Wild Olive Trees Research Articles

Temporal matching among diurnal photosynthetic patterns within the crown of the evergreen sclerophyll Olea europaea L.

Trees are modular organisms that adjust their within-crown morphology and physiology in response to within-crown light gradients. However, whether within-plant variation represents a strategy for optimizing light absorption has not been formally tested. We investigated the arrangement of the photosynthetic surface throughout one day and its effects on the photosynthetic process, at the most exposed and most sheltered crown layers of a wild olive tree (Olea europaea L.). Similar measurements were made for cuttings taken from this individual and grown in a greenhouse at contrasted irradiance-levels (100 and 20% full sunlight). Diurnal variations in light interception, carbon fixation and carbohydrate accumulation in sun leaves were negatively correlated with those in shade leaves under field conditions when light intensity was not limiting. Despite genetic identity, these complementary patterns were not found in plants grown in the greenhouse. The temporal disparity among crown positions derived from specialization of the photosynthetic behaviour at different functional and spatial scales: architectural structure (crown level) and carbon budget (leaf level). Our results suggest that the profitability of producing a new module may not only respond to construction costs or light availability, but also rely on its spatio-temporal integration within the productive processes at the whole-crown level.

Microbial communities associated with the root system of wild olives (Olea europaea L. subsp. europaea var. sylvestris) are good reservoirs of bacteria with antagonistic potential against Verticillium dahliae

Wild olive trees, namely oleaster, are considered the ancestor of cultivated olive and a unexplored source of genetic variability that might contain important traits of agronomic and biotechnological interest. The longevity and genetic diversity of oleasters may have favoured selection of specific and well adapted rhizosphere microbial populations that can constitute unique reservoirs of microbial antagonists of Verticillium dahliae, the main soilborne fungal pathogen of olive worldwide. The objective of this present study was to determine the structure and diversity of bacterial communities in the rhizosphere and endosphere of oleaster from 11 havens in Cadiz and Cordoba provinces of Andalusia, southern Spain. To carry out the study we used a multiphasic approach. First, the occurrence and diversity of rhizosphere bacteria was monitored by a cultivation-independent-approach, using fluorescent terminal restriction fragment length polymorphism (FT-RFLP) analyses of amplified 16S rDNA sequences. FT-RFLP patterns revealed a high heterogeneity in the composition of the sampled rhizosphere bacterial communities and suggested the existence of plant genotype-site-specific communities, with each oleaster haven being a unique reservoir of bacterial diversity. Secondly, to investigate the antagonistic potential of these root-associated bacterial populations, a total of 675 bacterial isolates obtained from oleaster rhizosphere and endosphere were screened by dual testing for inhibition of in vitro growth of the highly virulent, olive defoliating pathotype of V. dahliae. Out of 675 tested bacterial isolates, 94 (14%) showed a strong antagonistic activity against a defoliating V. dahliae pathotype. Of the antagonistic bacteria, a slightly lower proportion (12.9% of total bacteria) were inhabitant of the oleaster rhizosphere compared to that in the endosphere (16.5%). The biotechnological potential of those isolates was assessed by in vitro production of different hydrolytic enzymes, indole-1.3-acetic acid (IAA), siderophores, and antimicrobial compounds. Overall, most of bacterial antagonists (58.5 to 78.3%) showed proteolytic, lipolytic, and chitinolytic activity, and produced IAA and siderophores. Finally, analysis of the 16S rDNA gene sequence indicated that most of the 94 bacterial antagonists belong to genera Bacillus (56.4%), Pseudomonas (27.7%), and Paenibacillus (7.4%). Overall, the rhizosphere and endosphere of wild olives were proved as a good reservoir of bacteria antagonists against V. dahliae. Several of those bacteria showing high and broad antagonism potential may therefore be considered for further analyses as promising biocontrol agents against V. dahliae in olive.

Comparison of the Chemical Composition and the Organoleptic Profile of Virgin Olive Oil from Two Wild and Two Cultivated Tunisian Olea europaea

With the aim to select new olive cultivars with superior physical and chemical properties than the cultivar Chemlali Sfax, the present study focused on the comparison of the chemical composition and the sensory profile of the virgin olive oils (VOOs) of two wild olive trees (Oleasters K and M) with those of VOOs obtained from Chemlali Sfax and Neb Jmel olive cultivars, all growing in the coastal region of Tunisia. Despite the variability in the chemical composition (fatty acids, pigments, and phenolic and volatile compounds) and the organoleptic profile of the VOOs of the oleasters and the cultivars, the quality indices (free fatty acids, peroxide value, and spectrophotometric indices K232 and K270) as well as the fatty acid composition of all VOOs studied met the commercial standards. Both the α-tocopherol and phenol contents varied between the genotypes. The Neb Jmel and Oleaster K VOOs had more than two times higher total phenol levels than the Chemlali Sfax and Oleaster M VOOs. Also the contents of volatile compounds differed between the olive oils studied. Chemlali Sfax and Oleaster K oils were more abundant in aldehydes, whereas Oleaster M VOO had higher contents of alcohols. These results were confirmed by a sensorial analysis showing that the later oil was deprived for consumption despite its abundance in α-tocopherol. In conclusion, the oleasters studied revealed to be interesting, since they produced oils with good quality characteristics in terms of minor compounds (phenols and volatiles) compared to the Chemlali Sfax cultivar.

Genomic profiling of plastid DNA variation in the Mediterranean olive tree

BackgroundCharacterisation of plastid genome (or cpDNA) polymorphisms is commonly used for phylogeographic, population genetic and forensic analyses in plants, but detecting cpDNA variation is sometimes challenging, limiting the applications of such an approach. In the present study, we screened cpDNA polymorphism in the olive tree (Olea europaea L.) by sequencing the complete plastid genome of trees with a distinct cpDNA lineage. Our objective was to develop new markers for a rapid genomic profiling (by Multiplex PCRs) of cpDNA haplotypes in the Mediterranean olive tree.ResultsEight complete cpDNA genomes of Olea were sequenced de novo. The nucleotide divergence between olive cpDNA lineages was low and not exceeding 0.07%. Based on these sequences, markers were developed for studying two single nucleotide substitutions and length polymorphism of 62 regions (with variable microsatellite motifs or other indels). They were then used to genotype the cpDNA variation in cultivated and wild Mediterranean olive trees (315 individuals). Forty polymorphic loci were detected on this sample, allowing the distinction of 22 haplotypes belonging to the three Mediterranean cpDNA lineages known as E1, E2 and E3. The discriminating power of cpDNA variation was particularly low for the cultivated olive tree with one predominating haplotype, but more diversity was detected in wild populations.ConclusionsWe propose a method for a rapid characterisation of the Mediterranean olive germplasm. The low variation in the cultivated olive tree indicated that the utility of cpDNA variation for forensic analyses is limited to rare haplotypes. In contrast, the high cpDNA variation in wild populations demonstrated that our markers may be useful for phylogeographic and populations genetic studies in O. europaea.

Intracanopy plasticity under strong wind conditions in the wild olive tree (Olea europaea L.): a conserved response between closely related taxa?

Trees have the ability to respond to local environmental cues by expressing particular phenotypes across their canopy through a mechanism known as intracanopy plasticity. In this study, intracanopy plasticity of Olea europaea subsp. europaea was analyzed by sampling leeward and windward canopy exposures of individuals occurring in an area with sustained strong wind conditions. A suite of morphofunctional and reproductive traits was measured at these contrasting canopy positions and, for comparison, also in wind-protected trees. Furthermore, the pattern of intracanopy plasticity of these plants was compared to that previously documented in a closely related species, Olea europaea subsp. guanchica. Plants exposed to strong winds displayed substantial differences between leeward and windward exposures in most of the study traits. Leeward exposures experienced a mean reduction of 73% in wind speed as compared to windward ones, and displayed a modular phenotype matching that observed in wind-protected plants. Wind-exposed plants, however, were comparatively smaller and had fewer and smaller inflorescences, since inflorescence size was positively associated with crown size. The two closely related species showed similar crown and leaf sizes between populations exposed to strong winds, and intracanopy responses were comparable for most traits. These observations suggest that intracanopy plasticity resulted in the expression of contrasting phenotypes within individuals, which allowed trees to persist under sustained wind stress, although at the cost of a reduced reproductive fitness. In addition, this study gives support to the idea that intracanopy responses are conserved among closely related taxa evolving in different habitats, but experiencing a comparable limiting factor.

Effect of substrate treatments on survival and growth of Mediterranean shrubs in a revegetated quarry: An eight-year study

Loss of soil is a major problem that restoration actions must solve in Mediterranean quarries. Soil replacement by a low-quality substrate is often inadequate for revegetation and limits plant establishment and growth in these degraded areas, which are also subject to drought recurrent stress.In 1998, a field experiment was performed in a limestone quarry at Outão (Serra da Arrábida, SW Portugal) to test different types of substrate improvement: NPK fertilizer, water-holding polymer, mycorrhizal inoculum, and combinations of these. Two-year old plants of three native woody species were planted – carob (Ceratonia siliqua), wild-olive tree (Olea europaea var. sylvestris) and mastic (Pistacia lentiscus). Reports concerning the short-term results showed some differential effects of the assayed treatments on plant growth and physiology. The monitoring program was maintained, and here we report on the survival and growth of the introduced plants over eight years after planting. This prolonged study showed that, with the limited exception of fertilization, none of the assayed treatments added major advantages for plant survival or growth. Regardless of the tested substrate treatment, mortality was low and these native species became established in the revegetated area.

WILD OLIVE (OLEA EUROPAEA L.) SELECTION FOR QUALITY OIL PRODUCTION

ABSTRACT One hundred and fifty accessions belong to six wild olive populations from Tunisia were analyzed using fruit pomological traits and oil chemical characteristics. Analysis of variance revealed significant differences among olive accessions for the main studied parameters, indicating a high degree of variability among oleasters. Cluster analysis revealed no clear relationships between accessions according to their geographical origin. Among the screened oleasters, three stand out for their interesting oil parameters, as compared to traditional varieties with high oleic and low linoleic acid contents, high triolein level, large total phenol content, high oxidative stability and high oil content. ZI2 had the highest values of oleic acid (78.5%), whereas H3 was noteworthy for its higher content of phenolic compounds (435.3 mg/kg). Accordingly, oil from H3 presented the highest oxidative stability (83.5 h). A significant relationship was observed between phenolic compounds and oxidative stability. PRACTICAL APPLICATIONS The identification of new cultivars and their oil composition is a key point in the improvement of olive oil production. This is the first study of chemical composition of virgin olive oils from wild olives. The paper is a study on the quality characteristics of olive oils obtained from wild olive trees growing in Tunisia to define new cultivars well adapted to Tunisian environment and yielding high quality oils. Although the study was carried out in Tunisia, it might be applied to other countries with wild olive trees, in order to contrast productivity and oil quality with those resulting from native cultivars and transplanted foreign varieties.

Somatic embryogenesis induction in leaves and petioles of a mature wild olive

We describe a protocol for somatic embryogenesis (SE) induction from an adult wild olive tree (Oleaeuropaea ssp. europaea var. sylvestris. The protocol used confirms for the first time that there is no need to use juvenile or rejuvenated material for SE induction. For SE induction, petiole and leaf (proximal, intermediary and distal zones) explants were grown on Murashige and Skoog (MS) or Olive Medium (OM) media with different combinations of plant growth regulators (PGR): α- naphthaleneacetic acid (NAA), Zeatin (Zea), indole-3-butyric acid (IBA), 2-isopentyl adenine (2iP), thidiazuron (TDZ) and 6-benzylaminopurine (BAP). All media had 30 g/l sucrose and 7 g/l agar, and the pH was adjusted to 5.8. Cultures were incubated in the dark and, after 3 months, they were transferred to MS medium without PGR for expression. Petiole explants gave the highest callus production, while for SE induction and expression distal blade leaf and petiole explants gave the highest rates. The best medium for SE induction was MS with 12.25 μM IBA plus 4.56 μM Zea. Histological analyses confirmed the individuality of globular somatic embryos. This is the first report of SE expression in explants without rejuvenation in Olea genus, and opens perspectives for using this strategy in SE protocols both for this wild genotype and for commercial genotypes.

Genetic diversity and relationships of wild and cultivated olives at regional level in Spain

Genetic diversity and relationships between local cultivars and wild olive trees from three important Spanish olive-growing regions, Andalusia (South), Catalonia and Valencia (from Eastern Mediterranean Coastal area), were studied by means of eight SSR loci. Distinct allelic composition and heterozygosity levels were found in wild olive populations and cultivars. The observed patterns of genetic variation revealed: a) the independent clustering of Andalusian wild olives in a separate gene pool, b) the belonging of wild populations and most cultivars from Catalonia to another gene pool, c) the joined clustering of Andalusian and a set of Valencian cultivars in a third gene pool, and d) clustering of wild individuals from Valencia to the three different gene pools. These results suggest that wild populations of Andalusia may represent true oleasters, the ones from Catalonia may be feral forms derived from cultivar seed spreading, while the population of Valencia seems to be the most admixed one. The significant differentiation between Andalusian and most Catalonian cultivars is indicating an independent selection of olive cultivars in the two regions. The detection of a certain wild genetic background in some Catalonian and Valencian cultivars and the similarity found between wild and cultivated forms may suggest the use of local wild trees in olive domestication. The proposed scenario for the development of olive cultivars in Andalusia includes an empirical selection of outstanding local wild genotypes followed by various generations of crosses and various replanting campaigns, as well as possible introductions of ancestral cultivars. Therefore, our findings would lead us to support the hypothesis that the current diversity found in Spanish olive cultivars may be regionally differentiated and due to both, autochthonous and allochthonous origin. The information obtained in this work gives insights into the genetic resources of the main olive producing country, demonstrating that wild olive populations and local cultivars both represent potential sources of useful variability for olive breeding programs.

Genetic Relationships between Cultivated and Wild Olive Trees (Olea Europaea L. Var. Europaea and Var. Sylvestris) Based on Nuclear and Chloroplast SSR Markers

The olive is widely cropped in Tunisia where also oleaster trees thrive all around orchards and in natural sites. Little is known on the genetic relationships between the olive crop and oleaster trees in Tunisia. Fifty-two oleaster trees and fifteen cultivars were sampled from Tunisia. SSR genotyping was performed in polyacrylamide gels after fluorescent labeling. We used seven nuclear and two chloroplast SSR markers. AFC analyses showed close genetic relationships between cultivated and oleaster trees. Genetic relationships were also displayed in a dendrogram based on Unweighted Pair Group Method (UPGMA). Five clusters were defined mixing cultivar and oleaster trees suggesting close relationship between some cultivar and some oleaster trees. One oleaster is single in a cluster. The chlorotype SSR markers show probably three olive origins. Some cultivars have the CE chlorotype originates from the East of the Mediterranean basin, the CCK haplotype originates from Maghreb and the COM chlorotype originates from West Mediterranean. The cultivars were 1) introduced from the East; 2) selected in the West; 3) or selected in the North Africa region. The Tunisian oleaster trees carry eastern and western Mediterranean chlorotype CCK, COM and CE.

RAPD assay of wild-type olives in Turkey

Genetic similarities and distances between wild-type olives in Turkey were studied using an RAPD-PCR assay. Seven wild olive tree samples were collected from villages in Manisa and Izmir provinces. Genomic DNA was extracted from young leaves and the RAPD-PCR assay was used to generate RAPD markers. Sixty-five random primers obtained from Operon Technologies were tested for the assay (OP-A 1-20, OP-I 1-20, OP-Q 1-20, and OP-J 1-5). Thirty-two of these primers yielded 115 highly polymorphic bands. The mean number of usable bands per primer for all the samples was 3.59. The genetic distance values ranged from 0.1498 to 0.6845, and genetic similarity values varied from 0.8609 to 0.5043. We found that the closest samples based on their genetic distance and similarity values were from Harlak and Sabancilar; the most distant samples were from Bornova and Bademli.

Genetic analysis on wild olives by using RAPD markers

The aim of this study was to detect genetic similarities and distances among wild olive trees by RAPD-PCR technique. Olives are raised at a high range from Artvin on the north, up to Hatay on the south and up to Mardin on the Southeast part of Turkey. The Aegean, Mediterranean, Marmara and Black Sea regions of Turkey have suitable climatic conditions for olive breeding. Olive breeding had a rapid increase in the Aegean Region during recent years, among the agricultural products. Therefore, it is required to know the genetic relationships between wild olives and their cultural relatives in order to improve genetic resources and our knowledge of their evolutionary background. In this study, samples were mainly collected from Manisa province and additionally, samples were taken from Izmir and Mugla provinces for comparison. Genomic DNA’s were extracted from young leaves and PCR was used to generate RAPD bands. 60 random primers obtained from Operon Tech. were tested by RAPD-PCR. Total of 38 primers were given highly polymorphic and continuous scorable bands. 167 bands were found and each primer was screened at an average of 4.4 polymorphic bands. Key words: RAPD, PCR, wild olive.

Nutritional Status of Mediterranean Trees Growing in a Contaminated and Remediated Area

Soil contamination may contribute to forest decline, by altering nutrient cycling and acquisition by plants. This may hamper the establishment of a woody plant cover in contaminated areas, thus limiting the success of a restoration program. We studied the nutritional status of planted saplings of Holm oak (Quercus ilex subsp. ballota (Desf.) Samp.), white poplar (Populus alba L.), and wild olive tree (Olea europaea var. sylvestris Brot.) in the Guadiamar Green Corridor (SW Spain) and compared it with established adult trees. Soils in this area were affected by a mine-spill in 1998 and a subsequent restoration program. The spill resulted in soil acidification, due to pyrite oxidation, and deposited high concentrations of some trace elements. In some sites, we detected a phosphorus deficiency in the leaves of Q. ilex and O. europaea saplings, as indicated by a high N:P ratio (>16). For O. europaea, soil contamination explained 40% of the variability in leaf P and was negatively related to chlorophyll content. Soil pH was a significant factor predicting the variability of several nutrients, including Mg, P, and S. The uptake of Mg and S by P. alba was greater in acidic soils. The monitoring of soil pH is recommended since long-term effects of soil acidification may negatively affect the nutritional status of the trees.

The distinctive population structure ofColletotrichumspecies associated with olive anthracnose in the Algarve region of Portugal reflects a host–pathogen diversity hot spot

Anthracnose (Colletotrichum spp.) is an important disease of olive fruits. Diversity and biogeographic relationships of the olive anthracnose pathogens in the Algarve (Portugal) were investigated, along with host association patterns and disease levels during 2004-2007, to test the hypothesis that this region is a host-pathogen diversity hot spot. Diverse Colletotrichum acutatum and Colletotrichum gloeosporioides populations were identified based on rRNA-internal transcribed spacer and partial beta-tubulin 2 gene sequences of 95 isolates. Spatial and temporal variations in the occurrence of the eight genetic entities of the pathogens were linked to olive biogeography. Disease occurrence patterns suggest that C. acutatum populations are more stable pathogens, while C. gloeosporioides populations appear to be more influenced by favourable conditions. Three unique C. acutatum populations were identified, but none of the eight populations were dominant, with the most frequent type representing only 27%. Thus, the population structure of olive anthracnose pathogens in the Algarve is distinct from other parts of Portugal and other world locations, where only one or two genetic entities are dominant. This pattern and level of genetic diversity in a restricted area, where oleaster (wild olive tree), ancient landraces and modern cultivars of olive occur in close proximity, suggests the Algarve as a centre of diversity of the anthracnose pathogens and corroborates recent work suggesting western Mediterranean as an important centre of olive diversity and domestication.

Composition, quality and oxidative stability of virgin olive oils from some selected wild olives (<i>Olea europaea</i> L. subsp. <i>oleaster</i>)

A study on the characterization of virgin olive oils from wild olives ( Olea europaea L. subsp. oleaster ) was conducted in order to define new cultivars which are welladapted to the Tunisian environment and yield high quality oils. The study was done during the crop years 2003/04, 2004/05 and 2005/06. The main analytical parameters of the oils were evaluated: fatty acid compositions, chlorophylls, carotenoids, tocopherols and phenolic compounds as well as their relationship with oxidative stability. The fatty acid composition of all the wild olive trees tested produced virgin olive oil which complies with commercial standards, as well as for their appreciable amounts of tocopherols and phenolic compounds. Tocopherol analysis by HPLC revealed the presence of α, β, γ and δ tocopherols in all the studied olive oils. Total tocopherol content was significantly influenced by the varietal factor. It ranged from 310 (SB12) to 780 mg/kg (H3). As for total tocopherols, the amount of each tocopherol varied according to genotype. α tocopherol is the most prominent, whereas β, γ and δ tocopherols are less represented. Results showed a clear influence of total phenols and o-diphenols on virgin olive oil stability (R = 0.905, 0.963 P <10–3, respectively), and a much lower contribution of tocopherols (R = 0.568) and acidic composition (R = 0,563).

The Cultural Use of the Wild Olive Tree by the amaXhosa People in the Eastern Cape Province of South Africa

The cultural meanings of harvested plants have for the most part been ignored in academic research on non-timber forest products (NTFPs) in southern Africa. Historically scientists have tended to ignore the complex relationships between nature and culture. Given the country’s unique political and economic past and the current search for sustainable use of natural resources, a focus on the convergence of natural science and cultural diversity is important at this time. Empirical data on cultural practices is being collected in order to develop fresh and relevant insights into the complex relationships between culture and biodiversity. The purpose of this paper is to demonstrate that the concept of culture needs to be brought into our understanding of the role of NTFPs. We document the use and value of a specific tree, Olea europaea L. subsp. africana (Mill.) P.S. Green, called Umnquma in the Xhosa language, for cultural purposes, by both rural and urban households.

Genetic Structure of Wild and Cultivated Olives in the Central Mediterranean Basin

Olive cultivars and their wild relatives (oleasters) represent two botanical varieties of Olea europaea subsp. europaea (respectively europaea and sylvestris). Olive cultivars have undergone human selection and their area of diffusion overlaps that of oleasters. Populations of genuine wild olives seem restricted to isolated areas of Mediterranean forests, while most other wild-looking forms of olive may include feral forms that escaped cultivation. The genetic structure of wild and cultivated olive tree populations was evaluated by amplified fragment length polymorphism (AFLP) markers at a microscale level in one continental and two insular Italian regions. The observed patterns of genetic variation were able to distinguish wild from cultivated populations and continental from insular regions. Island oleasters were highly similar to each other and were clearly distinguishable from those of continental regions. Ancient cultivated material from one island clustered with the wild plants, while the old plants from the continental region clustered with the cultivated group. On the basis of these results, we can assume that olive trees have undergone a different selection/domestication process in the insular and mainland regions. The degree of differentiation between oleasters and cultivated trees on the islands suggests that all cultivars have been introduced into these regions from the outside, while the Umbrian cultivars have originated either by selection from local oleasters or by direct introduction from other regions.

Extensive gene flow blurs phylogeographic but not phylogenetic signal in Olea europaea L.

Genetic structure and evolutionary patterns of the wild olive tree (Olea europaea L.) were investigated with AFLP fingerprinting data at three geographic levels: (a) phylogenetic relationships of the six currently recognized subspecies in Eurasia and Africa; (b) lineage identification in subsp. europaea of the Mediterranean basin; and (c) phylogeography in the western Mediterranean. Two statistical approaches (Bayesian inference and analysis of molecular variance) were used to analyse the AFLP fingerprints. To determine the congruency and transferability of results across studies previous RAPD and ISSR data were analysed in a similar manner. Comparisons proved that qualitative results were mostly congruent but quantitative values differed, depending on the method of analysis. Neighbour-Joining analysis of AFLP phenotypes supported current classification of subspecies. At a Mediterranean scale no clear cut phylogeographic pattern was recovered, likely due to extensive gene flow between populations of subsp. europaea. Gene flow estimates calculated with conventional F-statistics showed that reproductive barriers separated neither populations nor lineages of O. europaea. Genetic divergence between eastern and western parts of the Mediterranean basin was observed only when geographical and population information were incorporated into the analyses through hierarchical analysis of molecular variance (AMOVA). Within the western Mediterranean, the highest genetic diversity was found in two regions: on both sides of the Strait of Gibraltar and in the Balearic archipelago. Additionally, long-lasting isolation of the northern-most populations of the Iberian Peninsula appeared to be responsible for a significant divergence.

Biomonitoring of trace elements in the leaves and fruits of wild olive and holm oak trees

Biomonitoring of trace elements is essential to assess ecosystem health, in particular in landscapes influenced by human activity. The Guadiamar Valley (SW Spain) was polluted in 1998 by a spill from an open-pit pyrite mine affecting about 55 km 2. In this paper, we used two common species of tree, namely wild olive and holm oak, to biomonitor the concentration of nine trace elements–As, Cd, Cu, Fe, Mn, Ni, Pb, Tl and Zn–in this spill-affected area over the 3-year period 1999–2001. We analysed the leaves and fruits of trees growing in the spill-affected terraces, and compared them with adjacent trees in the non-affected upper terraces. The main trace elements polluting the soil were Zn, As, Pb and Cu. In general, the oak leaves were richer in trace elements than the olive leaves, reaching phytotoxic levels for As and Pb, while the olive fruits (pulp) were more polluted than the oak seeds (protected inside a hard pericarp), reaching toxic values for Cd and Pb. The concentration of trace elements in the leaves and fruits decreased with time and, in consequence, the toxicity risk to the food web diminished.

First Report of Botryosphaeria ribis as a Branch Dieback Pathogen of Olive Trees in Spain.

Botryosphaeria spp. are known to produce cankers and dieback in several woody hosts. Since 2001, a severe disease resulting in the dieback of branches was observed in olive (Olea europaea) orchards in Andalucía, southern Spain, where the only cultivar affected was Gordal de Sevilla. The affected trees showed abundant dead twigs, and wilted leaves remained attached. These symptoms were similar to those caused by Resseliella oleisuga, a bark borer insect common in the region. Closer inspection showed no evidence of insects, but revealed reddish brown bark lesions that girdled the affected branches. When the outer bark was removed, the affected tissue appeared dark brown, in contrast to the yellowish green of healthy inner bark. A Fusicoccum sp. was consistently isolated from the necrotic bark. Conidia produced in black pycnidia were hyaline, aseptate, fusoid, with a truncate base averaging 19.9 × 6.7 μm, becoming pale brown with all cells the same color, and developing one or two septa when germinated. These characteristics conform to Fusicoccum ribis (1), the anamorph of B. ribis. Pathogenicity of two isolates from symptomatic branches was determined by stem inoculations of 4-year-old cv. Gordal de Sevilla grafted on wild olive trees growing in plastic pots containing a sand/lime/peat soil mixture. Both isolates were inoculated by three techniques. In the first, 7-mm-diameter agar plugs bearing actively growing mycelium were applied to 7-mm-diameter bark wounds made with a cork borer on the middle point of stems (averaging 10 mm in diameter). In the second technique, the same type of inoculum was applied directly to the bark without any wounding. For the third technique, 100 μl of conidial suspension in water (2 × 105 conidia per ml) was applied to bark wounds. The inoculated stem sections were wrapped in Parafilm to retain moisture. All inoculated and control plants (only sterile agar medium or sterile water added) were kept in a greenhouse and watered as needed. There were three replicate plants per isolate and inoculation technique. After 3 months, all plants inoculated with mycelium applied to wounds showed girdling and apical death with browning and wilting of leaves above the inoculation point. In the treatments where mycelium was applied to unwounded bark or where conidial suspensions were applied to wounds, small bark cankers developed to 56 mm long, but neither girdling nor foliar wilting were observed. No symptoms developed in the control plants. There were no significant differences in pathogenicity between the two F. ribis isolates. F. ribis was recovered from necrotic bark from every inoculated plant. These results demonstrate that B. ribis is pathogenic on cv. Gordal de Sevilla olives, and they indicate that bark wounding favors lesion development but is not needed for infection. Reference: (1) B. Slippers et al. Mycologia 96:83, 2004.