Effect of Genotype on the Sprouting of Pomegranate (Punica granatum L.) Seeds as a Source of Phenolic Compounds from Juice Industry by-Products.

Pomegranate (Punica granatum L.) fruits are used mainly by the juice industry, for which seeds are a by-product to be disposed of, though they could potentially be a source of bioactive compounds. In this work, germination (total germination percentage, G; mean germination time, MGT; time to reach 80% of germination, TG80; seedling shoot length, fresh weight and dry matter), and nutritional value (total phenolics, TP; total flavonoids, TF; total non-tannins, TNT; antioxidant activities) of pomegranate seeds and sprouts were determined on four commercial pomegranate cultivars (Akko, Dente di Cavallo, Mollar de Elche and Wonderful). Seeds were removed from ripe fruits and incubated in plastic trays containing sterile cotton wetted with distilled water. Sprout shoots were harvested when they reached the complete cotyledon expansion, i.e., the ready-to-eat stage. Akko showed the best germination performance (G = 98%; MGT = 14 days after sowing, DAS; TG80 = 16 DAS), followed by Mollar de Elche. Sprouting dramatically increased TP, TF, TNT and antioxidant activity in all genotypes, with the highest values recorded in Mollar de Elche and Dente di Cavallo. Overall, based on germination performance, Akko and Mollar de Elche would be the best cultivars for sprouting. Sprouting pomegranate seeds appears to be a suitable way of utilizing by-products of the juice industry to obtain bioactive compounds.

Cultivar ideotype for intensive olive orchards: plant vigor, biomass partitioning, tree architecture and fruiting characteristics.

In order to achieve higher and earlier yield, modern olive orchards are increasingly intensified, with tree densities up to > 1500 trees hectare-1. With increasing tree densities, individual-tree canopy volume must be proportionally reduced. Not all cultivars are adaptable to high and very high orchard densities, because of excessive vigor and/or insufficient bearing when the canopy is pruned to a small volume. However, what makes an olive cultivar suitable for intensive and super intensive orchards is not clear. Recently, few studies have addressed this topic, suggesting that tree architecture and early bearing are essential traits. Yet, what architectural and productive features are important, how they work and whether they are interrelated remains elusive. This review summarizes and interprets the literature on olive, as well as the more abundant literature available for other fruit species, aiming to provide a comprehensive knowledge framework for understanding how tree architectural characteristics, plant vigor, and fruiting vary across olive genotypes, and how they are interconnected. It is concluded that, among the architectural characteristics, greater branching and smaller diameters of woody structures are particularly important features for cultivar suitability to intensive and super intensive olive orchards. Greater branching allows to produce more fruiting sites in the small volume of canopy allowed in these systems. It also reduces investments in woody structures, liberating resources for fruiting. Additional resources are liberated with smaller structure diameters. Greater branching also increases resources by increasing biomass partitioning into leaves (i.e. the photosynthetic organs), relative to wood. Since yield is affected by the competition for resources with vegetative growth, reducing resource investments in woody structures and/or increasing resource directly, increases yield. Yield, in turn, depresses vegetative growth, reducing vigor and the need for pruning. High yields also produce short shoots which have relatively greater investments in leaf mass and area, and lower in the woody stem, making them more suitable than long shoots to support concurrent fruit growth. This single framework of interpretation of how the different architectural and fruiting characteristics work and interact with one-another, will provide guidance for cultivar selection and breeding for intensive and super intensive olive orchards.

Canopy Architectural Characteristics of Ten New Olive (Olea europaea L.) Genotypes and Their Potential for Cultivation in Super-High-Density Orchards.

In recent years, there has been growing interest in olive genotypes (Olea europaea L.) suitable for super-high-density (SHD > 1200 trees/hectare) orchards. To date, only a few cultivars are considered fitting for such cultivation system. In this study, the first results on the architectural characteristics of the canopy of ten new olive genotypes are presented. Their suitability for SHD orchards was evaluated and compared with the cultivar 'Arbequina', which is considered suitable for SHD olive orchards and, for this reason, was used as the control. Several canopy measurements were taken, and some architectural parameters, such as branching frequency, branching density, and branch diameter/stem diameter ratio were calculated. The branching frequency value was greater than 0.20 in 'Arbequina' and in only four of the genotypes. The branching density in five genotypes was similar to 'Arbequina'. 'Arbequina' had the lowest value for the branch diameter/stem diameter ratio, and only three genotypes had similar values. These initial results showed that only one genotype has all canopy architectural characteristics comparable to those of the cv. 'Arbequina'. Further studies are needed to evaluate the production traits of these new genotypes and complete their characterization.

Dodine an effective alternative to copper for controlling Venturia oleaginea, the causal agent of pea-cock eye disease, in highly infected olive trees.

A trial was carried out in central Italy in an olive orchard of cultivar Moraiolo, highly infected by Venturia oleaginea. The aim of the investigation was to evaluate the effects of autumn and spring applications of copper oxychloride or dodine to control the disease. Non treated trees were used as the control. The effects of the fungal attacks on leaves and inflorescence development confirmed the high susceptibility of the cultivar Moraiolo to the disease. The results show that in trees heavily infected, but with most of the infected leaves at the early stage of the disease (asymptomatic phase), treatments with dodine had a curative effect, with consequent reduction in the appearance of symptomatic leaves and defoliation with respect to the control or copper-treated trees. The use of dodine against the autumnal attacks of V. oleaginea allowed most of the old leaves to be maintained until the new ones had formed, which is important for the growth processes during the early part of the growing season. Overall, the results indicate that to efficiently control the pathogen using copper compounds, treatments must start soon after the beginning of the attack and be repeated in order to maintain the infection at a low level. Dodine can be efficiently used if there is a great increase in infected leaves. The use of dodine to solve particular situations and not for normal repeated use is regulated by the fact that in some countries, Italy included, protocols for integrated pest management allow only one dodine treatment/year.

From flower to fruit: fruit growth and development in olive (Olea europaea L.)-a review.

The olive (Olea europaea L.) is the most cultivated tree crop in the Mediterranean and among the most cultivated tree crops worldwide. Olive yield is obtained by the product of fruit number and fruit size; therefore, understanding fruit development, in terms of both number and size, is commercially and scientifically relevant. This article reviews the literature on fruit development, from the flower to the mature fruit, considering factors that affect both fruit size and number. The review focuses on olive but includes literature on other species when relevant. The review brings the different factors affecting different phases of fruit development, addressed separately in the literature, under a single frame of interpretation. It is concluded that the different mechanisms regulating the different phases of fruit development, from pistil abortion to fruit set and fruit size, can be considered as different aspects of the same overall strategy, that is, adjusting fruit load to the available resources while striving to achieve the genetically determined fruit size target and the male and female fitness targets.

Olive phenolic compounds: metabolic and transcriptional profiling during fruit development.

BACKGROUND: Olive (Olea europaea L.) fruits contain numerous secondary metabolites, primarily phenolics, terpenes and sterols, some of which are particularly interesting for their nutraceutical properties. This study will attempt to provide further insight into the profile of olive phenolic compounds during fruit development and to identify the major genetic determinants of phenolic metabolism. RESULTS: The concentration of the major phenolic compounds, such as oleuropein, demethyloleuropein, 3-4 DHPEA-EDA, ligstroside, tyrosol, hydroxytyrosol, verbascoside and lignans, were measured in the developing fruits of 12 olive cultivars. The content of these compounds varied significantly among the cultivars and decreased during fruit development and maturation, with some compounds showing specificity for certain cultivars. Thirty-five olive transcripts homologous to genes involved in the pathways of the main secondary metabolites were identified from the massive sequencing data of the olive fruit transcriptome or from cDNA-AFLP analysis. Their mRNA levels were determined using RT-qPCR analysis on fruits of high- and low-phenolic varieties (Coratina and Dolce d'Andria, respectively) during three different fruit developmental stages. A strong correlation was observed between phenolic compound concentrations and transcripts putatively involved in their biosynthesis, suggesting a transcriptional regulation of the corresponding pathways. OeDXS, OeGES, OeGE10H and OeADH, encoding putative 1-deoxy-D-xylulose-5-P synthase, geraniol synthase, geraniol 10-hydroxylase and arogenate dehydrogenase, respectively, were almost exclusively present at 45 days after flowering (DAF), suggesting that these compounds might play a key role in regulating secoiridoid accumulation during fruit development. CONCLUSIONS: Metabolic and transcriptional profiling led to the identification of some major players putatively involved in biosynthesis of secondary compounds in the olive tree. Our data represent the first step towards the functional characterisation of important genes for the determination of olive fruit quality.

Effects of cultivar, fruit presence and tree age on whole-plant dry matter partitioning in young olive trees.

This study investigated the effects of cultivar, fruit presence and tree age on whole-plant partitioning of dry matter and energy equivalents (i.e., glucose equivalents). Young trees of two cultivars characterized by different vigor (i.e., Arbequina, low vigor and Frantoio, high vigor) were either completely deflowered from 2014 to 2017 or never, providing two contrasting levels of cumulated reproductive growth over the following 4 years. Total vegetative dry matter growth over the 4 years was assessed by destructive samplings (whole tree). Plant growth was inversely correlated to reproductive efforts, with Arbequina producing more and growing less than Frantoio. Deflowered trees grew similarly across cultivars, although deflowered Arbequina grew statistically less than deflowered Frantoio by the fourth year, due to abundant flower production. Total reproductive (flowers + fruit) and vegetative biomass production were the same for all cultivars and treatments. Arbequina had a greater distribution of dry matter in directly productive structures (current and one-year-old shoots) and in leaves. This allows it to increase the number of current and following-year production sites, and to save in the resources invested in non-productive sinks (roots, trunk and branches), thus liberating resources for reproductive growth. Greater investments in leaves allow it to intercept more light and thus to increase assimilation. Increased assimilation and increased partitioning towards productive structures, and decreased competition by non-productive structures might contribute to explain the greater early bearing attitude of this cultivar.

Fruit growth and sink strength in olive (Olea europaea) are related to cell number, not to tissue size.

The relationship between tissue (mesocarp and endocarp) growth and either tissue initial (i.e. in the ovary at bloom) size or cell number was studied using the olive cultivar Leccino (L) and its mutated clone (LC), which produces tetraploid fruits. LC ovaries were 2.7 times the volume of L ovaries, but contained an overall similar number of much larger cells. This allowed decoupling cell number and ovary size, which are normally closely correlated. With this decoupling, cell number in the ovary correlated with tissue growth in the fruit while tissue size in the ovary did not. Cell size in the ovary was inversely correlated with the tissue relative growth from bloom to harvest (i.e. the ratio between final and initial tissue size). These results support the hypothesis that cell number and not tissue size are related to fruit growth and sink strength, and that cell size in the ovary tissues is a good predictor of tissue growth, across cultivars and tissues, even when cell size is strongly affected by ploidy.

Stone Fruits: Growth and Nitrogen and Organic Acid Metabolism in the Fruits and Seeds-A Review.

Stone fruits of the Rosaceae family consist of several distinct parts, and these include the flesh, woody endocarp, and seed. To understand the metabolism of these fruits, it is necessary to have knowledge of both their structure and growth characteristics. The nitrogen metabolism of the different tissues of stone fruits is interlinked. For example, there is an import and storage of nitrogenous compounds in the endocarp that are then exported to the seed. Moreover, there are links between the metabolism of nitrogen and that of malic/citric acids. In this article, the structure and growth characteristics, together with the import/export, contents, metabolism, and functions of nitrogenous compounds and organic acids in the different parts of stone fruits and their seeds are reviewed.

Changes in Absolute Contents of Compounds Affecting the Taste and Nutritional Properties of the Flesh of Three Plum Species Throughout Development.

The characteristics of plum fruits of three different species were investigated throughout their development (including over-ripening). The content of primary and secondary metabolites was expressed as amount per gram DW (dry weight) and per fruit in order to obtain information about the balance between their synthesis and dissimilation at different stages of fruit development. In all the plums, during the first stages of development, glucose was the most abundant sugar, whereas sucrose increased during ripening. There was no decrease in malate content per fruit before the commercial harvesting time of any of the plums, whereas a decrease was observed during over-ripening. In general, both the total phenol content and the contents of individual phenols in the flesh expressed on gram DW decreased throughout development, whereas their content per fruit increased, indicating that these decreases were due to a dilution effect arising from the expansion of the flesh. During the development of the flesh, the increase in the contents of the investigated metabolites per fruit shows that there was no net dissimilation of malate up to commercial harvest and of phenols throughout fruit development. Good correlations between the content of phenols to antioxidant activity were found. Shiro flesh, during the last part of fruit development, had lower total carbohydrate and polyphenol contents, lower antioxidant activities, and a higher malate content than the flesh of the other two genotypes.

Fruit production and branching density affect shoot and whole-tree wood to leaf biomass ratio in olive.

The amount of shoot stem (i.e., woody part of the shoot) dry matter per unit shoot leaf dry matter (i.e., the shoot wood to leaf biomass ratio) has been reported to be lower in short shoots than in long ones, and this is related to the greater and earlier ability of short shoots to export carbon. This is important in fruit trees, since the greater and earlier carbon export ability of shoots with a lower wood to leaf biomass ratio improves fruit production. This ratio may vary with cultivars, training systems or plant age, but no study has previously investigated the possible effect of fruit production. In this study on two olive cultivars (i.e., Arbequina, with low growth rate, and Frantoio, with high growth rate) subject to different fruit production treatments, we found that at increasing fruit production, shoot length and shoot wood to leaf biomass ratio were proportionally reduced in the new shoots growing at the same time as the fruit. Specifically, fruit production proportionally reduced total new-shoot biomass, length, leaf area and average shoot length. With decreasing shoot length, shoot diameter, stem mass, internode length, individual leaf area and shoot wood to leaf biomass ratio also decreased. This may be viewed as a plant strategy to better support fruit growth in the current year, given the greater and earlier ability of short shoots to export carbon. Moreover, at the whole-tree level, the percentage of total tree biomass production invested in leaves was closely correlated with branching density, which differed significantly across cultivars. By branching more, Arbequina concentrates more shoots (thus leaves) per unit of wood (trunk, branches and root) mass, decreasing wood to leaf biomass ratio at the whole-tree level. Therefore, while, at the shoot level, shoot length determines shoot wood to leaf biomass ratio, at the canopy level branching density is also an important determinant of whole-tree wood to leaf biomass ratio. Whole-tree wood to leaf biomass ratio is likely to affect the canopy's ability to export carbon (i.e., towards fruits), as shoot wood to leaf biomass ratio affects the carbon export ability of the shoot.

Resource investments in reproductive growth proportionately limit investments in whole-tree vegetative growth in young olive trees with varying crop loads.

It has long been debated whether tree growth is source limited, or whether photosynthesis is adjusted to the actual sink demand, directly regulated by internal and environmental factors. Many studies support both possibilities, but no studies have provided quantitative data at the whole-tree level, across different cultivars and fruit load treatments. This study investigated the effect of different levels of reproductive growth on whole-tree biomass growth across two olive cultivars with different growth rates (i.e., Arbequina, slow-growing and Frantoio, fast-growing), over 2 years. Young trees of both cultivars were completely deflowered either in 2014, 2015, both years or never, providing a range of levels of cumulated reproductive growth over the 2 years. Total vegetative dry matter growth over the 2 years was assessed by destructive sampling (whole tree). Vegetative growth increased significantly less in fruiting trees, however, the total of vegetative and reproductive growth did not differ significantly for any treatment or cultivar. Vegetative growth over the 2 years was closely (R2 = 0.89) and inversely related to reproductive growth across all treatments and cultivars. When using data from 2015 only, the regression improved further (i.e., R2 = 0.99). When biomass was converted into grams of glucose equivalents, based on the chemical composition of the different parts, the results indicated that for every gram of glucose equivalent invested in reproductive growth, vegetative growth was reduced by 0.73-0.78 g of glucose equivalent. This indicates that competition for resources played a major role in determining tree growth, but also that photosynthesis was probably also enhanced at increasing fruit load (or downregulated at decreasing fruit load). The leaf area per unit of trunk cross sectional area increased with deflowering (i.e., decreased with reproductive growth), suggesting that water relations might have limited photosynthesis in deflowered plants, which had much greater canopies. Net assimilation rate (NAR) increased with reproductive growth and decreased with plant size. Net assimilation rate was also negatively correlated with the leaf area per unit of trunk cross sectional area, suggesting that water relations might have contributed to decreasing NAR at increasing plant size.

Effect of agronomical practices on carpology, fruit and oil composition, and oil sensory properties, in olive (Olea europaea L.).

We examined whether some agronomical practices (i.e. organic vs. conventional) affect olive fruit and oil composition, and oil sensory properties. Fruit characteristics (i.e. fresh and dry weight of pulp and pit, oil content on a fresh and dry weight basis) did not differ. Oil chemical traits did not differ except for increased content of polyphenols in the organic treatments, and some changes in the acidic composition. Sensory analysis revealed increased bitterness (both cultivars) and pungency (Frantoio) and decreased sweetness (Frantoio) in the organic treatment. Fruit metabolomic analysis with HRMAS-NMR indicated significant changes in some compounds including glycocholate, fatty acids, NADPH, NADP+, some amino acids, thymidine, trigonelline, nicotinic acid, 5,6-dihydrouracil, hesanal, cis-olefin, ß-D-glucose, propanal and some unassigned species. The results suggest that agronomical practices may have effects on fruit composition that may be difficult to detect unless a broad-spectrum analysis is used.

Tissue size and cell number in the olive (Olea europaea) ovary determine tissue growth and partitioning in the fruit.

The relationship between tissue size and cell number in the ovary and tissue size in the fruit, was studied in eight olive (Olea europaea L.) cultivars with different fruit and ovary size. All tissues in the ovary increased in size with increasing ovary size. Tissue size in the fruits correlated with tissue size in the ovary for both mesocarp and endocarp, but with different correlations: the mesocarp grew about twice as much per unit of initial volume in the ovary. Tissue size in the fruit also correlated with tissue cell number in the ovary. In this case, a single regression fitted all data pooled for both endocarp and mesocarp, implying that a similar tissue mass was obtained in the fruit per initial cell in the ovary, independent of tissues and cultivars. Tissue relative growth from bloom to harvest (i.e. the ratio between final and initial tissue size) differed among cultivars and tissues, but correlated with tissue cell size at bloom, across cultivars and tissues. These results suggest that in olive, tissue growth and partitioning in the fruit is largely determined by the characteristics of the ovary tissues at bloom, providing important information for plant breeding and crop management.

Effects of kaolin application on light absorption and distribution, radiation use efficiency and photosynthesis of almond and walnut canopies.

BACKGROUND AND AIMS: Kaolin applied as a suspension to plant canopies forms a film on leaves that increases reflection and reduces absorption of light. Photosynthesis of individual leaves is decreased while the photosynthesis of the whole canopy remains unaffected or even increases. This may result from a better distribution of light within the canopy following kaolin application, but this explanation has not been tested. The objective of this work was to study the effects of kaolin application on light distribution and absorption within tree canopies and, ultimately, on canopy photosynthesis and radiation use efficiency. METHODS: Photosynthetically active radiation (PAR) incident on individual leaves within the canopy of almond (Prunus dulcis) and walnut (Juglans regia) trees was measured before and after kaolin application in order to study PAR distribution within the canopy. The PAR incident on, and reflected and transmitted by, the canopy was measured on the same day for kaolin-sprayed and control trees in order to calculate canopy PAR absorption. These data were then used to model canopy photosynthesis and radiation use efficiency by a simple method proposed in previous work, based on the photosynthetic response to incident PAR of a top-canopy leaf. KEY RESULTS: Kaolin increased incident PAR on surfaces of inner-canopy leaves, although there was an estimated 20 % loss in PAR reaching the photosynthetic apparatus, due to increased reflection. Assuming a 20 % loss of PAR, modelled photosynthesis and photosynthetic radiation use efficiency (PRUE) of kaolin-coated leaves decreased by only 6.3 %. This was due to (1) more beneficial PAR distribution within the kaolin-sprayed canopy, and (2) with decreasing PAR, leaf photosynthesis decreases less than proportionally, due to the curvature of the photosynthesis response-curve to PAR. The relatively small loss in canopy PRUE (per unit of incident PAR), coupled with the increased incident PAR on the leaf surface on inner-canopy leaves, resulted in an estimated increase in modelled photosynthesis of the canopy (+9 % in both walnut and almond). The small loss in PRUE (per unit of incident PAR) resulted in an increase in radiation use efficiency per unit of absorbed PAR, which more than compensated for the minor (7 %) reduction in canopy PAR absorption. CONCLUSIONS: The results explain the apparently contradictory findings in the literature of positive or no effects of kaolin applications on canopy photosynthesis and yield, despite the decrease in photosynthesis by individual leaves when measured at the same PAR.