The impact of wounding and postharvest storage conditions on retention of soluble protein in sugar beet leaves.

Sugar beet leaves can be a viable and economically interesting source of high-quality protein for the food industry. We investigated how storage conditions and leaf wounding at harvest affect the content and quality of the soluble protein. After collection, leaves were either stored intact or shredded to mimic wounding induced by commercial leaf harvesters. Leaf material was stored in small volumes at different temperatures to assess leaf physiology or in larger volumes to assess temperature development at different locations in the bins. Protein degradation was more pronounced at higher storage temperatures. Wounding accelerated the degradation of soluble protein at all temperatures. Both wounding and storage at higher temperatures greatly stimulated respiration activity and heat production. At temperatures below 5°C, ribulose-1,5-biphosphate carboxylase oxygenase (RuBisCO) in intact leaves was preserved for up to 3 weeks. At temperatures of 30-40°C, RuBisCO degradation occurred within 48 h. Degradation was more pronounced in shredded leaves. In 0.8-m3 storage bins at ambient temperature, core temperatures rapidly increased, up to 25°C in intact leaves and up to 45°C in shredded leaves within 2-3 days. Immediate storage at 5°C greatly suppressed the temperature increase in intact but not in shredded leaves. The indirect effect of excessive wounding, that is, heat production, is discussed as the pivotal factor responsible for increased degradation of protein. For optimal retention of soluble protein levels and quality in harvested sugar beet leaves, it is advised to minimize wounding and to store the material at temperatures around -5°C. PRACTICAL APPLICATION: To preserve the soluble protein content and quality for at least 3 weeks, sugar beet leaves should be harvested with minimal wounding and stored at temperatures between 1 and 5°C. When aiming to store minimally wounded leaves in larger volumes, it must be ensured that the product temperature in the core of the biomass meets the temperature criterium or the cooling strategy must be adjusted. The principles of minimal wounding and low temperature storage are transferable to other leafy crops that are harvested for food protein.

The association between the susceptibility to Botrytis cinerea and the levels of volatile and non-volatile metabolites in red ripe strawberry genotypes.

The relations between physical and chemical characteristics (e.g., color, firmness, volatile and non-volatile metabolites) of red ripe strawberry fruit and the natural spoilage caused by Botrytis cinerea were investigated. The spoilage rates differed between genotypes, and this was highly correlated over two successive years. Among seventeen genotypes, a more intense red coloration of the fruit skin was associated with a lower spoilage rate (r = -0.63). Additionally, weakly negative correlations were found between the levels of anthocyanins, ascorbic acid, malic acid and spoilage rates. No clear correlations were found between spoilage rates and soluble sugars, most volatiles, firmness and dry weight percentage. High levels of two volatile compounds, ethyl butanoate (r = 0.55) and 1-hexanol (r = 0.61), were correlated to high spoilage rates. These characteristics may assist strawberry breeders in selecting for genotypes with reduced susceptibility to B. cinerea.

High CO2 Reduces Spoilage Caused by Botrytis cinerea in Strawberry Without Impairing Fruit Quality.

High CO2 (> 20 kPa) conditions are beneficial for suppressing spoilage caused by Botrytis cinerea in strawberry fruit; however, these conditions are often accompanied by discoloration, off-flavors, and faster softening. Stepwise increments of CO2 concentrations have been proposed to alleviate injuries in fruits caused by high CO2. In this study, we investigated whether stepwise increments of CO2, up to 30 kPa and under a reduced O2 concentration, are beneficial for reducing fungal spoilage without inducing CO2 injury symptoms in strawberry fruit. Based on recommended settings (5-10 kPa O2 with 15-20 kPa CO2), we first selected optimal O2 and CO2 concentrations that best-reduced spoilage caused by B. cinerea in red ripe "Sonsation" strawberry fruit. We found that higher O2 (10 kPa) and CO2 (20 kPa) concentrations were most beneficial for prolonging strawberry fruit shelf life. Subsequently, we studied the performance of red ripe "Arabella" strawberry fruit stored at 5°C under different controlled atmosphere (CA) conditions (10 kPa O2 with either 0, 20, or 30 kPa CO2). The CO2 concentrations were achieved either within 8 h or in a stepwise manner within the first 4 days of storage. As a control, 21 kPa O2 and 0 kPa CO2 were used. Following storage for up to 11 days, the spoilage incidence was assessed at 12°C for 5 days. The application of high CO2 (20 and 30 kPa) combined with 10 kPa O2 greatly suppressed fruit spoilage during storage and subsequent shelf life. High CO2 suppressed respiration as well as maintained a higher pH and firmness in treated fruit. The level of total sugars did not change, but during storage, a substantial part of sucrose was converted into glucose and fructose, especially under high CO2 conditions. High CO2 did not affect ascorbic acid and anthocyanin levels. The stepwise increments of CO2 did not result in beneficial effects compared to the static application of high CO2. Our results show that "Arabella" strawberry fruit are highly tolerant to elevated CO2 and can be stored under 30 kPa CO2 to prolong the shelf life.

Lack of Blue Light Regulation of Antioxidants and Chilling Tolerance in Basil.

Blue light, measuring from 400 to 500 nm, is generally assumed to increase the content of antioxidants in plants independent of the species. Blue light stimulates the biosynthesis of phenolic compounds such as flavonoids and their subclass anthocyanins from the phenylpropanoid pathway. Flavonoids, anthocyanins, and phenolic acids are strong reactive oxygen species (ROS) scavengers and may lessen the symptoms of abiotic stresses such as chilling. We tested the hypothesis that a high percentage of blue light induces the accumulation of antioxidants and that this effect depends on the photosynthetic photon flux density (PPFD, 400-700 nm). The effect may be more pronounced at a lower PPFD. We investigated the changes in primary and secondary metabolites of basil in response to the percentage of blue light (9, 33, 65, and 100%) applied either as a 5-day End-Of-Production (EOP) treatment or continuous throughout the growth cycle in the green cv. Dolly. We also studied if the response to the percentage of blue light (9 or 90%) was dependent on the total PPFD (100 or 300 µmol m-2 s-1 PPFD) when applied as a 5-day EOP treatment in the green cv. Dolly and the purple cv. Rosie. For both green and purple basil, it was found that the percentage of blue light had little effect on the levels of antioxidants (rosmarinic acid, total ascorbic acid, total flavonoids, and total anthocyanins) at harvest and no interactive effect with PPFD was found. Antioxidants generally decreased during postharvest storage, wherein the decrease was more pronounced at 4 than at 12°C. Chilling injury, as judged from a decrease in F v /F m values and from the occurrence of black necrotic areas, was not affected by the percentage of blue light. Particularly, chilling tolerance in the purple cultivar was increased in plants grown under higher PPFD. This may be related to the increased levels of soluble sugar and starch in leaves from high PPFD treated plants.

Additional Blue LED during Cultivation Induces Cold Tolerance in Tomato Fruit but Only to an Optimum.

Tomato is a chilling-sensitive fruit. The aim of this study is to examine the role of preharvest blue LED lighting (BL) to induce cold tolerance in 'Foundation' tomatoes. Blue and red supplemental LED light was applied to achieve either 0, 12 or 24% additional BL (0B, 12B and 24B). Mature green (MG) or red (R) tomatoes were harvested and cold stored at 4 °C for 0, 5, 10, 15 and 20 d, and then stored for 20 d at 20 °C (shelf life). Chilling injury (CI) indices, color and firmness, hydrogen peroxide, malondialdehyde, ascorbic acid and catalase activity were characterized. At harvest, R tomatoes cultivated at 12B were firmer and showed less coloration compared to fruit of other treatments. These fruits also showed higher loss of red color during cold storage and lower CI symptoms during shelf-life. MG tomatoes cultivated at 12B showed delayed coloring (non-chilled) and decreased weight loss (long cold stored) during shelf life compared to fruit in the other treatments. No effects of light treatments, both for MG and R tomatoes, were observed for the selected antioxidant capacity indicators. Improved cold tolerance for R tomatoes cultivated at 12B points to lycopene having higher scavenging activity at lower concentrations to mitigate chilling injury.

High light intensity at End-Of-Production improves the nutritional value of basil but does not affect postharvest chilling tolerance.

Basil suffers from chilling injury (CI) when stored at temperatures below 10-12 °C which seems related to the imbalance between reactive oxygen species (ROS) and antioxidants. We hypothesized that increased light intensity applied shortly before harvest (EOP, End-Of-Production) increases nutritional value i.e. carbohydrates and antioxidants and could improve the chilling tolerance. Two basil cultivars were grown in a vertical farming set-up at a light intensity of 150 µmol m-2 s-1. During the last 5 days of growth, EOP light treatments ranging from 50 to 600 µmol m-2 s-1 were applied. After harvest the leaves were stored at 4 or 12 °C in darkness. Higher EOP light intensity increased the antioxidant (total ascorbic acid, rosmarinic acid) and carbohydrate contents at harvest. During storage antioxidants decreased more rapidly at 4 than at 12 °C. However, increased EOP light intensity did not alleviate chilling symptoms suggesting a minor role of antioxidants studied against chilling stress.

Low Oxygen Storage Improves Tomato Postharvest Cold Tolerance, Especially for Tomatoes Cultivated with Far-Red LED Light.

We investigated the effects of low oxygen storage on chilling injury development, colour development, respiration and H2O2 levels of 'Merlice' tomatoes cultivated with and without far red (FR) LED lighting during 20 days of shelf-life. Mature green (MG) and red (R) tomatoes were stored at 2 °C in combination with 0.5, 2.5, 5 and 21 kPa O2 for 15 days (experiment 1). MG tomatoes cultivated under either white LED or white LED light with FR LED light were stored at 2 °C in combination with 1, 5 and 21 O2 kPa for 14 days (experiment 2). Chilled MG and R tomatoes from experiment 1 showed decay, firmness loss and higher weight loss during shelf-life which were reduced under low oxygen conditions. FR during cultivation improved chilling tolerance of MG tomatoes. Fastest colour development and lowest respiration rate during shelf-life were observed for MG fruit cultivated with FR lighting prior to storage at 1 kPa O2/0 kPa CO2. H2O2 levels during the shelf-life were not affected during cold storage. The improved cold tolerance of MG tomatoes cultivated with FR lighting is likely due to lower oxygen uptake that led to both higher lycopene synthesis and less softening.

Blue Light Improves Photosynthetic Performance during Healing and Acclimatization of Grafted Watermelon Seedlings.

To investigate the importance of light on healing and acclimatization, in the present study, grafted watermelon seedlings were exposed to darkness (D) or light, provided by blue (B), red (R), a mixture of R (68%) and B (RB), or white (W; 35% B, 49% intermediate spectra, 16% R) LEDs for 12 days. Survival ratio, root and shoot growth, soluble carbohydrate content, photosynthetic pigments content, and photosynthetic performance were evaluated. Seedling survival was not only strongly limited in D but the survived seedlings had an inferior shoot and root development, reduced chlorophyll content, and attenuated photosynthetic efficiency. RB-exposed seedlings had a less-developed root system. R-exposed seedlings showed leaf epinasty, and had the smallest leaf area, reduced chlorophyll content, and suppressed photosynthetic apparatus performance. The R-exposed seedlings contained the highest amount of soluble carbohydrate and together with D-exposed seedlings the lowest amount of chlorophyll in their scions. B-exposed seedlings showed the highest chlorophyll content and improved overall PSII photosynthetic functioning. W-exposed seedling had the largest leaf area, and closely resembled the photosynthetic properties of RB-exposed seedlings. We assume that, during healing of grafted seedlings monochromatic R light should be avoided. Instead, W and monochromatic B light may be willingly adopted due to their promoting effect on shoot, pigments content, and photosynthetic efficiency.

High Light Intensity Applied Shortly Before Harvest Improves Lettuce Nutritional Quality and Extends the Shelf Life.

The effect of light intensity applied shortly before harvest on the nutritional quality, postharvest performance, and shelf life of loose-leaf lettuce (Lactuca sativa L. cv. Expertise RZ Salanova®) was investigated. Lettuce was grown either in a greenhouse with supplemental high-pressure sodium light (Experiment 1, EXP 1) or in a climate room under white LED light (Experiment 2, EXP 2). In both experiments full grown plants were transferred to a climate room for the End of Production (EoP) light treatments during the last week of cultivation. During EoP lighting plants were exposed to different intensities (0, 110, and 270 µmol m-2 s-1 in EXP 1; 50, 210, and 470 µmol m-2 s-1 in EXP 2) from white-red LEDs for 6 (EXP 2) or 7 days (EXP 1). Mature leaves were then harvested and stored in darkness at 10°C to study the postharvest performance. Changes in dry matter content, total ascorbic acid, and carbohydrates (including glucose, fructose sucrose, and starch) levels were determined during EoP lighting and during the subsequent shelf life as indicators of lettuce nutritional quality. Quality aspects (appearance, texture, and odor) were accessed during the shelf life as indicators of postharvest performance. In both experiments, high light intensities applied in EoP lighting increased dry matter percentage and contents of ascorbic acid (AsA) and carbohydrates at harvest and these increased levels were maintained during the shelf life. Increased light intensity in EoP treatment also extended the shelf life. The levels of AsA and carbohydrates at harvest correlated positively with the subsequent shelf life, indicating that the prolonged shelf life relies on the improved energy and antioxidant status of the crop at harvest.

Identifying key wavenumbers that improve prediction of amylose in rice samples utilizing advanced wavenumber selection techniques.

This study utilizes advanced wavenumber selection techniques to improve the prediction of amylose content in grounded rice samples with near-infrared spectroscopy. Four different wavenumber selection techniques, i.e. covariate selection (CovSel), variable combination population analysis (VCPA), bootstrapping soft shrinkage (BOSS) and variable combination population analysis-iteratively retains informative variables (VCPA-IRIV), were used for model optimization and key wavenumbers selection. The results of the several wavenumber selection techniques were compared with the predictions reported previously on the same data set. All the four wavenumber selection techniques improved the predictive performance of amylose in rice samples. The best performance was obtained with VCPA, where, with only 11 wavenumbers-based model, the prediction error was reduced by 19% compared to what reported previously on the same data set. The selected wavenumbers can help in development of low-cost multi-spectral sensors for amylose prediction in rice samples.

Cell death associated release of volatile organic sulphur compounds with antioxidant properties in chemical-challenged tobacco BY-2 suspension cultured cells.

The production of volatile organic compounds (VOCs) during programmed cell death (PCD) is still insufficiently studied and their implication in the process is not well understood. The present study demonstrates that the release of VOSCs with presumed antioxidant capacity (methanethiol, dimethylsulfide and dimethyldisulfide) accompanies the cell death in chemical-stressed tobacco BY-2 suspension cultured cells. The cells were exposed to cell death inducers of biotic nature mastoparan (MP, wasp venom) and camptothecin (CPT, alkaloid), and to the abiotic stress agent CdSO4. The VOCs emission was monitored by proton-transfer reaction mass spectrometry (PTR-MS). The three chemicals induced PCD expressing apoptotic-like phenotype. The identified VOSCs were emitted in response to MP and CPT but not in presence of Cd. The VOSCs production occurred within few hours after the administration of the elicitors, peaked up when 20-50 % of the cells were dead and further levelled off with cell death advancement. This suggests that VOSCs with antioxidant activity may contribute to alleviation of cell death-associated oxidative stress at medium severity of cell death in response to the stress factors of biotic origin. The findings provide novel information about cell death defence mechanisms in chemical-challenged BY-2 cells and show that PCD related VOSCs synthesis depends on the type of inducer.

Modulation of the Tomato Fruit Metabolome by LED Light.

Metabolic profiles of tomatoes change during ripening and light can modulate the activity of relevant biochemical pathways. We investigated the effects of light directly supplied to the fruits on the metabolome of the fruit pericarp during ripening. Mature green tomatoes were exposed to well-controlled conditions with light as the only varying factor; control fruits were kept in darkness. In experiment 1 the fruits were exposed to either white light or darkness for 15 days. In experiment 2, fruits were exposed to different light spectra (blue, green, red, far-red, white) added to white background light for seven days. Changes in the global metabolome of the fruit pericarp were monitored using LCMS and GCMS (554 compounds in total). Health-beneficial compounds (carotenoids, flavonoids, tocopherols and phenolic acids) accumulated faster under white light compared to darkness, while alkaloids and chlorophylls decreased faster. Light also changed the levels of taste-related metabolites including glutamate and malate. The light spectrum treatments indicated that the addition of blue light was the most effective treatment in altering the fruit metabolome. We conclude that light during ripening of tomatoes can have various effects on the metabolome and may help with shaping the levels of key compounds involved in various fruit quality characteristics.

Response of Basil Growth and Morphology to Light Intensity and Spectrum in a Vertical Farm.

Vertical farming is becoming increasingly popular for production of leafy vegetables and herbs, with basil (Ocimum basilicum L.) as one of the most popular herbs. In basil most research has focused on increasing secondary metabolites with light spectra. However, knowledge about the effect of light intensity (photosynthetic photon flux density, PPFD) and spectra on growth and morphology is key for optimizing quality at harvest. The impact of PPFD and spectrum on plant growth and development is species dependent and currently few studies in basil are available. Understanding the response to End-Of-Production (EOP) light of growth and morphology is important for successful vertical farming. We performed a comprehensive series of experiments, where the effects of EOP PPFD, fraction of blue and their interaction on the growth and morphology were analyzed in two green and one purple basil cultivar. In addition, the impact of different EOP intensities and duration of far-red were investigated. We found that increasing the PPFD increased fresh mass, dry matter content and plant height in all three cultivars. The responses were linear or quadratic depending on the cultivar. A high fraction of blue (>90%) increased plant height and decreased the dry mass partitioning to the leaves. The only interaction found between the fraction of blue and overall PPFD was on plant height in the green cultivar whereas other growth parameters and morphology responded stronger to PPFD than to the fraction of blue light. Plant dry matter production was increased with the addition of far-red. Far-red EOP intensity treatments enhanced the fraction of dry mass partitioned to the leaves, whereas a prolonged far-red treatment enhanced partitioning to the stem. Both plant fresh mass and dry matter content were improved by applying high PPFD shortly before harvest. Light spectra were found to be of less importance than PPFD with respect to plant dry matter content. Light use efficiency (LUE) based on fresh mass decreased with increasing PPFD whereas LUE based on dry mass increased with increasing PPFD, when given as EOP treatments. The overall physiological mechanisms of the light intensity and spectral effects are discussed.

Mango Firmness Modeling as Affected by Transport and Ethylene Treatments.

More and more, tropical fruit are subjected to accelerated ripening at receiving markets until "ready to eat." We propose a kinetic model that incorporates the effects of temperature and ethylene on the firmness behavior of "Keitt" and "Kent" mangoes. Stiffness of individual mangoes, as measured by the acoustic firmness tester, was measured repeatedly over time. The firmness model assumes fixed levels of ethylene, established after the climacteric peak, that steadily induces production of softening enzymes that subsequently denaturalize. The initial level of these enzymes is assumed to be zero due to either the tree factor for freshly harvested mangoes, or due to chilling injury for reefer transported mangoes. The kinetic parameter set for "Keitt" mangoes was estimated based on a Spanish batch, freshly harvested and ripened under dynamic temperature scenarios, combined with a reefer transported Brazilian batch stored at four constant temperatures. Firmness data from reefer transported batches, from Brazil, Ivory Coast and Mali, stored at four constant temperatures were used to estimate a set of kinetic parameters for the "Kent" mangoes. Only a partial set of "Kent" kinetic parameters could be established due to the often already advanced stage of softening at the time of arrival. The effect of ethylene was investigated by applying external ethylene levels, varying from 0 to 100 µL L-1. The effect of external application of ethylene was modeled by estimating EF, the ethylene factor, being a reflection of the internal ethylene level and ethylene sensitivity. The effect of ethylene application on softening was sometimes huge. For an Israeli "Keitt" batch a fifty times higher EF was found when the firmness behavior of low- (without ethylene application) and high temperature (with ethylene application) stored sub-batches were compared. However, this effect was sometimes also small, especially for reefer transported mangoes. For commercial application, a reliable prediction of the time until "ready to eat" is not possible because of the current inability to assess EF. Nevertheless, the proposed model described mango softening accurately, irrespective of the sourcing area and includes the effects of storage temperature and ethylene application.

The wound response in fresh-cut lettuce involves programmed cell death events.

In this work, the involvement of programmed cell death (PCD) in the wound-induced postharvest browning disorder and senescence in butterhead lettuce (Lactuca sativa L.) fresh-cuts was studied. At the wounded (cut, bruised) sites, rapid browning, loss of chlorophyll and massive cell death, accompanied with accumulation of reactive oxygen species and increased electrolyte leakage occurred in a narrow strip of tissue adjacent the injury. The dead cell morphology (protoplast and nuclei shrinkage) together with the biochemical and physiological changes resembled necrotic PCD type. With a slight delay post-wounding, senescence associated with similar cell death features was initiated in distant non-wounded sites. In addition to necrotic PCD, both in wounded and senescing tissue, the appearance of empty cell corpses was observed, indicating that part of the cells might undergo vacuolar PCD (self-digestion of cellular content after vacuole collapse). The wounding-induced local cell death at the primary site of damage suggested that PCD may serve as a mechanism to seal-off the wound by building a physical barrier of dead cells. However, the cell death at sites remote from the wound suggests the distribution of long-distance senescence-inducing wound messengers. Trichomes in unwounded tissue often were the first to show H2O2 accumulation and dead cells; thereafter, the elevated H2O2 and cell death appeared in connecting cells and senescence progressed over larger areas. This suggests that trichomes may contribute to mediating the wound signalling leading to subsequent senescence. Our findings demonstrate that PCD is an integral part of the wound syndrome in fresh-cut lettuce.

Xylogenesis in zinnia (Zinnia elegans) cell cultures: unravelling the regulatory steps in a complex developmental programmed cell death event.

MAIN CONCLUSION: Physiological and molecular studies support the view that xylogenesis can largely be determined as a specific form of vacuolar programmed cell death (PCD). The studies in xylogenic zinnia cell culture have led to many breakthroughs in xylogenesis research and provided a background for investigations in other experimental models in vitro and in planta . This review discusses the most essential earlier and recent findings on the regulation of xylem elements differentiation and PCD in zinnia and other xylogenic systems. Xylogenesis (the formation of water conducting vascular tissue) is a paradigm of plant developmental PCD. The xylem vessels are composed of fused tracheary elements (TEs)-dead, hollow cells with patterned lignified secondary cell walls. They result from the differentiation of the procambium and cambium cells and undergo cell death to become functional post-mortem. The TE differentiation proceeds through a well-coordinated sequence of events in which differentiation and the programmed cellular demise are intimately connected. For years a classical experimental model for studies on xylogenesis was the xylogenic zinnia (Zinnia elegans) cell culture derived from leaf mesophyll cells that, upon induction by cytokinin and auxin, transdifferentiate into TEs. This cell system has been proven very efficient for investigations on the regulatory components of xylem differentiation which has led to many discoveries on the mechanisms of xylogenesis. The knowledge gained from this system has potentiated studies in other xylogenic cultures in vitro and in planta. The present review summarises the previous and latest findings on the hormonal and biochemical signalling, metabolic pathways and molecular and gene determinants underlying the regulation of xylem vessels differentiation in zinnia cell culture. Highlighted are breakthroughs achieved through the use of xylogenic systems from other species and newly introduced tools and analytical approaches to study the processes. The mutual dependence between PCD signalling and the differentiation cascade in the program of TE development is discussed.

Chilling-Induced Changes in Aroma Volatile Profiles in Tomato.

Fruit and vegetables are regularly stored by consumers in the refrigerator at temperatures that may be well below the recommended storage temperatures. Apart from causing visible symptoms such as watery, sunken areas on the skin, chilling may also induce changes in fruit textural properties and flavour. The aim of this research was to investigate the effect of low temperature storage on tomato flavour and off-flavour production. To more closely mimic the real-consumer aroma perception while eating, in addition to the standard solid-phase microextraction gas chromatography-mass spectrometry (SPME/GC-MS) analysis, volatiles were also measured using a chewing device connected to a proton-transfer reaction-mass spectrometer (PTR-MS). Aroma volatiles were assessed in red ripe tomatoes of the cvs Cappricia RZ (round truss) and Amoroso RZ (cocktail truss) stored at refrigerator temperature (4 °C) and at higher temperatures (16 and 22 °C) for 20 days. The changes in aroma production were also monitored when the fruit was brought from room to refrigerator temperature and vice versa. After bringing the fruit from room to refrigerator temperature, the abundance of most volatiles was greatly reduced within 3 to 5 h, closely following the decrease in fruit temperature. When temperature was restored to room temperature following varying times of cold storage, the abundance of most volatiles increased again, but generally not to the original levels. Overall, the effects of low temperature storage on the decrease in volatile abundance were more pronounced in cv Cappricia RZ than in cv Amoroso RZ. On the contrary, the production of off flavours following prolonged cold storage was more pronounced in cv Amoroso RZ than in cv Cappricia RZ. Apart from changes in the overall abundance of the volatiles, marked changes in the volatile profile were observed in fruit stored for longer times in the cold and this may at least in part explain the negative effect of cold storage on overall tomato flavour.

Involvement of phospholipase D-related signal transduction in chemical-induced programmed cell death in tomato cell cultures.

Phospholipase D (PLD) and its product phosphatidic acid (PA) are incorporated in a complex metabolic network in which the individual PLD isoforms are suggested to regulate specific developmental and stress responses, including plant programmed cell death (PCD). Despite the accumulating knowledge, the mechanisms through which PLD/PA operate during PCD are still poorly understood. In this work, the role of PLDα1 in PCD and the associated caspase-like proteolysis, ethylene and hydrogen peroxide (H(2)O(2)) synthesis in tomato suspension cells was studied. Wild-type (WT) and PLDα1-silenced cell lines were exposed to the cell death-inducing chemicals camptothecin (CPT), fumonisin B1 (FB1) and CdSO(4). A range of caspase inhibitors effectively suppressed CPT-induced PCD in WT cells, but failed to alleviate cell death in PLDα1-deficient cells. Compared to WT, in CPT-treated PLDα1 mutant cells, reduced cell death and decreased production of H(2)O(2) were observed. Application of ethylene significantly enhanced CPT-induced cell death both in WT and PLDα1 mutants. Treatments with the PA derivative lyso-phosphatidic acid and mastoparan (agonist of PLD/PLC signalling downstream of G proteins) caused severe cell death. Inhibitors, specific to PLD and PLC, remarkably decreased the chemical-induced cell death. Taken together with our previous findings, the results suggest that PLDα1 contributes to caspase-like-dependent cell death possibly communicated through PA, reactive oxygen species and ethylene. The dead cells expressed morphological features of PCD such as protoplast shrinkage and nucleus compaction. The presented findings reveal novel elements of PLD/PA-mediated cell death response and suggest that PLDα1 is an important factor in chemical-induced PCD signal transduction.

Morphological and biochemical characterization of Erwinia amylovora-induced hypersensitive cell death in apple leaves.

In attached apple leaves, spot-inoculated with Erwinia amylovora, the phenotypic appearance of the hypersensitive response (HR) and the participation of ethylene, reactive oxygen species (ROS) and of vacuolar processing enzyme (VPE) (a plant caspase-1-like protease) were analysed. The HR in both the resistant and susceptible genotypes expressed a similar pattern of distinguishable micro HR lesions that progressed into confined macro HR lesions. The HR symptoms in apple were compared to those in non-host tobacco. The morphology of dead cells (protoplast shrinkage and retraction from cell wall) in apple leaves resembled necrotic programmed cell death (PCD). Lesion formation in both cv. Free Redstar (resistant) and cv. Idared (highly susceptible) was preceded by ROS accumulation and elevation of ethylene levels. Treatment of infected leaves with an inhibitor of ethylene synthesis led to a decrease of ethylene emission and suppression of lesion development in both cultivars. In the resistant but not in the susceptible apple cultivar an early and late increase in VPE gene expression was detected. This suggests that VPE might be an underlying component of the response to E. amylovora in resistant apple cultivars. The findings show that in the studied pathosystem the cell death during the HR proceeds through a signal transduction cascade in which ROS, ethylene and VPE pathways play a role.

Mastoparan-induced programmed cell death in the unicellular alga Chlamydomonas reinhardtii.

BACKGROUND AND AIMS: Under stress-promoting conditions unicellular algae can undergo programmed cell death (PCD) but the mechanisms of algal cellular suicide are still poorly understood. In this work, the involvement of caspase-like proteases, DNA cleavage and the morphological occurrence of cell death in wasp venom mastoparan (MP)-treated Chlamydomonas reinhardtii were studied. METHODS: Algal cells were exposed to MP and cell death was analysed over time. Specific caspase inhibitors were employed to elucidate the possible role of caspase-like proteases. YVADase activity (presumably a vacuolar processing enzyme) was assayed by using a fluorogenic caspase-1 substrate. DNA breakdown was evaluated by DNA laddering and Comet analysis. Cellular morphology was examined by confocal laser scanning microscopy. KEY RESULTS: MP-treated C. reinhardtii cells expressed several features of necrosis (protoplast shrinkage) and vacuolar cell death (lytic vesicles, vacuolization, empty cell-walled corpse-containing remains of digested protoplast) sometimes within one single cell and in different individual cells. Nucleus compaction and DNA fragmentation were detected. YVADase activity was rapidly stimulated in response to MP but the early cell death was not inhibited by caspase inhibitors. At later time points, however, the caspase inhibitors were effective in cell-death suppression. Conditioned medium from MP-treated cells offered protection against MP-induced cell death. CONCLUSIONS: In C. reinhardtii MP triggered PCD of atypical phenotype comprising features of vacuolar and necrotic cell deaths, reminiscent of the modality of hypersensitive response. It was assumed that depending on the physiological state and sensitivity of the cells to MP, the early cell-death phase might be not mediated by caspase-like enzymes, whereas later cell death may involve caspase-like-dependent proteolysis. The findings substantiate the hypothesis that, depending on the mode of induction and sensitivity of the cells, algal PCD may take different forms and proceed through different pathways.