First Report of Impatiens Necrotic Spot Virus (INSV) Infecting Lettuce in Mexico.

Impatiens necrotic spot virus (INSV; Order Bunyavirales, Family Tospoviridae) is transmitted by several thrips species and has emerged as an important pathogen of lettuce (Lactuca sativa) in several countries (Beris et al., 2020 and Hasegawa & Del Pozo-Valdivia, 2023). In 2023, a total of 22,092 hectares of lettuce were planted in Mexico, with a total production of 523,739 tons (Agri-Food and Fisheries Information Service, 2024). In the last several years, increased outbreaks of a disease showing virus-like symptoms have emerged in lettuce in central Mexico. In all cases, plants exhibited symptoms of yellowing, brown necrotic spots and ringspots on the leaves and midribs, and reduced growth. In fall 2023, symptoms were observed in 12 iceberg lettuce fields, with incidences between 5-70% in the municipality of Tenango del Valle, in the state of Mexico. 18 plants collected from 6 fields were initially tested for the presence of two thrips-transmitted viruses, INSV and tomato spotted wilt virus (TSWV) using ImmunoStrips (Agdia Inc., Elkhart, Indiana), which indicated positive results for INSV in 16 plants and TSWV in 2 plants. None of the plants tested positive for both viruses. Follow up sampling was conducted, which included a total of 17 symptomatic lettuce plants from three fields in October 2023, while an additional 11 lettuce plants were collected from Tepeaca, in the neighboring state of Puebla in March 2024. DAS-ELISA confirmed the presence of INSV in 13/17 plants and TSWV in 2/17 plants from Tenango del Valle, while only INSV was detected in the 11 plants from Tepeaca. Based on ELISA, none of the lettuce samples were co-infected by INSV and TSWV, which was similarly observed in California and Greece (Koike et al., 2008 and Beris et al., 2020). Initially, RNA was purified from 3 plants (two from Tenango del Valle, one from Tepeaca) reverse-transcribed, and PCR amplified with primers to the N gene of the INSV S RNA, as previously described (Hasegawa et al., 2022). All three reactions produced a single expected amplicon of 524 bp and were confirmed by bi-directional Sanger sequencing (MCLab, South San Francisco, CA). To obtain the full-length sequences for the N and NSm genes, RNA from a fourth sample (Tenango del Valle) was amplified with primers (Kuo et al., 2014), and Sanger sequenced. The 789 bp N gene (PP726902) shared >99% nucleotide and amino acid identity to the corresponding region of the INSV isolate from orchid in California (KF926828), while the 912 bp NSm gene (PP726901) shared >98% nucleotide and amino acid identity to the INSV isolate from basil in Washington (KX790322). Additionally, all four samples showed >99.5% similarity to one another. INSV was previously reported to affect other crops in Mexico, including tomatillo (Physalis ixocarpa) and pepper (Capsicum spp.) (González-Pacheco and Silva-Rosales, 2013), while TSWV has been reported to infect lettuce (Moreno et al., 2016). To our knowledge, this is the first report of INSV infecting lettuce in Mexico and was the dominant orthotospovirus in lettuce samples that were tested. INSV should be closely monitored throughout central Mexico, where majority of lettuce production occurs. Additional studies are warranted to identify the thrips vector species that are present, and to understand the role of crop and non-crop hosts in the epidemiology of INSV throughout the region.

Modifying the Ambient Light Spectrum Using LED Lamps Alters the Phenolic Profile of Hydroponically Grown Greenhouse Lettuce Plants without Affecting Their Agronomic Characteristics.

The growth and development of green lettuce plants can be modulated by the prevailing light conditions around them. The aim of this study was to evaluate the effect of ambient light enrichment with different LED light spectra on agronomic characteristics, polyphenol concentration and relative gene expression of enzymes associated with polyphenol formation in 'Levistro' lettuce grown hydroponically in a Nutrient Film Technique (NFT) system for 28 days in a greenhouse. The spectra (blue:green:red:far-red) and red:blue (R:B) ratios obtained by enriching ambient light with Blue (B), White (W), Blue-Red (BR) and Red (R) LED light were B: 47:22:21:10, 0.5:1; W: 30:38:23:9, 0.8:1; BR: 33:15:44:8, 1.3:1 and R: 16:16:60:8, 3.8:1, respectively, and photosynthetically active radiation (PAR) under the different treatments, measured at midday, ranged from 328 to 336 µmoles m-2 s-1. The resulting daily light integral (DLI) was between 9.1 and 9.6 mol m-2 day-1. The photoperiod for all enrichment treatments was 12 h of light. The control was ambient greenhouse light (25:30:30:15; R:B = 1.2:1; PAR = 702 µmoles m-2 s-1; DLI = 16.9 mol m-2 day-1; photoperiod = 14.2 h of light). Fresh weight (FW) and dried weight percentage (DWP) were similar among the enrichment treatments and the control. The leaf number increased significantly under BR and R compared to B lights. The relative index of chlorophyll concentration (RIC) increased as plants grew and was similar among the enrichment treatments and the control. On the other hand, the concentration of chlorogenic acid and chicoric acid increased under BR and B lights, which was consistent with the higher relative expression of the coumarate 3-hydroxylase enzyme gene. In view of the results, it is inferred that half of the PAR or DLI is sufficient to achieve normal growth and development of 'Levistro' lettuce plants, suggesting a more efficient use of light energy under the light enrichment treatments. On the other hand, the blue and combined blue-red lights promoted the accumulation of phenolic compounds in the leaves of 'Levistro' lettuce plants.

Microbial Biocontrol Agents and Natural Products Act as Salt Stress Mitigators in Lactuca sativa L.

One of the major problems related to climate change is the increase in land area affected by higher salt concentrations and desertification. Finding economically and environmentally friendly sustainable solutions that effectively mitigate salt stress damage to plants is of great importance. In our work, some natural products and microbial biocontrol agents were evaluated for their long-term effectiveness in reducing salt stress in lettuce (Lactuca sativa L. var. romana) plants. Fourteen different treatments applied to soil pots, with and without salt stress, were analyzed using biometric (leaf and root length and width), physiological (chlorophyll and proline content), and morphological (microscopic preparations) techniques and NGS to study the microbial communities in the soil of plants subjected to different treatments. Under our long-term experimental conditions (90 days), the results showed that salt stress negatively affected plant growth. The statistical analysis showed a high variability in the responses of the different biostimulant treatments. Notably, the biocontrol agents Papiliotrema terrestris (strain PT22AV), Bacillus amyloliquefaciens (strain B07), and Rahnella aquatilis (strain 36) can act as salt stress mitigators in L. sativa. These findings suggest that both microbial biocontrol agents and certain natural products hold promise for reducing the adverse effects of salt stress on plants.

Pharmaceutical metabolite identification in lettuce (Lactuca sativa) and earthworms (Eisenia fetida) using liquid chromatography coupled to high-resolution mass spectrometry and in silico spectral library.

Pharmaceuticals released into the aquatic and soil environments can be absorbed by plants and soil organisms, potentially leading to the formation of unknown metabolites that may negatively affect these organisms or contaminate the food chain. The aim of this study was to identify pharmaceutical metabolites through a triplet approach for metabolite structure prediction (software-based predictions, literature review, and known common metabolic pathways), followed by generating in silico mass spectral libraries and applying various mass spectrometry modes for untargeted LC-qTOF analysis. Therefore, Eisenia fetida and Lactuca sativa were exposed to a pharmaceutical mixture (atenolol, enrofloxacin, erythromycin, ketoprofen, sulfametoxazole, tetracycline) under hydroponic and soil conditions at environmentally relevant concentrations. Samples collected at different time points were extracted using QuEChERS and analyzed with LC-qTOF in data-dependent (DDA) and data-independent (DIA) acquisition modes, applying both positive and negative electrospray ionization. The triplet approach for metabolite structure prediction yielded a total of 3762 pharmaceutical metabolites, and an in silico mass spectral library was created based on these predicted metabolites. This approach resulted in the identification of 26 statistically significant metabolites (p < 0.05), with DDA + and DDA - outperforming DIA modes by successfully detecting 56/67 sample type:metabolite combinations. Lettuce roots had the highest metabolite count (26), followed by leaves (6) and earthworms (2). Despite the lower metabolite count, earthworms showed the highest peak intensities, closely followed by roots, with leaves displaying the lowest intensities. Common metabolic reactions observed included hydroxylation, decarboxylation, acetylation, and glucosidation, with ketoprofen-related metabolites being the most prevalent, totaling 12 distinct metabolites. In conclusion, we developed a high-throughput workflow combining open-source software with LC-HRMS for identifying unknown metabolites across various sample types.

Effects of vegetal protein hydrolysate application method, nitrogen level, and nitrate-to-ammonium ratio on growth and composition of hydroponic lettuce.

BACKGROUND: Vegetal-derived protein hydrolysates (PHs) have been recognized as sustainable biostimulant products due to their beneficial effects on crops. However, most studies on PHs have been conducted at a fixed ratio of nitrate-to-ammonium (NO3 -:NH4 +) without considering other N application scenarios, leading to inconsistent results among the studies. This study compared the influences of N levels (2 or 10 mM N), NO3:NH4 ratios (100:0, 75:25, 50:50, or 25:75), and PH application methods - control, foliar spray (PH-F) or root application (PH-R) - on the yield, morphology, nutrients, and nutraceutical quality of hydroponic lettuce. RESULTS: Nitrogen level, NO3:NH4 ratio, and PH application affected plant growth, morphology, and quality significantly, highlighting the importance of the interactions among these factors. Shoot growth was influenced by NO3:NH4 ratios, PH, and their interactions. Similar trends were observed in chlorophyll content. The interactions among all three factors significantly influenced root growth and morphology. Root application (PH-R) protected lettuce from yield loss caused by low NO3:NH4 ratios and from reduced antioxidant compounds caused by high N levels. Vegetal-derived protein hydrolysates improved nutrient uptake through two-way and three-way interactions although neither PH nor any interactions affected nitrate concentrations. CONCLUSION: This study demonstrated that PH interacts with N level and NO3:NH4 ratio, affecting hydroponic lettuce yield and quality. In particular, the root application of PH was the most effective method for enhancing yield (shoot fresh weight), quality (chlorophylls, carotenoids, flavonoids, and phenols), and nutrient uptake in hydroponically grown lettuce in relation to N form and level. © 2024 Society of Chemical Industry.

Phyto-cytogenotoxic potential assessment of two medicinal plants: Davilla nitida (Vahl) Kubitzki and Davilla elliptica (A. St.-Hill) (Dilleniaceae).

Humans have been using plants in the treatment of various diseases for millennia. Currently, even with allopathic medicines available, numerous populations globally still use plants for therapeutic purposes. Although plants constitute a safer alternative compared to synthetic agents, it is well established that medicinal plants might also exert adverse effects. Thus, the present investigation aimed to assess the phytotoxic, cytotoxic, and genotoxic potential of two plants from the Brazilian Cerrado used in popular medicine, Davilla nitida (Vahl) Kubitzki, and Davilla elliptica (A. St.-Hil.). To this end, germination, growth, and cell cycle analyses were conducted using the plant model Lactuca sativa. Seeds and roots were treated with 0.0625 to 1 g/L for 48 hr under controlled conditions. The germination test demonstrated significant phytotoxic effects for both species at the highest concentrations tested, while none of the extracts produced significant effects in the lettuce growth test. In the microscopic analyses, the aneugenic and cytotoxic action of D. elliptica was evident. In the case of D. nitida greater clastogenic action and induction of micronuclei, (MN) were noted suggesting that the damage initiated by exposure to these extracts was not repaired or led to apoptosis. These findings indicated that the observed plant damage was transmitted to the next generation of cells by way of MN. These differences in the action of the two species may not be attributed to qualitative variations in the composition of the extracts as both are similar, but to quantitative differences associated with synergistic and antagonistic interactions between the compounds present in these extracts.

Supplementary Far-Red Light for Photosynthetic Active Radiation Differentially Influences the Photochemical Efficiency and Biomass Accumulation in Greenhouse-Grown Lettuce.

Adding far-red (FR, 700-800 nm) light to photosynthetic active radiation (400-700 nm) proved to be a possible approach to increasing plant biomass accumulation for lettuce production in indoor vertical farms with artificial lighting as a sole-source lighting. However, how FR light addition influences plant growth, development, and metabolic processes and the optimal value of FR photon flux density for greenhouse-grown lettuce under sunlight are still unclear. This work aims to quantify the value of supplementary FR light with different intensities on lettuce morphological and physiological characteristics in a greenhouse. Lettuce 'Dasusheng' (Lactuca sativa L.) was grown in a greenhouse under seven light treatments, including white plus red LEDs with FR photon flux density at 0, 10, 30, 50, 70, and 90 µmol m-2 s-1 (WR, WR + FR10, WR + FR30, WR + FR50, WR + FR70, and WR + FR90, respectively), and lettuce grown with sunlight only was marked as natural light (NL). FR light addition improved the electron transport flux per cross section and performance index (PIabs, PItotal) and decreased the changes in relative variable fluorescence of lettuce leaves compared to plants under NL. Specifically, the PIabs of lettuce leaves were 41%, 41%, 38%, 33%, 26%, and 25% lower under control than in plants under treatments WR + FR90, WR + FR70, WR + FR50, WR + FR30, WR + FR10, and WR, respectively. Leaf number, leaf area, and biomass accumulation of lettuce followed a quadratic function with increasing FR light intensity and were the highest under treatment WR + FR50. The shoot fresh weight and dry weight of lettuce were increased by 111% and 275%, respectively, under treatment WR + FR50 compared to NL. The contents of vitamin C, reducing sugar, total soluble sugar, and starch in lettuce showed a similar trend with biomass accumulation. In conclusion, with commonly used photosynthetic photon flux density (PPFD, 400-700 nm) around 200 µmol m-2 s-1, supplementary FR light intensity of 30~50 µmol m-2 s-1 was suggested to enhance the photochemistry efficiency, biomass accumulation, and carbohydrates' contents in greenhouse-grown lettuce.

LsRTDv1, a reference transcript dataset for accurate transcript-specific expression analysis in lettuce.

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 role of nano-chelated iron on anatomical and biochemical characteristics and concentration of mineral nutrients in lettuce (Lactuca sativa L.) under cadmium toxicity.

Cadmium is one of the most hazardous environmental pollutants for plants due to its mobility and high toxicity. One effective method that may be utilized to decrease heavy metal pollution in the soil is the use of nano-chelated iron. In the present study, lettuce plants were treated with four different concentrations of cadmium chloride, two different concentrations of nano-chelated iron, and six combinations of cadmium chloride+nano-chelated iron. Application of 0.5 and 1 g/L nano-chelated iron reduced the adverse effects of cadmium on photosynthetic pigments and growth parameters. Combined application of cadmium chloride and nano-chelated iron (90 µg CdCl2/g perlite+0.5 g/L nano-chelated iron) led to an increase in soluble sugar content compared to the control lettuce plants. Lettuce had a high capacity to absorb cadmium from the contaminated medium. Interestingly, the levels of cadmium that accumulated in the roots (1.641 mg/g DW) were much higher than in the aerial parts of the plant (0.998 mg/g DW). The results showed that there was a decline in the mineral content of lettuce treated with cadmium, while the application of nano-chelated iron led to its increase. This study suggests that the application of nano-chelated iron is a cost-effective and practical method that can be used in the agricultural soil systems to enhance crop tolerance in cadmium-polluted soil.

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.

The nitrogen-sulfur ratio of acid rain modulates the leaf- and root-mediated co-allelopathy of Solidago canadensis.

The majority of allelopathic studies on invasive plants have focused primarily on their leaf-mediated allelopathy, with relatively little attention paid to their root-mediated allelopathy, especially co-allelopathy mediated by both leaves and roots. It is conceivable that the diversified composition of acid rain may influence the allelopathy of invasive plants. This study aimed to evaluate the leaf and root-mediated co-allelopathy of the invasive plant Solidago canadensis L. under acid rain with different nitrogen-sulfur ratios (N/S) on Lactuca sativa L. via a hydroponic incubation. The root-mediated allelopathy of S. canadensis was found to be more pronounced than the leaf-mediated allelopathy of S. canadensis with nitric acid at pH 4.5, but the leaf-mediated allelopathy of S. canadensis was observed to be more pronounced than the root-mediated allelopathy of S. canadensis with sulfuric-rich acid at pH 4.5. The leaf and root-mediated co-allelopathy of S. canadensis was more pronounced than that of either part alone with sulfuric acid at pH 5.6 and nitric acid at pH 4.5, but not with nitric-rich acid at pH 4.5 and sulfuric-rich acid at pH 4.5. Sulfuric acid and sulfuric-rich acid with stronger acidity intensified the leaf-mediated allelopathy of S. canadensis. Nitric acid and nitric-rich acid attenuated the leaf-mediated allelopathy of S. canadensis, and most types of acid rain (especially nitric acid and nitric-rich acid) also attenuated the root-mediated allelopathy of S. canadensis and the leaf and root-mediated co-allelopathy of S. canadensis. Sulfuric acid and sulfuric-rich acid produced a more pronounced effect than nitric acid and nitric-rich acid. Hence, the N/S ratio of acid rain influenced the allelopathy of S. canadensis under acid rain with multiple N/S ratios.

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.

Hyperspectral imaging of liverwort Marchantia polymorpha identifies MpWRKY10 as a key regulator defining Foliar pigmentation patterns.

Foliar pigmentation patterns vary among plant species and growth conditions. In this study, we utilize hyperspectral imaging to assess foliar pigmentation in the bryophyte Marchantia polymorpha under nutrient stress and identify associated genetic factors. Using singular value decomposition (SVD) for feature selection, we quantitate color variations induced by deficiencies in phosphate, nitrate, magnesium, calcium, and iron. Pseudo-colored thallus images show that disrupting MpWRKY10 causes irregular pigmentation with auronidin accumulation. Transcriptomic profiling shows that MpWRKY10 regulates phenylpropanoid pathway enzymes and R2R3-MYB transcription factors during phosphate deficiency, with MpMYB14 upregulation preceding pigment accumulation. MpWRKY10 is downregulated in older, pigmented thalli under phosphate deficiency but maintained in young thalli, where it suppresses pigmentation genes. This downregulation is absent in pigmented thalli due to aging. Comparative transcriptome analysis suggests similar WRKY and MYB roles in nutrient response and pigmentation in red-leaf lettuce, alluding to conserved genetic factors controlling foliar pigmentation patterns under nutrient deficiency.

Analysis of lettuce transcriptome reveals the mechanism of different light/dark cycle in promoting the growth and quality.

Light/dark (L/D) cycle plays a crucial role in controlling the production and quality of vegetables. However, the mechanism of L/D cycle on vegetable growth and quality is scarce studied. To investigate the impact of L/D cycle on lettuce growth and quality, we designed three diel scenarios, including 16 hours of light and 8 hours of darkness (L16/D8), 12 hours of light and 6 hours of darkness (L12/D6), and 8 hours of light and 4 hours of darkness (L8/D4). By phenotypic analysis, we found that lettuce grew taller under the L8/D4 scenario than under L16/D8 light cycle scenarios. The physiological indexes showed that the lettuce leaves grown in the L8/D4 scenario exhibited greater enhancements in the levels of soluble protein, soluble sugar, and carotenoid content compared to the other scenarios. By comparing the expression levels under different diel scenarios (L16/D8 vs L12/D6, L16/D8 vs L8/D4, and L12/D6 vs L8/D4), we identified 7,209 differentially expressed genes (DEGs). Additionally, 3 gene modules that were closely related to L/D cycle of lettuce were selected by WGCNA analysis. The eigengenes of three gene modules were enriched in plant hormone signal transduction, sphingolipid metabolism, and nucleocytoplasmic transport pathways. Through network analysis, we identified six hub genes (CIP1, SCL34, ROPGEF1, ACD6, CcmB, and Rps4) in the three gene modules, which were dominant in plant circadian rhythms and greatly affected lettuce growth. qRT-PCR analysis confirmed the diurnal response patterns of the 6 hub genes in different treatments were significant. This study intensively enhanced our comprehension of the L/D cycle in the growth morphology, nutritional quality, and metabolic pathways of lettuce.

Lettuce (Lactuca sativa L.) Cultures and the Bioactivity of Their Root Microflora Are Affected by Amended Soil.

This study aimed to highlight the positive effects of various recycled organic substrates on lettuce plants (Lactuca sativa L.) and to promote sustainable waste management practices, contributing to the concept of a circular economy. Over a two-month period, the growth potential and rhizosphere microflora of lettuce plants grown in soil amended with different recycled substrates were investigated. All data were compared, and the effects of the culture substrates were evaluated. All groups containing soil improvers offered a significant increase in the number of leaves per plant and, in two cases, an increase in dry biomass as well as an increase in the concentration of all leaf pigments. Both MDA and H2O2 concentrations were the lowest in two groups containing soil improvers (VG 5% and PLUS 10%). At the end of the culture period, isolation and culture of bacteria from the plant rhizosphere were performed. Different bacterial strains were isolated and tested for the production of antimicrobial agents against six microbial indicators (B. subtilis, E. coli, S. aureus, S. cerevisiae, C. albicans, and P. aeruginosa). The greater percentage of the isolated strains showed an ability to inhibit the growth of the B. subtilis index. Most of the strains with antimicrobial activity were isolated from the soil samples of the plain soil group and the soil amended with the commercial fertilizer. Three of the isolated strains originating from the Ginagro 5% group are multiproducers as they inhibit the growth of three microbial indicators or more.

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.

Lettuce Genotype-Dependent Effects of Temperature on Escherichia coli O157:H7 Persistence and Plant Head Growth.

Lettuce has been commonly associated with the contamination of human pathogens, such as Escherichia coli O157:H7 (hereafter O157:H7), which has resulted in serious foodborne illnesses. Contamination events may happen throughout the farm-to-fork chain, when O157:H7 colonizes edible tissues and closely interacts with the plant. Environmental conditions have a significant impact on many plant-microbe interactions; however, it is currently unknown whether temperature affects O157:H7 colonization of the lettuce phyllosphere. In this study, we investigated the relationship between elevated growth temperatures, O157:H7 persistence, and lettuce head growth using 25 lettuce genotypes. Plants were grown under optimal or elevated temperatures for 3.5 weeks before being inoculated with O157:H7. The bacterial population size in the phyllosphere and lettuce head area was estimated at 0- and 10-days postinoculation (DPI) to assess bacterial persistence and head growth during contamination. We found that growing temperature can have a positive, negative, or no effect on O157:H7 persistence depending on the lettuce genotype. Furthermore, temperature had a greater effect on head area size than the presence of O157:H7. The results suggested that the combination of plant genotype and temperature level is an important factor for O157:H7 colonization of lettuce and the possibility to combine desirable food safety traits with heat tolerance into the lettuce germplasm.

Stability Evaluation for Heat Tolerance in Lettuce: Implications and Recommendations.

Lettuce is an important cool-temperature crop, and its principal abiotic stress is low heat tolerance. Lettuce production has become more challenging in the context of global warming changes. Hence, the main objective of this research was to investigate the relationship between stability and lettuce heat tolerance. Field and greenhouse trials were run in 2015 (summer) and 2016 (fall and spring). The environments were composed of a combination of season and place (field, glass, and plastic greenhouse), and the assessed genotypes were BRS Leila and Mediterrânea, Elisa, Everglades, Simpson, and Vanda. Statistical analysis showed a significant effect (p < 0.05) of environments (E), genotypes (G), and the GEI. BRS Leila, Elisa, and BRS Mediterrânea showed the greatest means to the first anthesis in suitable environments (milder temperatures). Among these cultivars, BRS Mediterrânea was the most stable and adapted to hot environments. The environmental conditions studied in this research, mainly high temperatures, could become a reality in many lettuce-producing areas. Therefore, the results can help indicate and develop lettuce varieties with greater heat tolerance.