobar izbor za različite svrhe, budući da su specifični za određeni lokus, kodominantni, vrlo polimorfni i ponovljivi. Kod nekih biljnih vrsta kao što su kukuruz, riža ili Arabidopsis, prikupljena je velika kočina podataka o sekvencama DNA tako da se mikrosateliti mogu lako identificirati pretraživanjem baza podataka. No, budući da su mikrosateliti uglavnom specifični za određenu vrstu, za vrste kod kojih nema informacija o mikrosatelitnim početnicama, razvitak novih SSR analizom genomskih knjižnica DNA obogaćenima s jednim ili više ponavljajućih motiva iziskuje naporan rad uz relativno visoke troškove. U ovom je radu dan pregled osnovnih statističkih metoda analize mikrosatelitnih podataka. Raspravljena je informativnost mikrosatelitnih biljega kao i upotreba metoda genetske udaljenosti temeljenih na različitim modelima mutacije mikrosatelita. Dani su primjeri klasifikacije kultivara kod kukuruza, hmelja i maslina koji predstavljaju različite pristupe analizi mirkosatelitnih podataka.

RAPD markers were used for the study of 19 Albanian olive cultivars and two wild olives (oleasters). A total of 76 polymorphic bands (4.8 polymorphic markers per primer) out of 107 reproducible were obtained using 16 primers. The number of bands per primer ranged from 4 to 10, whereas the number of polymorphic bands ranged from 1 to 9, corresponding to 71% of the total amplification products. All the accessions could be identified by the combination of four primers: OPA-19 ; OPA-02 ; OPK-16 and OPP-19. The dendrogram, based on Jaccard's index, included three major groups according to their origin: 1) most of the cultivars from the area of Berat (South of Albania) 2) cultivars from the Centre and Centre-North of Albania and 3) cultivars from the Centre and North-West of Albania along with the oleaster from Elbasan. In order to evaluate the origin of Albanian cultivars they were compared to those diffused in other countries like Greece, Italy and Turkey, due to geographical and historical affinity among these countries, by using a one way AMOVA. Although most of the genetic diversity was attributable to differences among cultivars within each country (91.47%) significant φ -values among countries (φ st = 0.085 ; p < 0.001) suggested the existence of RAPD phenotypic differentiation. Significant φ -values in all pairs formed by Albania with the other countries were observed. These results are consistent with the autochthonous origin of Albanian cultivars.

El olivo (Olea europaea L.) es uno de los cultivos más antiguos de la cuencamediterránea y cuenta con un gran patrimonio genético. Sin embargo, latendencia actual en la mayoría de los países olivareros es hacia la utilizaciónde pocas variedades muy populares. Por tanto la conservación de estadiversidad genética resulta de vital importancia para evitar su pérdida. Durantelas últimas décadas, se han realizado importantes trabajos de prospección,recolección, caracterización y evaluación de las variedades más importantes deolivo. La conservación y el estudio de los recursos genéticos del olivo implicanel establecimiento de Bancos de Germoplasma, basados en colecciones encampo, como ocurre en el resto de especies frutales. El primer aspecto a teneren cuenta en el manejo de estas colecciones es la correcta identificación delmaterial existente, así como de las entradas que se van incorporando demanera sucesiva. En el caso del olivo, la presencia recurrente de sinonimias (lamisma variedad con distintos nombres) y homonimias (distintas variedades conel mismo nombre), hace que se acentúe la necesidad de una correctaidentificación varietal.El Banco de Germoplasma Mundial de Olivo (BGMO) de Córdoba es uno de losdos bancos de referencia Internacional. Incluye más de 450 cultivares de 17países distintos, lo que representa un porcentaje elevado de la variabilidad dela especie en la cuenca mediterránea. La correcta identificación de ésta y otrascolecciones de germoplasma de olivo así como el estudio de la variabilidadgenética son principalmente realizadas por marcadores moleculares. Tambiéntiene un creciente interés la utilidad de los mismos en la mejora genética. En elpresente trabajo se han utilizado tres tipos de marcadores moleculares (SSRs,SNPs y DArTs) para la identificación, estudios de variabilidad genética y mapeoen germoplasma asociado al BGMO.En un primer apartado se ha realizado la identificación de 14 nuevasvariedades argelinas, recientemente introducidas en el BGMO, con marcadoresmoleculares (SSRs y SNPs). La comparación de los perfiles de estosmarcadores con 5 variedades de Argelia, Túnez y Marruecos, previamente incluidas en el BGMO, indicó que estas variedades difieren de ellas y del restoy su introducción enriquecerá el BGMO con material vegetal de este país.Aunque se han detectado algunos casos de homonimias en las variedadesargelinas, la alta variabilidad genética observada es un indicio de la riqueza delsu germoplasma.Posteriormente se ha descrito la puesta a punto de los primeros marcadoresmoleculares de alto rendimiento para el olivo, los denominados DArTs. Estosmarcadores no necesitan un conocimiento previo de la secuencia de ADN.Aunque son dominantes, tienen una alta reproductividad (99,8%) y un bajocoste. Se han utilizado para identificar de manera inequívoca una muestra de62 variedades pertenecientes al BGMO. El dendograma construido en base aello ha confirmado su agrupación por origen geográfico, previamenteobservada con otros marcadores moleculares. También se ha demostrado lautilidad de estos marcadores de alto rendimiento en la construcción de unmapa de ligamiento en una progenie de las variedades ‘Picual’ y ‘Arbequina’.Se han obtenido un total de 23 grupos de ligamiento para cada parental.Además algunos microsatélites, también incluidos en dicho mapa, hanpermitido establecer relaciones entre grupos de ligamiento de ambosparentales.Por último, la variabilidad existente en el BGMO se ha evaluado conmarcadores DArT, microsatélites, SNPs y características agronómicas, paraconstruir la primera colección nuclear de dicho Banco. Se han obtenido cincocolecciones nucleares, que contenían desde 18 hasta 68 variedades. De todasellas, la colección nuclear de 68 variedades parece la más indicada paraestudios de conservación, dado que retenía todos los alelos y caracteresanalizados. Por otro lado, la colección nuclear de 36 variedades se reveló comola más adecuada para estudios de mejora, dada la elevada distancia genéticamedia y la buena representación de las distintas regiones del Mediterráneo enun número relativamente pequeño de variedades, Olive (Olea europaea L.) is one of the oldest trees cultivated in theMediterranean Basin and it includes a high genetic patrimony. However, in mostolive growing countries, olive orchards are composed by a very reducednumber of well known cultivars. In this sense, the preservation of olive geneticpatrimony is of vital importance for avoiding the possible erosion. During the lastdecades, considerable prospections, recollections, characterization andevaluation surveys, have been performed on the main olive cultivars.Similarly to other fruir species, the conservation and the study of olive geneticresources implies, the establishment of ‘ex situ’ collections (Germplasm Banks).The identification of existing plant material as well as of new accessionscontinously introduced in the collections is a priority task for their correctmanagement. In olive, the presence of many synonyms (the same variety withdifferent names) and homonyms (different varieties with the same name) makesthe identification highly needed.The World Olive Germplasm Bank (WOGB) in Cordoba, is one of the twointernational collections of reference. It includes more than 450 cultivars from 17different countries thus representing a high percentage of the variability ofspecies in the Mediterranean Basin.Molecular markers have mostly been used in olive for correct identification ofgermplasm collections as well as for genetic diversity studies. Their utility forolive breeding has being of great interest. In the present work, three types ofmolecular markers (SSRs, SNPs and DarTs) have been used for identification,mapping studies and genetic variability of plant material related to the WOGB.The first part of this work deals with the identification of 14 new Algeriancultivars, recently introduced into the WOGB, by means of SSR and SNPmarkers. The comparison of their molecular profiles with 5 existing cultivars ofAlgerian, Tunisian and Marocco origin showed that they were different,enriching so the WOGB with plant material from this country. Our study showedthe great variability of Algerian germplasm as well as some cases ofhomonyms.The setting up of the first high throughput marker for olive (DArTs) has beenfurther described. These markers do not require prior sequence information.They are dominant makers, with a high reproducibility (99,8%) and lower costcompared to other markers. The use of these markers for the correctidentification of 62 cultivars belonging to WOGB has been evaluated. In additionthese markers have been differentiated 62 cultivars according to theirgeographic origin, separating Eastern and Western varieties from theMediterranean ones.The utility of DArT markers for the development of a linkage map in oneprogeny of Picual’ x ‘Arbequina’cultivars has been confirmed. A total of 23linkage groups have been obtained per each parent. Besides, some of the SSRused allowed the stablishment of some connections with several linkage groupsof both parents.Finally, the existing genetic variability in the WOGB has been evaluated bymeans of DarTs, SSRs, SNPs and agronomical traits in order to develop thefirst core collection. Five core collections including from 18 to 68 cultivars wereobtained. From these core sets, the one with 68 cultivars seems the mostappropriate for conservation studies as it retained all the alleles and traits understudy. On the other hand, the core 36 was found to be the most indicated onefor breeding given the high genetic distance and the good representativeness ofthe Mediterranean regions in a realitvely small number of cultivars.