Bioactivity of hydroalcoholic extracts from tropaeolum majus L. (tropaeolaceae) on the germination, initial plant development and cell cycle of Lactuca sativa L.

Natural products are usually considered harmless; however, these substances need to be consumed with caution. Biological assays with plant models are a suitable alternative for prospective studies to assess natural product-initiated toxicity. The aim of this study was to examine the toxic potential of leaf and flower extracts derived from Tropaeolum majus L. a widely used plant in traditional medicine. Seeds of Lactuca sativa L. were exposed to T. majus extracts and based upon the seedling growth curve values, the 50% Inhibition Concentration (IC50) was calculated and applied for cell cycle analysis exposure. Both extracts contained organic acids, proteins, amino acids, and terpene steroids. Sesquiterpene lactones and depside were detected in leaf extracts. The higher concentration tested exhibited a marked phytotoxic effect. The extracts induced clastogenic, aneugenic cytotoxic, and potential mutagenic effects. The possible relationships between the classes of compounds found in the extracts and effects on cells and DNA were determined.

Effects of a S-metolachlor based herbicide on two plant models: Zea mays L. and Lactuca sativa L.

Corn is the second most cultivated crop in Brazil, the number-one country in pesticide consumption. Chemical control of weeds is performed using herbicides such as S-metolachlor with pre- and post-emergence action and thus the toxicity of herbicides constitutes a matter of great concern. The present investigation aimed to examine the effects of an S-metolachlor-based herbicide on Lactuca sativa L. (lettuce) and Zea mays L. (maize) utilizing various bioassays. The test solutions were prepared from commercial products containing the active ingredient. Seeds from the plant models were exposed in petri dishes and maintained under biochemical oxygen demand (BOD) at 24°C. Distilled water was negative and aluminium positive control. Macroscopic analyses (germination and growth) were conducted for both plant species, and microscopic analysis (cell cycle and chromosomal alterations) were performed for L. sativa root tip cells. Detrimental interference of S-metolachlor-based herbicide was noted with lettuce for all parameters tested reducing plant germination by over 50% and the germination speed by over 45% and showing a significant decrease in mitotic index, from 16.25% to 9,28% even on the lowest concentration tested. In maize, there was no significant interference in plant germination; however, speed of germination was significantly hampered, reaching a 51.22% reduction for the highest concentration tested. Data demonstrated that the herbicide was toxic as evidenced by its phyto- and cytotoxicity in L. sativa L. and Z. mays L.

Comparison of Anticancer, Antioxidant, Enzyme Inhibitory Effects and Phytochemical Contents between Edible Lettuce (Lactuca sativa) and A New Wild Species (Lactuca anatolica).

In this study, the bioactive components, enzyme inhibitory, antioxidant and anticancer potentials of edible (L. sativa) and a new species (L. anatolica) of Lactuca were evaluated and compared. The quantitative analyzes of the bioactive components of L. sativa (LS) and L. anatolica (LA) were analyzed quantitatively by GC-MS and Orbitrab HPLC-HRMS. Antioxidant, enzyme inhibitory and anticancer properties were analyzed by various assays. In general, LA exhibited more stronger antioxidant properties compared to LS. The extracts showed similar inhibitory effects on these enzymes. It was determined that LS was dominant in terms of linoleic acid (23.71%), while LA contained a high level of α-linolenic acid (31.70%). LA and LS inhibited the viability of A549 and MCF-7 cells in a dose-dependent manner. IC50 values for LA, LS and cisplatin were determined as 120.3, 197.5, 4.3 µg/mL in A549 cell line and 286.2, 472.8, 7.2 µg/mL in MCF-7 cell line, respectively. It was revealed that LA and LS treatment at 50 µg/mL concentrations in A549 cells completely suppressed the colony forming capacity, and treatment with IC50 doses inhibited cell migration, and triggered apoptosis by regulating caspase-3, cPARP, p53 and p21. The findings of this study suggested that these species have significant pharmacological potential.

First report of Fusarium oxysporum f. sp. lactucae causing wilt on greenhouse-grown lettuce in North Carolina.

In August 2023, butterhead lettuce (Lactuca sativa L.) presented wilting, chlorosis, and about 2 cm of reddish-tan internal discoloration in the crown from a commercial greenhouse in Orange County, North Carolina. Plant collapse beginning with the outer leaves near the soil surface was observed with 25% disease incidence. Symptomatic lettuce plants were submitted to North Carolina State University's Plant Disease and Insect Clinic. Vascular tissue from symptomatic crowns were cut into pieces, and surface-sterilized in 10% NaOCl for two minutes. The tissue was rinsed in sterile deionized water three times, blotted dry, and placed on acidified potato dextrose agar (APDA). Three isolates, each from a different symptomatic plant, were transferred to APDA and Spezieller Nährstoffarmer agar (SNA) with pieces of sterile filter paper on the surface of the SNA media and incubated for 14 days at 23°C in the dark. Each isolate produced micro and macroconidia consistent with the morphological description of Fusarium oxysporum Schlechtendahl emend. Snyder & Hansen (Leslie and Summerell 2006). DNA was extracted from 15-day-old fungal colonies grown on APDA media using the DNeasy Plant Mini Kit (Qiagen, Germantown, MD, U.S.A.). The intergenic spacer locus was amplified using two primer pairs, iNL11/CNSa and iCNS1/NLa, and sequences were aligned together to form a single contig (O'Donnell et al., 2009). Primers EF1/EF2 were used to amplify the elongation factor 1-alpha region (O'Donnell et al., 1998). Each isolate was deposited into GenBank with accession numbers PP216479, PP216480, PP216481, PP235836, PP235837, and PP235838. Individual isolates revealed a 100% query cover and identity match with sequences of F. oxysporum f. sp. lactucae (FOL) and >99% identity with F. oxysporum CBS 144134 type material accessions in GenBank using BLASTn. A comparison with previously described lettuce isolates showed a homologous match with FOL race 1 isolates from California (MH412701), Arizona (DQ837658), and Greece (OQ466116), and race 4 isolates from Italy (MK801787) and Spain (OP903519). Each isolate was verified as FOL using specific primers FLA0001F/FLA0001R for FOL based on sequence tagged site markers designed by Shimazu et al. (2005). To confirm Koch's postulates, fifteen 21-day-old lettuce cv. Red Tide plants were inoculated with FOL (isolate FOLNC_660). During transplanting, lettuce roots were submerged in a 1 × 105 conidia/mL suspension for five minutes, following an inoculation protocol from Schmale and Gordon (2003). The lettuce plants were placed separately in 8.9 × 8.3 cm pots containing potting soil and maintained in a greenhouse with 31°C daytime and 25°C nighttime temperature, relative humidity of 60%, and 12-hour photoperiod. After 15 days, 80% and 86.7% of infected plants exhibited wilting, chlorosis, and vascular discoloration. The fifteen control plants remained symptomless for both experimental runs. FOL was recovered from the vascular tissue of all symptomatic plants. To our knowledge, this constitutes the first report of FOL infecting lettuce in North Carolina. Fusarium wilt of lettuce has been reported in California (Hubbard and Gerik 1993), Arizona (Matheron and Koike 2003), and most recently in Florida (Murray et al., 2020). The presence of FOL in North Carolina may result in significant crop loss for commercial growers. One of the most effective management strategies is to plant lettuce cultivars that are resistant against FOL.

Distribution of Three Verticillium dahliae Races in Coastal California and Evaluation of Resistance in Lettuce.

Verticillium wilt, caused by Verticillium dahliae, is one of the most devastating soilborne diseases of lettuce (Lactuca sativa L.). There are three races of V. dahliae, and each race has been characterized by markers representing race-specific effectors. Race 1 is differentiated by the presence of the functional secretory Ave1 effector. Similarly, races 2 and 3 are differentiated by effectors VdR2e and VdR3e, respectively. Although the presence of race 1 in coastal California was well established, the presence of effector-based races 2 and 3 was uncertain. This study therefore focused on characterizing 727 isolates collected from 142 ranches of symptomatic lettuce and other crops from coastal California. Based on this evaluation, 523 isolates were designated as race 1, 20 isolates as race 2, 23 isolates as race 3, and 17 as race undefined. Isolates representing other Verticillium species totaled 110, and 34 were non-Verticillium fungal species. Because the use of resistant cultivars is a key strategy to manage this disease, we evaluated 48 lettuce germplasm lines and 1 endive (Cichorium endivia L.) line, comprising commercial cultivars and breeding lines, including the race 1-resistant heirloom cultivar La Brillante and the susceptible cultivar Salinas as controls. Resistance against races 1, 2, and 3 along with VdLs17, a virulent isolate of V. dahliae from lettuce that is currently not assigned to a race, was evaluated in replicated greenhouse experiments. Two crisphead lettuce lines, HL28 and HL29, exhibited resistance against race 1 and a partial resistance against race 2, whereas all other lines were highly susceptible to races 1 and 2 and VdLs17. The majority of lines exhibited higher resistance to race 3 relative to the other two races. This study documents the current distribution of the different races in coastal California. In addition, the sources of resistance currently being developed should be effective or partially effective against these races for targeted deployment as soon as they are available.

First Report of Collar and Root Rot of Lettuce Caused by Plectosphaerella cucumerina in Serbia.

In March 2021, unusual plant stuning, collar, and wet root rot of lettuce (Lactuca sativa L.) during the rosette stage was observed in two commercial fields in Serbia (44°58'N, 20°32'E; 44°45'N, 20°43'E). Disease incidence in the fields (≈ 0.9 ha each) was approximately 15 and 20%, respectively. Initial above-ground symptoms were yellowing and wilting of leaves, while below-ground symptoms were collar, wet root rot, and lesions becoming necrotic. Eventually, whole plants wilted, collapsed, and died. A total of 35 symptomatic plants were collected from the fields, and diseased tissues were cut into small pieces, surface sterilized, and plated on potato dextrose agar (PDA). Isolation resulted in 20 morphologically uniform monoconidial isolates. The isolates formed white to creamy colonies, gradually becoming salmon pink, slimy, or moist in appearance, with sparse aerial mycelia. Numerous hyphal coils with conidiophores and hyaline, smooth-surfaced, ellipsoid to ovoid, septate or aseptate conidia were formed (4.5 to 10.1×1.2 to 3.7 µm (n = 100)). To confirm the species identity, the internal transcribed spacer (ITS) region and the D1/D2 region of a selected representative isolate 13-3-c were amplified and sequenced by using primer pairs ITS1/ITS4 (White et al. 1990) and N1/N2 (O'Donnell and Gray 1995), respectively. The sequences were deposited in GenBank (ITS: OR880564 and D1/D2: OR880567). Sequence analysis revealed 100% nucleotide identity with P. cucumerina isolates from different countries deposited in the NCBI GenBank, including isolate MH860704 (Vu et al. 2019) (ITS region) and isolate KY662256 (Su et al. 2017) (D1/D2 region). Neighbor-joining analysis was conducted based on the combined ITS and D1/D2 regions, and the tree was constructed with the substitution models (1,000 bootstrap). The combined phylogeny confirmed that the sequences shared a common clade with P. cucumerina. Hence, morphological, microscopic, and molecular characterization confirmed the pathogen as P. cucumerina (Palm et al., 1995; Carlucci et al., 2012). In a pathogenicity assay, 10 isolates were tested. Five 30-day-old lettuce plants (cv. Majska Kraljica) per isolate were root-dipped in the conidial suspensions (1×105 conidia/ml). The 10 inoculated plants were transplanted into 1 L pots containing sterile substrate (Floragard, Germany). Plants treated with sterile distilled water were used as controls. Plants were maintained in a greenhouse at 25 to 28°C under a 12-hour photoperiod (Cai et al., 2021). Four weeks after inoculation, stunting, chlorosis, and wilting of plants were observed, while collars and roots exhibited typical decaying symptoms. No symptoms were observed on the control plants. The pathogen was reisolated from symptomatic tissue as previously described. Koch's postulates were completed by confirming the identity of reisolates based on morphological features. To our knowledge, this is the first report of P. cucumerina on lettuce or any other crop in Serbia. P. cucumerina is already known as a pathogen of lettuce and other hosts grown in many countries worldwide, as well as in some European countries (Belgium, England, Italy, the Netherlands, and Switzerland) (Zhang et al. 2019). This emerging pathogen may cause significant economic losses in lettuce production in Serbia and in the entire Balkan region. Our results may help to develop effective management strategies based on accurate and timely identification and regular pathogen monitoring.

Effective, consistent, and rapid non-contact application methods for seedling basal stem infection by Sclerotinia sclerotiorum.

Sclerotinia sclerotiorum (Lib.) de Bary, an economically devastating soil-borne fungal pathogen known to cause disease across a wide range of plants, produces long-term inoculum called sclerotia that can either germinate carpogenically by ascospores infecting above-ground plant parts or myceliogenically to infect stem base and roots. Typically for research purposes S. sclerotiorum diseases are initiated by direct contact methods, using S. sclerotiorum mycelium agar plugs wrapped around the stem or sclerotia placed directly beneath root mass. However, reproducible non-contact methods leading to basal stem infection are not currently available. Therefore, the objective of this study was to develop effective non-contact protocols that consistently generate basal plant stem infection from S. sclerotiorum in the soil. Using three host plant species (canola, lupin, and lettuce) we determined two methods that reliably produced basal stem infection. The first method, where mycelial agar plugs were positioned just below the soil surface at a distance of 5 mm from each seedling, led to 100% infection in all plants. The second method used pathogen-infested soil by mixing the soil with dry inoculum in the form of a powder prepared from mycelium-colonized organic substrates. Four substrates consistently produced 100% seedling infection at four days after inoculation (DAI); wheat bran, wheat grain, red rice, and hulled millet. In contrast, chia, canary, sesame, and ryegrass seed substrates resulted in less than 50% seedling infection at 10 DAI and infection levels did not progress further. The two soil inoculation methods outlined in this study will enhance future research on the progression of S. sclerotiorum diseases, with the potential to screen disease-resistant host genotypes to basal S. sclerotiorum infection, and in particular to test the effectiveness of soil applications of fungicides or biocontrol agents against S. sclerotiorum basal infection.

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.

Automated Imaging to Evaluate the Exogenous Gibberellin (Ga3) Impact on Seedlings from Salt-Stressed Lettuce Seeds.

Salinity stress is a common challenge in plant growth, impacting seed quality, germination, and general plant health. Sodium chloride (NaCl) ions disrupt membranes, causing ion leakage and reducing seed viability. Gibberellic acid (GA3) treatments have been found to promote germination and mitigate salinity stress on germination and plant growth. 'Bauer' and 'Muir' lettuce (Lactuca sativa) seeds were soaked in distilled water (control), 100 mM NaCl, 100 mM NaCl + 50 mg/L GA3, and 100 mM NaCl + 150 mg/L GA3 in Petri dishes and kept in a dark growth chamber at 25 °C for 24 h. After germination, seedlings were monitored using embedded cameras, capturing red, green, and blue (RGB) images from seeding to final harvest. Despite consistent germination rates, 'Bauer' seeds treated with NaCl showed reduced germination. Surprisingly, the 'Muir' cultivar's final dry weight differed across treatments, with the NaCl and high GA3 concentration combination yielding the poorest results (p < 0.05). This study highlights the efficacy of GA3 applications in improving germination rates. However, at elevated concentrations, it induced excessive hypocotyl elongation and pale seedlings, posing challenges for two-dimensional imaging. Nonetheless, a sigmoidal regression model using projected canopy size accurately predicted dry weight across growth stages and cultivars, emphasizing its reliability despite treatment variations (R2 = 0.96, RMSE = 0.11, p < 0.001).

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.

Application of Chlorophyll Fluorescence Analysis Technique in Studying the Response of Lettuce (Lactuca sativa L.) to Cadmium Stress.

To reveal the impact of cadmium stress on the physiological mechanism of lettuce, simultaneous determination and correlation analyses of chlorophyll content and photosynthetic function were conducted using lettuce seedlings as the research subject. The changes in relative chlorophyll content, rapid chlorophyll fluorescence induction kinetics curve, and related chlorophyll fluorescence parameters of lettuce seedling leaves under cadmium stress were detected and analyzed. Furthermore, a model for estimating relative chlorophyll content was established. The results showed that cadmium stress at 1 mg/kg and 5 mg/kg had a promoting effect on the relative chlorophyll content, while cadmium stress at 10 mg/kg and 20 mg/kg had an inhibitory effect on the relative chlorophyll content. Moreover, with the extension of time, the inhibitory effect became more pronounced. Cadmium stress affects both the donor and acceptor sides of photosystem II in lettuce seedling leaves, damaging the electron transfer chain and reducing energy transfer in the photosynthetic system. It also inhibits water photolysis and decreases electron transfer efficiency, leading to a decline in photosynthesis. However, lettuce seedling leaves can mitigate photosystem II damage caused by cadmium stress through increased thermal dissipation. The model established based on the energy captured by a reaction center for electron transfer can effectively estimate the relative chlorophyll content of leaves. This study demonstrates that chlorophyll fluorescence techniques have great potential in elucidating the physiological mechanism of cadmium stress in lettuce, as well as in achieving synchronized determination and correlation analyses of chlorophyll content and photosynthetic function.

Ultrasound-Assisted Extraction of Phenolic Compounds from Celtuce (Lactuca sativa var. augustana) Leaves Using Natural Deep Eutectic Solvents (NADES): Process Optimization and Extraction Mechanism Research.

Natural deep eutectic solvents (NADESs), as emerging green solvents, can efficiently extract natural products from natural resources. However, studies on the extraction of phenolic compounds from celtuce (Lactuca sativa var. augustana) leaves (CLs) by NADESs are still lacking. This study screened the NADES L-proline-lactic acid (Pr-LA), combined it with ultrasound-assisted extraction (UAE) to extract phenolic compounds from CLs, and conducted a comparative study on the extraction effect with traditional extraction solvents. Both SEM and FT-IR confirmed that Pr-LA can enhance the degree of fragmentation of cell structures and improve the extraction rate of phenolic compounds. Molecular dynamics simulation results show that Pr-LA can improve the solubility of phenolic compounds and has stronger hydrogen bonds and van der Waals interactions with phenolic compounds. Single-factor and Box-Behnken experiments optimized the process parameters for the extraction of phenolic compounds from CLs. The second-order kinetic model describes the extraction process of phenolic compounds from CLs under optimal process parameters and provides theoretical guidance for actual industrial production. This study not only provides an efficient and green method for extracting phenolic compounds from CLs but also clarifies the mechanism of improved extraction efficiency, which provides a basis for research on the NADES extraction mechanism.

Alternative splicing triggered by the insertion of a CACTA transposon attenuates LsGLK and leads to the development of pale-green leaves in lettuce.

Lettuce (Lactuca sativa) is one of the most important vegetable crops in the world. As a leafy vegetable, the polymorphism of lettuce leaves from dark to pale green is an important trait. However, the genetic and molecular mechanisms underlying such variations remain poorly understood. In this study, one major locus controlling the polymorphism of dark- and pale-green leaves in lettuce was identified using genome-wide association studies (GWAS). This locus was then fine mapped to an interval of 5375 bp on chromosome 4 using a segregating population containing 2480 progeny. Only one gene, homologous to the GLK genes in Arabidopsis and other plants, is present in the candidate region. A complementation test confirmed that the candidate gene, LsGLK, contributes to the variation of dark- and pale-green leaves. Sequence analysis showed that a CACTA transposon of 7434 bp was inserted 10 bp downstream of the stop codon of LsGLK, followed by a duplication of a 1826-bp fragment covering exons 3-6 of the LsGLK gene. The transposon insertion did not change the expression level of the LsGLK gene. However, because of alternative splicing, only 6% of the transcripts produced from the transposon insertion were wild-type transcripts, which led to the production of pale-green leaves. An evolutionary analysis revealed that the insertion of the CACTA transposon occurred in cultivated lettuce and might have been selected in particular cultivars to satisfy the diverse demands of consumers. In this study, we demonstrated that a transposon insertion near a gene may affect its splicing and consequently generate phenotypic variations.

Genome-wide identification and characterization of the lettuce GASA family in response to abiotic stresses.

BACKGROUND: Lettuce is one of the most extensively farmed vegetables in the world, and it prefers cool growing conditions. High temperatures promote premature bolt formation, reducing quality and yield. The gibberellic acid-stimulated Arabidopsis (GASA) family genes play critical roles in plant growth, development, and stress responses. However, the biological functions of GASA proteins in lettuce have yet to be thoroughly investigated. RESULTS: Using genome-wide analysis, 20 GASAs were identified in lettuce including, three groups of LsGASA proteins based on the phylogenetic analysis. Except for one, all GASA proteins included a conserved GASA domain with 12 cysteine residues. Cis-element analysis showed that LsGASAs were closely associated with light, phytohormones, and stress resistance. Five segmental and three tandem duplication events were observed in the LsGASA family based on duplication analysis. GASA synteny analysis among lettuce, Arabidopsis, tobacco, and rice revealed that LsGASA5 is highly collinear with all species. Six of the 20 LsGASA showed increased expression patterns at specific time points in the shoot apical meristem when subjected to heat stress. According to gene expression analysis, the majority of GASA were highly expressed in flowers compared to other organs, and six GASA exhibited highly increased expression levels in response to NaCl, abscisic acid, and gibberellin treatment. Furthermore, LsGASA proteins are predominantly found in the plasma membrane and/or the cytosol. CONCLUSIONS: This study provides a comprehensive characterization of LsGASA genes for their diversity and biological functions. Moreover, our results will be useful for further studies on the function of lettuce GASA in abiotic stress- and heat-induced bolting signaling.

Leaf chlorophyll fluorescence and reflectance of oakleaf lettuce exposed to metal and metal(oid) oxide nanoparticles.

BACKGROUND: Most nanoparticles (NPs) have a significant impact on the structure and function of the plant photosynthetic apparatus. However, their spectrum of action varies significantly, from beneficial stimulation to toxicity, depending on the type of NPs, the concentration used and plant genotypic diversity. Photosynthetic performance can be assessed through chlorophyll a fluorescence (ChlF) measurements. These data allow to indirectly obtain detailed information about primary light reactions, thylakoid electron transport reactions, dark enzymatic stroma reactions, slow regulatory processes, processes at the pigment level. It makes possible, together with leaf reflectance performance, to evaluate photosynthesis sensitivity to stress stimuli. RESULTS: We investigated effects of different metal and metal(oid) oxide nanoparticles on photosynthesis of oakleaf lettuce seedlings by monitoring the chlorophyll a fluorescence light radiation and reflectance from the leaves. Observations of ChlF parameters and changes in leaf morphology were carried out for 9 days in two-day intervals. Spectrophotometric studies were performed at 9th day. Suspensions of NPs with the following concentrations were used: 6% TiO2, SiO2; 3% CeO2, SnO2, Fe2O3; 0.004% (40 ppm) Ag; 0.002% (20 ppm) Au. Nanoparticles were applied directly on the leaves which caused small symptoms of chlorosis, necrosis and leaf veins deformation, but the plants fully recovered to the initial morphological state at 9th day. Leaf reflectance analysis showed an increase in FRI for SiO2-NPs and CeO2-NPs treatments and ARI2 for Fe2O3, however, WBI and PRI coefficients for the latter nanoparticle were lower than in control. Chlorophyll a fluorescence parameters have changed due to NPs treatment. Fe2O3-NPs caused an increase in Fv/F0, PIABS, ET0/RC, DI0/RC, ABS/RC in different time points in comparison to control, also Ag, Au and SnO2 treatment caused an increase in Fv/F0, PIABS or ET0/RC, respectively. On the other hand, TiO2-NPs caused a decrease in Fv/Fm and Fv/F0 parameters, but an increase in DI0/RC value was observed. SnO2-NPs decreased PIABS, but increased ET0/RC than compared to control. Nanoparticles affected the shape of the O-J-I-P curve in slight manner, however, further analyses showed unfavourable changes within the PSII antenna, manifested by a slowdown in the transport of electrons between the Chl molecules of the light-harvesting complex II and the active center of PSII due to NPs application. CONCLUSION: Changes in ChlF parameters and leaf reflectance values clearly proved the significant influence of NPs on the functioning of the photosynthetic apparatus, especially right after NPs application. The nature of these changes was strictly depended on the type of nanoparticles and sometimes underwent very significant changes over time. The greatest changes in ChlF parameters were caused by Fe2O3 nanoparticles, followed by TiO2-NPs. After slight response of O-J-I-P curves to treatment of the plants with NPs the course of the light phase of photosynthesis stabilized and at 9th day were comparable to the control curve.

Deciphering transcriptomic signatures explaining the phenotypic plasticity of nonheading lettuce genotypes under artificial light conditions.

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.

In situ control of root-bacteria interactions using optical trapping in transparent soil.

Bacterial attachment on root surfaces is an important step preceding the colonization or internalization and subsequent infection of plants by pathogens. Unfortunately, bacterial attachment is not well understood because the phenomenon is difficult to observe. Here we assessed whether this limitation could be overcome using optical trapping approaches. We have developed a system based on counter-propagating beams and studied its ability to guide Pectobacterium atrosepticum (Pba) cells to different root cell types within the interstices of transparent soils. Bacterial cells were successfully trapped and guided to root hair cells, epidermal cells, border cells, and tissues damaged by laser ablation. Finally, we used the system to quantify the bacterial cell detachment rate of Pba cells on root surfaces following reversible attachment. Optical trapping techniques could greatly enhance our ability to deterministically characterize mechanisms linked to attachment and formation of biofilms in the rhizosphere.

Controlling root zone temperature improves plant growth and pigments in hydroponic lettuce.

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.

New Insights into the Accumulation, Transport, and Distribution Mechanisms of Hexafluoropropylene Oxide Homologues, Important Alternatives to Perfluorooctanoic Acid, in Lettuce (Lactuca sativa L.).

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.

Spring water quality monitoring using multiple bioindicators from multiple collection sites.

The aim of this study was to examine the water quality of the Extrema River spring in a Brazilian Cerrado area. Three collection sites (P1 - P3) were sampled in the dry and rainy seasons, which are close to industries from different sectors. In the physicochemical analysis, a decrease in dissolved oxygen levels (<5 mg/L) and pH (< 6) at P3 was detected. An increase in heterotrophic bacteria count was recorded at all sites (> 500 colonies/ml). In ecotoxicological analyses, P2 and P3 exhibited toxicity using Vibrio fischeri (> 20%). In evaluating toxicity, the reduction in seed germination was significant utilizing Lactuca sativa at all locations and with Allium cepa only at P2; rootlet length was decreased at P3 on L. sativa and at all sites with A. cepa. In contrast, loss of membrane integrity and mitochondrial function of meristems was adversely affected at all locations using both L. sativa and A. cepa assays. Principal components analysis (PCA) approach indicated that seasonality apparently did not markedly interfere with the obtained data, but it is important to include more collection locations to be evaluated with multiple bioindicators in the spring region. Our data indicate the urgent need for more rigorous programs to monitor the discharge of effluents into water springs.