Pistil abortion is related to ovary mass in olive (Olea europaea L.)

Pistil abortion is considered to be an evolutionary adaptation to save resources in andromonoecious species, balancing the number of pistils with the resources available. It follows that large-fruited species/cultivars should have greater levels of pistil abortion. Working in olive, we found that pistil abortion, expressed as percentage of staminate flowers, was positively correlated to the average ovary mass at bloom, across cultivars with different ovary/fruit mass. Since ovary mass correlated with fruit mass, pistil abortion also correlated with fruit mass. The absolute number of perfect flowers per inflorescence was negatively correlated with both ovary mass and pistil abortion, while the number of staminate flowers per inflorescence increased with both parameters. The total number of flowers (i.e. staminate + perfect) was not correlated to ovary mass or pistil abortion, suggesting that male function was not affected. The results support the hypothesis that pistil abortion is related to competition for resources among ovaries, and suggest that genetic differences in pistil abortion among olive cultivars may be explained by their different pistil mass and sink strength.     

Fruit weight is related to ovary weight in olive (Olea europaea L.)

Fruit size is an important parameter both for scientific understanding and for commercial purposes. In many species, mature fruit size is often related to floral ovary size, but no literature exists in olive that demonstrates such a relationship. Previous work suggests that olive cultivars with different fruit sizes have similar cell number and size in the ovary transectional area, but ovary and fruit dry weight was not measured. In the present study, ovary dry weight and fruit dry weight during the whole fruit development season until harvest were measured in olive cultivars with different fruit size, over three years. Flower dry weight was also measured. Fruit weight at harvest was strongly correlated to ovary weight at bloom, both in single-year data and when data from three years were pooled. Flower dry weight, excluding the ovary, was also correlated to ovary dry weight. Ovary dry weight was strongly correlated not only to the fruit dry weight at maturity, but also at any date during fruit development. The mature fruit/ovary dry weight ratio ranged between 1000 and 4000 among cultivars, but was not correlated to the fruit dry weight at maturity. These results suggest that, in olive, fruit weight is genetically controlled through the ovary weight at bloom. This knowledge may have implications in the understanding of fruit set and source-sink relationships in olive.     

Microsatellite markers based assessment of genetic diversity in Iranian olive (Olea europaea L.) collections

To study the genetic variation in Iranian olive collections and some foreign olive cultivars, 47 accessions of 18 local cultivars from 6 olive collections of Iran (Roudbar, Zanjan, Ahvaz, Dezful, Kazeroon and Shiraz), were analyzed along with 30 imported cultivars using 16 microsatellite primer pairs. All the used microsatellite loci revealed polymorphism in the studied genotypes, except GAPU14 and GAPU113 markers. Fourteen microsatellite primers amplified 126 polymorphic alleles in the 87 selected olive accessions. The average number of alleles per locus was 9, ranging from 3 to 14. Polymorphic information content (PIC) was 0.85. The genetic similarity based on Jaccard coefficient ranged from 0.15 to 1. The genetic relationships among accessions were investigated using cluster analysis and principal component analysis (PCA). Most of the accessions with the same name were grouped together; some exceptions were also observed. As expected, close relationship was observed among accessions within same cultivar. Most of the Iranian olive accessions were clustered to a main distinct group. Two-dimensional scatter plot of principal component analysis revealed a clear separation of most of the Iranian olives from Syrian and other introduced cultivars. These suggest that Iranian cultivars have different origin related to West Mediterranean basin cultivars and have evolved independently from the others. Between and within Iranian and foreign cultivars (cultivars including three or more accessions) genetic diversity was analyzed using analysis of molecular variance (AMOVA). AMOVA revealed higher within cultivar genetic variation (62.76%) as compare to that between cultivar variations (37.24%). The intra- and inter-cultivar variance tested by permutation test showed significant genetic variation at both levels. The high level within cultivar genetic variance could be due to mislabeling and presence of homonyms in cultivars produced by vegetative propagation from original plants.     

Impacts of water stress on gas exchange,water relations,chlorophyll content and leaf structure in the two main Tunisian olive (Olea europaea L.) cultivars

Leaf structural adaptations for the reduction of water loss were examined in two olive (Olea europaea L.) cultivars (Chemlali and Chétoui) growing under water stress conditions. Leaf measurements included leaf tissue thickness, stomatal density, trichome density, specific leaf area, leaf density, water relations, and gas exchange. We found considerable genotypic differences between the two cultivars. Chemlali exhibited more tolerance to water stress, with a thicker palisade parenchyma, and a higher stomatal and trichome density. Chemlali leaves also revealed lower specific leaf area and had higher density of foliar tissue and lower reduction in net CO₂ assimilation rate. The mechanisms employed by these two cultivars to cope with water deficit are discussed at the morpho-structural level. The morphological and structural characteristics of the leaves are in accordance with physiological observations and contribute to the interpretation of why the olive cv. Chemlali is more drought-tolerant than cv. Chetoui. Furthermore, from the behaviour of Chemlali plants we consider this cultivar very promising for cultivation in semi-arid areas.     

Differences in ovary size among olive (Olea europaea L.) cultivars are mainly related to cell number,not to cell size

Differences in mature fruit size among olive cultivars are related to differences in ovary size at bloom, but it is not known whether cell number or size determines the variation in ovary size. In this study we measured cell size and number in equatorial cross-sectional areas of the principal ovary tissues (mesocarp and endocarp) in eight cultivars with very different fruit and ovary size. The results showed that cell number explained most of the differences in ovary size, while cell size was not related to ovary size, except for a weak (R² = 0.33) correlation in the endocarp. Since sink strength is thought to be related to cell number, this finding supports the hypothesis that larger ovaries represent stronger sinks. The implications of greater sink strength with bigger ovaries are discussed. Within cultivars, while the cross-sectional areas of mesocarp and endocarp were similar, cell number was higher in the mesocarp but cell size was smaller compared to the endocarp.     

Timing of fruit removal affects concurrent vegetative growth and subsequent return bloom and yield in olive (Olea europaea L.)

Olive (Olea europaea) demonstrates a high tendency toward alternate fruit production, with significant negative consequences on the industry. Fruit load is one of the main cause-and-effect factors in the phenomenon of biennial bearing, often disrupting the balance between reproductive and vegetative processes. The objectives of the present study were to identify the time range during which heavy fruit load reversibly interrupts the reproductive processes of the following year. The linkage between timing of fruit removal, vegetative growth, return bloom, and fruit yield was studied. Complete fruit removal in cv. Coratina until about 120 days after full bloom (August 15) caused an immediate resumption of vegetative growth. The new shoots grew to twice the length of those on trees that underwent later fruit removal. Moreover, a full return bloom, corresponding with high subsequent yields, was obtained by early fruit removal, while poor or no bloom developed on late-defruited or control trees. Thus, the critical time to affect flowering and subsequent fruiting in the following year by fruit thinning occurs in olive trees even weeks after pit hardening—much later than previously suggested. Furthermore, the data indicate that flowering-site limitation, due to insufficient or immature vegetative growth during the On-year, is a primary factor inducing alternate bearing in olive.     

Encapsulation of in vitro-derived explants of olive (Olea europaea L. cv. Moraiolo): II. Effects of storage on capsule and derived shoots performance

The experiments described in this paper were designed to examine the effect of storage at different temperatures for various periods on regrowth and proliferation ability of encapsulated olive microcuttings (capsules). The proliferation and rooting ability of the shoots derived from the capsules were compared with those maintained in normal proliferation by 45 days period of subculturing. A simple method of encapsulated microcuttings storage in plastic cuvettes was used. Interesting feature was the ability of the capsules to retain their viability and regrowth after storage at room temperature as well as at 4 °C for 15 and 30 days and both axillary buds on the nodes of almost all capsules sprouted successfully. Improvement in proliferation and rooting abilities were found in subsequent culturing of the shoots developed from the capsules. The study indicates bright prospects of commercial application of alginate nutrient capsule technology in various situations like supply of certified plant material from both official or private organizations to the plant nurseries and/or exchange of germplasm at international level without refrigerated containers just in plastic cuvettes.     

Coconut water and BAP successfully replaced zeatin in olive (Olea europaea L.) micropropagation

The data presented report on trials conducted during 24 months using the Portuguese olive cultivar ‘Galega vulgar’. The effectiveness of coconut water, BAP, or kinetin, as possible zeatin substitutes in olive micropropagation protocols, was investigated. In all stages of the micropropagation process, the mineral and vitamin formulation of olive medium (OM) was used. Regarding culture establishment the best results were achieved when 50 ml l⁻¹ coconut water and 2.22 μM BAP were used as medium supplements. For the in vitro multiplication stage, the highest proliferation rates with an average of 3.4 new explants on each 30 days were achieved maintaining the coconut water concentration at 50 ml l⁻¹ and increasing BAP up to 8.87 μM. The effects of IBA and activated charcoal on the in vitro root induction were also studied. Rooting rates of over 85% were obtained by basal immersion of the explants in IBA solution at 3 g l⁻¹ for 10 s, followed by inoculation in the OM culture medium, added with 2 g l⁻¹ of activated charcoal and without growth regulators. All in vitro rooted plants were transferred into Jiffy-Pots filled with vermiculite–perlite 3:1 (v/v) substrate. Those were subsequently wetted with the OM mineral solution, placed into polystyrene plates each one with 100 Jiffy-Pots capacity, which were transferred to traditional rooting mist benches, on a water-cooling equipped greenhouse. Such a simple acclimatization procedure allowed for 95% of plants survival.     

Growth,biomass allocation and leaf ion concentration of seven olive (Olea europaea L.) cultivars under increased salinity

Olive (Olea europaea L.) is the major fruit tree in the Mediterranean region, often grown in locations where plants are exposed to increased salinity. To determine the effect of NaCl on shoot and root growth, dry matter allocation, leaf Na⁺ and K⁺ concentration, electrolyte (EL) and K⁺ leakage (KL), seven olive cultivars of different origins were grown in nutrient solution containing 0, 33, 66, 100 or 166 mM NaCl for three months. The general effect of salinity was linear and quadratic decrease of observed plant growth parameters. Different responses of tested cultivars to applied levels of salinity were found for stem dry weight, shoot length and number of leaves. As salinity increased, growth of ‘Manzanillo’ declined sharply, whereas ‘Frantoio’ was the most tolerant to growth reduction in most of the observed growth parameters. Allometric analysis showed that biomass allocation under salinity stress was similar in all cultivars, but the slope between shoot weight and total plant weight decreased as salinity increased. Since the higher allocation in roots was not found, it seems that salinity only slowed the above ground plant canopy growth. Sodium concentration in leaves of all cultivars increased as salinity increased with the highest increment reached when the salinity of nutrient solution was raised from 100 to 166 mM NaCl. Significant differences among genotypes were found in leaf Na⁺ and K⁺ concentration and K⁺:Na⁺ ratio, but they were not related to the growth rate. Generally, ‘Frantoio’ and ‘Oblica’ accumulated less Na⁺ and were able to maintain higher K⁺:Na⁺ ratios as compared to other genotypes. Electrolyte leakage and KL linearly increased with increasing salinity and the magnitude of the response depended upon the olive cultivar.     

Cultivar-based fruit size in olive depends on different tissue and cellular processes throughout growth

In drupe fruits, in addition to fruit size, the proportions of mesocarp and endocarp tissues are critical objectives for fruit quality, crop production and management. The olive fruit is a typical drupe, with cultivars which show a wide range in both fruit size and the proportions of mesocarp and endocarp. Characterizing the roles of tissue and cellular processes in producing genetically based fruit size variability is necessary for crop improvement, as well as deepening our understanding of fruit developmental physiology. This study used microscope image analysis to evaluate cell number and size, the growth of mesocarp and endocarp tissues, and their developmental timing in producing fruit size among six olive cultivars with a large range of fruit size. We found that cultivar mesocarp and endocarp size increased linearly with fruit size, with larger sizes favoring an increasingly greater mesocarp/endocarp ratio. Within the mesocarp, cultivar-based fruit size related directly to cell number and was established soon after bloom by cell division rate. In spite of different cell division rates, all cultivars showed similar timing of cell division activity, with the majority of cells produced in the two months after bloom but, surprisingly, a substantial number of cells formed during the following 6 months. Cell expansion was high throughout fruit growth and an important factor in achieving final fruit size, but cell size did not differ among cultivars at any time. We can conclude that fruit size differences among olive cultivars are due at the tissue level to both mesocarp and endocarp sizes and at the cellular level to cell division throughout fruit growth. Furthermore, since cell size is consistent among cultivars in spite of variable cell division, it is likely that cultivar differences in cell expansion accompany those in cell division.     

Effects of water deficit on the vegetative response,yield and oil quality of olive trees (Olea europaea L., cv Coratina) grown under intensive cultivation

An experiment was carried out in a young high-density olive grove (556 plants ha⁻¹—Olea europaea L., cv Coratina) located in Southern Italy to evaluate the effect of different soil water availability on the vegetative and productive performances of olive trees also looking into the quality of the resulting oils. Trials were carried out over a 3-year period on trees subjected to irrigation and grown under rainfed conditions. Vegetative tree response, as shoot elongation and trunk diameter, was evaluated. Yield per plant, fruit characteristics, oil quality indices (free fatty acid content, peroxide value, UV adsorption at 232 and 270 nm, total phenols, α-tocopherol content) were determined for both irrigated and non-irrigated treatments in the ‘on’ years 1997 and 1999 (6th and 8th year after planting, respectively).     

Relation of carbohydrate reserves with the forthcoming crop,flower formation and photosynthetic rate,in the alternate bearing ‘Salustiana’ sweet orange (Citrus sinensis L.)

The aim of this work was to assess the relation between carbohydrate levels and flower and fruit production, as well as the role of carbohydrates on CO₂ fixation activity, by analysis of leaves, twigs and roots from the alternate bearing ‘Salustiana’ cultivar of sweet orange (Citrus sinensis [L.] Osbeck). A heavy crop load (on year) did not affect photosynthesis activity when compared to non-fruiting trees (off year). Fruiting trees accumulated most of the fixed carbon in mature fruits, whilst no accumulation was observed in roots before harvest. Non-fruiting trees transported part of the fixed carbon to the roots and mobilize it for growth processes and, at the end of the season (December), store it as reserves. Reserve carbohydrates accumulation in leaves started by early December for both tree types, showing the same levels in on and off trees until spring bud sprouting. A heavy flowering after an off year caused the rapid mobilization of the stored reserves, which were exhausted at full bloom. We found no evidence on carbon fixation regulation by either fruit demand or carbohydrate levels in leaves. Carbohydrate reserves played little or no role over fruit set, which actually relied on current photosynthesis.     

Effects of cadmium and copper on pollen germination and fruit set in pea (Pisum sativum L.)

In this work, pea (Pisum sativum) plants exposed to increasing cadmium and copper concentrations were tested for heavy metals accumulation in flowers and for ‘in vivo’ pollen germination. Based on the Cd and Cu accumulation amounts in the flowers, an evaluation of the same metals effects on ‘in vitro’ pollen germination was achieved. Moreover, the effects of both metals on fruits number and weight and on seed set and yield at individual plant level were examined. While cadmium concentrations did not affect ‘in vivo’ pollen germination, only higher copper concentrations rendered a significant reduction. This is in contrast with the clear negative effect on pollen germination in vitro and might be explained by the different dynamic and bioavailability of both metals. A clear effect of Cd and Cu was observed on two important yield components ie, fruit weight and seed set. Although results obtained herein cannot give a clear cut relationship between the effect of Ca and Cu on reproductive development and its consequences on yields, they represent emerging results on the potential consequences of metals contamination on reproductive development in plants.     

Phenological,morphological and functional indicators of genetic variability and their implication in the sexual reproductive system of Olea europaea L. (Oleaceae)

The floral biology of ‘Koroneiki’, ‘Kalamata’ and ‘Mastoidis’ was studied for three consecutive years to elucidate the potential physiological and genetic controls of inflorescence architecture and phenology and to identify potential genotype-distinguishing characters that could be employed for morphological cultivar discrimination. The first open flowers were mainly observed at position I for ‘Koroneiki’ (83%) while they were mainly located at position II for ‘Mastoidis’ (40%) and ‘Kalamata’ (63%). The last open flowers were principally located at the apical position for ‘Koroneiki’ (46%) and ‘Kalamata’ (68%) and at position II for ‘Mastoidis’ (75%). Most of the first open flowers were perfect for ‘Koroneiki’ (89%) and ‘Mastoidis’ (97%) while lower percentages were observed in the last open flowers (67% and 80%, respectively). Higher percentages of perfect flowers were observed in ‘Mastoidis’ compared to ‘Koroneiki’ in both cases. The average flowering shoots of ‘Kalamata’ produced significantly more flowers (1523/m) compared to ‘Koroneiki’ (1139/m) and ‘Mastoidis’ (1044/m). The half of the flowers in ‘Mastoidis’ were located at position I while the 38% was located at position II and only the 12% at the apex. By contrast, approximately the 80% of flowers in ‘Koroneiki’ and ‘Kalamata’ panicles was equally distributed at positions I and II, while the 18% and 19%, respectively, was located at the apex. A marked relationship was also observed between flower position in the panicle and flower gender.     

Immunolocalization and quantification of IAA after self- and free-pollination in Olea europaea L.

In order to characterise the self-compatibility in ‘Pisa 9’, that is an improved clone of cultivar Leccino (Olea europaea L.), biological, histological and biochemical analyses were carried out. Observations were also focused on ‘Pendolino’, the main pollinator for Italian cultivars considered as being self-incompatible. After self-pollination, in ‘Pisa 9’ the majority of the pollen tubes reached the distal part of the style, entered the ovule leading to a high fruit-set percentage. These same processes were also noted in the Pendolino cultivar, though in a lesser degree. The indole acetic acid (IAA) concentration and its immunocytochemical localization were determined in pollinated flowers after self- and free-pollination and in unpollinated flowers. In both cultivars the highest IAA levels and signal intensity were observed in free-pollinated pistils, whereas after self-pollination a different behaviour emerged. In ‘Pendolino’ the immunostaining was not uniformly distributed and stopped at the style level. This finding sustained the low percentage of fruit-set obtained in the cultivar. In ‘Pisa 9’ the auxin distribution was similar to the free-pollinated pistils where the immunolocalization staining reached the ovary. The correspondence between the IAA signal in the ovary and the high percentage of fruit-set in ‘Pisa 9’ self-pollinated pistils, confirms the role of this hormone in the induction and regulation of the fertilisation process.     

Analysis of highbush blueberry (Vaccinium corymbosum L.) fruit growth with exponential mixed models

Blueberry fruit growth commonly exhibits a double-sigmoid pattern. The goal of this work was to characterize and compare fruit growth of cultivars differing in ripening time using exponential non-linear mixed models. Mixed-effects five-parameter exponential (Gompertz I and II; logistic; monomolecular) models were fitted to fruit diameter data from 2 years and three cultivars grown in the field in a cool temperate environment. Gompertz II mixed model provided the best fit to fruit growth data and was used for further analysis. In later ripening cultivars ‘Cape Fear’ and ‘Herbert’, clear-cut double-sigmoid patterns were observed, absolute growth rate models exhibited two marked peaks, and relative growth rate showed an initial decreasing trend, and a subsequent peak. The earlier cv. ‘O’Neal’ did not exhibit a defined double-sigmoid pattern. The time between relative extremes of absolute growth acceleration is proposed as an objective criterion for fixing stage boundaries within growth curves. Exponential mixed models accurately fitted blueberry growth patterns. These equations highlighted differences in fruit growth patterns between early and late ripening highbush blueberry cultivars.     

Effect of pollination intensity,frequency and pollen source on fig (Ficus carica L.) productivity and fruit quality

Yield and fruit quality in fig (Ficus carica L.) are highly dependent on cultural practices especially caprification (pollination). However, this technique remains not well controlled in Tunisia. This study was conducted during two successive years, 2009 and 2010, to investigate different pollination conditions, i.e. number of caprifigs and repetitions of caprification on fruiting of two cultivars Bouhouli (San Pedro type) and Zidi (Smyrna type). In addition, the efficiency of four pollen sources was evaluated to identify the most effective pollinator for female cultivar Zidi. The following parameters were recorded: fruit number per shoot, fruit set, productivity, fruit characteristics and vegetative growth.     

Seasonal changes of mineral nutrients and phenolics in pomegranate (Punica granatum L.) fruit

Pomegranate fruit is an important source of potentially healthy bioactive compounds and mineral nutrients. Changes in total phenolic compound, concentrations, and levels of macronutrients (P, K, N, Mg, Ca and Na) and micronutrients (Zn, Cu, Mn, Fe and B) in arils and peel of pomegranate fruit were recorded from 10 days after full bloom until harvest. Total phenolics levels increased at early stage of growth both in peel and arils of fruit, but thereafter generally decreased during maturation and reached to 3.70 and 50.22 mg g⁻¹ of dry weight in arils and peel, respectively, at harvest. The amount of total phenolics in peel was markedly higher than arils of pomegranate fruit. The concentration of most elements in arils and peel decreased during fruit growth and development. At harvest the relative order of concentration of macronutrients both in arils and peel was K > N > Ca > P > Mg > Na. The concentration of most micronutrients was greater in the arils than in the peel especially in early season. The relative order of concentration of micronutrients in arils was B > Fe > Zn > Cu > Mn. The accumulation of all the macro- and microelement within the fruit also increased during fruit growth and development. These results provide important data on total phenolics and macro- and micronutrient changes during fruit growth and development, emphasizing that pomegranate fruit can be a good source of bioactive compounds and minerals.