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At least 19 recordsLinked to original sources
PubMed2025

Biosynthetic potential of the culturable foliar fungi associated with field-grown lettuce.

Fungal endophytes and epiphytes associated with plant leaves can play important ecological roles through the production of specialized metabolites encoded by biosynthetic gene clusters (BGCs). However, their functional capacity, especially in crops like lettuce (Lactuca sativa L.), remains poorly understood. We sequenced the genomes of nine fungal isolates, representing Fusarium sp., Fulvia sp., Alternaria alternata, and Alternaria postmessia, from leaves of lettuce grown under field conditions in Arizona, USA. We used antibiotics and secondary metabolite analysis shell (antiSMASH) and the database for automated carbohydrate-active enzyme annotation (dbCAN3), to predict BGCs and carbohydrate-active enzymes (CAZymes) for each strain, and then compared them to conspecific strains from other environments and substrates. Foliar lettuce-associated fungi featured 39-95 BGCs per genome, with substantial overlap between isolates occurring in association with lettuce leaves vs. from other substrates. Species identity was a significant determinant of BGC count, while host type, isolation source, and lifestyle were not. Several BGCs, including those for alternariol and 1,3,6,8-Tetrahydroxynaphthalene (T4HN), showed 100% similarity to characterized minimum information about a biosynthetic gene cluster (MIBiG) clusters based on antiSMASH predictions. Although analysis by biosynthetic gene similarity clustering and prospecting engine (BiG-SCAPE) identified gene cluster families (GCFs) across the dataset, these reference-matching clusters were not always grouped, reflecting methodological differences in how the tools assess similarity. Comparative CAZyme analysis in a focal species (Fulvia sp.) revealed higher gene counts in a foliar lettuce-derived isolate than in tomato (Solanum lycopersicum)-associated strains, challenging assumptions about host chemical complexity. These results highlight the importance of phylogenetic context in shaping fungal functional potential and suggest that selection on microbial traits in edible leafy crops may be more subtle and species-specific than previously assumed. KEY POINTS: • Lettuce-associated fungi feature diverse biosynthetic potential • Phylogeny predicts fungal BGC content more strongly than ecological lifestyle • Findings support genome-informed microbiome strategies for leafy crops.

Lactuca↗
PubMed2025

Diversity of leaf- and root microbiomes among genotypes and market classes of desert-grown lettuce (Lactuca sativa L.).

Plant microbiomes are increasingly acknowledged both as extensions of plant characteristics and as biological factors that influence plant traits important for nutrition and resilience. In the context of global change, manipulation of microbiomes has the potential to complement genetic approaches to enhance crop health and productivity under rising heat and drought stress. Understanding the factors that influence microbial communities and their variation across plant genotypes is essential for developing such capabilities. We employed metabarcoding via the Illumina sequencing platform to investigate microbial communities that occur within healthy leaves and roots of 12 lettuce genotypes (Lactuca sativa L.) grown in a desert agriculture environment. We detected diverse foliar- and root-endophytic fungi and bacteria in field-grown lettuce at the Maricopa Agricultural Center (Arizona, USA). The composition of microbial community structure varied with foliar chemistry and root traits. Notably, levels of zinc and other beneficial nutrients in the leaves were strongly linked with specific endophytes. These results document the lettuce microbiome in desert farming and provide insights into endophytes in lettuce leaves, which are noteworthy because they remain after washing and are regularly ingested.

Lactuca↗
PubMed1977

Folic acid compounds in romaine lettuce.

The composition of folate coenzymes in romaine lettuce was studied. Lettuce extract was purified on QAE-Sephadex A-25 and folate compounds were separated into a monoglutamate fraction and a polyglutamate fraction by chromatography on Sephadex G-15. Both the mono- and poly-glutamate fractions were resolved on DEAE-cellulose. Positive identification of DEAE peaks was made by further cochromatography with high specific activity radioactive marker folate compounds and with differential microbiological assay. The distribution of folate compounds in lettuce is as follows: 32% 5-CH3-H4PteGlu; 1% 5-CHO-H4PteGlu; 3% 5-CHO-H4PteGlu4; 9% 5-CH3-H4PteGlu4; 13% 5-CHO-H4PteGlu5; and 31% 5-CH3-H4PteGlu5.

Biological Assay↗
PubMed1978

Evaluation of potential for aflatoxin occurrence on celery, cauliflower, lettuce, and taro root inoculated with Aspergillus flavus and A. parasiticus.

Culture samples of lettuce, cauliflower, celery, and taro root (Colocasia esculenta) were assayed for the presence of aflatoxin after inoculation with Aspergillus flavus and A. parasiticus. Cultures of A. flavus produced both aflatoxins B1 and G1 on taro root, but produced by B1 on lettuce, cauliflower, and celery. For taro root, the percentage of aflatoxin G1 produced was considerably greater than that of B1. While A. parasiticus did produce mycelia and spores on the lettuce and taro root samples, there were not detectable levels of any aflatoxin produced. All the samples studied were successfully extracted and analyzed qualitatively and quantatively for the presence of aflatoxin by using official AOAC thin layer chromatographic procedures. There is sufficient evidence that Aspergilli can grow on some leafy produce and one strain produced aflatoxins.

Aflatoxins↗
PubMed1978

Tentoxin. An uncompetitive inhibitor of lettuce chloroplast coupling factor 1.

The interaction of tentoxin [cyclo-(-L-leucyl-N-methyl-(Z)-dehydrophenylalanyl-glycyl-N-methyl-L-alanyl-)] with solubilized lettuce chloroplast coupling factor 1 was characterized by direct binding studies, measurement of the time course of ATPase inhibition, and steady-state enzyme kinetics. Neither substrates, products or Ca2+ competed with the tentoxin binding site, nor did they induce any large change in tentoxin affinity. The inhibition of lettuce chloroplast coupling factor 1 ATPase was found to be the time dependent, and at equilibrium the affinities estimated by equilibrium ultrafiltration and enzyme inhibition were similar (1.8 . 10(8) M-1). The steady-state kinetics best fit an uncompetitive pattern suggesting that the inhibited steps follow an irreversible step occurring after ATP binding.

Adenosine Diphosphate↗
PubMed2026

Cerium dioxide nanoparticle exposure attenuates mobility-linked antibiotic resistome signatures across the soil-lettuce continuum.

Antibiotic resistance genes (ARGs) are contaminants of emerging concern in agricultural microbiomes. Their association with mobile genetic elements (MGEs) can enhance dissemination across soil-plant interfaces, creating potential environmental and food-chain exposure risks. However, how engineered nanoparticles modulate relative ARG abundance and mobility-linked resistome features in plant-associated microbiomes remains poorly understood. Here, we examined the effects of graded, experimentally elevated cerium dioxide nanoparticle (CeO2 NP) loadings in a soil-lettuce system by integrating compartment-resolved metagenomics, ARG-MGE co-occurrence analysis, putative host-reservoir profiling, transcriptomics, and functional assays. Metagenomic profiling identified 16 ARG types and 125 subtypes and revealed niche-dependent microbiome restructuring under CeO2 NP exposure. Rhizosphere relative ARG abundance showed a negative dose-associated trend, although overall inter-group differences were not significant, whereas leaf endophytes showed a weaker response. Relative MGE abundance decreased significantly in both compartments, and lower assembly-level ARG-MGE co-occurrence reflected fewer ARGs detected in MGE-associated genomic contexts, whereas fewer multi-ARG contigs suggested reduced ARG clustering and potential co-selection. Putative host-reservoir analysis associated key efflux determinants with bacterial families whose relative representation declined following CeO2 NP exposure. Transcriptomic profiling of representative putative ARG hosts revealed host-specific responses, including downregulation of genes involved in central metabolism and Sec-dependent trafficking. Complementary host assays showed reduced apparent envelope permeability and lower recovery of tetracycline-resistant recipient-identity colonies in the plasmid-associated host system. Together, under the tested elevated-loading conditions, CeO2 NP exposure was associated with lower relative ARG signals and weaker mobility-linked resistome features across the soil-lettuce continuum, providing mechanistic insight into nanoparticle-resistome interactions in soil-plant systems.

ARG dissemination↗
PubMed2025

Differential methylation of a retrotransposon upstream of a MYB gene causes variegation of lettuce leaves, which is abolished by the presence of an (AT)5 repeat in the promoter.

Variegation, a common phenomenon in plants, can be the result of several genetic, developmental, and physiological factors. Leaves of some lettuce cultivars exhibit dramatic red variegation; however, the genetic mechanisms underlying this variegation remain unknown. In this study, we cloned the causal gene for variegation on lettuce leaves and elucidated the underlying molecular mechanisms. Genetic analysis revealed that the polymorphism of variegated versus uniformly red leaves is caused by an "AT" repeat in the promoter of the RLL2A gene encoding a MYB transcription factor. Complementation tests demonstrated that the RLL2A allele (RLL2AV) with (AT)n repeat numbers other than five led to variegated leaves. RLL2AV was expressed in the red spots but not in neighboring green regions. This expression pattern was in concert with a relatively low level of methylation in a retrotransposon inserted in -761 bp of the gene in the red spots compared to high methylation of the retrotransposon in the green region. The presence of (AT)5 in the promoter region, however, stabilized the expression of RLL2A, resulting in uniformly red leaves. In summary, we identified a novel promoter mechanism controlling variegation through inconsistent levels of methylation and showed that the presence of a simple sequence repeat of specific size could stabilize gene expression.

Promoter Regions, Genetic↗
PubMed2019

Cold chain and virus-free oral polio booster vaccine made in lettuce chloroplasts confers protection against all three poliovirus serotypes.

To prevent vaccine-associated paralytic poliomyelitis, WHO recommended withdrawal of Oral Polio Vaccine (Serotype-2) and a single dose of Inactivated Poliovirus Vaccine (IPV). IPV however is expensive, requires cold chain, injections and offers limited intestinal mucosal immunity, essential to prevent polio reinfection in countries with open sewer system. To date, there is no virus-free and cold chain-free polio vaccine capable of inducing robust mucosal immunity. We report here a novel low-cost, cold chain/poliovirus-free, booster vaccine using poliovirus capsid protein (VP1, conserved in all serotypes) fused with cholera non-toxic B subunit (CTB) expressed in lettuce chloroplasts. PCR using unique primer sets confirmed site-specific integration of CTB-VP1 transgene cassettes. Absence of the native chloroplast genome in Southern blots confirmed homoplasmy. Codon optimization of the VP1 coding sequence enhanced its expression 9-15-fold in chloroplasts. GM1-ganglioside receptor-binding ELISA confirmed pentamer assembly of CTB-VP1 fusion protein, fulfilling a key requirement for oral antigen delivery through gut epithelium. Transmission Electron Microscope images and hydrodynamic radius analysis confirmed VP1-VLPs of 22.3&#xa0;nm size. Mice primed with IPV and boosted three times with lyophilized plant cells expressing CTB-VP1co, formulated with plant-derived oral adjuvants, enhanced VP1-specific IgG1, VP1-IgA titres and neutralization (80%-100% seropositivity of Sabin-1, 2, 3). In contrast, IPV single dose resulted in <50% VP1-IgG1 and negligible VP1-IgA titres, poor neutralization and seropositivity (<20%, <40% Sabin 1,2). Mice orally boosted with CTB-VP1co, without IPV priming, failed to produce any protective neutralizing antibody. Because global population is receiving IPV single dose, booster vaccine free of poliovirus or cold chain offers a timely low-cost solution to eradicate polio.

Animals↗
PubMed1976

Contributions to ecological chemistry CVII1. Fate of lindane-14C in lettuce, endives and soil under outdoor conditions.

In seven successive outdoor experiments, lindane-14C was applied to lettuce or endive leaves as an aqueous formulation (about 12 mg on 20 plants for each experiment). The growing periods varied between 21 and 37 days. After this time, between 4.5% and 13.9% of the applied radiocarbon was recovered from the plants. Conversion rates to soluble metabolites as well as to unextractable residues appeared to be dependent on weather conditions. During the summer months, the radiocarbon in plants consisted of 36% soluble metabolites and of 30% unextractable residues (average of 4 experiments); in autumn, the conversion rates were much lower. The following metabolites were identified in both plant species by gas chromatography/mass spectrometry: a polar group (a free trichlorophenol, 2,3,4,6-tetrachlorophenol, pentachlorophenol, conjugates of the latter two compounds, and unidentified water-soluble products) amounting to 35% of the radioactivity in plants cultivated in summer, and a nonpolar group (a dichlorobenzene, 1,2,3-trichlorobenzene, 1,2,4-trichlorobenzene, 1,2,3,5, and/or 1,2,4,5-tetrachlorobenzene, pentachlorobenzene, hexachlorobenzene, and gamma-pentachlorocyclohexene) amounting to 1% of the radioactivity in plants cultivated in summer. The 20 cm top-soil layer had about 14% of the total radioactivity applied to all plants. Six % of the radioactivity recovered from the soil was soluble metabolites and about 50% was not extractable. The soluble metabolites comprised a polar group (free and conjugated 2,3,4,6-tetrachlorophenol, pentachlorophenol, and unidentified water soluble products) amounting to 5% of the radioactivity in the soil as well as a nonpolar group (1,2,3-trichlorobenzene, 1,2,3,4-tetrachlorobenzene, 1,2,3,5 and/or 1,2,4,5-tetrachlorobenzene, pentachlorobenzene, hexachlorobenzene, and gamma-pentachlorocyclohexene) amounting to 1% of the radioactivity in the soil.

Biodegradation, Environmental↗
PubMed2026

Co-occurrence of biofilm formation, acid tolerance, and antibiotic resistance in environmental Escherichia coli associated with lettuce.

BACKGROUND: Environmental niches represent important reservoirs of Escherichia coli with stress-adaptation traits that support persistence outside the host. Contaminated irrigation water and soils can facilitate transfer to fresh produce, where bacterial survival may reduce the effectiveness of downstream control measures. This study investigated the co-occurrence of biofilm formation, acid tolerance, and antibiotic resistance (AR) in environmental E. coli and their contribution to persistence along the farm-to-produce continuum. RESULTS: Eighteen E. coli isolates recovered from irrigation water, soil, and lettuce were characterized using phenotypic assays and genome-based analyses. Most isolates remained susceptible to the majority of tested antibiotics, with multidrug resistance observed in only 11.1% of isolates. In contrast, moderate-to-strong biofilm formation was widespread (83.3%), and several isolates exhibited reduced susceptibility to acetic acid at concentrations relevant to household washing practices. Genotypic screening revealed a broad distribution of adhesion, iron acquisition, biofilm-associated, and plasmid-borne resistance determinants, indicating substantial functional diversity. Significant positive associations were observed between acid tolerance, biofilm formation, and antibiotic resistance, suggesting co-occurrence of stress-adaptation phenotypes rather than definitive evolutionary convergence. While antibiotic resistance phenotypes showed strong concordance with corresponding resistance genes, biofilm formation and acid tolerance were not associated with specific genetic determinants, supporting a multifactorial basis of these traits. CONCLUSIONS: These findings demonstrate that environmental E. coli can combine multiple stress-adaptation mechanisms that enhance persistence across agricultural and food-associated environments, even in the absence of high-risk resistance profiles. The observed co-occurrence of phenotypic traits highlights the potential for co-selection under environmental pressures and underscores the limitations of relying solely on downstream decontamination strategies. Effective risk mitigation requires integrated, preventive approaches targeting pre-harvest contamination and environmental reservoirs.

Biofilms↗
PubMed1979

Differential survival of Salmonella typhi, Escherichia coli, and Enterobacter aerogenes on lettuce in the field.

Artificial contamination of young lettuce plants with Salmonella typhi (ST), Escherichia coli (EC), and Enterobacter aerogenes (EA) in the field in winter and summer resulted in presence of the pathogen and increased density of the two indicator bacteria (IB) in harvested produce. Viable counts of the three bacteria/g fresh weight declined at a decreasing rate with increasing time after contamination. The overall pattern of variation of the ST/IB ratio was correlated more closely with changes in the ST/EA than in the ST/EC ratio values. When viable counts were expressed as a proportion of the contaminating dose at different times after contamination, however, a closer similarity existed between ST and EC than between ST and EA values throughout winter and through the early part of summer experiments.

Enterobacter↗
PubMed1977

Occupational dermatitis from Lactuca sativa (lettuce) and Cichorium (endive). Simultaneous occurrence of immediate and delayed allergy as a cause of contact dermatitis.

Four patients with occupational contact dermatitis to Lactuca sativa had cross-sensitivity to Cichorium endivia. One of the patients also had contact urticaria to Lactuca and Cichorium, and another reacted positively to scratch tests with these plants as a sign of immediate allergy. In two cases such immediate allergy was considered the cause of a vesicular, intense itching eruption within a few minutes of contact with fresh leaves of Lactuca on previously eczematous skin. The severe chronic dermatitis of the hands of these patients is ascribed to combined delayed and immediate allergy.

Adult↗
PubMed2025

From aerial drone to quantitative trait locus: leveraging next-generation phenotyping to reveal the genetics of color and height in field-grown Lactuca sativa.

In recent years, accurate and low-cost variant calling has enabled the genotyping of large diversity panels for genome-wide association studies. As a result, phenotyping rather than genotyping is now the rate-limiting step, especially in field experiments. This has created a strong need for high-throughput, accurate, and low-cost in-field phenotyping. Here, we present a genome-wide association study (GWAS) study on 194 field-grown accessions of lettuce (Lactuca sativa). These accessions were non-destructively phenotyped at two time points 15&#x2009;days apart using a drone equipped with an RGB and multispectral (MSP) camera. Our high-throughput phenotyping approach integrates an RGB- and MSP camera to measure the color and height of lettuce in this large-scale field experiment. We used the mean and other summary statistics, such as median, quantiles, skewness, kurtosis, minimum, and maximum to quantify different aspects of color and height variation in lettuce from the drone images. Using these summary statistics as traits for GWAS, we confirm several previously described genetic associations, now under field conditions, and identify additional novel associations for color and height traits in lettuce.

Lactuca↗
PubMed1978

Evidence for a catalytic function of the coupling factor 1 protein reconstituted with chloroplast thylakoid membranes.

The effects of tentoxin on the ATPase activities of coupling factor 1 proteins (CF1) and photophosphorylation with isolated chloroplasts and chloroplasts reconstituted with coupling factor proteins have been examined. 1. The calcium-dependent ATPase activities of coupling factors isolated from spinach, lettuce and Nicotiana otophora are completely inhibited by tentoxin. The ATPase activities of coupling factors isolated from Nicotiana tabacum and Nicotiana knightiana are not affected by tentoxin. 2. Phenazine methosulfate-catalyzed cyclic photophosphorylation with chloroplasts isolated from spinach, lettuce and N. otophora is completely inhibited by tentoxin, whereas chloroplasts isolated from N. knightiana and N. tabacum are relatively insensitive to tentoxin. 3. Spinach chloroplasts, partially depleted in CF1, can be reconstituted with coupling factors isolated from a wide variety of plants including lettuce, radish, N. tabacum, N. knightiana and N. otophora. 4. Spinach chloroplasts reconstituted with spinach, lettuce and N. otophora CF1 retain their sensitivity to tentoxin; however, when reconstituted with N. knightiana and N. tabacum coupling factor proteins, a significant fraction of the reconstituted rate remains tentoxin insensitive. These data are interpreted as evidence that coupling factors that reconstitute with spinach thylakoid membranes have both a catalytic and structural function.

Adenosine Triphosphatases↗