Biochemical dynamics during postharvest: Highlighting the interplay of stress during storage and maturation of fresh produce.

The lifecycle of fresh produce involves a sequence of biochemical events during their ontology, and these events are particularly significant for climacteric fruits. A high demand during ripening is observed in these plant products, which is reflected in a high rate of respiration and ethylene production. Increased respiratory demand triggers the activation of secondary pathways such as alternate oxidase, which do not experience critical increases in energy consumption in non-climacteric fruit. In addition, biochemical events produced by external factors lead to compensatory responses in fresh produce to counteract the oxidative stress caused by the former. The dynamics of these responses are accompanied by signaling, where reactive oxygen species play a pivotal role in fresh product cell perception. This review aims to describe the protection mechanisms of fresh produce against environmental challenges and how controlled doses of abiotic stressors can be used to improve quality and prolong their shelf-life through the interaction of stress and defense mechanisms.

Phytochemical Enhancement in Broccoli Florets after Harvest by Controlled Doses of Ozone.

The objective of this work was to examine the effect of controlled doses of O3 (0, 5 µL L-1 of O3 for 60 min, and 5 µL L-1 of O3 for 720 min) on the quality and phytochemical content of broccoli florets during postharvest storage. The optimal dose was found at 5 µL L-1 of O3 for 60 min, from the color retention of broccoli florets exposed to the gas treatment. Overall, the antioxidant capacity of the florets was significantly affected by both doses of O3 compared to the non-exposed florets. The profile of glucosinolates was determined for up to 14 days in broccoli florets stored at 4 °C by LC-MS. The amount of total glucobrassicins and total hydroxy-cinnamates in florets significantly (p ≤ 0.05) improved by the application of 5 µL L-1 of O3 for 60 min compared to non-treated florets. The up-regulation of genes of the tryptophan-derived glucosinolate pathway was observed immediately after both treatments. The gene expression of CYP79A2 and CYP79B3 in broccoli was significantly higher in broccoli florets exposed to 5 µL L-1 of O3 for 720 min compared to non-exposed florets. Although enhancement of secondary metabolites can be achieved by the fumigation of broccoli florets with low doses of ozone, quality parameters, particularly weight loss, can be compromised.

Abiotic stress hormesis: An approach to maintain quality, extend storability, and enhance phytochemicals on fresh produce during postharvest.

Postharvest losses of whole and fresh-cut fruits and vegetables cause significant reductions in food availability and an increase in economic losses/damages. Additionally, regulatory agencies are increasingly restricting the postharvest use of synthetic chemicals. This has strengthened the need to develop environmentally friendly approaches to postharvest management, such as utilization of natural compounds, antagonist microorganisms, and treatments with abiotic stresses, among others. The current review focuses on the potential of low doses of abiotic stresses to extend the shelf life, increase the amount of health beneficial phytochemicals, and reduce postharvest losses of fresh produce. The positive effects of the responses to low doses of abiotic stresses are based on a biological phenomenon termed hormesis. Research to develop new technologies to improve postharvest handling of fresh fruit and vegetables as well as minimally processed products is critical. The phenomenon of abiotic stress hormesis in fresh fruit and vegetables shows the potential not only to enhance defense compounds that could reduce diseases during postharvest storage and extend shelf life but also to elevate the content of health-promoting substances. The beneficial effects of UV-C hormesis have been extensively investigated in numerous types of fresh produce. However, our knowledge on hormesis exhibited by other abiotic stresses is still limited. Hence, the objective of this review is to discuss the relevance of hormesis for postharvest research by examining whether all abiotic stresses exhibit the phenomenon, its biological significance, the potential application in various commodities, and how it may direct the future of postharvest research.

Quality and PR gene expression of table grapes treated with ozone and sulfur dioxide to control fungal decay.

BACKGROUND: Gaseous fumigants are commonly employed to control fungal decay of cold-stored grapes. So far it is not clear if these fumigants, besides the direct interaction against fungal structures, induce transcriptional responses of defensive markers. In order to contribute to understanding the mechanisms by which these fumigants exert their effect, we studied the influence of ozone (O3) and sulfur dioxide (SO2) on the decay caused by Botrytis cinerea, and the quality and expression of the defense-related genes chitinase, ß-1,3-glucanase and phenylalanine ammonia-lyase (PAL) in the table grape cultivars 'Redglobe' and 'Sugraone'. RESULTS: The application of SO2 or O3 delayed decay of both table grape cultivars caused by B. cinerea compared with the inoculated control. O3 treatments altered weight loss, firmness and shatter in both cultivars. Significant upregulation of chitinase and ß-1,3-glucanase were observed in SO2 -treated 'Redglobe' berries stored at 2 °C. O3 treatment transiently increased the expression of chitinase and PAL in 'Redglobe' and 'Sugraone' berries, respectively. CONCLUSION: Ozone and sulfur dioxide treatments can influence the expression patterns of PAL, chitinase and ß-1,3-glucanase to different extents in different grape cultivars and under different exposure conditions. The upregulation of these genes may be involved in the mechanism by which these fumigants inhibit the decay caused by pathogenic fungi.

D3 dopamine receptors interact with dopamine D1 but not D4 receptors in the GABAergic terminals of the SNr of the rat.

The firing rate of substantia nigra reticulata (SNr) neurons is modulated by GABA release from striatonigral and pallidonigral projections. This release is, in turn, modulated by dopamine acting on dopamine D1 receptors at striatonigral terminals and D4 receptors at pallidonigral terminals. In addition, striatal neurons that express D1 receptors also express D3 receptors. In this study we analyzed the possible significance of D3 and D1 receptor colocalization in striatonigral projections. We found that these receptors coprecipitate in SNr synaptosomes suggesting their close association in this structure. D1 agonist SKF 38393 administered alone increased mIPSC frequency in SNr slices and cAMP production in SNr synaptosomes, however, the selective D3 agonist PD 128,907 increased mIPSC frequency and cAMP production only when D1 receptors were concurrently stimulated. The D1 antagonist SCH 23390 blocked completely the effects of the concurrent administration of these agonists while the selective D3 antagonist GR 103691 blocked only the potentiating effects of PD 128,907. These findings further indicate that D1 and D3 receptors are localized in the same structure. The D4 agonist PD 168,077 decreased mIPSCs frequency without changing amplitude, an effect that was blocked by the selective D4 antagonist L 745,870. The effects of D4 receptor stimulation disappeared after lesioning the globus pallidus. D3 agonist PD 128,907 did not reduce mIPSC frequency even in neurons that responded to D4 agonist. In sum, activation of D3 receptors in SNr potentiates the stimulation of transmitter release and cAMP production caused by D1 receptor activation of striatonigral projections while it is without effects in terminals, probably of pallidal origin, that are inhibited by activation of D4 receptors.

D4 and D1 dopamine receptors modulate [3H] GABA release in the substantia nigra pars reticulata of the rat.

Neurons of the globus pallidus express dopamine D4 receptors that can modulate transmitter release by their axon terminals. Indeed, GABA release from pallidal terminals in the subthalamic nucleus and in the reticular nucleus of the thalamus is inhibited by activation of D4 receptors. Here we investigated whether GABA release by pallidal projections to the substantia nigra reticulate (SNr) is also modulated by D4 receptors. Dopamine-stimulated depolarization-induced GABA release in slices of the SNr; however, after selective blockade of D1 receptors, dopamine inhibited release. The selective D4 agonist PD 168,077 (IC(50) = 5.30 nM) mimicked the inhibition of release while the selective D4 antagonist L-745,870 blocked the inhibition. To identify the source of D1 and D4 modulated terminals, we unilaterally injected kainic acid in either the GP or the striatum. After lesions of the pallidum, the D4 induced inhibition of release was blocked while the D1 induced stimulation was still significant. Lesions of the striatum had the converse effects. We conclude that release of dopamine in the SNr enhances GABA release mainly through activation of D1 receptors in striatonigral projections and inhibits release mainly through activation of D4 receptors in pallidonigral projections. Because deficient D4 receptor signaling in globus pallidus terminals will lead to disinhibition of impulse traffic through the thalamus we speculate that the D4 abnormalities observed in ADHD patients may be important in the generation of the syndrome.

Presynaptic D1 dopamine receptors facilitate glutamatergic neurotransmission in the rat globus pallidus.

The effects of D1/5 dopamine agonists on spontaneous excitatory postsynaptic currents (sEPSCs) were studied in neurons of the rat globus pallidus using whole-cell recordings in the presence of TTX and bicuculline. In this condition, CNQX abolished the sEPSCs, indicating that they were solely mediated by AMPA receptors. SKF 38393, a D1-like agonist, increased the frequency but not the amplitude of the sEPSCs, suggesting a presynaptic site of action. The increase in frequency was blocked by SCH 23390, a D1/5 antagonist. Quinpirole, a D2-like agonist, decreased the frequency but did not affect the amplitude of the synaptic currents. SKF 38393 increased the frequency of sEPSCs currents, even in presence of quinpirole, indicating that D1/5- and D2-like receptors independently modulate glutamate release upon a single neuron. The results suggest that the dopaminergic control of the glutamate transmission in the globus pallidus may play a role in processing cortical information in the indirect pathway of the basal ganglia.

Control of the subthalamic innervation of the rat globus pallidus by D2/3 and D4 dopamine receptors.

The effects of activating dopaminergic D(2/3) and D(4) receptors during activation of the subthalamic projection to the globus pallidus (GP) were explored in rat brain slices using the whole cell patch-clamp technique. Byocitin labeling and both orthodromic and antidromic activation demonstrated the integrity of some subthalamopallidal connections in in vitro parasagittal brain slices. Excitatory postsynaptic currents (EPSCs) that could be blocked by CNQX and AP5 were evoked onto pallidal neurons by local field stimulation of the subthalamopallidal pathway in the presence of bicuculline. Bath application of dopamine and quinpirole, a dopaminergic D(2)-class receptor agonist, reduced evoked EPSCs by about 35%. This effect was only partially blocked by sulpiride, a D(2/3) receptor antagonist. The sulpiride-sensitive reduction of the subthalamopallidal EPSC was associated with an increase in the paired-pulse ratio (PPR) and a reduction in the frequency but not the mean amplitude of spontaneous EPSCs (sEPSCs), indicative of a presynaptic site of action, which was confirmed by variance-mean analysis. The sulpiride-resistant EPSC reduction was mimicked by PD 168,077 and blocked by L-745,870, selective D(4) receptor agonist and antagonist, respectively, suggesting the involvement of D(4) receptors. The reduction of EPSCs produced by PD 168,077 was not accompanied by changes in PPR or the frequency of sEPSCs; however, it was accompanied by a reduction in mean sEPSC amplitude, indicative of a postsynaptic site of action. These results show that dopamine modulates subthalamopallidal excitation by presynaptic D(2/3) and postsynaptic D(4) receptors. The importance of this modulation is discussed.

Noradrenaline increases the firing rate of a subpopulation of rat subthalamic neurones through the activation of alpha 1-adrenoceptors.

In the rat subthalamic nucleus, which plays a critical role in the control of motor behaviour, specific binding of [3H]-prazosin was detected by radioligand binding to homogenates and by autoradiography in slices. [3H]-Prazosin binding to homogenates (Bmax 71 +/- 5 fmol/mg protein; Kd 0.27 +/- 0.05 nM) was competed for by alpha1-antagonists. In subthalamic nucleus slices and in the presence of 10 mM LiCl, noradrenaline (100 microM) produced a modest, but consistent, stimulation of [3H]-inositol phosphate accumulation (146 +/- 6% of basal), reversed by the alpha1-antagonist prazosin (1 microM). Extracellular single-unit recordings in slices showed that in a subpopulation (61 out of 94 cells) of rat subthalamic neurones with regular, single-spike firing pattern, noradrenaline induced a concentration-dependent increase in the firing rate (EC50 2.5 +/- 0.2 microM, maximum effect 272 +/- 33% of basal). The action of noradrenaline was mimicked by the selective alpha1-agonist phenylephrine but not by selective alpha2- or beta-agonists, and was blocked by the alpha1-antagonist prazosin but not by alpha2- or beta-antagonists. The excitatory effect of noradrenaline was not prevented by perfusion with low Ca2+/high Mg2+ solution. In four out of 11 neurones perfusion with 3 microM noradrenaline resulted in a shift from bursting to regular firing. Taken together, our results indicate that rat subthalamic neurones express alpha1-adrenoceptors responsible for noradrenaline-induced stimulation of the firing rate of a subpopulation of neurones. By modulating the spontaneous activity of STN neurones, noradrenergic pathways might have a significant role in regulating basal ganglia function and thus motor activity.