Non-steroidal anti-inflammatory drugs, diclofenac (DCF) and naproxen (NPX), represent a group of environmental contaminants often detected in various water and soil samples. This work aimed to assess possible phytotoxic effects of DCF and NPX in concentrations 0.1, 1 and 10 mg/L, both individually and in binary mixtures, on the seed germination and primary root elongation of crops, monocots Allium porrum and Zea mays, and dicots Lactuca sativa and Pisum sativum. Results proved that the seed germination was affected by neither individual drugs nor their mixture. The response of primary root length in monocot and dicot species to the same treatment was different. The Inhibition index (%) comparing the root length of drug-treated plants to controls proved to be approximately 10% inhibition in the case of dicots lettuce and pea, and nearly 20% inhibition in monocot leek, but almost 20% stimulation in monocot maize. Assessment of the binary mixture effect confirmed neither synergistic nor antagonistic interaction of DCF and NPX on early plant development in the applied concentration range.

Cadmium (Cd) is a harmful heavy metal that is highly toxic to plants and animals. Expansins are cell wall proteins inducing cell wall loosening and participate in all plant growth and development processes which are associated with cell wall modifications. We investigated lettuce\'s expansin gene LsEXPA6 and found that LsEXPA6 overexpression Arabidopsis lines were much more resistant to cadmium stress. Our results revealed that the root system of the expa6 mutant was suppressed under cadmium stress, resulting in shorter plant height, reduced biomass, and a significant increase in cadmium content in the plants compared with wild-type plants, whereas LsEXPA6 overexpression lines had a well-developed root system and reduced cadmium accumulation in the roots and shoots of the plants. The above results indicated that overexpression of LsEXPA6 affected root development and reduced Cd absorption in Arabidopsis. In addition, the higher absorption capacity of nutrients, increased antioxidant enzymes activities, improved chlorophyll and photosynthetic function in the overexpression Arabidopsis plants, supported the Cd stress tolerance mechanism. Taken together, these results provided a new insight on the role of expansin proteins in the tolerance of plants to Cd stress by root cell elongation.

Cyanobacteria represent a promising resource for sustainable agriculture, as they have demonstrated the ability to restore soil fertility even after death and decay. However, several cyanobacteria can also release secondary metabolites, such as cyanotoxins, which may compromise the quality of agricultural products and pose a potential risk to human health. Depending on the concentration of exposure, few studies reported deleterious effects on plant species when irrigated with cylindrospermopsin (CYN) contaminated water, impairing plant growth and leading to food product contamination, while other studies show promoting effects on plant yield. To evaluate the potential of cyanobacteria biomass (cyanotoxin-containing or not) as a sustainable resource for soil amendment, biostimulants or fertilizers for lettuce cultivation, a study was carried out that consisted of the culture of lettuce plants under controlled conditions, in soil: (1) with no extra nutrient addition (control) and supplemented with 0.6 g of freeze-dried Raphidiopsis raciborskii biomass of (2) a non-CYN producer strain, (3) a CYN producer strain, and (4) the same CYN producer strain pasteurized. Results showed no significant differences in photosystem II efficiency with the amendment addition. On the contrary, shoot fresh weight significantly increased in lettuce plants grown with the cyanobacterial biomass addition, especially in condition (3). In addition, there were significant differences in mineral concentrations in lettuce leaves after the cyanobacterial biomass addition, such as K, Na, Ca, P, Mg, Mn, Zn, Cu, Mo, and Co. CYN accumulation was detected under conditions (3) and (4), with concentrations observed in descending order from roots > soil > shoot. Nevertheless, the CYN concentration in edible tissues did not exceed the WHO-proposed tolerable daily intake of 0.03 μg/kg/day. These findings suggest that incorporating cyanobacterial biomass as a soil amendment, biostimulant or fertilizer for lettuce cultivation, even with trace amounts of CYN (1∼40 μg/g), may enhance plant yield without leading to cyanotoxin accumulation in edible tissues above the WHO-recommended tolerable daily intake.

The accuracy of predictive microbial models used in quantitative microbial risk assessment (QMRA) relies on the relevancy of conditions influencing growth or inactivation. The continued use of log-linear models in studies remains widespread, despite evidence that they fail to accurately account for biphasic kinetics or include parameters to account for the effect of environmental conditions within the model equation. Although many experimental studies detail conditions of interest, studies that do not do so lead to uncertainty in QMRA modeling because the applicability of the predictive microbial models to the conditions in the risk scenarios is questionable or must be extrapolated. The current study systematically reviewed 65 articles that provided quantitative data and documented the conditions influencing the inactivation or growth of Shiga toxin-producing Escherichia coli (STEC) O157:H7 in leafy greens. The conditions were identified and categorized as environmental, biological, chemical, and/or processing. Our study found that temperature (n = 37 studies) and sanitizing and washing procedures (n = 12 studies) were the most studied conditions in the farm-to-table continuum of leafy greens. In addition, relative humidity was also established to affect growth and inactivation in more than one stage in the continuum. This study proposes the evaluation of the interactive effects of multiple conditions in processing and storage stages from controlled experiments as they relate to the fate of STEC O157:H7 in leafy greens for future quantitative analysis.

Lettuce is an established food commodity in the UK increasingly facing supply challenges in winter due to adverse weather events and rising energy costs. We investigate whether an agroecologically grown salad mix of lettuce and underutilised leafy greens produced in the Azores, Portugal, could be part of a sustainable solution. We performed a Life Cycle Assessment to compare the environmental impacts of this salad mix with four other value chains for winter salad supply to the UK: conventional open-field lettuce production chains in Spain using (1) current irrigation practices; (2) 100 % desalinated irrigation water; or hydroponic controlled environment agriculture within the UK powered by (3) the national electricity mix; (4) 100 % wind-generated electricity. Results indicated that the leafy-greens agroecological value chain incurred the smallest environmental burdens across 7 to 11 of 16 impact categories studied. Substituting Spanish winter salad supply with agroecological leafy green production in the Azores, if well managed, could reduce many environmental burdens whilst diversifying leafy greens intake. Nevertheless, all winter value chains were associated with larger environmental burdens than conventional open-field production of lettuce in the UK summer, pointing to the importance of seasonal consumption and wider adoption of agroecological techniques to effectively reduce environmental impact.

Organic farming methods, including the use of organic substrates, fertilizers, pesticides, and biological control, are gaining popularity in controlled environment agriculture (CEA) due to economic benefits and environmental sustainability. However, despite several studies focusing on the preparation and evaluation of liquid organic fertilizers, none have explored the compatibility of these fertilizers with different hydroponic systems. Therefore, the objective of this study was to evaluate lettuce production using a liquid organic fertilizer under different hydroponic systems. Four distinct hydroponic methods were selected: nutrient film technique (NFT), deep water culture (DWC) (liquid culture systems), and Dutch bucket (DB), regular plastic container (RPC) (substrate-based systems). \'Green Butter\' lettuce was grown using a liquid organic fertilizer (Espartan) for four weeks. Shoot growth parameters (e.g., shoot width, number of leaves, leaf area, foliar chlorophyll content, fresh weight, and dry weight) and root growth parameters (e.g., root length, fresh weight, and dry weight) were measured. The growth difference of lettuce under the DB and RPC systems was negligible, but the growth in RPC was 29% to 60% and 15% to 44% higher than the NFT and DWC systems, respectively, for shoot width, number of leaves, leaf area, shoot fresh weight and dry weight. Root parameters were nearly identical for the NFT and DWC systems but significantly lower (21% to 94%) than the substrate-based DB and RPC systems. Although lettuce grown in the NFT system showed the least growth, its mineral content in the leaf tissue was comparable or sometimes higher than that of substrate-based hydroponic systems. In conclusion, the tested liquid organic fertilizer is suitable for substrate-based hydroponic systems; however, further evaluation of different liquid organic fertilizers, and crop species is required.

Accurate quantification of gene and transcript-specific expression, with the underlying knowledge of precise transcript isoforms, is crucial to understanding many biological processes. Analysis of RNA sequencing data has benefited from the development of alignment-free algorithms which enhance the precision and speed of expression analysis. However, such algorithms require a reference transcriptome. Here we generate a reference transcript dataset (LsRTDv1) for lettuce (cv. Saladin), combining long- and short-read sequencing with publicly available transcriptome annotations, and filtering to keep only transcripts with high-confidence splice junctions and transcriptional start and end sites. LsRTDv1 identifies novel genes (mostly long non-coding RNAs) and increases the number of transcript isoforms per gene in the lettuce genome from 1.4 to 2.7. We show that LsRTDv1 significantly increases the mapping rate of RNA-seq data from a lettuce time-series experiment (mock- and Botrytis cinerea-inoculated) and enables detection of genes that are differentially alternatively spliced in response to infection as well as transcript-specific expression changes. LsRTDv1 is a valuable resource for investigation of transcriptional and alternative splicing regulation in lettuce.

The irrigation of soils with reclaimed contaminated wastewater or its amendment with sewage sludge contributes to the uptake of pharmaceuticals by vegetables growing in the soil. A multiresidue method has been devised to determine five pharmaceuticals and nine of their main metabolites in leafy and root vegetables. The method employs ultrasound-assisted extraction, clean-up via dispersive solid-phase extraction, and analysis through liquid chromatography-tandem mass spectrometry. Box-Behnken design was used to refine variables such as extraction solvent volume, time of extraction, number of extraction cycles, and the type and amount of d-SPE sorbent. The method achieved linearity (R2) greater than 0.994, precision (relative standard deviation) under 16% for most compounds, and detection limits ranging from 0.007 to 2.25 ng g-1 dry weight. This method was applied to a leafy vegetable (lettuce) and to a root vegetable (carrot) sourced from a local market. Parent compounds were detected at higher concentrations than their metabolites, with the exception of carbamazepine-10,11-epoxide.

Systemic plant protection products, such as neonicotinoids (NIs), are capable of being translocated throughout a plant. Although NIs are less toxic to mammals, fish, and birds, their impact on microbial and non-target insects is of concern. This study investigates the uptake, translocation, and accumulation of the NI, imidacloprid (IMI), in romaine lettuce (Lactuca sativa L. var. longipolia). Exposing 15-day-old seedlings to \"10 mg/L\" of IMI, the effects on microbial communities in both cultivated (CS) and non-cultivated soil (NCS) were studied along with IMI translocation within plant tissues. The concentrations of IMI in soil varied temporally and between soil types after initial application, with a decrease from 2.0 and 7.7 mg/kg on the first day of sampling to 0.5 and 2.6 mg/kg on the final sampling day (day 35) for CS and NCS, respectively. The half-life of IMI soil was 10.7 and 72.5 days in CS and NCS, respectively, indicating that IMI degraded more quickly in CS, possibly due to smaller grain size, aeration, microbial degradation, and water flow. The accumulated concentrations of IMI in lettuce tissues ranged from 12.4 ± 0.2 and 18.7± 0.9 mg/kg in CS and NCS, respectively. The highest concentration of IMI was found in the shoots, followed by the roots, whereas the soil showed the lowest IMI residuals at the end of the trial. Soil bacteria and fungi were altered by the application of IMI, with a lower abundance index within the bacterial community, indicating a negative impact on the distribution of bacteria in the soil.

Biofortification of green leafy vegetables with pro-vitamin A carotenoids, such as β-carotene, has remained challenging to date. Here, we combined two strategies to achieve this goal. One of them involves producing β-carotene in the cytosol of leaf cells to avoid the negative impacts on photosynthesis derived from changing the balance of carotenoids and chlorophylls in chloroplasts. The second approach involves the conversion of chloroplasts into non-photosynthetic, carotenoid-overaccumulating chromoplasts in leaves agroinfiltrated or infected with constructs encoding the bacterial phytoene synthase crtB, leaving other non-engineered leaves of the plant to sustain normal growth. A combination of these two strategies, referred to as strategy C (for cytosolic production) and strategy P (for plastid conversion mediated by crtB), resulted in a 5-fold increase in the amount of β-carotene in Nicotiana benthamiana leaves. Following several attempts to further improve β-carotene leaf contents by metabolic engineering, hormone treatments and genetic screenings, it was found that promoting the proliferation of plastoglobules with increased light-intensity treatments not only improved β-carotene accumulation but it also resulted in a much higher bioaccessibility. The combination of strategies C and P together with a more intense light treatment increased the levels of accessible β-carotene 30-fold compared to controls. We further demonstrated that stimulating plastoglobule proliferation with strategy P, but also with a higher-light treatment alone, also improved β-carotene contents and bioaccessibility in edible lettuce (Lactuca sativa) leaves.