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Avelumab is an IgG1 anti–programmed death ligand 1 (anti–PD-L1) monoclonal antibody that has been approved as a monotherapy for metastatic Merkel cell carcinoma and advanced urothelial carcinoma, and in combination with axitinib for advanced renal cell carcinoma. Avelumab is cleared faster and has a shorter half-life than other anti–PD-L1 antibodies, such as atezolizumab and durvalumab, but the mechanisms underlying these differences are unknown. IgG antibodies can be cleared through receptor-mediated endocytosis after binding of the antibody Fab region to target proteins, or via Fcγ receptor (FcγR)-mediated endocytosis. Unlike other approved anti–PD-L1 antibodies, avelumab has a native Fc region that retains FcγR binding capability. We hypothesized that the rapid clearance of avelumab might be due to the synergistic effect of both FcγR-mediated and PD-L1 target–mediated internalization. To investigate this, we performed in vitro and in vivo studies that compared engineered variants of avelumab and atezolizumab to determine mechanisms of cellular internalization. We found that both FcγR and PD-L1 binding contribute to avelumab internalization. While FcγR binding was the dominant mechanism of avelumab internalization in vitro, with CD64 acting as the most important FcγR, studies in mice and cynomolgus monkeys showed that both FcγR and PD-L1 contribute to avelumab elimination, with PD-L1 binding playing a greater role. These studies suggest that the rapid internalization of avelumab might be due to simultaneous binding of both PD-L1 and FcγR in trans. Our findings also provide a basis to alter the clearance and half-life of monoclonal antibodies in therapeutic development.

In 2019–2020, trials were carried out with the aim of evaluating the possibilities of using an organic fertilizer, also having a biostimulant action, for promoting the growth of young olive trees. The experiments were implemented using both potted and field-grown trees. The effects of the organic fertilizer were evaluated with respect to trees treated with a chemical fertilizer, for potted trees a control without any fertilization was also used. Therefore, the compared treatments were: organic fertilization vs. chemical fertilization or no fertilization in the experiment with potted trees, organic fertilization vs. chemical fertilization in the experiment with field-grown trees. Non-fertilized potted trees had the lowest growth. The organic fertilizer, with respect to the chemical one, determined higher growth in both potted and field-grown trees (+22–29% of the increment of the trunk cross sectional area). In potted trees, it determined a rapid elongation of the stem (+30% of the increment of tree height with respect to chemically fertilized trees) and this likely favored the development of a higher number of leaves, which, together with an increase in their photosynthetic activity (in August, +27% with respect to chemical fertilized trees), created conditions for higher assimilate production and in turn greater tree growth. Trees in the field showed that the greater growth was not obtained at the expenses of reproductive growth, which increased to the same degree as the vegetative growth. Overall, the results support the biostimulant action of the organic fertilizer and indicate the possibility of its use to improve the growth of young olive trees.

Appropriate selection of conjugation sites and conjugation technologies is now widely accepted as crucial for the success of antibody-drug conjugates (ADCs). Herein, we present ADCs conjugated by different conjugation methods to different conjugation positions being systematically characterized by multiple in vitro assays as well as in vivo pharmacokinetic (PK) analyses in transgenic Tg276 mice. Conjugation to cysteines, genetically introduced at positions N325, L328, S239, D265, and S442, was compared to enzymatic conjugation via microbial transglutaminase (mTG) either to C-terminal light (LC) or heavy chain (HC) recognition motifs or to endogenous position Q295 of a native antibody. All conjugations yielded homogeneous DAR 2 ADCs with similar hydrophobicity, thermal stability, human neonatal Fc receptor (huFcRn) binding, and serum stability properties, but with pronounced differences in their PK profiles. mTG-conjugated ADC variants conjugated either to Q295 or to LC recognition motifs showed superior PK behavior. Within the panel of engineered cysteine variants L328 showed a similar PK profile compared to previously described S239 but superior PK compared to S442, D265, and N325. While all positions were first tested with trastuzumab, L328 and mTG LC were further evaluated with additional antibody scaffolds derived from clinically evaluated monoclonal antibodies (mAb). Based on PK analyses, this study confirms the newly described position L328 as favorable site for cysteine conjugation, comparable to the well-established engineered cysteine position S239, and emphasizes the favorable position Q295 of native antibodies and the tagged LC antibody variant for enzymatic conjugations via mTG. In addition, hemizygous Tg276 mice are evaluated as an adequate model for ADC pharmacokinetics, facilitating the selection of suitable ADC candidates early in the drug discovery process.

Experiments were carried out in 2018 and 2019 to evaluate the effects of a biostimulant of animal origin, with a hormone-like action, on the growth and physiology of young olive trees. The biostimulant, consisting of a complex of natural amino acids (glycine, proline, and hydroxyproline, etc.), was applied by fertigation to both potted (one-year-old) and field-grown (2 years after transplanting) young olive trees. The control consisted of trees treated with an amount of urea nitrogen equal to the total nitrogen supplied with the biostimulant. Potted trees treated with the biostimulant showed greater growth than the control soon after treatment and up to the end of the vegetative season. Generally, this was associated with higher leaf photosynthetic rates and stomatal conductance. At the end of the experiment, all tree parts (roots, stem, shoots, and leaves) of the treated trees had greater biomass than the control. The trunks of field-grown trees treated with the biostimulant had greater growth with respect to the control. Overall, the results indicate the possibility of using the biostimulant based on complexes of natural amino acids to promote the growth of young olive trees.

Blocking Fibroblast Growth Factor Receptor 1 (FGFR1) is an attractive therapeutic option for treatment of cancer subtypes with amplification and over-expression of FGFR1. Selective targeting of FGFR1 can be achieved using an antibody-based approach, as small molecule inhibitors may not discriminate between FGFR1, 2, 3 and 4 due to their highly homologous kinase domain. However, development of classical bivalent FGFR1 directed antibodies has failed due to non-tolerated body weight decreases in preclinical species. M6123 is a novel IgG-like monovalent FGFR1 specific binder with enhanced Antibody-Dependent Cellular Cytotoxicity (ADCC) effector function and inhibits tumor growth significantly in FGFR1-dependent human xenograft models without reduced body weight in tumor-bearing mice. Toxicology studies reported here characterized the safety profile of M6123 in mouse, rat, and monkey. There were significant differences among animal species under similar M6123 exposure levels. Rats showed metastatic mineralization with an imbalance in serum phosphate at low doses, while mineralization was not found in mice or monkeys, even though hyperphosphatemia was detected in mice. Subtle differences in calcium/phosphate homoeostasis feedback loops may trigger the susceptibility to mineralization among animal species; nevertheless, the exact mechanism remains unknown. Monkeys showed marked, but reversible, decreases in peripheral blood NK cells and neutrophils. The latter was associated with considerably increased neutrophilic infiltrates in the liver sinusoids and red pulp of the spleen. These effects in monkeys are likely related to the enhanced ADCC activity of M6123. Overall, M6123 showed a superior safety profile in animals compared to bivalent FGFR1 antagonists or pan-FGFR inhibitors.

Low vigor and early and abundant production are desirable traits for modern tree crops. In olive, most cultivars are too vigorous and cannot be successfully constrained in the small volume allowed by the straddle harvester used in the so-called superhigh-density (SHD) orchards. Only few cultivars appear to have sufficiently low vigor to be suitable for this system. These cultivars combine low vigor with earlier and higher yield. This study investigated the hypothesis that differences in vigor between Arbequina, a low vigor and the most commonly used cultivar in SHD orchards, and Frantoio, a highly vigorous cultivar not suitable for such orchards, are related to their differences in early bearing and consequent differences in dry matter partitioning into fruit. Young trees of both cultivars were deflowered either in 2014, 2015, both years, or neither one, resulting in a range of cumulative yields over the 2 years. Tree trunk cross-sectional area (TCSA) was measured at the beginning of each year. This was closely related to total tree mass, as assessed at the beginning and at the end of the experiment. Cumulative yield, in terms of fruit dry matter, was also assessed. TCSA increased less in fruiting trees in both years. As expected, when not deflowered, ‘Frantoio’ was less productive and more vigorous than ‘Arbequina’. However, there was no difference in TCSA increment when both cultivars were completely deflowered. TCSA increments were closely inversely related to yield across all treatments and cultivars (R2 = 0.90). The regressions improved further when data from 2015 only were used (R2 = 0.99). The results represent the first quantitative report showing that differences in vigor among cultivars can be completely explained in terms of different dry matter partitioning into fruit, supporting the hypothesis that early bearing is a major cause, rather than merely a consequence, of lower vigor in young ‘Arbequina’ trees. These results provide new understanding on vigor differences across cultivars, which will be useful for breeding and selection of new genotypes.

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.

T cell immunoglobulin and mucin domain-3 (TIM-3) is an immune checkpoint that regulates normal immune responses but can be exploited by tumor cells to evade immune surveillance. TIM-3 is primarily expressed on immune cells, particularly on dysfunctional and exhausted T cells, and engagement of TIM-3 with its ligands promotes TIM-3-mediated T cell inhibition. Antagonistic ligand-blocking anti-TIM-3 antibodies have the potential to abrogate T cell inhibition, activate antigen-specific T cells, and enhance anti-tumor immunity. Here we describe M6903, a fully human anti-TIM-3 antibody without effector function and with high affinity and selectivity to TIM-3. We demonstrate that M6903 blocks the binding of TIM-3 to three of its ligands, phosphatidylserine (PtdSer), carcinoembryonic antigen cell adhesion-related molecule 1 (CEACAM1), and galectin 9 (Gal-9). These results are supported by an atomic resolution crystal structure and functional assays, which demonstrate that M6903 monotherapy enhanced T cell activation. This activation was further enhanced by the combination of M6903 with bintrafusp alfa, a bifunctional fusion protein that simultaneously blocks the transforming growth factor-β (TGF-β) and programmed death ligand 1 (PD-L1) pathways. M6903 and bintrafusp alfa combination therapy also enhanced anti-tumor efficacy in huTIM-3 knock-in mice, relative to either monotherapy. These in vitro and in vivo data, along with favorable pharmacokinetics in marmoset monkeys, suggest that M6903 as a monotherapy warrants further pre-clinical assessment and that M6903 and bintrafusp alfa may be a promising combination therapy in the clinic.

Seeds from Citrus species represent a relevant by-product of the juice industry and a potential source of bioactive compounds such as phenols and other antioxidants. Sprouting could be an intriguing idea to enhance the content of these compounds, as explored for other fruittree species. In this experiment, the sprouting performance, the concentration of total phenols and phenolic acids, and the antioxidant activity of seeds and sprouts were evaluated for bitter orange (Citrus aurantium L. seedlings), blonde orange (Citrus sinensis (L.) Osbeck cv.Biondocomune), sweet orange (Citrus sinensis (L.) Osbeck seedlings), lemon (Citrus limon (L.) Osbeck cv.Femminello), and mandarin (Citrus reticulata Blanco cv.Tardivo di Ciaculli). The germination was high for all genotypes except for mandarin, but it took 4&ndash, 8 weeks. Sprouts did not differ among genotypes for size and generally had hard consistency of cotyledons and a bitter taste. The concentrations of total phenols and phenolic acids of seeds and sprouts varied with the genotype, while the antioxidant activity was not statistically different among treatments. Sprouting increased both the concentration of phenolic compounds and antioxidant activity but no correlation was found between them, suggesting that other antioxidants, besides phenols, are present. Given the slow germination and the bitter taste, Citrus sprouts appear unsuitable for homemade production aimed at direct consumption, while they may have perspectives for extraction of food additives, cosmetics, and pharmaceutics.

This study evaluated the effects of different olive harvesting systems and fruit storage durations on quality of oils from both super-intensive (cv. Arbequina) and intensive (cv. Frantoio) olive groves. Four harvesting systems were used: gentle manual harvesting, manual harvesting with hand-held combs, facilitated harvesting with hand-held pneumatic combs, and mechanical harvesting with a straddle machine (in super-intensive grove) or a trunk shaker + reversed umbrella (in intensive grove). Gentle hand harvesting caused the least damage to the fruits, whereas the trunk shaker and the straddle machine caused the greatest damage. The damage increased with olive storage. Total polyphenols and single secoiridoid were all negatively correlated with the degree of fruit damage. Volatile compounds (aldehydes, alcohols, esters and ketones) were strongly affected by both the harvesting system and the olive storage. In both cultivars, increasing the mechanization level (from gentle hand harvest to the straddle machine or the trunk shaker) caused a decrease of total aldehydes and total esters, and an increase of total alcohols and total ketones. Moreover, there were negative relationships between the degree of fruit damage and the total C5 and C6 aldehydes and total esters and positive relationships between the degree of fruit damage and the total ketones. Overall, the results indicate that fruit damage explains most of the worsening of oil quality across harvesting systems and storage duration. This is the first study reporting a clear quantitative/linear correlation between fruit damage and important components of oil quality, such as polyphenols and volatile compounds deriving from the lipoxygenase pathway.

Total phenolic content (TPC) and antioxidant activity (AOXA) of olive (Olea europaea L.) sprouts were measured in three cultivars (Arbequina, Moraiolo and Leccino). Olive seeds, obtained by manually breaking whole stones derived from de-stoning ripe fruits, were sown in plastic trays and subjected to cold stratification for 30 days. The trays were then moved to 25 °C and 16:8 light:dark regime. Total germination was 27% in Arbequina, 18% in Leccino, 14% in Moraiolo. Mean germination time (MGT) was 10 days in Leccino and Moraiolo and 14 days in Arbequina. Shoots were progressively harvested once the cotyledons were completely unfolded, i.e., between 8 and 16 days after germination. Sprouting dramatically increased TPC and AOXA in all cultivars (on average, 14 and 30 times higher, respectively, compared to seeds), with Moraiolo and Leccino showing higher values than Arbequina. Sprouting olive seeds appears to be a valid way to obtain antioxidants for food supplementation or pharmaceutical purposes.

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., dry matter partitioning, harvest index, harvest increment, growth, canopy to root ratio, Olea europaea

N and K fertilization usually increases olive fruit oil content as well as fruit size and pulp to pit ratio. The objective of this work was to investigate whether the increase in fruit oil content occurs via increased oil concentration in the pulp, or via increased fraction of pulp in the fruit. In 2009 Leccino olive trees were fertilized with the equivalent of 180 kg of urea-N, split in three equal doses (April, June, September). In April, K2O fertilizer (potassium sulphate) was also applied for an equivalent amount of 100 kg of K2O per hectare. Control trees were not fertilized. Fresh and dry weight of fruit, pulp and pit, fruit and pulp oil and water content, pulp firmness and fruit maturity index were measured throughout fruit growth, on fruit samples from both fertilized and control trees. At all sampling dates, fertilized plants had significantly greater fruit weight, both on a fresh (+23% on average) and dry (+17% on average) basis. Pulp/pit ratio also increased significantly with fertilization (+23%, both on fresh and dry basis). Fruits of fertilized plants also had significantly higher oil content on dry matter basis (+9% on average). However, pulp oil content did not differ significantly between treated and control plants. The same was found for water content, which was significantly higher in the fruit of fertilized plants (+5% on average), but not in the pulp. Fertilization did not affect fruit maturation (i.e., color and pulp firmness).

The occurrence of phosphoenolpyruvate carboxykinase (PEPCK) and pyruvate orthophosphate dikinase (PPDK) was determined in the ripe pericarps of several grape genotypes. Moreover, the abundance pattern of PEPCK during the development of grape berries was studied in a range of cultivars. In addition, the presence of PEPCK in grape leaves and the location/abundance of PEPCK in developing grape seeds were investigated. PEPCK was present in the pericarp of all the cultivars of Vitis vinifera studied and in that of the hybrid 420A (Vitis berlandieri × Vitis riparia). By contrast, PPDK was either not present or present at very low abundance. The results strongly indicated that the PEPCK and not the PPDK pathway is the predominant one utilised in gluconeogenesis in the berry pericarp of grape genotypes. Further, the results showed that the abundance of PEPCK increased greatly at the start of ripening in all the cultivars, showing a similar pattern in all of them. PEPCK was not present (or at very low abundance) in grape leaves throughout their development. In developing grape seeds, the results indicated a similar abundance of PEPCK in both the seed coat and the tissues enclosed by it.

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.

Analyzing plant architecture is important for the understanding of plant growth, branching pattern, yield, and for the development of crop models. In other species tree architecture has been studied in detail, but not in olive. In this species, super high density (SHD) orchards (>1500 trees per hectare) have been extensively planted in recent years, using only a few cultivars (i.e. Arbequina, Arbosana and Koroneiki) considered suitable for this system. What makes these cultivars suitable for SHD orchards is not clear. In this work it is hypothesized that tree architecture plays an important role in this respect. We studied several tree architectural and reproductive parameters on different olive cultivars, including Arbequina, Arbosana and 19 others, mostly Italian. The impact of the architectural parameters on tree productivity was also studied. The architectural parameters studied were: diameter and node number of the trunk; number, diameter and angle of insertion of the lateral branches departing from the central leader; diameter, length, node number and average angle of fruiting shoots. On fruiting shoots, inflorescence and fruit number were also recorded respectively at bloom and at harvest. Branching frequency was calculated as the average number of lateral branches per bud. Arbequina and Arbosana differed from other cultivars for a greater branching frequency. The latter was negatively correlated to the diameter of lateral branches as well as to the diameter of one-year-old shoots. Consequently, Arbequina and Arbosana produced a greater number of smaller lateral branches and shoots, both parameters being desirable for SHD olive orchards. Combining branching and fruiting characteristics, we developed yield indices which segregated these cultivars from most others. The results suggest that high branching and small diameters are important architectural characteristics to increase yield efficiency, and affect the cultivar suitability for SHD orchards.

The modern olive industry is increasingly interested in olive cultivars that start producing early and remain relatively small, because they are suitable for super high-density orchards. Some cultivars are better suited to this than others but it is not clear why. Understanding the mechanisms that lead to early production and reduced canopy size is therefore important. The object of this study was to investigate whether differences in vigor across olive cultivars are related to earliness and abundance of bearing. We analyzed tree growth and productivity in young coetaneous trees of 12 olive cultivars, grown together in the same orchard. Trunk diameter increased over the observation period, reaching significantly different values across cultivars. Canopy volume also increased, reaching 2-fold differences between the minimum and the maximum values. Cumulative yield increased, reaching up to 3-fold differences. When the cumulative yield at the end of the experiment was plotted against the final trunk diameter, no correlation was found. A significant correlation was found when cumulative yield was plotted against the increment in trunk diameter during the observation period for which yield data were collected. This relationship improved (i.e., R2 rose from 0.57 to 0.83) when yield efficiency [i.e., cumulative yield per unit of final trunk cross-sectional area (TCSA) or per unit of canopy volume] was used instead of yield. These results clearly showed that trees that produced proportionally more (i.e., higher yield efficiencies) grew less. We conclude that, in young olive trees, vigor is inversely related to early bearing efficiency, which differs significantly across cultivars. The results support the hypothesis that early and abundant bearing is a major factor in explaining differences in vigor across olive cultivars.

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. © 2011 Elsevier B.V., The study was funded by the Italian Ministry for Agricultural, Food and Forestry Politics (MiPAAF) RIOM project, and Spanish Ministry of Science and Innovation Grant AGL2009-07248.

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 strongly correlated to ovary weight in olive as big-fruited cultivars have larger ovaries and flowers at bloom. We tested the hypothesis that larger ovaries imply stronger sinks and therefore reduce fruit set, expressed as number of fruits. Flower/fruit load per centimeter of shoot was assessed every month from flowering to harvest, on sample shoots of many different olive cultivars differing in fruit size. Data were taken for two years: 2007 and 2008. Additionally, in 2008, a fruit thinning experiment was carried out by leaving a single fruit per shoot, to assess the maximum potential fruit weight achievable by each cultivar. Results indicated that fruit size was mostly genetically determined as thinned fruits of small-fruited cultivars, although bigger than control fruits, never got as big as fruits from large-fruited cultivars, confirming that large ovaries are a prerequisite for large fruits. Among all cultivars, the number of flowers per inflorescence, the number of inflorescences, and the number of flowers per centimeter of shoot were not correlated to average flower weight while total flower mass per centimeter of shoot was positively correlated to average flower weight (i.e. large flowered/fruited cultivars had greater total flower mass). However, one month after bloom and thereafter, all parameters were negatively and exponentially correlated with fruit average weight across cultivars, except fruit mass per centimeter of shoot which was generally not correlated to fruit average weight, except in one year, at harvest, when fruit mass per centimeter of shoot was positively correlated to fruit average weight. These results suggest that fruit set is a consequence of, and inversely proportional to flower/fruit size in olive.

In this study the occurrence of a number of enzymes involved in gluconeogenesis was investigated in both tomato fruits and leaves during their development and senescence and in some other fruits. The enzymes studied were phosphoenolpyruvate carboxykinase (PEPCK), pyruvate orthophosphate dikinase (PPDK) and glyoxysomal isocitrate lyase (ICL). PPDK was detected in the ripe flesh of tomato, and much smaller amounts were detected in the flesh of both peach and pepper, whereas it was not detected (not present or at very low abundance) in the other fruits which were investigated (apricot, aubergine, blackberry, blueberry, cherry, grape, plum, raspberry and red current). By contrast PEPCK was present in the flesh of all the fruits investigated. Very small amounts of ICL were detected in ripe tomato flesh. PEPCK was present in the skin, flesh, locular gel and columella of tomato fruit, and in these its abundance increased greatly during ripening. PPDK showed a similar distribution, however, its abundance did not increase during ripening. PEPCK was not detected in tomato leaves at any stage of their development or senescence. The content of PPDK g(-1) fresh weight (FW) increased in tomato leaves as they matured, however, it declined during their senescence. In tomato leaves the content of ICL g(-1) FW increased until the mid-stage of development, then decreased as the leaf matured, and then increased during the latter stages of senescence. In the flesh of tomato fruits the contents of PPDK and PEPCK g(-1) FW decreased during senescence. The results suggest that in fruits other than tomato the bulk of any gluconeogenic flux proceeds via PEPCK, whereas in tomato both PEPCK and PPDK could potentially be utilised. Further, the results indicate that the conversion of pyruvate/acetyl-CoA to malate by the glyoxylate cycle, for which ICL is necessary, is not a major pathway utilised by gluconeogenesis in fruits under normal conditions of growth. Finally, the results contribute to our understanding of the role of several enzymes in the senescence of both leaves and fruits.

SUMMARY The aim of the present study was to analyze the effect of range enrichment (trees or tall grass stand) on the performance and herbage intake of free-range chicken. This trial was conducted in 2 different seasons (winter and summer) comparing a standard free-range system with 2 kinds of range enrichment: a stand of sorghum (a tall grass) and olive trees. Two hundred fifty male naked neck chickens were used in each season and system; performance, behavior, forage intake, and body lesions were recorded. Productive performance was not affected by range enrichment. However, in the presence of trees or tall grass stand, no predation losses were experienced; whereas, in the standard paddock, cases of predation by raptors or crows were registered, mainly during the first period of rearing. Consequently, mortality rate was significantly lower in the enriched environment. Control chickens stayed indoors more rather than forage in the pasture, whereas, in enriched environments, birds spent more time outdoors and widely exploited the available pasture. Forage intake was significantly influenced by rearing system and season; chickens reared under olive trees had higher herbage ingestion, exploring the available area up to almost 50 m from the hut. With the olive trees, chickens also showed the lowest frequency of foot and breast damage.

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, β-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.

Floral biology has important practical implications, in addition to its scientific relevance, given that flower characteristics and bloom affect fruit characteristics and yield. Yield derives from fruit quality (e.g. weight) and quantity (i.e. number), which, in turns, depend on flower quantity and quality: flowers must be suitable to become fruits, and then must be pollinated and fertilized, and must set fruits, which must then grow. Not all flowers can do all of this: some flowers, for instance, have aborted ovaries which are partially developed or absent at bloom, depending on when the abortion occurred. Even when still present, these aborted ovaries are not capable of becoming fruits. Normal pistils, may not be pollinated or fertilized, but also fertilized ovaries may drop after some growth, resulting in fruit drop. From 100 flowers, in olive, all the above phenomena result in one to few fruits (Hartmann, 1950). Because of this low fruit set, it is often believed that cultural practices aimed at improving pollination, increasing fruit set or reducing ovary abortion or fruit drop, may lead to increased olive yields.

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. © 2012 CSIRO., The study was funded by the Italian Ministry for Agricultural, Food and Forestry Policies (MiPAAF), RIOM project, Progetto Strategico OLEA–Genomica e Miglioramento Genetico dell’Olivo’, D.M. 27011/7643/10 and by the Spanish Ministry of Science and Innovation ERDF co-financed funds project AGL2009–07248.

The estimates of potential biomass production from pruning in olive often follow generic indications not considering varietal differences, training and pruning systems. The main limitation to the valorization of pruning concerns the collection-logistics chain which requires precise starting data, often linked to specific production areas. Traditional groves still represent the majority of the Italian olive cropping systems. Information about intensity and type of rejuvenation pruning is still lacking for these orchards and data from different climatic conditions are important for further planification of new power plants for energy valorization. This study evaluates the biomass produced from two rejuvenation pruning intensities (i.e. intense and light) in five Italian cropping systems in two consecutive years. In the first year, the amount of removed material per tree was correlated to the initial canopy volume within treatment so that the biomass produced with intense pruning in the cultivars with bigger tree size passed 30 t ha-1. In both years, shreddable material was the prevalent fraction (>50%) in the tested cultivars and for both pruning treatments. Even adopting the lowest calorific value of 14.4 MJ kg-1 of dry matter, the energy potential appears promising for a possible economic valorization of the biomass., Proceedings of the 29th European Biomass Conference and Exhibition, 26-29 April 2021, Online, pp. 273-278

In Italy, the transport cost is a bottleneck for most biomass agricultural products. The increase in by-product density without deteriorating the quality of the final product is a fundamental aspect for the spreading of bioenergy crops in agricultural areas. In wheat chaff samples harvested in the Emilia-Romagna region (Italy), the present work investigates the briquettes techniques for biomass volume reduction and density increase. Regardless of the type of enhancement, the increase of density with a specific and versatile system would be an immediate advantage, especially in logistics and transport costs, which would benefit almost linearly. Nonetheless, the first experiences aim also at avoiding the adoption of specific and expensive industrial technologies that could initially limit the resulting advantages. In this regard, the Council for Agricultural Research and Economics (CREA) has been undertaking experimental activities aimed at the valorization (not only for energy purposes) of residual lignocellulosic materials of various origins. The threshing residues resulted in being viable as long as there is adequate logistical support. The present work investigates applying a professional briquetting machine (not expressly developed for agricultural by-products) to wheat threshing samples for biomass volume reduction and density increase. The preliminary results were of interest: the volume of the processed biomass was 19 times smaller than that of the raw one with an increase of density from 33 kg m-3 of the bulk product to about 626 kg m-3 of the briquettes., Proceedings of the 29th European Biomass Conference and Exhibition, 26-29 April 2021, Online, pp. 178-181