Global identification of LIM genes in response to different heat stress regimes in Lactuca sativa.

BACKGROUND: LIM (Lineage-11 (LIN-11), Insulin-1 (ISL-1), and Mechanotransduction-3 (MEC-3)) genes belong to a family that hold ubiquitous properties contributing to organ, seed, and pollen development as well as developmental and cellular responses to biotic and abiotic stresses. Lettuce (Lactuca sativa) is a highly consumed vegetable crop susceptible heat stress. High temperatures limit lettuce's overall yield, quality and marketability. Lettuce LIM genes have not been identified and their role in response to high temperatures is not known. Aiming to identify potential new targets for thermoresilience, we searched for LIM genes in lettuce and compared them with orthologous of several dicotyledons and monocotyledons plant species. RESULTS: We identified fourteen lettuce LIM genes distributed into eight different subgroups using a genome-wide analysis strategy. Three belonging to DAR (DA means "large" in Chinese) class I, two DAR class II, one in the WLIM1, two in the WLIM2, one in the PLIM1, two in the PLIM2 class, one ßLIM and two δLIMs. No DAR-like were identified in any of the species analyzed including lettuce. Interestingly, unlike other gene families in lettuce which underwent large genome tandem duplications, LIM genes did not increase in number compared to other plant species. The response to heat stress induced a dynamic transcriptional response on LsLIM genes. All heat stress regimes, including night stress, day stress and day and night stress were largely responsible for changes in LIM transcriptional expression. CONCLUSIONS: Our global analysis at the genome level provides a detailed identification of LIM genes in lettuce and other dicotyledonous and monocotyledonous plant species. Gene structure, physical and chemical properties as well as chromosomal location and Cis-regulatory element analysis together with our gene expression analysis under different temperature regimes identified LsWLIM1, LsWLIM2b, LsDAR3 and LsDAR5 as candidate genes that could be used by breeding programs aiming to produce lettuce varieties able to withstand high temperatures.

The use of biostimulants as a key to sustainable hydroponic lettuce farming under saline water stress.

BACKROUND: The utilization of high-quality water in agriculture is increasingly constrained by climate change, affecting availability, quality, and distribution due to altered precipitation patterns, increased evaporation, extreme weather events, and rising salinity levels. Salinity significantly challenges salt-sensitive vegetables like lettuce, particularly in a greenhouse. Hydroponics water quality ensures nutrient solution stability, enhances nutrient uptake, prevents contamination, regulates pH and electrical conductivity, and maintains system components. This study aimed to mitigate salt-induced damage in lettuce grown via the floating culture method under 50 mM NaCl salinity by applying biostimulants. RESULTS: We examined lettuce's physiological, biochemical, and agronomical responses to salt stress after applying biostimulants such as amino acids, arbuscular mycorrhizal fungi, plant growth-promoting rhizobacteria (PGPR), fulvic acid, and chitosan. The experiment was conducted in a greenhouse with a randomized complete block design, and each treatment was replicated four times. Biostimulant applications alleviated salt's detrimental effects on plant weight, height, leaf number, and leaf area. Yield increases under 50 mM NaCl were 75%, 51%, 31%, 34%, and 33% using vermicompost, PGPR, fulvic acid, amino acid, and chitosan, respectively. Biostimulants improved stomatal conductance (58-189%), chlorophyll content (4-10%), nutrient uptake (15-109%), and water status (9-107%). They also reduced MDA content by 26-42%. PGPR (1.0 ml L‒1), vermicompost (2 ml L‒1), and fulvic acid (40 mg L‒1) were particularly effective, enhancing growth, yield, phenol, and mineral content while reducing nitrate levels under saline conditions. CONCLUSIONS: Biostimulants activated antioxidative defense systems, offering a sustainable, cost-effective solution for mitigating salt stress in hydroponic lettuce cultivation.

LsFUL-LsSMU2 module positively controls bolting time in leaf lettuce (Lactuca sativa L.) under high temperature.

High temperature (HT) is an environmental factor that considerably affects plant physiology, development, crop yield, and economic value. HT can cause diseases and early bolting of leaf lettuce, thereby reducing the yield and quality of leaf lettuce. Herein, we used two leaf lettuce (Lactuca sativa L.) cultivars (bolting-resistant 'S24' and bolting-sensitive 'S39') to investigate the key factors and molecular mechanism impacting bolting. We found that 14 MADS-box genes implicated in bolting and flowering, LsMADS54 (also referred to as L. sativa FRUITFULL, LsFUL), was significantly up-regulated 1000 times after 5-d HT treatment and that HT-induced up-regulation of LsFUL was higher in bolting-sensitive than in resistant cultivars. The overexpression lines of LsFUL exhibited an earlier bolting time than that in the non-transformed 'S39'(CK). However, the RNA interference, and CRISPR-Cas9-mediated knockout lines of LsFUL exhibited a later bolting time than that in CK. In addition, we found that L. sativa SUPPRESSORS OF MEC-8 AND UNC-52 PROTEIN 2 (LsSMU2) and L. sativa CONSTANS-LIKE PROTEIN 5 (LsCOL5) interact with LsFUL, and these interactions could stimulate or prevent bolting. We observed that elevated temperature stimulated the abundance of LsSMU2 in the stem, which collaborated with LsFUL to accelerate bolting. Conversely, room temperature (RT) condition led to relatively more stable LsCOL5, which worked with LsFUL to postpone bolting. In summary, our findings demonstrate a molecular regulatory module of LsSMU2-LsFUL associated with HT-induced premature bolting, which serves as a reference for understanding HT-induced premature bolting phenomenon in leaf lettuce.

Low-Cost Imaging to Quantify Germination Rate and Seedling Vigor across Lettuce Cultivars.

The survival and growth of young plants hinge on various factors, such as seed quality and environmental conditions. Assessing seedling potential/vigor for a robust crop yield is crucial but often resource-intensive. This study explores cost-effective imaging techniques for rapid evaluation of seedling vigor, offering a practical solution to a common problem in agricultural research. In the first phase, nine lettuce (Lactuca sativa) cultivars were sown in trays and monitored using chlorophyll fluorescence imaging thrice weekly for two weeks. The second phase involved integrating embedded computers equipped with cameras for phenotyping. These systems captured and analyzed images four times daily, covering the entire growth cycle from seeding to harvest for four specific cultivars. All resulting data were promptly uploaded to the cloud, allowing for remote access and providing real-time information on plant performance. Results consistently showed the 'Muir' cultivar to have a larger canopy size and better germination, though 'Sparx' and 'Crispino' surpassed it in final dry weight. A non-linear model accurately predicted lettuce plant weight using seedling canopy size in the first study. The second study improved prediction accuracy with a sigmoidal growth curve from multiple harvests (R2 = 0.88, RMSE = 0.27, p < 0.001). Utilizing embedded computers in controlled environments offers efficient plant monitoring, provided there is a uniform canopy structure and minimal plant overlap.

The Use of Imaging to Quantify the Impact of Seed Aging on Lettuce Seed Germination and Seedling Vigor.

The decline in seed quality over time due to natural aging or mishandling requires assessing seed vigor for resilience in adverse conditions. Accelerated aging (AA) methods simulate seed deterioration by subjecting seeds to high temperatures and humidity. Saturated salt accelerated aging (SSAA) is an AA method adopted for small seeds like lettuce (Lactuca sativa). In this study, we subjected seeds of two lettuce cultivars ('Muir' and 'Bauer') to SSAA by sealing them in a box containing 40 g/100 mL of a sodium chloride (NaCl) solution in a dark growth chamber at 41 °C for 24, 48, and 72 h with a control. We monitored their vigor using embedded computer cameras, tracking the projected canopy size (PCS) daily from sowing to harvest. The cultivar 'Muir' exhibited consistent PCS values across the treatments, while 'Bauer' showed PCS variations, with notable declines after prolonged aging. The germination rates dropped significantly after 48 and 72 h of SSAA. A nonlinear regression model revealed a strong relationship between PCS and shoot dry weight across harvests and cultivars (R2 = 0.93, RMSE = 0.15, p < 0.001). The research found that the projected canopy size and shoot dry weight increased over time with significant differences in treatments for the cultivar 'Bauer' but not for 'Muir,' with the canopy size being a strong predictor of dry weight and no significant impact from the SSAA treatments. This study highlights cultivar-specific responses to aging and demonstrates the efficacy of our imaging tool in predicting lettuce dry weight despite treatment variations. Understanding how aging affects different lettuce varieties is crucial for seed management and crop sustainability.

Integrated transcriptomic and metabolomic analysis provides insights into cold tolerance in lettuce (Lactuca sativa L.).

The popular leafy vegetable lettuce (Lactuca sativa L.) is susceptible to cold stress during the growing season, which slows growth rate, causes leaf yellowing and necrosis, and reduced yield and quality. In this study, transcriptomic and metabolomic analyses of two cold-resistant lettuce cultivars (GWAS-W42 and F11) and two cold-sensitive lettuce cultivars (S13K079 and S15K058) were performed to identify the mechanisms involved in the cold response of lettuce. Overall, transcriptome analysis identified 605 differentially expressed genes (DEGs), including significant enrichment of genes involved in the flavonoid and flavonol (CHS, CHI, F3H, FLS, CYP75B1, HCT, etc.) biosynthetic pathways related to oxidation-reduction and catalytic activity. Untargeted metabolomic analysis identified fifteen flavonoid metabolites and 28 other metabolites potentially involved in the response to cold stress; genistein, quercitrin, quercetin derivatives, kaempferol derivatives, luteolin derivatives, apigenin and their derivatives accumulate at higher levels in cold-resistant cultivars. Moreover, MYBs, bHLHs, WRKYs and Dofs also play positive role in the low temperature response, which affected the expression of structural genes contributing to the variation of metabolites between the resistant and sensitive. These results provide valuable evidence that the metabolites and genes involved in the flavonoid biosynthetic pathway play important roles in the response of lettuce to cold stress.

Low frequency of the wild-type freezing-tolerance LsCBF7 allele among lettuce population suggests a negative selection during domestication and breeding.

KEY MESSAGE: Sustainable winter production in lettuce requires freezing tolerant varieties. This study identified a wild-type allele of LsCBF7 that could contribute to freezing tolerance improvement in lettuce. Lettuce is one of the most consumed vegetables globally. While ideally grown in 13-21 °C, its cultivation extends into winter in milder climates. However, occasional freezing temperatures can significantly reduce yields. Therefore, the development of freezing-tolerant lettuce varieties has become a long-term goal of lettuce breeding programs. Despite its significance, our understanding of freezing tolerance in lettuce remains limited. Plants have evolved a coping mechanism against freezing, known as cold acclimation, whereby they can increase freezing tolerance when pre-exposed to low nonfreezing temperatures. The CBF pathway is well-known for its central role in cold acclimation. Previously, we identified 14 CBF genes in lettuce and discovered that one of them, LsCBF7, had a loss-of-function mutation. In this study, we uncovered that accessions from colder regions carried the wild-type allele of LsCBF7 and this allele likely contributed to increased freezing tolerance, with 14% of the lettuce population carrying this allele. Interestingly, in wild lettuce (L. serriola) that is considered a progenitor of cultivated lettuce, this wild-type allele was much more common, with a frequency of 90%. This finding suggests that this wild-type allele may have undergone negative selection during the domestication or breeding of lettuce. Our data strongly indicate that this allele could be linked to early bolting, an undesirable trait in lettuce, which may have driven the negative selection. While this wild-type allele shows promise for improving freezing tolerance in lettuce, it is crucial to decouple it from the early bolting trait to fully harness its potential in lettuce breeding.

Sulfate supply decreases barium availability, uptake, and toxicity in lettuce plants grown in a tropical Ba-contaminated soil.

Barium (Ba) is a non-essential element that can cause toxicity in living organisms and environmental contamination. Plants absorb barium predominantly in its divalent cationic form Ba2+. Sulfur (S) can decrease the availability of Ba2+ in the soil by causing its precipitation as barium sulfate, a compound known for its very low solubility. The objective of this study was to evaluate the effect of soil sulfate supply in soil Ba fractions, as well as on plant growth, and Ba and S uptake by lettuce plants grown in artificially Ba-contaminated soil under greenhouse conditions. The treatments consisted of five Ba doses (0, 150, 300, 450, and 600 mg kg-1 Ba, as barium chloride) combined with three S doses (0, 40, and 80 mg kg-1 S, as potassium sulfate). The treatments were applied to soil samples (2.5 kg) and placed in plastic pots for plant cultivation. The Ba fractions analyzed were extractable-Ba, organic matter-Ba, oxides associated-Ba, and residual-Ba. The results indicate that the extractable-Ba fraction was the main one responsible for Ba bioavailability and phytotoxicity, probably corresponding to the exchangeable Ba in the soil. The dose of 80 mg kg-1 of S reduced extractable-Ba by 30% at higher Ba doses while it increased the other fractions. Furthermore, S supply attenuated the growth inhibition in plants under Ba exposure. Thus, S supply protected the lettuce plants from Ba toxicity by reduction of Ba availability in soil and plant growth enhancement. The results suggest that sulfate supply is a suitable strategy for managing Ba-contaminated areas.

Design and test of a laser lighting device for plant growth.

Aiming at the problems of high energy consumption, complex wiring, high layout cost, limited use environment, and limited function of conventional plant lighting equipment such as fluorescent lamps, sodium lamps, etc., this paper develops a type of laser device for plant growth with nanometer lasers based on the design of an intelligent control system of an immune algorithm, constant current driving circuit of the laser, pulse power supply, and rotatable intelligent platform to make the device more stable, reliable, practical, and energy efficient, and provides a useful reference for the innovation and application of materials, processes, and methods of plant lighting. The effects of nanometer laser light supplementation on the growth of purple lettuce, romaine lettuce, Chinese cabbage, and you-mai vegetable have been studied with the vegetables mentioned above as experimental materials and with natural light as the control sample. The results show that the nanometer laser device significantly increases stem height, stem thickness, leaf area, leaf number, and chlorophyll content, effectively promotes plant growth, and achieves efficient cultivation. In the future, studies of the effects of laser treatment on plant physiology and biochemistry will be sped up to explore the molecular biological mechanism of lasers to promote application and technological innovation of lasers in lighting for plant growth and the laser device in productivity.

Chronic toxicity, bioavailability and bioaccumulation of Zn, Cu and Pb in Lactuca sativa exposed to waste from an abandoned gold mine.

Abandoned mines with untreated waste cause environmental pollution. The complex mixture of mining waste includes high metal content, anthropogenic chemicals and sterile rocks. Adverse effects of contaminated soils have been widely assessed by the use of plants. The aim of this study was to assess the chronic toxicity of a contaminated soil by waste from an abandoned gold mine on Lactuca sativa and its relationship with the bioavailability and bioaccumulation of Zn, Cu and Pb. Soil samples were taken from the site of mining waste stacking and a reference site in La Planta (Argentina). Contamination indices were calculated and acute and chronic exposures on L. sativa were carried out. Phytotoxicity indices, morphological and biochemical parameters, and concentrations of Zn, Cu and Pb in pseudo total and bioavailable soil fractions and in plant tissue were determined. Concentration- and time-dependent toxicity effects were observed, especially on plant width, fresh aerial biomass, leaf area and percentage of plants with completely necrotic aerial biomass. High levels of Zn (1453.3 ± 220.3 µg g-1) were found in plant tissue compared to Pb (277.2 ± 18.0 µg g-1) and Cu (255.3 ± 25.6 µg g-1). Toxicological endpoints correlated with metal uptake and mining waste concentration. In addition, bioaccumulation factors correlated with mobilisable and water soluble fractions. The concentration of Pb in aerial biomass surpassed the permissible concentrations in leaf vegetables, even at the reference site, indicating that lettuce crop consumption could be risky for the local population's health. This study demonstrated a strong relationship between metal bioavailability, toxicity endpoints and bioaccumulation, contributing with novel information to future ecotoxicological risk assessments and remediation plans.

Similarity assessment of metallic nanoparticles within a risk assessment framework: A case study on metallic nanoparticles and lettuce.

Similarity assessment is one of the means of optimally using scarcely available experimental data on the fate and hazards of nanoforms (NFs) for regulatory purposes. For a set of NFs that are shown to be similar it is allowed in a regulatory context to apply the information available on any of the NFs within the group to the whole set of NFs. Obviously, a proper justification for such a similarity assessment is to be provided. Within the context of exemplifying such a justification, a case study was performed aimed at assessing the similarity of a set of spherical metallic NFs that different with regard to chemical composition (three metals) and particle size (three different sizes). The endpoints of assessment were root elongation and biomass increase of lettuce (Lactuca sativa L.) seedlings and exposure assessment was performed in order to express the actual exposure concentration in terms of time-weighted average particle concentrations. The results of the study show that for the specific endpoints assessed, chemical composition is driving NF toxicity and this is mostly due to impacts on the fate of the NFs. On the other hand, particle size of Cu NFs had a negligible impact on the dose-response relationships for the specific endpoints assessed. It is thus concluded that hazard data available on spherical Cu NF tested in our case can be used to inform on the hazards of any spherical Cu NF within the size range of 25-100 nm, but only applies for the certain endpoints. Also, toxicity data for the Cu2+-ion are suited for such a similarity assessment.

A Physiological behavior and tolerance of Lactuca sativa to lead nitrate and silver nitrate heavy metals / Comportamento fisiológico e tolerância de Lactuca sativa aos metais pesados nitrato de chumbo e nitrato de prata

This study goal to evaluate the effects of different concentrations of lead (Pb) and silver (Ag) on germination, initial growth and anatomical alterations of Lactuca sativa L. Plants use various mechanisms to reduce the impacts caused by anthropic action, such as xenobiotic elements of soils and water contaminated by heavy metals. These metals were supplied as lead nitrate and silver nitrate and the following treatments were established: control for both metals, maximum dose of heavy metals, for arable soils, allowed by the National Council of the Environment (Ag = 25 mg. Kg-¹, Pb = 180 mg. Kg-¹), double (Ag = 50 mg. Kg-¹, Pb = 360 mg. Kg-¹) and triple (Ag = 75 mg. Kg-¹, Pb = 540 mg. Kg-¹) of this dosage. Vigor and germination tests of the seeds and possible anatomical changes in the leaves and roots of lettuce plants were performed. The species showed a high capacity to germinate under Pb and Ag stress, and the germination was never completely inhibited; however, the germination decreased with increasing Pb concentrations, but not under Ag stress. The use of increasing doses of metals reduced seed vigor and increased chlorophyll content. An increase in biomass was also observed in plants from treatments submitted to Pb. The phytotoxic effects of metals were more pronounced at 15 days after sowing. Anatomically, L. sativa was influenced by metal concentrations, and had a reduction of up to 79.9% in root epidermis thickness at the highest Pb concentration, although some structures did not suffer significant changes. The results suggest that L. sativa presents tolerance to high concentrations of heavy metals, showing possible mechanisms to overcome the stress caused by these metals. In this research lettuce possibly used the mechanism of exclusion of metals retaining Pb and Ag in the roots preserving the photosynthetic apparatus in the aerial part of the plants. In general, the chemical element Pb was more toxic than Ag, in these experimental conditions.

Este estudo teve como objetivo avaliar os efeitos de diferentes concentrações de chumbo (Pb) e prata (Ag) na germinação, crescimento inicial e alterações anatômicas de Lactuca sativa L. As plantas utilizam vários mecanismos para reduzir os impactos causados pela ação antrópica, como elementos xenobióticos de solos e água contaminada por metais pesados. Esses metais foram fornecidos como nitrato de chumbo e nitrato de prata e foram estabelecidos os seguintes tratamentos:controle para ambos os metais, dose máxima de metais pesados, para solos cultiváveis, permitida pelo Conselho Nacional do Meio Ambiente (Ag = 25mg.Kg-¹, Pb = 180mg.Kg-¹), dobro (Ag = 50mg.Kg-¹, Pb = 360mg.Kg-¹) e triplo (Ag = 75mg.Kg-¹, Pb = 540 mg.Kg-¹) desta dosagem. Foram realizados testes de vigor e germinação das sementes e possíveis alterações anatômicas nas folhas e raízes das plantas de alface. A espécie apresentou alta capacidade de germinar sob estresse de Ag e Pb, e a germinação nunca foi completamente inibida; entretanto, a germinação diminuiu com o aumento das concentrações de Pb, mas não sob estresse de Ag. O uso de doses crescentes dos metais, reduziu o vigor das sementes e aumentou o teor de clorofila. Também foi observado aumento da biomassa nas plantas a partir dos tratamentos submetidos ao Pb. Os efeitos fitotóxicos dos metais foram mais acentuados aos 15 dias após a semeadura. Anatomicamente, L. sativa foi influenciada pelas concentrações de metais, e teve uma redução de até 79,9% na espessura da epiderme radicular na maior concentração de Pb, embora algumas estruturas não tenham sofrido alterações significativas. Os resultados sugerem que L. sativa apresenta tolerância a altas concentrações de metais pesados, mostrando possíveis mecanismos para superar o estresse causado por esses metais. Nesta pesquisa a alface possivelmente utilizou o mecanismo de exclusão de metais retendo Pb e Ag nas raízes preservando o aparato [...].

A digital sensor to measure real-time leaf movements and detect abiotic stress in plants.

Plant and plant organ movements are the result of a complex integration of endogenous growth and developmental responses, partially controlled by the circadian clock, and external environmental cues. Monitoring of plant motion is typically done by image-based phenotyping techniques with the aid of computer vision algorithms. Here we present a method to measure leaf movements using a digital inertial measurement unit (IMU) sensor. The lightweight sensor is easily attachable to a leaf or plant organ and records angular traits in real-time for two dimensions (pitch and roll) with high resolution (measured sensor oscillations of 0.36 ± 0.53° for pitch and 0.50 ± 0.65° for roll). We were able to record simple movements such as petiole bending, as well as complex lamina motions, in several crops, ranging from tomato to banana. We also assessed growth responses in terms of lettuce rosette expansion and maize seedling stem movements. The IMU sensors are capable of detecting small changes of nutations (i.e. bending movements) in leaves of different ages and in different plant species. In addition, the sensor system can also monitor stress-induced leaf movements. We observed that unfavorable environmental conditions evoke certain leaf movements, such as drastic epinastic responses, as well as subtle fading of the amplitude of nutations. In summary, the presented digital sensor system enables continuous detection of a variety of leaf motions with high precision, and is a low-cost tool in the field of plant phenotyping, with potential applications in early stress detection.

Quantitative Trait Loci and Candidate Genes Associated with Photoperiod Sensitivity in Lettuce (Lactuca spp.).

KEY MESSAGE: A population of lettuce that segregated for photoperiod sensitivity was planted under long-day and short-day conditions. Genetic mapping revealed two distinct sets of QTLs controlling daylength-independent and photoperiod-sensitive flowering time. The molecular mechanism of flowering time regulation in lettuce is of interest to both geneticists and breeders because of the extensive impact of this trait on agricultural production. Lettuce is a facultative long-day plant which changes in flowering time in response to photoperiod. Variations exist in both flowering time and the degree of photoperiod sensitivity among accessions of wild (Lactuca serriola) and cultivated (L. sativa) lettuce. An F6 population of 236 recombinant inbred lines (RILs) was previously developed from a cross between a late-flowering, photoperiod-sensitive L. serriola accession and an early-flowering, photoperiod-insensitive L. sativa accession. This population was planted under long-day (LD) and short-day (SD) conditions in a total of four field and screenhouse trials; the developmental phenotype was scored weekly in each trial. Using genotyping-by-sequencing (GBS) data of the RILs, quantitative trait loci (QTL) mapping revealed five flowering time QTLs that together explained more than 20% of the variation in flowering time under LD conditions. Using two independent statistical models to extract the photoperiod sensitivity phenotype from the LD and SD flowering time data, we identified an additional five QTLs that together explained more than 30% of the variation in photoperiod sensitivity in the population. Orthology and sequence analysis of genes within the nine QTLs revealed potential functional equivalents in the lettuce genome to the key regulators of flowering time and photoperiodism, FD and CONSTANS, respectively, in Arabidopsis.

The genetic basis of water-use efficiency and yield in lettuce.

BACKGROUND: Water supply limits agricultural productivity of many crops including lettuce. Identifying cultivars within crop species that can maintain productivity with reduced water supply is a significant challenge, but central to developing resilient crops for future water-limited climates. We investigated traits known to be related to water-use efficiency (WUE) and yield in lettuce, a globally important leafy salad crop, in a recombinant inbred line (RIL) lettuce mapping population, produced from a cross between the cultivated Lactuca sativa L. cv. Salinas and its wild progenitor L. serriola L. RESULTS: Wild and cultivated lettuce differed in their WUE and we observed transgressive segregation in yield and water-use traits in the RILs. Quantitative trait loci (QTL) analysis identified genomic regions controlling these traits under well-watered and droughted conditions. QTL were detected for carbon isotope discrimination, transpiration, stomatal conductance, leaf temperature and yield, controlling 4-23 % of the phenotypic variation. A QTL hotspot was identified on chromosome 8 that controlled carbon isotope discrimination, stomatal conductance and yield under drought. Several promising candidate genes in this region were associated with WUE, including aquaporins, late embryogenesis abundant proteins, an abscisic acid-responsive element binding protein and glutathione S-transferases involved in redox homeostasis following drought stress were also identified. CONCLUSIONS: For the first time, we have characterised the genetic basis of WUE of lettuce, a commercially important and water demanding crop. We have identified promising candidate genomic regions determining WUE and yield under well-watered and water-limiting conditions, providing important pre-breeding data for future lettuce selection and breeding where water productivity will be a key target.

Lighting Direction Affects Leaf Morphology, Stomatal Characteristics, and Physiology of Head Lettuce (Lactuca sativa L.).

Plants are exposed to numerous biotic and abiotic stresses, and light is one of the most important factors that influences the plant morphology. This study was carried out to examine how the lighting direction affected the plant morphology by investigating the growth parameters, epidermal cell elongation, stomatal properties, and physiological changes. Seedlings of two head lettuce (Lactuca sativa L.) cultivars, Caesar Green and Polla, were subjected to a 12 h photoperiod with a 300 µmol·m-2·s-1 photosynthetic photon flux density (PPFD) provided by light emitting diodes (LEDs) from three directions: the top, side, and bottom, relative to the plants. Compared with the top and side lighting, the bottom lighting increased the leaf angle and canopy by stimulating the epidermal cell elongation in leaf midrib, reduced the leaf number and root biomass, and induced large stomata with a low density, which is associated with reduced stomatal conductance and carbohydrate contents. However, the proline content and quantum yield exhibited no significant differences with the different lighting directions in both cultivars, which implies that the plants were under normal physiological conditions. In a conclusion, the lighting direction had a profound effect on the morphological characteristics of lettuce, where the plants adapted to the changing lighting environments.

Toxicogenetic of tebuconazole based fungicide through Lactuca sativa bioassays.

The rampant use of pesticides can cause serious environmental problems. They can be contaminating surface water and groundwater, affecting the surrounding micro and macro biota. In this sense, this work aimed to evaluate the effects of a tebuconazole-based fungicide through endpoints accessed in Lactuca sativa bioassays. Germinated-seeds with roots upon 2 mm were treated with a fungicide containing Tebuconazole (TBZ) as active compound. The final concentration of TBZ in the tested solutions were 0.025 (C1); 0.05 (C2); 0.1 (C3); 0.2 (C4) and 0.4 g/L (C5). L. sativa roots were exposed for 24 h to these solutions and Petri dishes containing the treated seeds were kept in incubation chamber at 24 °C. Two positive controls (PC,) the herbicide trifluralin (0.84 mg/L) and Methanesulfonate (4 ×10-4 mol/L), were applied. Distilled water was negative control (NC). The following endpoints were analyzed: root growth (RG), cytogenotoxic potential by cell cycle analysis, induction of DNA damage through TUNEL and comet assays. The obtained data were submitted to one-way variance analysis (ANOVA) and then to Tukey or Kruskal Wallis (P < 0.05) tests. The concentrations (C1, C2, C4 and C5) affected negatively the RG of L. sativa, in comparison with the NC. The mitotic index was reduced by 25% from NC to C1 and in the rest of treatments it did not present significant modifications. However, from C3 to C5 great amount of chromosome alterations were observed, in comparison with the NC. TBZ-based fungicide also induced DNA fragmentation as measured by TUNEL and comet assays. Thus, TBZ-based fungicide in some concentrations can have phytotoxic, cytotoxic and genotoxic effects in roots and meristematic cells of L. sativa.

Ozone uptake at night is more damaging to plants than equivalent day-time flux.

MAIN CONCLUSION: Plants exposed to equivalent ozone fluxes administered during day-time versus night-time exhibited greater losses in biomass at night and this finding is attributed to night-time depletion of cell wall-localised ascorbate. The present study employed Lactuca sativa and its closest wild relative, L. serriola, to explore the relative sensitivity of plants to ozone-induced oxidative stress during day-time versus night-time. By controlling atmospheric ozone concentration and measuring stomatal conductance, equivalent ozone uptake into leaves was engineered during day and night, and consequences on productivity and net CO2 assimilation rate were determined. Biomass losses attributable to ozone were significantly greater when an equivalent dose of ozone was taken-up by foliage at night compared to the day. Linkages between ozone impacts and ascorbic acid (AA) content, redox status and cellular compartmentation were probed in both species. Leaf AA pools were depleted by exposure of plants to darkness, and then AA levels in the apoplast and symplast were monitored on subsequent transfer of plants to the light. Apoplast AA appeared to be more affected by light-dark transition than the symplast pool. Moreover, equivalent ozone fluxes administered to leaves with contrasting AA levels resulted in contrasting effects on the light-saturated rate of CO2 assimilation (Asat) in both species. Once apoplast AA content recovered to pre-treatment levels, the same ozone flux resulted in no impacts on Asat. The results of the present investigation reveal that plants are significantly more sensitive to equivalent ozone fluxes taken-up at night compared with those during the day and were consistent with diel shifts in apoplast AA content and/or redox status. Furthermore, findings suggest that some thought should be given to weighing regional models of ozone impacts for extraordinary night-time ozone impacts.

Aquaporins and cation transporters are differentially regulated by two arbuscular mycorrhizal fungi strains in lettuce cultivars growing under salinity conditions.

The aim was to identify the effects of AM symbiosis on the expression patterns of genes associated with K+ and Na+ compartmentalization and translocation and on K+/Na+ homeostasis in some lettuce (Lactuca sativa) cultivars as well as the effects of the relative abundance of plant AQPs on plant water status. Two AM fungi species (Funneliformis mosseae and Claroideoglomus lamellosum) isolated from the hyper-arid Atacama Desert (northern Chile) were inoculated to two lettuce cultivars (Grand Rapids and Lollo Bionda), and watered with 0 and 60 mM NaCl. At 60 days of plant growth, the AM symbiotic development, biomass production, nutrient content (Pi, Na+, K+), physiological parameters, gene expressions of ion channels and transporters (NHX and HKT1), and aquaporins proteins abundance (phosphorylated and non-phosphorylated) were evaluated. Salinity increased the AM root colonization by both inocula. AM lettuce plants showed an improved growth, increased relative water content and improved of K/Na ratio in root. In Grand Rapids cultivar, the high efficiency of photosystem II was higher than Lollo Bionda cultivar; on the contrary, stomatal conductance was higher in Lollo Bionda. Nevertheless, both parameters were increased by AM colonization. In the same way, LsaHKT1;1, LsaHKT1;6, LsaNHX2, LsaNHX4, LsaNHX6 and LsaNHX8 genes and aquaporins PIP2 were up-regulated differentially by both AM fungi. The improved plant growth was closely related to a higher water status due to increased PIP2 abundance, as well as to the upregulation of LsaNHX gene expression, which concomitantly improved plant nutrition and K+/Na+ homeostasis maintenance.

Physiological and Oxidative Stress Responses of Lettuce to Cleomside A: A Thiohydroximate, as a New Allelochemical from Cleome arabica L.

The inclination toward natural products have led the onset for the discovery of new bioactive metabolites that could be targeted for specific therapeutic or agronomic applications. This study aimed to isolate bioactive compounds from Cleome arabica L., and subsequently determine the unexplored mechanism of action of the newly identified compounds on Lactuca sativa L. Chemical investigation of the ethyl acetate fraction of methanolic silique extract of C. arabica afforded seven secondary metabolites belonging to different classes such as flavonoids, triterpene, and a new thiohydroximate derivative, named cleomside A. Among phytotoxic assays, the growth of lettuce was totally inhibited by cleomside A compared to the other identified compounds. This effect was associated with the increased levels of electrolyte leakage, malondialdehyde, and hydrogen peroxide indicating disruption of membrane integrity and induction of oxidative stress. Activities of the antioxidant enzymes SOD, CAT, and APX were also elevated, thereby demonstrating the enhanced generation of reactive oxygen species upon identified allelochemical exposure. Thus, the changes caused by cleomside A described herein can contribute to better understanding the allelochemical actions of thiohydroximate and the potential use of these substances in the production of natural herbicides compared to the other identified flavonoids and triterpene.