Non-Thermal Technologies as Tools to Increase the Content of Health-Promoting Compounds in Whole Fruits and Vegetables While Retaining Quality Attributes

Fruits and vegetables contain health-promoting compounds. However, their natural concentration in the plant tissues is low and in most cases is not sufficient to exert the expected pharmacological effects. The application of wounding stress as a tool to increase the content of bioactive compounds in fruits and vegetables has been well characterized. Nevertheless, its industrial application presents different drawbacks. For instance, during the washing and sanitizing steps post-wounding, the primary wound signal (extracellular adenosine triphosphate) that elicits the stress-induced biosynthesis of secondary metabolites is partially removed from the tissue. Furthermore, detrimental reactions that affect the quality attributes of fresh produce are also activated by wounding. Therefore, there is a need to search for technologies that emulate the wound response in whole fruits and vegetables while retaining quality attributes. Herein, the application of non-thermal technologies (NTTs) such as high hydrostatic pressure, ultrasound, and pulsed electric fields are presented as tools for increasing the content of health-promoting compounds in whole fruits and vegetables by inducing a wound-like response. The industrial implementation and economic feasibility of using NTTs as abiotic elicitors is also discussed. Whole fruits and vegetables with enhanced levels of bioactive compounds obtained by NTT treatments could be commercialized as functional foods.

Shade suppresses wound-induced leaf repositioning through a mechanism involving PHYTOCHROME KINASE SUBSTRATE (PKS) genes

Shaded plants challenged with herbivores or pathogens prioritize growth over defense. However, most experiments have focused on the effect of shading light cues on defense responses. To investigate the potential interaction between shade-avoidance and wounding-induced Jasmonate (JA)-mediated signaling on leaf growth and movement, we used repetitive mechanical wounding of leaf blades to mimic herbivore attacks. Phenotyping experiments with combined treatments on Arabidopsis thaliana rosettes revealed that shade strongly inhibits the wound effect on leaf elevation. By contrast, petiole length is reduced by wounding both in the sun and in the shade. Thus, the relationship between the shade and wounding/JA pathways varies depending on the physiological response, implying that leaf growth and movement can be uncoupled. Using RNA-sequencing, we identified genes with expression patterns matching the hyponastic response (opposite regulation by both stimuli, interaction between treatments with shade dominating the wound signal). Among them were genes from the PKS (Phytochrome Kinase Substrate) family, which was previously studied for its role in phototropism and leaf positioning. Interestingly, we observed reduced shade suppression of the wounding effect in pks2pks4 double mutants while a PKS4 overexpressing line showed constitutively elevated leaves and was less sensitive to wounding. Our results indicate a trait-specific interrelationship between shade and wounding cues on Arabidopsis leaf growth and positioning. Moreover, we identify PKS genes as integrators of external cues in the control of leaf hyponasty further emphasizing the role of these genes in aerial organ positioning.

Leaf-Wounding Long-Distance Signaling Targets AtCuAOβ Leading to Root Phenotypic Plasticity

The Arabidopsis gene AtCuAOβ (At4g14940) encodes an apoplastic copper amine oxidase (CuAO) highly expressed in guard cells of leaves and flowers and in root vascular tissues, especially in protoxylem and metaxylem precursors, where its expression is strongly induced by the wound signal methyl jasmonate (MeJA). The hydrogen peroxide (H2O2) derived by the AtCuAOβ-driven oxidation of the substrate putrescine (Put), mediates the MeJA–induced early root protoxylem differentiation. Considering that early root protoxylem maturation was also induced by both exogenous Put and leaf wounding through a signaling pathway involving H2O2, in the present study we investigated the role of AtCuAOβ in the leaf wounding-induced early protoxylem differentiation in combination with Put treatment. Quantitative and tissue specific analysis of AtCuAOβ gene expression by RT-qPCR and promoter::green fluorescent protein-β-glucuronidase fusion analysis revealed that wounding of the cotiledonary leaf induced AtCuAOβ gene expression which was particularly evident in root vascular tissues. AtCuAOβ loss-of-function mutants were unresponsive to the injury, not showing altered phenotype upon wounding in comparison to wild type seedlings. Exogenous Put and wounding did not show synergy in inducing early root protoxylem maturation, suggesting their involvement in a shared signaling pathway.

Does long-distance GABA signaling via the phloem really occur?

The ubiquitous nonprotein amino acid known as γ-aminobutyrate (GABA) has been proposed as a mechanism whereby environmental and developmental cues are translocated in phloem. Information about the composition of translocation fluids from various species that bleed spontaneously reveals that GABA is always present in xylem at minor levels, but sometimes absent in phloem. In species that do not bleed spontaneously, GABA is also absent or present at minor levels in aphid stylet exudate. By contrast, GABA in ethylenediaminetetraacetate-facilitated phloem exudates is relatively abundant compared to other amino acids or the concentration of total amino acids present in xylem. Evidence indicates that xylem-borne GABA is primarily retrieved and metabolized by mature leaves, and tissue pools of GABA reflect in situ biosynthesis. Recovery of significant GABA from phloem might be an artifact of wounding and the use of ethylenediaminetetraacetate. Scarce data from aphid stylectomy experiments indicates that GABA is a wound signal. However, the composition of phloem sap collected by this method is not uniform, shedding doubt on some results in the scientific literature. Elevated GABA levels in the host plant are known to influence the success of biotic interactions; however, these outcomes could be attributed to the impact of wounding at local sites, rather than delivery of GABA via phloem. Further research is required to provide unambiguous support for long-distance γ-aminobutyrate signaling in phloem.

Wound-induced Phenolic Accumulation and Browning in Lettuce (Lactucasativa L.) Leaf Tissue is Reduced by Exposure to n-alcohols

A wound signal originates at the site of injury in lettuce [Lactucasativa (L.)] leaf tissue and propagates into adjacent tissue where it induces a number of physiological responses that include increased phenolic metabolism with the de novo synthesis of phenylalanine ammonia lyase (PAL, EC 4.3.1.5), the synthesis and accumulation of soluble phenolic compounds (e.g., chlorogenic acid), and subsequent tissue browning. Exposing excised mid-rib leaf tissue to vapors (20 μmol·g-1 FW) or aqueous solutions (100 mm) of n-alcohols inhibited this wound-induced tissue browning by 40% and 60%, respectively. Effectiveness of the alcohol increased linearly from ethanol to the seven-carbon heptanol, and then was lost for the longer n-alcohols 1-octanol and 1-nonanol. The 2- and 3-isomers of the effective alcohols did not significantly reduce wound-induced phenolic accumulation at optimal 1-alcohol concentrations, but significant reductions did occur at much higher concentrations (100 μmol·g-1 FW) of the 2-, and 3-isomers. The active n-alcohols were maximally effective when applied during the first 2 h after excision, and were ineffective if applied 12 h after excision. Phospholipase D (PLD) and its product phosphatidic acid (PA) are thought to initiate the oxylipin pathway that culminates in the production of jasmonic acid, and PLD is specifically inhibited by 1-butanol, but not by 2- or 3-butanol. These results suggest that PLD, PA, and the oxylipin pathway may be involved in producing the wound signal responsible for increased wound-induced PAL activity, phenolic accumulation, and browning in fresh-cut lettuce leaf tissue.