George A. Manganaris

Hydrogen sulfide induces systemic tolerance to salinity and non-ionic osmotic stress in strawberry plants through modification of reactive species biosynthesis and transcriptional regulation of multiple defence pathways

Hydrogen sulfide (H2S) has been recently found to act as a potent priming agent. This study explored the hypothesis that hydroponic pretreatment of strawberry (Fragaria × ananassa cv. Camarosa) roots with a H2S donor, sodium hydrosulfide (NaHS; 100 μM for 48h), could induce long-lasting priming effects and tolerance to subsequent exposure to 100mM NaCI or 10% (w/v) PEG-6000 for 7 d. Hydrogen sulfide pretreatment of roots resulted in increased leaf chlorophyll fluorescence, stomatal conductance and leaf relative water content as well as lower lipid peroxidation levels in comparison with plants directly subjected to salt and non-ionic osmotic stress, thus suggesting a systemic mitigating effect of H2S pretreatment to cellular damage derived from abiotic stress factors. In addition, root pretreatment with NaHS resulted in the minimization of oxidative and nitrosative stress in strawberry plants, manifested via lower levels of synthesis of NO and H2O2 in leaves and the maintenance of high ascorbate and glutathione redox states, following subsequent salt and non-ionic osmotic stresses. Quantitative real-time RT-PCR gene expression analysis of key antioxidant (cAPX, CAT, MnSOD, GR), ascorbate and glutathione biosynthesis (GCS, GDH, GS), transcription factor (DREB), and salt overly sensitive (SOS) pathway (SOS2-like, SOS3-like, SOS4) genes suggests that H2S plays a pivotal role in the coordinated regulation of multiple transcriptional pathways. The ameliorative effects of H2S were more pronounced in strawberry plants subjected to both stress conditions immediately after NaHS root pretreatment, rather than in plants subjected to stress conditions 3 d after root pretreatment. Overall, H2S-pretreated plants managed to overcome the deleterious effects of salt and non-ionic osmotic stress by controlling oxidative and nitrosative cellular damage through increased performance of antioxidant mechanisms and the coordinated regulation of the SOS pathway, thus proposing a novel role for H2S in plant priming, and in particular in a fruit crop such as strawberry.

A Knowledge Base for The Recovery of Natural Phenols with Different Solvents

The current study denotes the prediction of activity coefficients of fifteen natural phenols (tyrosol, hydroxytyrosol, oleuropein, caffeic, cinnamic, p-coumaric, ferulic, gallic, p-hydroxybenzoic, p-hydroxyphenyl acetic, protocatechuic, rosmarinic, sinapic, syringic, and vanillic acid) in seven solvents (water, ethanol, methanol, acetone, dichloromethane, ethyl acetate, and diethyl ether), and three extraction temperatures (298.15, 313.15, and 333.15 K), using the universal functional-group activity coefficient model. Solvents were classified for their ability to dissolve phenols and were compared with experimental data of the literature in order to observe if the solvent extraction of phenols in practice matches with the authors’ theoretical approach. Results indicated the superiority of alcohols and acetone for the recovery of phenols in line with experimental data of previous studies. Furthermore, activity coefficients’ values were found to increase with the increase of temperature. This study provided a knowledge base for the selection of the most appropriate solvents for a given phenolic compound.

Physiological and proteomic approaches to address the active role of ozone in kiwifruit post-harvest ripening

Post-harvest ozone application has recently been shown to inhibit the onset of senescence symptoms on fleshy fruit and vegetables; however, the exact mechanism of action is yet unknown. To characterize the impact of ozone on the post-harvest performance of kiwifruit (Actinidia deliciosa cv. ‘Hayward’), fruits were cold stored (0 °C, 95% relative humidity) in a commercial ethylene-free room for 1, 3, or 5 months in the absence (control) or presence of ozone (0.3 μl l−1) and subsequently were allowed to ripen at a higher temperature (20 °C), herein defined as the shelf-life period, for up to 12 days. Ozone blocked ethylene production, delayed ripening, and stimulated antioxidant and anti-radical activities of fruits. Proteomic analysis using 1D-SDS-PAGE and mass spectrometry identified 102 kiwifruit proteins during ripening, which are mainly involved in energy, protein metabolism, defence, and cell structure. Ripening induced protein carbonylation in kiwifruit but this effect was depressed by ozone. A set of candidate kiwifruit proteins that are sensitive to carbonylation was also discovered. Overall, the present data indicate that ozone improved kiwifruit post-harvest behaviour, thus providing a first step towards understanding the active role of this molecule in fruit ripening.

Nutritional Quality of fruits and vegetables

The nutritional value of fruits and vegetables depends on their composition, which shows a wide range of variation depending on the species, cultivar, and maturity stage. This chapter describes the general characteristics of the components of fruits and vegetables, related to their benefits as food sources. There are two types of acids, namely aliphatic (straight chain) and aromatic acids. The most abundant acids in fruits and vegetables are citric and malic (both aliphatic) acids. However, large amounts of tartaric acid occur in grapes. Malic acid is the major component in oranges and apples. The acid content of fruits and vegetables generally decreases during maturation. Aromatic organic acids occur in several fruits and vegetables, but in very low concentrations. Benzoic acid occurs in cranberries, quinic acid in bananas, and chlorogenic acid in potatoes. In general, vegetables are a richer source of minerals than fruits, but both vegetables and fruits are considered nutrient-dense foods in that they provide substantial amounts of micronutrients, such as minerals and vitamins, but relatively few calories. Minerals have both direct and indirect effects on human health. The direct effects of minerals focus on the consequences of their consumption on human nutrition, while the indirect effects refer to their incidence in fruit and vegetable quality and subsequent consumer acceptance. From a direct nutrition standpoint, potassium has the biggest presence in both fruits and vegetables, but nitrogen and calcium show major impacts on horticultural crop quality.

Towards an Efficient Protocol for the Determination of Triterpenic Acids in Olive Fruit: A Comparative Study of Drying and Extraction Methods

INTRODUCTION Triterpenic acids, such as maslinic acid and oleanolic acid, are commonly found in olive fruits and have been associated with many health benefits. The drying and extraction methods, as well as the solvents used, are critical factors in the determination of their concentration in plant tissues. Thus, there is an emerging need for standardisation of an efficient extraction protocol that determines triterpenic acid content in olive fruits. OBJECTIVE To evaluate common extraction methods of triterpenic acids from olive fruits and to determine the effect of the drying method on their content in order to propose an optimum protocol for their quantification. METHODOLOGY The efficacy of different drying and extraction methods was evaluated through the quantification of maslinic acid and oleanolic acid contents using the reversed-phase HPLC technique. RESULTS Data showed that ultrasonic assisted extraction with ethanol or a mixture of ethanol:methanol (1:1, v/v) resulted in the recovery of significantly higher amounts of triterpenic acids than other methods used. The drying method also affected the estimated triterpenic acid content; frozen or lyophilised olive fruit material gave higher yields of triterpenic acids compared with air-dried material at both 35°C and 105°C. CONCLUSION This study provides a rapid and low-cost extraction method, i.e. ultrasonic assisted extraction with an eco-friendly solvent such as ethanol, from frozen or lyophilised olive fruit for the accurate determination of the triterpenic acid content in olive fruit.

Establishment of a rapid, inexpensive protocol for extraction of high quality RNA from small amounts of strawberry plant tissues and other recalcitrant fruit crops

Strawberry plant tissues and particularly fruit material are rich in polysaccharides and polyphenolic compounds, thus rendering the isolation of nucleic acids a difficult task. This work describes the successful modification of a total RNA extraction protocol, which enables the isolation of high quantity and quality of total RNA from small amounts of strawberry leaf, root and fruit tissues. Reverse-transcription polymerase chain reaction (RT-PCR) amplification of GAPDH housekeeping gene from isolated RNA further supports the proposed protocol efficiency and its use for downstream molecular applications. This novel procedure was also successfully followed using other fruit tissues, such as olive and kiwifruit. In addition, optional treatment with RNase A following initial nucleic acid extraction can provide sufficient quality and quality of genomic DNA for subsequent PCR analyses, as evidenced from PCR amplification of housekeeping genes using extracted genomic DNA as template. Overall, this optimized protocol allows easy, rapid and economic isolation of high quality RNA from small amounts of an important fruit crop, such as strawberry, with extended applicability to other recalcitrant fruit crops.

Effect of post-harvest calcium treatments on the physicochemical properties of cell wall pectin in nectarine fruit during ripening after harvest or cold storage

Summary The effects of post-harvest dips with calcium salts on the quality attributes of nectarine fruits (Prunus persica var. nectarina Ait. Maxim, cv. ‘Caldesi 2000’) were determined after harvest or in cold storage for up to 6 weeks. Nectarine fruits were harvested at the firm-ripe stage (flesh firmness 70 ± 2 N; soluble solids content 10.5 ± 0.2%) and dipped in deionised water (control) or calcium salts (chloride, lactate or propionate) at two calcium concentrations (62.5 or 187.5 mM). The calcium content of nectarine fruits increased up to 240% in the peel, and up to 31% in the flesh, 1 d after immersion. The increase in calcium content in the flesh was accompanied by increased cell wall calcium, corresponding to a significant increase in the “insoluble” pectin fraction. The highest calcium accumulation in the “insoluble” pectin fraction was attained with 62.5 mM calcium which positively affected fruit firmness. The higher calcium concentration (187.5 mM) resulted in increased calcium in the “water-soluble” pectin fraction which did not appear to contribute to retention of tissue firmness. Additionally, still higher concentrations of calcium lactate and calcium propionate resulted in surface damage, leading to undesirable characteristics during fruit ripening.

REVIEW ARTICLE Hydrogen sulphide: a versatile tool for the regulation of growth and defence responses in horticultural crops

Summary The improvement of plant growth and productivity under biotic and abiotic stress conditions, as well as the reduction of post-harvest losses, are of paramount significance to meet the increasing demand for food by a growing World population. Hydrogen sulphide (H2S) has recently appeared as a key contributor towards this goal through its bioactive role in the regulation of plant defence responses. In this review, we provide an up-to-date overview of recent literature concerning the biosynthesis and regulation of H2S within plant cells, as well as its involvement in a series of plant physiological processes. More specifically, H2S is actively associated with several horticultural plant growth and developmental processes, tolerance responses following exposure to stress factors, as well as in post-harvest fruit physiology.The H2S-mediated enhancement of tolerance to several abiotic stress factors is highlighted, with particular emphasis on the priming effects of H2S on the anti-oxidant capacity, redox regulation and signalling, and transcriptional regulation of cellular defence components in plants. Moreover,H2S has been reported to be associated with seed germination, increased growth, and root organogenesis, while also enhancing the post-harvest performance and anti-oxidant capacity of fruit.The present review proposes a practical role for H2S in horticultural crop physiology, further supporting the notion that H2S acts as a key signalling molecule in plants, possibly acting by “cross-talk” with other secondary plant cell messengers.

Classification, Biotransformation and Antioxidant Activity of Olive Fruit Biophenols: A Review

Olive biophenols affect the organoleptic and nutraceutical properties of fruits; therefore, it is an emerging re- search field. In addition, a plethora of health-promoting properties have been attributed to the phenolic content of olive fruits. The main biophenols can be divided into different sub-groups such as phenolic acids and alcohols, flavonoids, lig- nans and secoiridoids. The latter sub-group of biophenols is typical in olive fruits, since oleuropein and the products of its biotransformation characterize olive fruit. An array of genetic, agronomic and technological factors that affect the rate of synthesis and biotransformation of secoiridoids in olive fruits is discussed. Furthermore, the relationship between antioxi- dant activity and structures of olive biophenols is explored.

1H NMR Metabolic Fingerprinting to Probe Temporal Postharvest Changes on Qualitative Attributes and Phytochemical Profile of Sweet Cherry Fruit

Sweet cherry fruits (Prunus avium cvs. ‘Canada Giant’, ‘Ferrovia’) were harvested at commercial maturity stage and analyzed at harvest and after maintenance at room temperature (storage at ∼20°C, shelf life) for 1, 2, 4, 6, and 8 days, respectively. Fruit were initially analyzed for respiration rate, qualitative attributes and textural properties: ‘Canada Giant’ fruit were characterized by higher weight losses and stem browning index, being more intense over the late stages of shelf life period; meanwhile ‘Ferrovia’ possessed appreciably better performance even after extended shelf life period. A gradual decrease of respiration rate was monitored in both cultivars, culminated after 8 days at 20°C. The sweet cherry fruit nutraceutical profile was monitored using an array of instrumental techniques (spectrophotometric assays, HPLC, 1H-NMR). Fruit antioxidant capacity was enhanced with the progress of shelf life period, concomitant with the increased levels of total anthocyanin and of phenolic compounds. ‘Ferrovia’ fruit presented higher contents of neochlorogenic acid and p-coumaroylquinic acid throughout the shelf life period. We further developed an 1H-NMR method that allows the study of primary and secondary metabolites in a single running, without previous separation and isolation procedures. Diagnostic peaks were located in the aliphatic region for sugars and organic acids, in the aromatic region for phenolic compounds and at 8.2–8.6 ppm for anthocyanins. This NMR-based methodology provides a unifying tool for quantitative and qualitative characterization of metabolite changes of sweet cherry fruits; it is also expected to be further exploited for monitoring temporal changes in other fleshy fruits.