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Indoor farming systems enable plant production in precisely controlled environments. However, implementing stable growth conditions and the absence of stress stimulants can weaken plants' defense responses and limit the accumulation of bioactive, health-beneficial phytochemicals. A potential solution is the controlled application of stressors, such as supplemental ultraviolet (UV) light. To this end, we analyzed the efficiency of short-term pre-harvest supplementation of the red-green-blue (RGB, LED) spectrum with ultraviolet B (UV-B) or C (UV-C) light to boost phytochemical synthesis. Additionally, given the biological harm of UV radiation due to high-energy photons, we monitored plants' photosynthetic activity during treatment and their morphology as well as sensory attributes after the treatment. Our analyses showed that UV-B radiation did not negatively impact photosynthetic activity while significantly increasing the overall antioxidant potential of lettuce through enhanced levels of secondary metabolites (total phenolics, flavonoids, anthocyanins), carotenoids, and ascorbic acid. On the contrary, UV-C radiation-induced anthocyanin accumulation in the green leaf cultivar significantly harmed the photosynthetic apparatus and limited plant growth. Taken together, we showed that short-term UV-B light supplementation is an efficient method for lettuce biofortification with healthy phytochemicals, while UV-C treatment is not recommended due to the negative impact on the quality (morphology, sensory properties) of the obtained leafy products. These results are crucial for understanding the potential of UV light supplementation for producing functional plants.

UV can serve as an effective light spectrum for regulating plant secondary metabolites, while relevant studies on UV-A are much less extensive than those on UV-B. A comprehensive understanding of the selective effects of UV-A on different secondary metabolites and the specific features of primary metabolism that drive these effects is still lacking. To address this knowledge gap, we conducted a study to analyze the dynamic changes in the metabolome and transcriptome of lettuce leaves irradiated with red plus UV-A light (monochromatic red light as control). Generally, UV-A promoted the synthesis of most phenylpropanoids and terpenoids originating from the shikimate and methylerythritol phosphate (MEP) pathway in plastids but sacrificed the synthesis of terpenoids derived from the mevalonate (MVA) pathway, particularly sesquiterpenes. Increased precursors supply for the shikimate and MEP pathway under UV-A was directly supported by the activation of the Calvin-Benson cycle and phosphoenolpyruvate transport. Whereas, along with phosphoenolpyruvate transport, the TCA cycle was restrained, causing deprivation of the MVA pathway precursor. In addition, UV-A also activated the plastidic oxidative branch of the pentose phosphate pathway, photorespiration, and malate shuttle, to ensure a sufficient supply of nitrogen, circulation homeostasis of the Calvin-Benson cycle, and energy balance, thus indirectly supporting UV-A-induced specific secondary metabolic output. This study provides a comprehensive framework for understanding the flexible primary-secondary metabolism interactions that are able to produce specific metabolites favorable for adaptation to environmental stimuli.

Background: Heat shock factors (Hsfs) and Heat shock proteins (Hsps) belong to an essential group of molecular regulators involved in controlling cellular processes under normal and stress conditions. The role of Hsfs and Hsps is well known in model plant species under diverse stress conditions. While plants Hsfs are vital components of the signal transduction response to maintain cellular homeostasis, Hsps function as chaperones helping to maintain folding of damaged and newly formed proteins during stress conditions. In lettuce (Lactuca sativa), a highly consumed vegetable crop grown in the field and in hydroponic systems, the role of these gene families in response to artificial light is not well characterized., Results: Using a genome-wide analysis approach, we identified 32 Hsfs and 22 small heat shock proteins (LsHsps) in lettuce, some of which do not have orthologs in Arabidopsis, poplar, and rice. LsHsp60s, LsHsp90s, and LsHsp100s are highly conserved among dicot and monocot species. Surprisingly, LsHsp70s have three times more members than Arabidopsis and two times more than rice. Interestingly, the lettuce genome triplication did not contribute to the increased number of LsHsp70s genes. The large number of LsHsp70s was the result of genome tandem duplication. Chromosomal distribution analysis shows larger tandem repeats of LsHsp70s genes in Chr1, Chr7, Chr8, and Chr9. At the transcriptional level, some genes of the LsHsfs, LsHsps, LsHsp60s, and LsHsp70s families were highly responsive to UV and high intensity light stress, in contrast to LsHsp90s and LsHsp100s which did not respond to a light stimulus., Conclusions: Our genome-wide analysis provides a detailed identification of Hsfs and Hsps in lettuce. Chromosomal location and syntenic region analysis together with our transcriptional analysis under different light conditions provide candidate genes for breeding programs aiming to produce lettuce varieties able to grow healthy under hydroponic systems that use artificial light.

Leaves of lettuce, pepper, tomato and grapevine plants grown in greenhouse conditions were exposed to UV-C light for either 60 s or 1 s, using a specific LEDs-based device, and wavelengths and energy were the same among different light treatments. Doses of UV-C light that both effectively stimulated plant defences and were innocuous were determined beforehand. Tomato plants and lettuce plants were inoculated with Botrytis cinerea, pepper plants with Phytophthora capsici, and grapevine with Plasmopara viticola. In some experiments we investigated the effect of a repetition of treatments over periods of several days. All plants were inoculated 48 h after exposure to the last UV-C treatment. Lesions on surfaces were measured up to 12 days after inoculation, depending on the experiment and the pathogen. The results confirmed that UV-C light stimulates plant resistance; they show that irradiation for one second is more effective than irradiation for 60 s, and that repetition of treatments is more effective than single light treatments. Moreover a systemic effect was observed in unexposed leaves that were close to exposed leaves. The mechanisms of perception and of the signalling and metabolic pathways triggered by flashes of UV-C light vs. 60 s irradiation exposures are briefly discussed, as well as the prospects for field use of UV-C flashes in viticulture and horticulture., Competing Interests: The authors have declared that no competing interests exist.

Main Conclusion: Environmentally induced variation and the genotypic differences in flavonoid and phenolic content in lettuce can be reliably detected using the appropriate parameters derived from the records of rapid non-invasive fluorescence technique. The chlorophyll fluorescence excitation ratio method was designed as a rapid and non-invasive tool to estimate the content of UV-absorbing phenolic compounds in plants. Using this technique, we have assessed the dynamics of accumulation of flavonoids related to developmental changes and environmental effects. Moreover, we have tested appropriateness of the method to identify the genotypic differences and fluctuations in total phenolics and flavonoid content in lettuce. Six green and two red genotypes of lettuce (Lactuca sativa L.) grown in pots were exposed to two different environments for 50 days: direct sunlight (UV-exposed) and greenhouse conditions (low UV). The indices based on the measurements of chlorophyll fluorescence after red, green and UV excitation indicated increase of the content of UV-absorbing compounds and anthocyanins in the epidermis of lettuce leaves. In similar, the biochemical analyses performed at the end of the experiment confirmed significantly higher total phenolic and flavonoid content in lettuce plants exposed to direct sun compared to greenhouse conditions and in red compared to green genotypes. As the correlation between the standard fluorescence indices and the biochemical records was negatively influenced by the presence of red genotypes, we proposed the use of a new parameter named Modified Flavonoid Index (MFI) taking into an account both absorbance changes due to flavonol and anthocyanin content, for which the correlation with flavonoid and phenolic content was relatively good. Thus, our results confirmed that the fluorescence excitation ratio method is useful for identifying the major differences in phenolic and flavonoid content in lettuce plants and it can be used for high-throughput pre-screening and phenotyping of leafy vegetables in research and breeding applications towards improvement of vegetable health effects.

The purpose of the present study was to evaluate the role of proline (Pro) in relieving UV-B radiation-induced oxidative stress in lettuce. Lettuce seedlings were exposed to 3.3 W m -2 UV-B radiation for 12 h after pre-treatment sprayed with 20 mM Pro. The data for malondialdehyde (MDA), hydrogen peroxide (H 2 O 2 ), endogenous Pro level, the activities of antioxidant enzymes [superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and peroxidase (POD)], total phenolic concentration, antioxidant capacity, expression of phenylalanine ammonia lyase (PAL), γ-tocopherol methyltransferase (γ-TMT) and proline dehydrogenase (ProDH) genes, phytohormone levels such as abscisic acid (ABA), gibberellic acid (GA), indole acetic acid (IAA) and salicylic acid (SA), soluble sugars and organic acids were recorded. It was found that Pro alleviated the oxidative damage in the seedlings of lettuce as demonstrated by lower lipid peroxidation and H 2 O 2 content, increasing the endogenous Pro level, the activity of antioxidant enzymes, total phenolic concentration and the antioxidant capacity. Additionally, it was revealed that exogenous application of Pro enhanced the levels of GA, IAA, the concentrations of soluble sugars and organic acids and expressions of PAL, γ-TMT and ProDH genes as compared to the control. The results obtained in this study suggest that pre-treatment with exogenous Pro provides important contributions to the increase in the UV-B tolerance of lettuce by regulating the biochemical mechanisms of UV-B response.

Background: The production of high-quality crops based on phytochemicals is a strategy for accelerating the practical use of plant factories. Previous studies have demonstrated that ultraviolet (UV) light is effective in improving phytochemical production. This study aimed to determine the effect of various UV wavelengths on growth and phenolic compound accumulation in lettuce (Lactuca sativa L.) grown in a closed-type plant production system., Results: Seven days, 1 day and 0.25 day were determined as the upper limit of the irradiation periods for UV-A, -B, and -C, respectively, in the lettuce based on physiological disorders and the fluorescence parameter F(v)/F(m). Continuous UV-A treatment significantly induced the accumulation of phenolic compounds and antioxidants until 4 days of treatment without growth inhibition, consistent with an increase in phenylalanine ammonia lyase (PAL) gene expression and PAL activity. Repeated or gradual UV-B exposure yielded approximately 1.4-3.6 times more total phenolics and antioxidants, respectively, than the controls did 2 days after the treatments, although both treatments inhibited lettuce growth. Repeated UV-C exposure increased phenolics but severely inhibited the growth of lettuce plants., Conclusion: Our data suggest that UV irradiation can improve the accumulation of phenolic compounds with antioxidant properties in lettuce cultivated in plant factories., (© 2013 Society of Chemical Industry.)

The effects of two non thermal disinfection processes, Ultraviolet light (UV 254 nm) and Ultrasound (US) on the inactivation of bacteria and color in two freshly cut produces (lettuce and strawberry) were investigated. The main scope of this work was to study the efficacy of UV and US on the decontamination of inoculated lettuce and strawberries with a cocktail of four bacteria, Escherichia coli, Listeria innocua, Salmonella Enteritidis and Staphylococcus aureus. Treatment of lettuce with UV reduced significantly the population of E. coli, L. innocua, S. Enteritidis and S. aureus by 1.75, 1.27, 1.39 and 1.21 log CFU/g, respectively. Furthermore, more than a 2-log CFU/g reduction of E. coli and S. Enteritidis was achieved with US. In strawberries, UV treatment reduced bacteria only by 1-1.4 log CFU/g. The maximum reductions of microorganisms, observed in strawberries after treatment with US, were 3.04, 2.41, 5.52 and 6.12 log CFU/g for E. coli, S. aureus, S. Enteritidis and L. innocua, respectively. Treatment with UV and US, for time periods (up to 45 min) did not significantly (p>0.05) change the color of lettuce or strawberry. Treatment with UV and US reduced the numbers of selected inoculated bacteria on lettuce and strawberries, which could be good alternatives to other traditional and commonly used technologies such as chlorine and hydrogen peroxide solutions for fresh produce industry. These results suggest that UV and US might be promising, non-thermal and environmental friendly disinfection technologies for freshly cut produce., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The effect of various conditions on inactivation of foodborne pathogens and quality of fresh-cut lettuce during ultraviolet (254 nm, UVC) radiation was investigated. Lettuce was inoculated with a cocktail of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes and treated at different temperatures (4 and 25 °C), distances between sample and lamp (10 and 50 cm), type of exposure (illuminated from one or two sides), UV intensities (1.36 to 6.80 mW/cm²), and exposure times (0.5 to 10 min), sequentially. UV treatment at 25 °C for 1 min achieved 1.45-, 1.35-, and 2.12-log reductions in surface-inoculated E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively, whereas the reduction of these pathogens at 4 °C was 0.31, 0.57, and 1.16 log, respectively. UV radiation was most effective when distance from UV lamp to the sample was minimal (10 cm) and radiation area was maximal (two-sided exposure). All UV intensities significantly (P<0.05) reduced the three pathogens after 10 min exposure, but the effect of treatment was correlated with UV intensity and exposure time. Color values and texture parameters of lettuce subjected to UV treatment under the optimum conditions (25 °C, 10 cm between sample and lamp, two-sided exposure, 6.80 mW/cm²) were not significantly (P>0.05) different from those of nontreated samples up to 5 min exposure. However, these qualities significantly (P<0.05) changed at prolonged treatment time. These results suggest that UV radiation under optimized conditions could reduce foodborne pathogens without adversely affecting color quality properties of fresh-cut lettuce., (© 2013 Elsevier B.V. All rights reserved.)

Seedlings of 10-day-old lettuce (Lactuca sativa L., cultivar Berlin) were preilluminated by low intensity red light (λmax=660 nm, 10 min, 5 μmol quanta m(-2) s(-1)) and far-red light (λmax=730 nm, 10 min, 5 μmol quanta m(-2) s(-1)) to study the effect of pre-treatment on photosynthesis, photochemical activity of photosystem II (PSII), the contents of photosynthetic and UV-A-absorbing pigments (UAPs) and H2O2, as well as total and ascorbate peroxidase activities in cotyledonary leaves of seedlings exposed to UV-A. UV radiation reduced the photosynthetic rate (Pn), the activity of PSII, and the contents of Chl a and b, carotenoids and UAPs in the leaves, but increased the content of H2O2 and the total peroxidase activity. Preillumination with red light removed these effects of UV. In turn, the illumination with red light, then far-red light removed the effect of the red light. Illumination with red light alone increased the content of UAPs, as well as peroxidase activity. It is suggested that higher resistance of the lettuce photosynthetic apparatus to UV-A radiation is associated with involvement of the active form of phytochrome B, thereby increasing peroxidase activities as well as UAPs and saving preservation of photosynthetic pigment contents due to pre-illumination with red light., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Solar ultraviolet (UV)-B radiation (280-315 nm) has a wide range of effects on terrestrial ecosystems, yet our understanding of how UV-B influences the complex interactions of plants with pest, pathogen and related microorganisms remains limited. Here, we report the results of a series of experiments in Lactuca sativa which aimed to characterize not only key plant responses to UV radiation in a field environment but also consequential effects for plant interactions with a sap-feeding insect, two model plant pathogens and phylloplane microorganism populations. Three spectrally modifying filters with contrasting UV transmissions were used to filter ambient sunlight, and when compared with our UV-inclusive filter, L. sativa plants grown in a zero UV-B environment showed significantly increased shoot fresh weight, reduced foliar pigment concentrations and suppressed population growth of green peach aphid (Myzus persicae). Plants grown under a filter which allowed partial transmission of UV-A radiation and negligible UV-B transmission showed increased density of leaf surface phylloplane microbes compared with the UV-inclusive treatment. Effects of UV treatment on the severity of two plant pathogens, Bremia lactucae and Botrytis cinerea, were complex as both the UV-inclusive and zero UV-B filters reduced the severity of pathogen persistence. These results are discussed with reference to known spectral responses of plants, insects and microorganisms, and contrasted with established fundamental responses of plants and other organisms to solar UV radiation, with particular emphasis on the need for future integration between different experimental approaches when investigating the effects of solar UV radiation., (Copyright © Physiologia Plantarum 2011.)

Reductions in leaf growth are a commonly observed response to ultraviolet radiation, but the underlying mechanisms remain poorly defined. This study examined the response of leaves exposed to a UV environment across a range of organizational scales, including leaf expansion rate, epidermal cell size and number, biomechanical properties, leaf-water relations and activity of cell-wall peroxidases. Two experimental approaches were used; Lettuce (Lactuca sativa L.) plants were propagated under (a) supplementary UV-B (9 kJ m(-2) day(-1)) in controlled environment (CE) conditions, and (b) field conditions, where plants were placed under three horticultural films with differing UV transmissions. In both experiments, UV-B caused the greatest reductions in leaf expansion and final leaf size, with some reductions attributable to UV-A wavelengths. In supplementary UV-B conditions, adaxial cell size was reduced, while in field plants, both cell size and cell number were lower in an increased UV environment, as was the case with abaxial cells in CE plants. Although leaf turgor and leaf extensibility were not affected by UV wavelengths, breaking strain of leaf tissue was decreased under supplementary UV-B. Cell-wall peroxidase activity was increased in both supplementary UV conditions and in the field, where only a zero UV environment showed no upregulation of cell-wall peroxidase.

Molecular analysis of gene expression differences between green and red lettuce leaves was performed using the SSH method. BlastX comparisons of subtractive expressed sequence tags (ESTs) indicated that 7.6% of clones encoded enzymes involved in secondary metabolism. Such clones had a particularly high abundance of flavonoid-metabolism proteins (6.5%). Following SSH, 566 clones were rescreened for differential gene expression using dot-blot hybridization. Of these, 53 were found to overexpressed during red coloration. The up-regulated expression of six genes was confirmed by Northern blot analyses. The expression of chalcone synthase (CHS), flavanone 3-hydroxylase (F3H), and dihydroflavonol 4-reductase (DFR) genes showed a positive correlation with anthocyanin accumulation in UV-B-irradiated lettuce leaves; flavonoid 3',5'-hydroxylase (F3',5'H) and anthocyanidin synthase (ANS) were expressed continuously in both samples. These results indicated that the genes CHS, F3H, and DFR coincided with increases in anthocyanin accumulation during the red coloration of lettuce leaves. This study show a relationship between red coloration and the expression of up-regulated genes in lettuce. The subtractive cDNA library and EST database described in this study represent a valuable resource for further research for secondary metabolism in the vegetable crops.

Field experiments assessing UV-B effects on plants have been conducted using two contrasting techniques: supplementation of solar UV-B with radiation from fluorescent UV lamps and the exclusion of solar UV-B with filters. We compared these two approaches by growing lettuce and oat simultaneously under three conditions: UV-B exclusion, near-ambient UV-B (control) and UV-B supplementation (simulating a 30% ozone depletion). This permitted computation of "solar UV-B" and "supplemental UV-B" effects. Microclimate and photosynthetically active radiation were the same under the two treatments and the control. Excluding UV-B changed total UV-B radiation more than did supplementing UV-B, but the UV-B supplementation contained more "biologically effective" shortwave radiation. For oat, solar UV-B had a greater effect than supplemental UV-B on main shoot leaf area and main shoot mass, but supplemental UV-B had a greater effect on leaf and tiller number and UV-B-absorbing compounds. For lettuce, growth and stomatal density generally responded similarly to both solar UV-B and supplemented UV-B radiation, but UV-absorbing compounds responded more to supplemental UV-B, as in oat. Because of the marked spectral differences between the techniques, experiments using UV-B exclusion are most suited to assessing effects of present-day UV-B radiation, whereas UV-B supplementation experiments are most appropriate for addressing the ozone depletion issue.