Population genetics of Mediterranean and Saharan olives: geographic patterns of differentiation and evidence for early generations of admixture

Background and Aims

The olive (Olea europaea subsp. europaea) was domesticated in the Mediterranean area but its wild relatives are distributed over three continents, from the Mediterranean basin to South Africa and south-western Asia. Recent studies suggested that this crop originated in the Levant while a secondary diversification occurred in most westward areas. A possible contribution of the Saharan subspecies (subsp. laperrinei) has been highlighted, but the data available were too limited to draw definite conclusions. Here, patterns of genetic differentiation in the Mediterranean and Saharan olives are analysed to test for recent admixture between these taxa.

Methods

Nuclear microsatellite and plastid DNA (ptDNA) data were compiled from previous studies and completed for a sample of 470 cultivars, 390 wild Mediterranean trees and 270 Saharan olives. A network was reconstructed for the ptDNA haplotypes, while a Bayesian clustering method was applied to identify the main gene pools in the data set and then simulate and test for early generations of admixture between Mediterranean and Saharan olives.

Key Results

Four lineages of ptDNA haplotypes are recognized: three from the Mediterranean basin and one from the Sahara. Only one haplotype, primarily distributed in the Sahara, is shared between laperrinei and europaea. This haplotype is detected once in ‘Dhokar’, a cultivar from the Maghreb. Nuclear microsatellites show geographic patterns of genetic differentiation in the Mediterranean olive that reflect the primary origins of cultivars in the Levant, and indicate a high genetic differentiation between europaea and laperrinei. No first-generation hybrid between europaea and laperrinei is detected, but recent, reciprocal admixture between Mediterranean and Saharan subspecies is found in a few accessions, including ‘Dhokar’.

Conclusions

This study reports for the first time admixture between Mediterranean and Saharan olives. Although its contribution remains limited, Laperrine''s olive has been involved in the diversification of cultivated olives.     

Occurrence of Plant-Parasitic Nematodes of Turfgrass in Korea

Plant-parasitic nematodes are not only an important constraint on agricultural crop production, but also cause both direct and indirect damage to turfgrass, which is a ground cover plant. However, studies on plant-parasitic nematodes of turfgrass in Korea are scarce. A survey for plant-parasitic nematodes was carried out on 13 golf courses in Korea. The results yielded 28 species/taxa belonging to 16 genera and 12 families of plant-parasitic nematodes. Among the isolated species, Helicotylenchus microlobus, Mesocriconema nebraskense, Tylenchorhynchus claytoni, Mesocriconema sp., and Meloidogyne graminicola were the most prevalent species in all management zones. Twelve species were new records of plant-parasitic nematodes in Korea. Highest maximum densities were showed by T. claytoni, Paratylenchus nanus, M. nebraskense, M. graminicola, and H. microlobus. Diversity (H′), was significantly higher in fairways compared to tees and greens, though species evenness (J′) and dominance (D) showed no statistically significant differences. This information is crucial in nematode problem diagnosis, and the subsequent formulation of management strategies.     

On the origins and spread of Olea europaea L. (olive) domestication: evidence for shape variation of olive stones at Ugarit,Late Bronze Age,Syria—a window on the Mediterranean Basin and on the westward diffusion of olive varieties

Charred archaeological stones of Olea europaea L. (olive) from Late Bronze Age Ugarit, Syria, were analyzed with geometric morphometry and compared with a morphological differentiation model established on the basis of analyses of modern spontaneous (uncultivated) olive populations and cultivated varieties of various origins within the Mediterranean Basin. The results allow a reinterpretation of the east–west morphological diversity previously observed in wild olives. The archaeobotanical data were compared in detail to the partly geographically structured modern morphological diversity of the cultivated olive. Ancient morphotypes could be distinguished, among which one is dominant in the assemblage. Their diffusion from east to west is shown, and their time of arrival in the northwestern Mediterranean can be evaluated by comparison to archaeological material from that area. Combining morphometric and genetic data, modern reference and archaeological material also guides us in understanding the mechanisms that prevailed in the long-term agrobiodiversity of the olive.     

Centennial olive trees as a reservoir of genetic diversity

Background and Aims

Genetic characterization and phylogenetic analysis of the oldest trees could be a powerful tool both for germplasm collection and for understanding the earliest origins of clonally propagated fruit crops. The olive tree (Olea europaea L.) is a suitable model to study the origin of cultivars due to its long lifespan, resulting in the existence of both centennial and millennial trees across the Mediterranean Basin.

Methods

The genetic identity and diversity as well as the phylogenetic relationships among the oldest wild and cultivated olives of southern Spain were evaluated by analysing simple sequence repeat markers. Samples from both the canopy and the roots of each tree were analysed to distinguish which trees were self-rooted and which were grafted. The ancient olives were also put into chronological order to infer the antiquity of traditional olive cultivars.

Key Results

Only 9·6 % out of 104 a priori cultivated ancient genotypes matched current olive cultivars. The percentage of unidentified genotypes was higher among the oldest olives, which could be because they belong to ancient unknown cultivars or because of possible intra-cultivar variability. Comparing the observed patterns of genetic variation made it possible to distinguish which trees were grafted onto putative wild olives.

Conclusions

This study of ancient olives has been fruitful both for germplasm collection and for enlarging our knowledge about olive domestication. The findings suggest that grafting pre-existing wild olives with olive cultivars was linked to the beginnings of olive growing. Additionally, the low number of genotypes identified in current cultivars points out that the ancient olives from southern Spain constitute a priceless reservoir of genetic diversity.     

High Genetic Diversity Detected in Olives beyond the Boundaries of the Mediterranean Sea

Background

Olive trees (Olea europaea subsp. europaea var. europaea) naturally grow in areas spanning the Mediterranean basin and towards the East, including the Middle East. In the Iranian plateau, the presence of olives has been documented since very ancient times, though the early history of the crop in this area is shrouded in uncertainty.

Methods

The varieties presently cultivated in Iran and trees of an unknown cultivation status, surviving under extreme climate and soil conditions, were sampled from different provinces and compared with a set of Mediterranean cultivars. All samples were analyzed using SSR and chloroplast markers to establish the relationships between Iranian olives and Mediterranean varieties, to shed light on the origins of Iranian olives and to verify their contribution to the development of the current global olive variation.

Results

Iranian cultivars and ecotypes, when analyzed using SSR markers, clustered separately from Mediterranean cultivars and showed a high number of private alleles, on the contrary, they shared the same single chlorotype with the most widespread varieties cultivated in the Mediterranean.

Conclusion

We hypothesized that Iranian and Mediterranean olive trees may have had a common origin from a unique center in the Near East region, possibly including the western Iranian area. The present pattern of variation may have derived from different environmental conditions, distinct levels and selection criteria, and divergent breeding opportunities found by Mediterranean and Iranian olives.These unexpected findings emphasize the importance of studying the Iranian olive germplasm as a promising but endangered source of variation.     

Agistemus aimogastaensis sp. n. (Acari,Actinedida, Stigmaeidae), a recently discovered predator of eriophyid mites Aceria oleae and Oxycenus maxwelli,in?olive orchards in Argentina

A new species, Agistemus aimogastaensis, is described with the aid of optical and Scanning Electron Microscopy. This mite is an important predator of two eriophyid mites (Aceria oleae and Oxycenus maxwelli) in olive orchards (Olea europaea, variety Arauco) in La Rioja Province. The problems related to eriophyids in olive orchards in Argentina are highlighted and photos of the damage on leaves and fruit are included.     

Soil Properties and Olive Cultivar Determine the Structure and Diversity of Plant-Parasitic Nematode Communities Infesting Olive Orchards Soils in Southern Spain

This work has studied for the first time the structure and diversity of plant-parasitic nematodes (PPNs) infesting olive orchard soils in a wide-region in Spain that included 92 locations. It aims at determining which agronomical or environmental factors associated to the olive orchards are the main drivers of the PPNs community structure and diversity. Classical morphological and morphometric identification methods were used to determine the frequency and densities of PPNs. Thirteen families, 34 genera and 77 species of PPNs were identified. The highest diversity was found in Helicotylenchus genus, with six species previously reported in Spain and with H. oleae being a first report. Neodolichorhynchus microphasmis and Diptenchus sp., Diphtherophora sp., and Discotylenchus sp., usually considered fungal feeders, were also reported for the first time associated to olive rhizosphere. PPNs abundance ranged from 66 to 16,288 individuals/500-cm³ of soil with Helicotylenchus digonicus being the most prevalent species, followed by Filenchus sp., Merlinius brevidens and Xiphinema pachtaicum. Nematode abundance and diversity indexes were influenced by olive cultivar, and orchard and soil management practices; while olive variety and soil texture were the main factors driving PPN community composition. Soil physicochemical properties and climatic characteristics most strongly associated to the PPN community composition included pH, sand content and exchangeable K, and maximum and minimum average temperature of the sampled locations. Our data suggests that there is a high diversity of PPNs associated to olive in Southern Spain that can exert different damage to olive roots depending on the olive variety and their abundance. Further analysis to determine the resistance levels of most common olive varieties to the prevalent PPNs in Spain will help to choose the most appropriate ones for the establishment of new plantations. This choice will take into consideration the specific soils and environments where those olive varieties will be established.     

UHPLC-DAD-FLD and UHPLC-HRMS/MS based metabolic profiling and characterization of different Olea europaea organs of Koroneiki and Chetoui varieties

The olive tree (Olea europaea L., Oleaceae) is one of the most important fruit trees in Mediterranean basin and has been associated with numerous biological assets. These effects have been mainly attributed to certain phenolic compounds found in fruits, olive oil and by-products of olive oil production. However, other Olea organs such as stems, roots and drupe stones have received little attention leading to limited knowledge about their phytochemical content. Thus, the main goal of the current study was the investigation of the chemical composition of diverse organs from two O. europaea varieties (i.e. Koroneiki and Chetoui) using combinations of modern analytical techniques. A fast UHPLC-DAD-FLD method was developed and applied for the profiling of different extracts of O. europaea organs as well as for the quantification of oleuropein. In addition, a dereplication strategy was developed using an Orbitrap platform (UHPLC-ESI-HRMS/MS) aiming to further characterization of the contained secondary metabolites. In total, 86 molecules were identified including compounds described for the first time in O. europaea such as coumarins. Some compounds were found to be organ specific such as nuzhenide derivatives in stone, flavonoids in leaves and oleuropein which was mainly found in Olea roots, in both varieties. Overall, it is noticeable that except olive oil, diverse organs of olive tree might comprise an alternative and valuable source of biologically active compounds.     

<Emphasis Type="Italic">Olea europaea</Emphasis> (Oleaceae) phylogeography based on chloroplast DNA polymorphism

Chloroplast DNA diversity in the olive (Olea europaea L.) complex was studied using PCR-RFLP and microsatellite markers. Fifteen chlorotypes were distinguished. We constructed a cpDNA phylogenetic tree in which five clades were recognised and located in distinct geographic areas: clade A in Central and Southern Africa, clade C in Asia, clade M in North-West Africa, clade E1 in the Mediterranean Basin and Sahara, and clade E2 in West Mediterranea. Cultivated olive clustered with Mediterranean and Saharan wild forms (clades E1 and E2). Strong genetic differentiation for cpDNA markers was observed between eastern and western Mediterranean olives, suggesting that these areas have represented different glacial refugia. Humans most likely spread one eastern chlorotype, preponderant in cultivars, across the western Mediterranean Basin. Its presence in O. e. subsp. laperrinei from the Sahara suggests a possible Mediterranean olive origin in an African population, which may have overlapped in the Southern Mediterranean during the Quaternary.     

Nondestructive determination of nitrogen and chlorophyll content in olive tree leaves and the relation with photosynthesis and fluorescence parameters

For Tunisian olive tree orchards, nitrogen deficiency is an important nutritional problem, in addition to the availability of water. Establishment of relationships between nutrients such as nitrogen and ecophysiological parameters is a promising method to manage fertilisation at orchard level. Therefore, a nitrogen stress experiment with one-year-old olive trees (Olea europaea L. ‘Koroneiki’ and ‘Meski’) was conducted with trees respectively subjected to four nitrogen supply regimes (23.96 meq l⁻¹, 9.58 meq l⁻¹, 4.79 meq l⁻¹ and 0 meq l⁻¹ NO₃ ⁻¹).     

Plant-parasitic Nematode Communities and Their Associations with Soil Factors in Organically Farmed Fields in Minnesota

A survey was conducted to determine the assemblage and abundance of plant-parasitic nematodes and their associations with soil factors in organically farmed fields in Minnesota. A total of 31 soil samples were collected from southeast (SE), 26 samples from southwest (SW), 28 from west-central (WC), and 23 from northwest (NW) Minnesota. The assemblage and abundance of plant-parasitic nematodes varied among the four regions. The soybean cyst nematode, Heterodera glycines, the most destructive pathogen of soybean, was detected in 45.2, 88.5, 10.7, and 0% of organically farmed fields with relative prominence (RP) values of 10.3, 26.5, 0.6, and 0 in the SE, SW, WC, and NW regions, respectively. Across the four regions, other common genera of plant-parasitic nematodes were Helicotylenchus (42.6, RP value, same below), Pratylenchus (26.9), Tylenchorhynchus and related genera (9.4), Xiphinema (5.6), and Paratylenchus (5.3). Aphelenchoides, Meloidogyne, Hoplolaimus, Mesocriconema, and Trichodorus were also detected at low frequencies and/or low population densities. The similarity index of plant-parasitic nematodes between two regions ranged from 0.44 to 0.71 and the similarity increased with decreasing distance between regions. The densities of most plant-parasitic nematodes did not correlate with measured soil factors (organic matter, pH, texture). However, the densities of Pratylenchus correlated negatively with % sand, and Xiphinema was correlated negatively with soil pH.     

The age of monumental olive trees (Olea europaea) in northeastern Spain

Trees can reach ages that in some cases amount to thousands of years. In the Mediterranean region, olive trees (Olea europaea) have traditionally been considered a particularly long-lived species. The main objective of this study was to assess the age of large olive trees considered to be millenarian and classified as monumental trees in northeastern Spain. We extracted cores of 14 individuals and obtained 8 sections of trees which had already been cut in the area where the largest olive trees in the northeastern Iberian Peninsula are found. The age of the sampled olive trees was assessed by counting the number of annual growth rings. Tree rings did not cross-date well, neither within nor between individuals, but boundaries between likely annual rings were clearly distinct. We found a linear relationship between DBH and tree age (in years) (Age = 2.11 × diameter(cm) + 88.93, R² = 0.80), which was used to estimate the age of unsampled olive trees. The maximum estimated age (627 ± 110 years) is among the greatest ages reported for olive trees around the world (700 years) and among the oldest trees in Mediterranean ecosystems.     

Substantial genetic diversity in cultivated Moroccan olive despite a single major cultivar: a paradoxical situation evidenced by the use of SSR loci

To assess the genetic diversity in Moroccan cultivated olive, Olea europaea L. subsp. europaea, we performed molecular analysis of olive trees sampled in four geographic zones representing all areas of traditional olive culture. The analysis of 215 trees using 15 simple sequence repeat (SSR) loci revealed 105 alleles distributed among 60 SSR profiles. The analysis of chloroplast deoxyribonucleic acid polymorphism for these 60 olive genotypes allowed to identify four chlorotypes: 42 CE1, one CE2, nine COM1 and eight CCK. Among the 60 SSR profiles, 52 corresponded to cultivated olive trees for which neither denomination nor characterisation is available. These local olive genotypes displayed a spatial genetic structuring over the four Moroccan geographic zones (northwest, north centre, Atlas and southwest), as pairwise Fst values ranged from 0.0394 to 0.1383 and varied according to geographic distance. As single alleles detected in local olive were also observed in Moroccan oleaster populations, results suggest that plant material was mainly selected from indigenous populations. The assumption that Picholine marocaine cultivar is a multi-clonal cultivar was not supported by our data because we found a single genotype for 112 olive trees representing 31 to 93% of the olives sampled locally in the 14 different areas. Picholine marocaine and the few other named cultivars do not seem to belong to the same gene pools as the unnamed genotypes cultivated only locally. The situation is paradoxical: a substantial genetic diversity in Moroccan olive germplasm, probably resulting from much local domestication, but a single cultivar is predominant.     

Management to control citrus greening alters the soil food web and severity of a pest–disease complex

Since 2005, the Florida citrus industry has faced the need to control the devasting bacterial disease huanglongbing (HLB). Advanced production systems (APS) were designed to grow citrus trees more quickly than conventional citriculture (CC) methods in order to mitigate the impact of HLB. Daily fertigation required by APS produces changes in the soil physical–chemical properties compared to those in conventionally managed orchards. We used real-time PCR in an ongoing field experiment to compare the effects of APS and CC on more than a dozen metazoan and microorganism species in soil food webs that affect larvae of a major arthropod pest of citrus, Diaprepes abbreviatus. Soil chemical properties, citrus performance, weevil occurrence, and abundance of free-living and plant-parasitic nematodes were also evaluated. The effects of polypropylene mulch that provides a barrier to soil entry by D. abbreviatus larvae were also investigated in each of the two cultural systems. Trees grew significantly larger in APS and mulching increased tree growth and reduced tree mortality, thereby increasing fruit yield per ha. APS increased the fruit yield in 2011; however, by 2013 the number of fruit per tree was not affected by any of the treatments. Root mass density increased in APS, but decreased under mulch. The numbers of plant-parasitic and free-living nematodes and some natural enemies of nematodes such as Catenaria sp. and Paecilomyces lilacinus were more abundant in the treatments with greater root mass density. Both organisms in the D. abbreviatus–Phytophthora nicotianae pest–disease complex were more abundant in APS than in CC, whereas fewer steinernematid entomopathogenic nematodes (EPNs) that prey on insect larvae occurred in APS. By contrast, heterorhabditid EPNs tended to be more numerous in APS than in CC, although they comprised <25% of the EPN communities in any treatment. Major differences between APS and CC in almost all of the measured physical and chemical soil properties provide a basis for controlled studies to understand why EPN taxa responded differently to these treatments and how APS and soils generally might be modified to conserve the beneficial activities of nematodes.     

Effect of AMF application on growth, productivity and susceptibility to Verticillium wilt of olives grown under desert conditions

The effect of arbuscular mycorrhizal fungi (AMF) on olive (Olea europaea) growth and development was followed for 4 years after transplanting in irrigated commercial orchards under arid conditions. Sites I and II were irrigated with saline water (EC?=?4.5 dS/m). In site I, the soil was infested with Verticillium dahliae and olive varieties ‘Picual’ (Verticillium susceptible) and ‘Barnea’ (relatively Verticillium tolerant) were tested. In site II, the soil was virgin soil (previously non-cultivated soil) and olive varieties ‘Souri’ and ‘Barnea’ were tested. Plants for all sites were inoculated in the nursery with Glomus intraradices alone or in a mixture with G. mosseae. Relative to non-inoculated trees, AMF colonization enhanced vegetative growth, expressed as tree height and trunk circumference, at all sites. At first commercial harvest, AMF-treated trees had higher fruit and oil yields than non-mycorrhitic controls. Under saline water irrigation, differences between inoculated and non-inoculated treatments were reduced in the slow-growing ‘Souri’ but remained apparent in the modern fast-growing ‘Barnea’. AMF colonization did not appear to improve tolerance of either ‘Picual’ or ‘Barnea’ to V. dahliae, and both were more susceptible than the non-inoculated controls. Thus inoculation of olive plants with AMF improves transplant growth and adaptation in arid areas during the first 3 years of growth and until the first commercial harvesting season.     

Host Suitability of the Olive Cultivars Arbequina and Picual for Plant-Parasitic Nematodes

Host suitability of olive cultivars Arbequina and Picual to several plant-parasitic nematodes was studied under controlled conditions. Arbequina and Picual were not suitable hosts for the root-lesion nematodes Pratylenchus fallax, P. thornei, and Zygotylenchus guevarai. However, the ring nematode Mesocriconema xenoplax and the spiral nematodes Helicotylenchus digonicus and H. pseudorobustus reproduced on both olive cultivars. The potential of Meloidogyne arenaria race 2, M. incognita race 1, and M. javanica, as well as P. vulnus and P. penetrans to damage olive cultivars, was also assessed. Picual planting stocks infected by root-knot nematodes showed a distinct yellowing affecting the uppermost leaves, followed by a partial defoliation. Symptoms were more severe on M. arenaria and M. javanica-infected plants than on M. incognita-infected plants. Inoculation of plants with 15,000 eggs + second-stage juveniles/pot of these Meloidogyne spp. suppressed the main height of shoot and number of nodes of Arbequina, but not Picual. Infection by each of the two lesion nematodes (5,000 nematodes/pot) or by each of the three Meloidogyne spp. suppressed (P < 0.05) the main stem diameter of both cultivars. On Arbequina, the reproduction rate of Meloidogyne spp. was higher (P < 0.05) than that of Pratylenchus spp.; on Picual, Pratylenchus spp. reproduction was higher (P < 0.05) than that of Meloidogyne spp.     

Bacterial and β-proteobacterial diversity in Olea europaea var. mastoidis- and O. europaea var. koroneiki-generated olive mill wastewaters: influence of cultivation and harvesting practice on bacterial community structure

Despite the fact that several approaches have been applied for the bioremediation of olive mill wastewaters, little information is available on bacteria inhabiting these agro-industrial effluents. In the present study, 16S rRNA gene clone libraries were constructed to identify bacterial diversity in olive-oil mill wastewaters generated by two olive varieties, Olea europaea var. mastoidis and O. europaea var. koroneiki. Due to chloroplast excess in wastewater produced from the processing of O. europaea var. koroneiki, a clone library using specific PCR primers for β-Proteobacteria was further constructed. The bacterial diversity in O. europaea var. mastoidis-generated olive mill wastewaters consisted mainly of members of Acetobacteriaceae, Prevotellaceae and Lactobacillaceae, while the majority of β-proteobacteria identified in O. europaea var. koroneiki-generated olive mill wastewaters were placed within the families Comamonadaceae, Oxalobacteraceae, Hydrogenophilaceae and Rhodocyclaceae. At least 17 novel phylogenetic linkages among Bacteria were identified. Olive-oil mill wastewaters microbiota appears to have originated from soil and freshwater environments, while the cultivation and harvesting practice highly influenced the bacterial community structure in olive mill wastewaters. The presence of fecal bacteria in O. europaea var. mastoidis-generated olive mill wastewaters, due to the long harvesting period, should be of concern.     

Arbuscular mycorrhizal fungus <Emphasis Type="Italic">Rhizophagus irregularis</Emphasis> influences key physiological parameters of olive trees (<Emphasis Type="Italic">Olea europaea</Emphasis> L.) and mineral nutrient profile

In this study, we hypothesized that colonization of olive trees (Olea europaea L.) with the arbuscular mycorrhizal fungus Rhizophagus irregularis could modify the profiles of rhizosphere microbial communities with subsequent effects on nutrient uptake that directly affects olive tree physiology and performance. In this context, a greenhouse experiment was carried out in order to study the effects of mycorrhizal colonization by R. irregularis on photosynthesis, pigment content, carbohydrate profile, and nutrient uptake in olive tree. After six months of growth, photosynthetic rate in mycorrhizal (M) plants was significantly higher than that of nonmycorrhizal plants. A sugar content analysis showed enhanced concentrations of mannitol, fructose, sucrose, raffinose, and trehalose in M roots. We also observed a significant increase in P, K, Ca, Mg, Zn, Fe, and Mn contents in leaves of the M plants. These results are important, since nutrient deficiency often occurs in Mediterranean semiarid ecosystems, where olive trees occupy a major place.     

Ruscus hypophyllum: A New Host for Aphelenchoides fragariae

Aphelenchoides fragariae was isolated from the phylloclades of the ornamental plant Ruscus hypophyllum (Liliaceae). Rotylenchus buxophilus, Scutellonema brachyurum, and Meloidogyne were identified as the most common plant-parasitic nematodes in the soil near the roots. The pathology and life history of A. fragariae were closely related to the climate. To our knowledge, this is the first report of R. hypophyllum as a host of plant-parasitic nematodes.