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While it is commonly understood that air temperature can greatly affect the process of photosynthesis and the growth of higher plants, the impact of root zone temperature (RZT) on plant growth, metabolism, essential elements, as well as key metabolites like chlorophyll and carotenoids, remains an area that necessitates extensive research. Therefore, this study aimed to investigate the impact of raising the RZT on the growth, metabolites, elements, and proteins of red leaf lettuce. Lettuce was hydroponically grown in a plant factory with artificial light at four different air temperatures (17, 22, 27, and 30°C) and two treatments with different RZTs. The RZT was raised 3°C above the air temperature in one group, while it was not in the other group. Increasing the RZT 3°C above the air temperature improved plant growth and metabolites, including carotenoids, ascorbic acids, and chlorophyll, in all four air temperature treatments. Moreover, raising the RZT increased Mg, K, Fe, Cu, Se, Rb, amino acids, and total soluble proteins in the leaf tissue at all four air temperatures. These results showed that raising the RZT by 3°C improved plant productivity and the metabolites of the hydroponic lettuce by enhancing nutrient uptake and activating the metabolism in the roots at all four air temperatures. Overall, this research demonstrates that plant growth and metabolites can be improved simultaneously with an increased RZT relative to air temperature. This study serves as a foundation for future research on optimizing RZT in relation to air temperature. Further recommended studies include investigating the differential effects of multiple RZT variations relative to air temperature for increased optimization, examining the effects of RZT during nighttime versus daytime, and exploring the impact of stem heating. This research has the potential to make a valuable contribution to the ongoing growth and progress of the plant factory industry and fundamental advancements in root zone physiology. Overall, this research demonstrates that plant growth and metabolites can be improved simultaneously with an increased RZT relative to air temperature. This study serves as a foundation for future research on optimizing RZT in relation to air temperature. Further recommended studies include investigating the differential effects of multiple RZT variations relative to air temperature for increased optimization, examining the effects of RZT during nighttime versus daytime, and exploring the impact of stem heating. This research has the potential to make a valuable contribution to the ongoing growth and progress of the plant factory industry and fundamental advancements in root zone physiology.

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Introduction: Water is one of the important factors affecting the yield of leafy vegetables. Lettuce, as a widely planted vegetable, requires frequent irrigation due to its shallow taproot and high leaf evaporation rate. Therefore, screening drought-resistant genotypes is of great significance for lettuce production. Methods: In the present study, significant variations were observed among 13 morphological and physiological traits of 42 lettuce genotypes under normal irrigation and water-deficient conditions. Results: Frequency analysis showed that soluble protein (SP) was evenly distributed across six intervals. Principal component analysis (PCA) was conducted to transform the 13 indexes into four independent comprehensive indicators with a cumulative contribution ratio of 94.83%. The stepwise regression analysis showed that root surface area (RSA), root volume (RV), belowground dry weight (BDW), soluble sugar (SS), SP, and leaf relative water content (RWC) could be used to evaluate and predict the drought resistance of lettuce genotypes. Furthermore, the drought resistance ranks of the genotypes were similar according to the drought resistance comprehensive evaluation value (D value), comprehensive drought resistance coefficient (CDC), and weight drought resistance coefficient (WDC). The cluster analysis enabled the division of the 42 genotypes into five drought resistance groups; among them, variety Yidali151 was divided into group I as a strongly drought-resistant variety, group II included 6 drought-resistant genotypes, group III included 16 moderately drought-resistant genotypes, group IV included 12 drought-sensitive genotypes, and group V included 7 highly drought-sensitive genotypes. Moreover, a representative lettuce variety was selected from each of the five groups to verify its water resistance ability under water deficit conditions. In the drought-resistant variety, it was observed that stomatal density, superoxide anion (O2.-wfi2) production rate, and malondialdehyde (MDA) content exhibited a low increase rate, while catalase (CAT), superoxide dismutase (SOD), and that peroxidase (POD) activity exhibited a higher increase than in the drought-sensitive variety. Discussion: In summary, the identified genotypes are important because their drought-resistant traits can be used in future drought-resistant lettuce breeding programs and water-efficient cultivation.

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BACKGROUND AND AIMS: Air and root zone temperatures are important environmental factors affecting plant growth and yield. Numerous studies have demonstrated that air temperature strongly affects plant growth and development. Despite the extensive literature on air temperature, comprehensive studies on the effects of root zone temperature (RZT) on plant growth, elemental composition, and pigments are limited. In this study, we carefully observed the effects of RZT in red leaf lettuce to understand its effect on lettuce growth and pigment content. METHODS: Lettuce (Lactuca sativa, red leaf cultivar 'Red Fire') was grown hydroponically in a plant factory with artificial light under three RZT treatments (15, 25, or 35 °C) for 13 days. We investigated the comprehensive effects of RZT on the production of red leaf lettuce by metabolome and ionome analyses. KEY RESULTS: The 25 °C RZT treatment achieved maximum shoot and root dry weight. The 35 °C RZT decreased plant growth but significantly increased pigment contents (e.g. anthocyanins, carotenoids). In addition, a RZT heating treatment during plant cultivation that changed from 25 to 35 °C RZT for 8 days before harvest significantly increased shoot dry weight compared with the 35 °C RZT and significantly increased pigments compared with the 25 °C RZT. The 15 °C RZT resulted in significantly less pigment content relative to the 35 °C RZT. The 15 °C RZT also resulted in shoot and root dry weights greater than the 35 °C RZT but less than the 25 °C RZT. CONCLUSIONS: This study demonstrated that plant growth and pigments can be enhanced by adjusting RZT during different stages of plant growth to attain enhanced pigment contents while minimizing yield loss. This suggests that controlling RZT could be a viable method to improve lettuce quality via enhancement of pigment content quality while maintaining acceptable yields.

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Three-dimensional measurement is a high-throughput method that can record a large amount of information. Three-dimensional modelling of plants has the possibility to not only automate dimensional measurement, but to also enable visual assessment to be quantified, eliminating ambiguity in human judgment. In this study, we have developed new methods that could be used for the morphological analysis of plants from the information contained in 3D data. Specifically, we investigated characteristics that can be measured by scale (dimension) and/or visual assessment by humans. The latter is particularly novel in this paper. The characteristics that can be measured on a scale-related dimension were tested based on the bounding box, convex hull, column solid, and voxel. Furthermore, for characteristics that can be evaluated by visual assessment, we propose a new method using normal vectors and local curvature (LC) data. For these examinations, we used our highly accurate all-around 3D plant modelling system. The coefficient of determination between manual measurements and the scale-related methods were all above 0.9. Furthermore, the differences in LC calculated from the normal vector data allowed us to visualise and quantify the concavity and convexity of leaves. This technique revealed that there were differences in the time point at which leaf blistering began to develop among the varieties. The precise 3D model made it possible to perform quantitative measurements of lettuce size and morphological characteristics. In addition, the newly proposed LC-based analysis method made it possible to quantify the characteristics that rely on visual assessment. This research paper was able to demonstrate the following possibilities as outcomes: (1) the automation of conventional manual measurements, and (2) the elimination of variability caused by human subjectivity, thereby rendering evaluations by skilled experts unnecessary.

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Elucidating the mechanisms and pathways involved in genotype-environment (G×E) interactions and phenotypic plasticity is critical for improving plant growth. Controlled environment agricultural systems allow growers to modulate the environment for particular genotypes. In this study, we evaluated the effects of interactions among 14 genotypes and four artificial light environments on leaf lettuce phenotypes and dissected the underlying molecular mechanism via transcriptome-based modeling. Variations in morphological traits and phytochemical concentrations in response to artificial light treatments revealed significant G×E interactions. The appropriate genotype and artificial light combinations for maximizing phenotypic expression were determined on the basis of a joint regression analysis and the additive main effect and multiplicative interaction model for these G×E interactions. Transcriptome-based regression modeling explained approximately 50%-90% of the G×E variations. Further analyzes indicated Red Lettuce Leaves 4 (RLL4) regulates UV-B and blue light signaling through the effects of the HY5-MBW pathway on flavonoid biosynthesis and contributes to natural variations in the light-responsive plasticity of lettuce traits. Our study represents an important step toward elucidating the phenotypic variations due to G×E interactions in nonheading lettuce under artificial light conditions.

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Hexafluoropropylene oxide (HFPO) homologues, which are important alternatives to perfluorooctanoic acid, have been frequently identified in crops. Although exposure to HFPO homologues via crops may pose non-negligible threats to humans, their impact on crops is still unknown. In this study, the accumulation, transport, and distribution mechanisms of three HFPO homologues in lettuce were investigated at the plant, tissue, and cell levels. More specifically, HFPO trimer acid and HFPO tetramer acid were primarily fixed in roots and hardly transported to shoots (TF, 0.06-0.63). Conversely, HFPO dimer acid (HFPO-DA) tended to accumulate in lettuce shoots 2-264 times more than the other two homologues, thus resulting in higher estimated daily intake values. Furthermore, the dissolved organic matter derived from root exudate enhanced HFPO-DA uptake by increasing its desorption fractions in the rhizosphere. The transmembrane uptake of HFPO homologues was controlled by means of a transporter-mediated active process involving anion channels, with the uptake of HFPO-DA being additionally facilitated by aquaporins. The higher accumulation of HFPO-DA in shoots was attributed to the larger proportions of HFPO-DA in the soluble fraction (55-74%) and its higher abundance in both vascular tissues and xylem sap. Our findings expand the understanding of the fate of HFPO homologues in soil-crop systems and reveal the underlying mechanisms of the potential exposure risk to HFPO-DA.

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Seed priming increases germination, yield, and resistance to abiotic factors and phytopathogens. Chitosan is considered an ecofriendly growth stimulant and crop protection agent. Chitosan hydrolysate (CH) is an unfractionated product of hydrolysis of high-molecular-weight crab shell chitosan with a molecular weight of 1040 kDa and a degree of deacetylation of 85% with nitric acid. The average molecular weight of the main fraction in CH was 39 kDa. Lettuce seeds were soaked in 0.01-1 mg/mL CH for 6 h before sowing. The effects of CH on seed germination, plant morphology, and biochemical indicators at different growth stages were evaluated. Under the 0.1 mg/mL CH treatment, earlier seed germination was detected compared to the control. Increased root branching was observed, along with 100% and 67% increases in fresh weight (FW) at the 24th and 38th days after sowing (DAS), respectively. An increase in the shoot FW was found in CH-treated plants (33% and 4% at the 24th and 38th DAS, respectively). Significant increases in chlorophyll and carotenoid content compared to the control were observed at the 10th DAS. There were no significant differences in the activity of phenylalanine ammonia-lyase, polyphenol oxidase, ß-1,3-glucanase, and chitinase at the 24th and 38th DAS. Seed priming with CH could increase the yield and uniformity of plants within the group. This effect is important for commercial vegetable production.

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Producing microalgae with agricultural drainage water (ADW) allows recycling water and nutrients, with the production of a biofertilizer, avoiding receiving waters' contamination. Chlorella vulgaris and Scenedesmus obliquus were cultivated using ADW and standard media supplementation and presented higher productivities, relatively to the control industrial growth medium (using freshwater). Selected strains were grown outdoors in pilot flat panel photobioreactors, reaching 2.20 g L-1 for S. obliquus and 1.15 g L-1 for C. vulgaris, and degrading herbicides in the ADW to non-quantifiable concentrations. The potential of the C. vulgaris and S. obliquus suspensions to replace 50% of nitrogen (N) mineral fertilization of lettuce (0.5 g pot-1) was evaluated through a pot trial, also using a 2-times (1.0 g pot-1) and 5-times (2.5 g pot-1) higher dose, applied 31 days before lettuce transplanting. Even the lower dose of N, applied via C. vulgaris or S. obliquus suspensions, was able to provide significantly higher lettuce fresh matter yield, relatively to the mineral fertilized control. Soil enzymatic activities were improved, with significantly higher dehydrogenase, ß-glucosidase, and acid phosphatase activities for the 2.5 g pot-1 dose, more marked for S. obliquus, which was also able to increase soil organic matter content. Both the non-fertilized control and microalgae fertilized pots led to similar soil electrical conductivities, 3-fold lower than in the N-mineral fertilized pots, evidencing the capacity of microalgae fertilizers to avoid soil secondary salinization. Results suggest benefits from using ADW from maize cultivation to produce C. vulgaris or S. obliquus suspensions, that can be further used as liquid organic slow-release fertilizer.

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BACKGROUND: High temperature induces early bolting in lettuce (Lactuca sativa L.), which affects both quality and production. However, the molecular mechanism underlying high temperature-induced bolting is still limited. RESULTS: We performed systematical analysis of morphology, transcriptome, miRNAs and methylome in lettuce under high temperature treatment. Through a comparison of RNA-Seq data between the control and the high temperature treated lettuces at different time points totally identified 2944 up-regulated genes and 2203 down-regulated genes, which cover three floral pathways including photoperiod, age and gibberellin (GA) pathways. Genome wide analysis of miRNAs and methylome during high temperature treatment indicated miRNAs and DNA methylation might play a role controlling gene expression during high temperature-induced bolting. miRNA targets included some protein kinase family proteins, which potentially play crucial roles in this process. CONCLUSIONS: Together, our results propose a possible regulation network involved in high temperature-induced bolting.

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Some cultivars of lettuce accumulate anthocyanins, which act as functional food ingredients. Leaf lettuce has been known to be erratic in exhibiting red color when grown under artificial light, and there is a need for cultivars that more stably exhibit red color in artificial light cultivation. In this study, we aimed to dissect the genetic architecture for red coloring in various leaf lettuce cultivars grown under artificial light. We investigated the genotype of Red Lettuce Leaf (RLL) genes in 133 leaf lettuce strains, some of which were obtained from publicly available resequencing data. By studying the allelic combination of RLL genes, we further analyzed the contribution of these genes to producing red coloring in leaf lettuce. From the quantification of phenolic compounds and corresponding transcriptome data, we revealed that gene expression level-dependent regulation of RLL1 (bHLH) and RLL2 (MYB) is the underlying mechanism conferring high anthocyanin accumulation in red leaf lettuce under artificial light cultivation. Our data suggest that different combinations of RLL genotypes cause quantitative differences in anthocyanin accumulation among cultivars, and some genotype combinations are more effective at producing red coloration even under artificial lighting.

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Green light, as part of the photosynthetically active radiation, has been proven to have high photosynthetic efficiency once absorbed by plant leaves and can regulate plant physiological activities. However, few studies have investigated the appropriate and efficient way of using the green light for plant production. Thus, the objective of this study was to investigate a moderate amount of green light, partially replacing red and blue light, for plant growth and development. In this experiment, four treatments were set up by adjusting the relative amount of green light as 0 (RB), 30 (G30), 60 (G60), and 90 (G90) µmol m-2 s-1, respectively, with a total photosynthetic photon flux density of 200 µmol m-2 s-1 and a fixed red-to-blue ratio of 4:1. Lettuce (Lactuca sativa cv. 'Tiberius') plant growth and morphology, stomatal characteristics, light absorptance and transmittance, photosynthetic characteristics, and nutritional quality were investigated. The results showed that: (1) shoot dry weight increased by 16.3 and 24.5% and leaf area increased by 11.9 and 16.2% under G30 and G60, respectively, compared with those under RB. Plant stem length increased linearly with increasing green-to-blue light ratio; (2) light transmittance of lettuce leaf under treatments employing green light was higher than that under RB, especially in the green region; (3) stomatal density increased, whereas stomatal aperture area decreased with the increase in the relative amount of green light; and (4) carbohydrate accumulation increased under G60 and G90. Soluble sugar contents under G60 and G90 increased by 39.4 and 19.4%, respectively. Nitrate contents under G30, G60, and G90 decreased by 26.2, 40.3, and 43.4%, respectively. The above results indicated that 15-30% green light replacing red and blue light effectively increased the yield and nutritional quality of lettuce plants.

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BACKGROUND: Salinity and drought stresses have become widespread in many regions of the world. Although there are several studies, their findings about the response of lettuce to water and salinity stresses are contradictory. This paper therefore aims to evaluate the effects of water deficit and irrigation water salinity on growth, yield, and water consumption of iceberg lettuce. For these purposes, two experiments were carried out under Mediterranean conditions. RESULTS: The water yield response factors (Ky ) determined for the lettuce plant grown under stress conditions caused by water (Ky  = 1.69) and irrigation water salinity (Ky  = 2.62) were quite different from each other. The classical salinity tolerance model did not reflect the results accurately because the fresh yield and plant water consumption of lettuce increased sharply with increasing soil salinity up to 2.17 dS m-1 and then decreased slightly after this value. Thus, a new model was created to reflect both the increase and decrease in fresh yield. The actual salt tolerance model for iceberg lettuce plant showed that the optimum salinity is 1.84 dS m-1 with relative yield decreases of 8.26 and 22.7% per unit salinity increase above and below the optimum salinity level, respectively. CONCLUSION: Lettuce fresh yield at soil salinity below the optimum salinity experienced greater reduction than at soil salinity above the optimum value. The results reveal that the use of low-salinity irrigation water should be preferred to increase fresh yield in iceberg lettuce cultivation. © 2021 Society of Chemical Industry.

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Currently, there is a lack of information about the influence of foliar-applied nanoplastics on crop growth and nutritional quality. To fill the knowledge gap, soil-grown lettuces (Lactuca sativa L.) were foliar-exposed to polystyrene nanoplastics (PSNPs) at 0, 0.1 and 1 mg/L for one month. Foliar exposure to PSNPs significantly decreased the dry weight, height, and leaf area of lettuce by 14.3%-27.3%, 24.2%-27.3%, and 12.7%-19.2%, respectively, compared with the control. Similarly, plant pigment content (chlorophyll a, b and carotenoid) was considerably reduced (9.1%, 8.7%, 12.5%) at 1 mg/L PSNPs. However, the significant increase in electrolyte leakage rate (18.6%-25.5%) and the decrease in total antioxidant capacity (12.4%-26%) were the key indicators of oxidative stress in lettuce leaves, demonstrating the phytotoxicity of PSNPs by foliar exposure. In addition, the remarkable reduction in micronutrients and essential amino acids demonstrated a decrease in nutritional quality of lettuce caused by PSNPs. Besides, SEM and TEM analysis indicated the possible absorption of PSNPs through leaves stoma and the translocation downwards to plant roots. This study provides new information about the interaction of airborne NPs with plants. It also warns against atmospheric NPs pollution that the adverse effects of airborne NPs on crop production and food quality should be assessed as a matter of urgency.

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BACKGROUND: Insufficient intake of zinc is associated with various diseases worldwide. To overcome this problem, we aimed to establish a method for cultivating leafy vegetables with high zinc content in hydroponics without inhibiting their growth. Furthermore, we evaluated the effectiveness of the cultivated leafy vegetables with high zinc content in zinc-deficient mice. RESULTS: By adjusting the zinc concentration in the hydroponic solution to 5 mg L-1 starting from 7 days before harvesting, the zinc content in leaf lettuce increased eight times of that in the control, without any inhibition of the growth. Furthermore, when oxidized glutathione (GSSG) was added simultaneously with zinc to the hydroponic solution, the zinc content further doubled (16 times of that in the control). Similar results were obtained with komatsuna and red leaf lettuce, although there was a difference in the effect of GSSG treatment. The effectiveness of leafy vegetables with high zinc content in ameliorating zinc deficiency was evaluated by feeding lettuce with high zinc content to zinc-deficient mice. High zinc content lettuce significantly increased the zinc content in the liver, kidneys, gastrocnemius, and tibia of these mice. CONCLUSION: We established a cultivation method for lettuce and komatsuna with high zinc content without inhibiting growth by adjusting the zinc concentration in the hydroponic solution at an appropriate concentration for an appropriate period. The result of feeding test indicates that the intake of leafy vegetables with high zinc content can ameliorate zinc deficiency and might be useful in protection from several diseases associated with this deficiency. © 2020 Society of Chemical Industry.

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SCOPE: Gut microbiota converts dietary phytochemicals into metabolites and modulates their health effects. The microbial metabolism of dietary terpenoids, as the sesquiterpene lactones of leafy vegetables, is unknown. METHODS AND RESULTS: In vitro fermentation of lactucopicrin, lactucin, and romaine lettuce with gut microbiota from independent donors, show their extensive metabolism through untargeted metabolomics of the fecal incubations. Dehydroxylations and double bond hydrogenations are the main catabolic reactions. Isomers of dihydrolactucopicrin, tetrahydrolactucopicrin, and deoxylactucin, are observed after lactucopicrin metabolism. Tetrahydrolactucin and hexahydrolactucin are also found after lactucin metabolism. Lettuce fermentation shows similar metabolic conversions. Phase II conjugates of most of these metabolites are detected in the urine of healthy volunteers after escarole salad intake. Glucuronides, and sulfates, of dihydrolactucopicrin, tetrahydrolactucopicrin, dihydrolactucin, and deoxylactucin, are detected in the urine although with large inter-subject variability. CONCLUSION: This is the first report on the gut microbiota metabolism of sesquiterpene lactones in humans, and one of the first reports to describe that dietary terpenoids of widely consumed leafy vegetables are extensively catabolized by human gut microbiota. A large inter-subject variation in the metabolism of sesquiterpene lactones also reflects differences in gut microbiota composition. It suggests that inter-individual differences in their health effects should be expected.

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Reactive oxygen species (ROS) are increasingly recognized as non-enzymatic players in the processes of radicle elongation growth and endosperm weakening during seed germination. NADPH oxidases (EC 1.6.3.1), also known as respiratory burst oxidase homologues (Rbohs), are key enzymes for the production of ROS. We previously reported that ROS are involved in the radicle elongation growth and endosperm weakening during lettuce seed germination. However, the function of the gene(s) encoding Rbohs during lettuce seed germination remains to be elucidated. In this study, one lettuce Rboh gene LsRbohC1 was cloned, and over-expression and RNAi-lines of this gene were generated. It was found that LsRbohC1 was abundantly expressed in germinating seeds, especially in the endosperm cap and hypocotyl. Over-expression/knock-down of this gene observably increased/decreased the production of superoxide radicals in the radicle and endosperm cap, and significantly promoted/delayed the germination process. The results suggest that LsRbohC1 plays a role in lettuce seed germination.

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Poly-γ-glutamic acid (γ-PGA) could efficiently stabilize heavy metals in the environment. This study characterized the effects of two plant growth-promoting and γ-PGA-producing bacteria Bacillus subtilis W7 and Bacillus amyloliquefaciens W25 on Cd immobilization and γ-PGA production in soil filtrate and on the biomass and Cd uptake by lettuce in Cd-contaminated soil, the impact of these strains on the rhizosphere soil bacterial community was also evaluated. The strains reduced Cd concentration (16-75 %) in soil filtrate and strain W25 had a higher ability of producing γ-PGA and immobilizing Cd than strain W7. Compared with the control, the strains significantly increased the biomass (41-85 %) and reduced Cd uptake (19-41 %) by lettuce, reduced available Cd content (25-37 %) and increased the relative abundance of γ-PGA-producing bacteria (24-30 %) in Cd-contaminated soil, among which the effects of strain W25 were better than that of strain W7. Besides, these isolates also increased soil pH value, urease activity and the relative abundance of plant growth-promoting and metal-immobilizing bacteria such as Sphingomonas and Bacillus. In summary, the two strains reduced soil available Cd and lettuce Cd uptake by increasing the pH value, urease activity and the abundance of γ-PGA-producing bacteria, and regulating bacterial community structure in rhizosphere soil.

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Contamination of romaine lettuce with human pathogens, antibiotic-resistant bacteria (ARB), and antibiotic resistance genes (ARGs) occurs during production. Post-harvest interventions are emplaced to mitigate pathogens, but could also mitigate ARB and ARGs on vegetables. The objective of this research was to determine changes to lettuce phyllosphere microbiota, inoculated ARB, and the resistome (profile of ARGs) following washing with a sanitizer, gamma irradiation, and cold storage. To simulate potential sources of pre-harvest contamination, romaine lettuce leaves were inoculated with compost slurry containing antibiotic-resistant strains of pathogenic (Escherichia coli O157:H7) and representative of spoilage bacteria (Pseudomonas aeruginosa). Various combinations of washing with sodium hypochlorite (50 ppm free chlorine), packaging under modified atmosphere (98% nitrogen), irradiating (1.0 kGy) and storing at 4°C for 1 day versus 14 days were compared. Effects of post-harvest treatments on the resistome were profiled by shotgun metagenomic sequencing. Bacterial 16S rRNA gene amplicon sequencing was performed to determine changes to the phyllosphere microbiota. Survival and regrowth of inoculated ARB were evaluated by enumeration on selective media. Washing lettuce in water containing sanitizer was associated with reduced abundance of ARG classes that confer resistance to glycopeptides, ß-lactams, phenicols, and sulfonamides (Wilcoxon, p < 0.05). Washing followed by irradiation resulted in a different resistome chiefly due to reductions in multidrug, triclosan, polymyxin, ß-lactam, and quinolone ARG classes (Wilcoxon, p < 0.05). Irradiation followed by storage at 4°C for 14 days led to distinct changes to the ß-diversity of the host bacteria of ARGs compared to 1 day after treatment (ANOSIM, R = 0.331; p = 0.003). Storage of washed and irradiated lettuce at 4°C for 14 days increased the relative abundance of Pseudomonadaceae and Carnobacteriaceae (Wilcoxon, p < 0.05), two groups whose presence correlated with detection of 10 ARG classes on the lettuce phyllosphere (p < 0.05). Irradiation resulted in a significant reduction (∼3.5 log CFU/g) of inoculated strains of E. coli O157:H7 and P. aeruginosa (ANOVA, p < 0.05). Results indicate that washing, irradiation and storage of modified atmosphere packaged lettuce at 4°C are effective strategies to reduce antibiotic-resistant E. coli O157:H7 and P. aeruginosa and relative abundance of various ARG classes.

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Lettuce (Lactuca sativa L.) constitutes one the most important vegetable crops worldwide. Poultry litter is being applied as an economically suitable alternative to nitrogen fertilizers in lettuce cultivation. However, little is known about the effects of this practice over this fresh product safety that is usually consumed as a salad. The aim of this work was to determine the microbiological quality and the nitrate content in lettuce produced, under field conditions, using either raw or composted poultry litter, coming from the same original batch. Two experiments were conducted in the experimental field of Facultad de Ciencias Agrarias (UNL, Santa Fe, Argentina) to assess the effects of recently extracted poultry litter that consisted of broiler chicken manure plus rice husk, or composted for 12 mo. The application amounts were: 20 T ha-1 (T1); 40 T ha-1 (T2); and no application of manure (T). Increasing the applied quantities had also increased the health risk associated with lettuce consumption, due to higher nitrate levels and microbial contamination. However, these risks were reduced by composting the material. Even when lettuce contamination with faecal bacteria was mainly due to the use of poultry litter, the number and incidence of pathogens were reduced when properly composted manure was applied instead of raw one. Increasing the dose of poultry litter applied also increases the health risk in lettuce. Though, when the material is properly composted, its fertilizing capacity is maintained, giving proper yields with lower nitrate levels and microbial contamination by enterobacteria.

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Perfluorooctanesulfonate (PFOS) is a toxic and persistent organic pollutant that can be widely detected in agricultural soils. In this study, two lettuce cultivars with low PFOS accumulation were screened out to reduce the exposure of PFOS to the human body via vegetable consumption. The screened low-PFOS cultivars may help to ensure food safety, despite planting in highly PFOS-polluted soils (1.0 mg/kg), due to their high tolerance to PFOS and 4.4-5.7 times lower shoot PFOS concentration than the high-PFOS cultivars. Protein content and protein-mediated transpiration played key roles in regulating PFOS accumulation in the lettuce cultivars tested. Lower protein content, lower stomatal conductance, and lower transpiration rate resulted in low PFOS accumulation. This study reveals the mechanism of forming low-PFOS accumulation of lettuce cultivars at physiological and biochemical levels and lays a foundation for developing a cost-effective and safe approach to grow vegetables in PFOS-polluted soils.

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