Boosting pro-vitamin A content and bioaccessibility in leaves by combining engineered biosynthesis and storage pathways with high-light treatments.

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

The genome of Lactuca saligna, a wild relative of lettuce, provides insight into non-host resistance to the downy mildew Bremia lactucae.

Lactuca saligna L. is a wild relative of cultivated lettuce (Lactuca sativa L.), with which it is partially interfertile. Hybrid progeny suffer from hybrid incompatibility (HI), resulting in reduced fertility and distorted transmission ratios. Lactuca saligna displays broad-spectrum resistance against lettuce downy mildew caused by Bremia lactucae Regel and is considered a non-host species. This phenomenon of resistance in L. saligna is called non-host resistance (NHR). One possible mechanism behind this NHR is through the plant-pathogen interaction triggered by pathogen recognition receptors, including nucleotide-binding leucine-rich repeat (NLR) proteins and receptor-like kinases (RLKs). We report a chromosome-level genome assembly of L. saligna (accession CGN05327), leading to the identification of two large paracentric inversions (>50 Mb) between L. saligna and L. sativa. Genome-wide searches delineated the major resistance clusters as regions enriched in NLRs and RLKs. Three of the enriched regions co-locate with previously identified NHR intervals. RNA-seq analysis of Bremia-infected lettuce identified several differentially expressed RLKs in NHR regions. Three tandem wall-associated kinase-encoding genes (WAKs) in the NHR8 interval display particularly high expression changes at an early stage of infection. We propose RLKs as strong candidates for determinants of the NHR phenotype of L. saligna.

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

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

Role of gamma-irradiated sodium alginate on growth, physiological and active components of iceberg lettuce (Lactuca sativa) plant.

BACKGROUND: One of the most widely recognized biostimulators of plant development; is oligoalginate, which regulates the biological processes of plants and was used in horticultural fields as a plant growth regulator. The plan of the current research was to study, however, the foliar application of un-irradiated and irradiated Na-alginate (UISA and ISA) to improve the growth, physiological activity, and other active components of the Egyptian iceberg lettuce plant. Degraded Na-alginate is equipped with exposure of sodium alginate in its solid state to gamma-rays at different dose levels (0.0, 25, 50, 75, and 100 kGy). The characterization of the oligo-alginates achieved by γ-radiation deprivation at different dose levels was performed by FTIR, XRD, TGA, SEM, and TEM. Different concentrations of irradiated sodium alginate at dose levels of 100 kGy (200, 400, 600, and 800 ppm, as well as deionized water used as a control) were sprayed with a hand sprayer every week after transplanting the iceberg lettuce seedlings in the field until the harvest stage. Morphological traits were evaluated, as well as pigments, ascorbic acid, phenols, flavonoids, soluble proteins, and antioxidant activity. RESULTS: Irradiated Na-alginate resulted in the depolymerization of Na-alginate into small molecular-weight oligosaccharides, and the best dose to use was 100 kGy. Certain chemical modifications in the general structure were observed by FTIR analysis. Two absorbed bands at 3329 cm-1 and 1599 cm-1, were recognized that are assigned to O-H and C-O stretching, respectively, and peaks achieved at 1411 cm-1 represent the COO-stretching group connected to the sodium ion. The peak obtained at 1028 cm-1 was owing to the stretching vibration of C-O. The results of TGA provided that the minimum weight reminder was in the ISA at 100 kGy (28.12%) compared to the UISA (43.39%). The images of TEM pointed out that the Na-alginate was globular in shape, with the particle distribution between 12.8 and 21.7 nm in ISA at 100 kGy. Irradiated sodium alginate caused a noteworthy enhancement in the vegetative growth traits (leaf area, stem length, head weight, and leaf number). By spraying 400 ppm, ISA showed a maximum increase in total pigments (2.209 mg/g FW), ascorbic acid (3.13 mg/g fresh weight), phenols (1.399 mg/g FW), flavonoids (0.775 mg/g FW), and antioxidant activities (82.14. %). Also, there were correlation coefficients (R values) between leaf area, stem length, head weight, and leaf number values with total pigment content, antioxidant activity, total soluble proteins, and ascorbic acid. CONCLUSIONS: The outcomes of the recent investigation demonstrated that the application of spraying irradiated Na-alginate (100 kGy) resulted in an improvement of the considered characters.

A WOX homolog disrupted by a transposon led to the loss of spines and contributed to the domestication of lettuce.

The loss of spines is one of the most important domestication traits for lettuce (Lactuca sativa). However, the genetics and regulation of spine development in lettuce remain unclear. We examined the genetics of spines in lettuce using a segregating population derived from a cross between cultivated and wild lettuce (Lactuca serriola). A gene encoding WUSCHEL-related homeobox transcription factor, named as WOX-SPINE1 (WS1), was identified as the candidate gene controlling the spine development in lettuce, and its function on spines was verified. A CACTA transposon was found to be inserted into the first exon of the ws1 allele, knocking out its function and leading to the lack of spines in cultivated lettuce. All lettuce cultivars investigated have the nonfunctional ws1 gene, and a selection sweep was found at the WS1 locus, suggesting its important role in lettuce domestication. The expression levels of WS1 were associated with the density of spines among different accessions of wild lettuce. At least two independent loss-of-function mutations in the ws1 gene caused the loss of spines in wild lettuce. These findings provide new insights into the development of spines and facilitate the exploitation of wild genetic resources in future lettuce breeding programs.

Genome assembly and transcriptomic analysis to elucidate the ability of Nasonovia ribisnigri to break host plant resistance.

Aphid genomic resources enable the study of complex life history traits and provide information on vector biology, host adaption and speciation. The currant-lettuce aphid (Nasonovia ribisnigri (Hemiptera: Aphididae) (Mosley)) is a cosmopolitan pest of outdoor lettuce (Lactuca sativa (Asterales: Asteraceae) (Linnaeus)). Until recently, the use of resistant cultivars was an effective method for managing N. ribisnigri. A resistant cultivar containing a single gene (Nr-locus), introduced in the 1980s, conferred complete resistance to feeding. Overreliance of this Nr-locus in lettuce resulted in N. ribisnigri's ability to break resistance mechanism, with first reports during 2003. Our work attempts to understand which candidate gene(s) are associated with this resistance-breaking mechanism. We present two de novo draft assembles for N. ribisnigri genomes, corresponding to both avirulent (Nr-locus susceptible) and virulent (Nr-locus resistant) biotypes. Changes in gene expression of the two N. ribisnigri biotypes were investigated using transcriptomic analyses of RNA-sequencing (RNA-seq) data to understand the potential mechanisms of resistance to the Nr-locus in lettuce. The draft genome assemblies were 94.2% and 91.4% complete for the avirulent and virulent biotypes, respectively. Out of the 18,872 differentially expressed genes, a single gene/locus was identified in N. ribisnigri that was shared between two resistant-breaking biotypes. This locus was further explored and validated in Real-Time Quantitative Reverse Transcription PCR (qRT-PCR) experiments and has predicted localisations in both the cytoplasm and nucleus. This is the first study to provide evidence that a single gene/locus is likely responsible for the ability of N. ribisnigri to overcome the Nr-locus resistance in the lettuce host.

Public perception of folate-biofortified genetically modified lettuce varieties in Brazil.

Lettuce is one of the most widely consumed vegetables in the world, commonly eaten fresh in salads, sandwiches, wraps, and as a garnish in various dishes. Consequently, it is a very promising vehicle to deliver vitamins, such as folate (vitamin B9), to a specific population using biofortified varieties generated by conventional or molecular breeding. A new genetically modified lettuce was generated with increased folate content. However, some issues related to public perception regarding this technology should still be evaluated. The aim of this study was to analyze whether consumers are willing to accept a folate-biofortified GM lettuce that could become available to the Brazilian market. A questionnaire involving several issues regarding lettuce consumption was answered by 2,391 people from almost all Brazilian states. When informed that the folic acid biofortified lettuce is a transgenic plant, 46.1% of respondents stated that they would eat it and 30.5% stated that it would be a possibility. This study demonstrated that if there is any explanation regarding the advantage in relation to the use of biotechnology, like enrichment with folic acid, the number of people who accept it increases.

Effects of dietary nitrate supplementation on oral health and associated markers of systemic health: a systematic review.

Poor oral health can impact an individual's ability to eat and has been associated with an increased risk of non-communicable diseases. While the benefits of nitrate consumption on oral health were first proposed more than 20 years ago, no systematic review has been published examining effects of dietary nitrate on oral health. This systematic review investigated the effects of dietary nitrate on markers of oral health in vivo in randomized controlled trials (RCTs). Five databases (PubMed, The Cochrane Library, CINAHL, MEDLINE, and SPORTDiscus) were searched from inception until March 2023. Nine articles reporting data on 284 participants were included. Dietary nitrate was provided via beetroot juice in most studies. The duration of the interventions ranged from one day to six weeks. Dietary nitrate supplementation increased the relative abundance of several individual bacterial genera including Neisseria and Rothia. Dietary nitrate supplementation increased salivary pH and decreased salivary acidification following consumption of a sugar-sweetened beverage. Furthermore, dietary nitrate supplementation resulted in a decrease in the gingival inflammation index. The results of this systematic review suggest that dietary nitrate could represent a potential nutritional strategy to positively modify oral health by impacting the oral microbiome, altering salivary pH, and minimizing gingival inflammation.

Arbuscular Mycorrhizal Fungus Alleviates Charged Nanoplastic Stress in Host Plants via Enhanced Defense-Related Gene Expressions and Hyphal Capture.

Contamination of small-sized plastics is recognized as a factor of global change. Nanoplastics (NPs) can readily enter organisms and pose significant ecological risks. Arbuscular mycorrhizal (AM) fungi are the most ubiquitous and impactful plant symbiotic fungi, regulating essential ecological functions. Here, we first found that an AM fungus, Rhizophagus irregularis, increased lettuce shoot biomass by 25-100% when exposed to positively and negatively charged NPs vs control, although it did not increase that grown without NPs. The stress alleviation was attributed to the upregulation of gene expressions involving phytohormone signaling, cell wall metabolism, and oxidant scavenging. Using a root organ-fungus axenic growth system treated with fluorescence-labeled NPs, we subsequently revealed that the hyphae captured NPs and further delivered them to roots. NPs were observed at the hyphal cell walls, membranes, and spore walls. NPs mediated by the hyphae were localized at the root epidermis, cortex, and stele. Hyphal exudates aggregated positively charged NPs, thereby reducing their uptake due to NP aggregate formation (up to 5000 nm). This work demonstrates the critical roles of AM fungus in regulating NP behaviors and provides a potential strategy for NP risk mitigation in terrestrial ecosystems. Consequent NP-induced ecological impacts due to the affected AM fungi require further attention.

Research on precise phenotype identification and growth prediction of lettuce based on deep learning.

In recent years, precision agriculture, driven by scientific monitoring, precise management, and efficient use of agricultural resources, has become the direction for future agricultural development. The precise identification and assessment of phenotypes, which serve as external representations of a crop's growth, development, and genetic characteristics, are crucial for the realization of precision agriculture. Applications surrounding phenotypic indices also provide significant technical support for optimizing crop cultivation management and advancing smart agriculture, contributing to the efficient and high-quality development of precision agriculture.This paper focuses on lettuce and employs common nutritional stress conditions during growth as experimental settings. By collecting RGB images throughout the lettuce's complete growth cycle, we developed a deep learning-based computational model to tackle key issues in the lettuce's growth and precisely identify and assess phenotypic indices. We discovered that some phenotypic indices, including custom ones defined in this study, are representative of the lettuce's growth status. By dynamically monitoring the changes in phenotypic traits during growth, we quantitatively analyzed the accumulation and evolution of phenotypic indices across different growth stages. On this basis, a predictive model for lettuce growth and development was trained.The model incorporates MSE, SSIM, and perceptual loss, significantly enhancing the predictive accuracy of the lettuce growth images and phenotypic indices. The model trained with the reconstructed loss function outperforms the original model, with the SSIM and PSNR improving by 1.33% and 10.32%, respectively. The model also demonstrates high accuracy in predicting lettuce phenotypic indices, with an average error less than 0.55% for geometric indices and less than 1.7% for color and texture indices. Ultimately, it achieves intelligent monitoring and management throughout the lettuce's life cycle, providing technical support for high-quality and efficient lettuce production.

Effects of vermicompost on soil microbiological properties in lettuce rhizosphere: An environmentally friendly approach for sustainable green future.

The aim of this study is to investigate the effects of vermicompost on the biological and microbial properties of lettuce rhizosphere in an agricultural field in Samsun, Turkey. The experiment was conducted in a completely randomised design (CRD) and included four vermicompost dosages (0%, 1%, 2%, and 4%) and two application methods (with and without plants). Batavia lettuce was selected as the test plant due to its sensitivity to environmental conditions and nutrient deficiencies. The study evaluated the changes in organic matter (OM), pH, electrical conductivity (EC), carbon dioxide (CO2), dehydrogenase activity (DHA), microbial biomass carbon (MBC), and catalase activity (CA) in the rhizosphere of lettuce plants treated with different vermicompost levels (0%, 1%, 2%, and 4%). The findings showed that vermicompost application significantly increased chlorophyll content in lettuce plants, with the highest content observed in plants treated with V1 compared to the control. Different vermicompost concentrations also influenced chlorophyll b and total chlorophyll levels, with positive effects observed at lower concentrations than the control. Plant height and fresh weight were highest in plants treated with V2, indicating the positive impact of vermicompost on plant growth. Additionally, vermicompost application increased plant dry weight and improved soil properties such as pH, organic matter content, and microbial activity. The findings showed that vermicompost increased the rhizosphere's microbial biomass and metabolic activity, which can be beneficial for plant growth and disease suppression. The study highlights the importance of understanding the effects of organic amendments on soil properties and the microbial community in the rhizosphere, which can contribute to sustainable agricultural practices. Overall, the results suggest that vermicompost can be used as an effective organic amendment for enhancing plant growth and improving soil properties in agricultural fields. Moreover, based on the data, it can be suggested that a dose between 1% and 2% vermicompost is beneficial for the overall growth of plants.

LsMybW-encoding R2R3-MYB transcription factor is responsible for a shift from black to white in lettuce seed.

KEY MESSAGE: We identified LsMybW as the allele responsible for the shift in color from black to white seeds in wild ancestors of lettuce to modern cultivars. Successfully selected white seeds are a key agronomic trait for lettuce cultivation and breeding; however, the mechanism underlying the shift from black-in its wild ancestor-to white seeds remains uncertain. We aimed to identify the gene/s responsible for white seed trait in lettuce. White seeds accumulated less proanthocyanidins than black seeds, similar to the phenotype observed in Arabidopsis TT2 mutants. Genetic mapping of a candidate gene was performed with double-digest RAD sequencing using an F2 population derived from a cross between "ShinanoPower" (white) and "Escort" (black). The white seed trait was controlled by a single recessive locus (48.055-50.197 Mbp) in linkage group 7. Using five PCR-based markers and numerous cultivars, eight candidate genes were mapped in the locus. Only the LG7_v8_49.251Mbp_HinfI marker, employing a single-nucleotide mutation in the stop codon of Lsat_1_v5_gn_7_35020.1, was completely linked to seed color phenotype. In addition, the coding region sequences for other candidate genes were identical in the resequence analysis of "ShinanoPower" and "Escort." Therefore, we proposed Lsat_1_v5_gn_7_35020.1 as the candidate gene and designated it as LsMybW (Lactuca sativa Myb White seeds), an ortholog encoding the R2R3-MYB transcription factor in Arabidopsis. When we validated the role of LsMybW through genome editing, LsMybW knockout mutants harboring an early termination codon showed a change in seed color from black to white. Therefore, LsMybW was the allele responsible for the shift in seed color. The development of a robust marker for marker-assisted selection and identification of the gene responsible for white seeds have implications for future breeding technology and physiological analysis.

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.

Molecular and proteomic response of Pseudomonas fluorescens biofilm cultured on lettuce (Lactuca sativa L.) after ultrasound treatment at different intensity levels.

Ultrasonic treatment is widely used for surface cleaning of vegetables in the processing of agricultural products. In the present study, the molecular and proteomic response of Pseudomonas fluorescens biofilm cultured on lettuce was investigated after ultrasound treatment at different intensity levels. The results show that the biofilm was efficiently removed after ultrasound treatment with intensity higher than 21.06 W/cm2. However, at an intensity of less than 18.42 W/cm2, P. fluorescens was stimulated by ultrasound leading to promoted bacterial growth, extracellular protease activity, extracellular polysaccharide secretion (EPS), and synthesis of acyl-homoserine lactones (AHLs) as quorum-sensing signaling molecules. The expression of biofilm-related genes, stress response, and dual quorum sensing system was upregulated during post-treatment ultrasound. Proteomic analysis showed that ultrasound activated proteins in the flagellar system, which led to changes in bacterial tendency; meanwhile, a large number of proteins in the dual-component system began to be regulated. ABC transporters accelerated the membrane transport of substances inside and outside the cell membrane and equalized the permeability conditions of the cell membrane. In addition, the expression of proteins related to DNA repair was upregulated, suggesting that bacteria repair damaged DNA after ultrasound exposure.

The attachment factors and attachment receptors of human noroviruses.

Human noroviruses (HuNoVs) are the leading etiological agent causing the worldwide outbreaks of acute epidemic non-bacterial gastroenteritis. Histo-blood group antigens (HBGAs) are commonly acknowledged as cellular receptors or co-receptors for HuNoVs. However, certain genotypes of HuNoVs cannot bind with any HBGAs, suggesting potential additional co-factors and attachment receptors have not been identified yet. In addition, food items, such as oysters and lettuce, play an important role in the transmission of HuNoVs. In the past decade, a couple of attachment factors other than HBGAs have been identified and analyzed from foods and microbiomes. Attachment factors exhibit potential as inhibitors of viral binding to receptors on host cells. Therefore, it is imperative to further characterize the attachment factors for HuNoVs present in foods to effectively control the spread of HuNoVs within the food chain. This review summarizes the potential attachment factors/receptors of HuNoVs in humans, foods, and microbiome.

Combined treatment of pulsed light and nisin-organic acid based antimicrobial wash for inactivation of Escherichia coli O157:H7 in Romaine lettuce, reduction of microbial loads, and retention of quality.

Microbial safety of fresh produce continues to be a major concern. Novel antimicrobial methods are needed to minimize the risk of contamination. This study investigated the antimicrobial efficacy of pulsed light (PL), a novel nisin-organic acid based antimicrobial wash (AW) and the synergy thereof in inactivating E. coli O157:H7 on Romaine lettuce. Treatment effects on background microbiota and produce quality during storage at 4 °C for 7 days was also investigated. A bacterial cocktail containing three outbreak strains of E. coli O157:H7 was used as inoculum. Lettuce leaves were spot inoculated on the surface before treating with PL (1-60 s), AW (2 min) or combinations of PL with AW. PL treatment for 10 s, equivalent to fluence dose of 10.5 J/cm2, was optimal and resulted in 2.3 log CFU/g reduction of E. coli O157:H7, while a 2 min AW treatment, provided a comparable pathogen reduction of 2.2 log CFU/g. Two possible treatment sequences of PL and AW combinations were investigated. For PL-AW combination, inoculated lettuce leaves were initially exposed to optimum PL dose followed by 2 min AW treatment, whereas for AW-PL combination, inoculated lettuce were subjected to 2 min AW treatment prior to 10 s PL treatment. Both combination treatments (PL-AW and AW-PL) resulted in synergistic inactivation as E. coli cells were not detectable after treatment, indicating >5 log pathogen reductions. Combination treatments significantly (P < 0.05) reduced spoilage microbial populations on Romaine lettuce and also hindered their growth in storage for 7 days. The firmness and visual quality appearance of lettuce were not significantly (P > 0.05) influenced due to combination treatments. Overall, the results reveal that PL and AW combination treatments can be implemented as a novel approach to enhance microbial safety, quality and shelf life of Romaine lettuce.

Therapeutic potential of hypnotic herbal medicines: A comprehensive review.

Insomnia affects millions of people worldwide, prompting considerable interest in herbal remedies for its treatment. This review aims to assess the therapeutic potential of such remedies for insomnia by analyzing current scientific evidence. The analysis identified several herbs, including Rosmarinus officinalis, Crocus sativus, Rosa damascena, Curcuma longa, Valeriana officinalis, Lactuca sativa, Portulaca oleracea, Citrus aurantium, Lippia citriodora, and Melissa officinalis, which show promise in improving overall sleep time, reducing sleep latency, and enhancing sleep quality. These plants act on the central nervous system, particularly the serotonergic and gamma-aminobutyric acid (GABA)ergic systems, promoting sedation and relaxation. However, further research is necessary to fully understand their mechanisms of action, optimal dosages, and treatment protocols. Combining herbal medicines with conventional treatments may offer an effective natural alternative for those seeking medication. Nevertheless, individuals should consult their healthcare provider before using herbal remedies for insomnia. While this review provides evidence supporting their use, additional high-quality studies are needed to firmly establish their clinical efficacy.

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.

Risk of Illness with Salmonella Spp. and Antibiotic-Resistant Salmonella sp. due to Consumption of Lettuce Irrigated with Water from La Ramada Irrigation District.

Despite heavy contamination of the Bogotá River with domestic and industrial waste, it remains vital for various purposes, including agricultural use at La Ramada Irrigation District. There are important concerns regarding pathogen concentrations in irrigation water at La Ramada, including the presence of antibiotic-resistant Salmonella spp. This study aimed to estimate the risk of Salmonella-related illness from consuming lettuce irrigated with Bogotá River water at La Ramada. We collected lettuce samples from 4 different sites, all irrigated with water from La Ramada. The methodology involved a process to detach Salmonella spp. from lettuce leaves, quantification through plate counts on SS agar, and establishment of antibiotic-resistant bacteria concentrations through growth on media supplemented with ampicillin or ciprofloxacin. The results showed concentrations of Salmonella spp. of 103.59,102.66, and 104.56 CFU/g lettuce at sites 1, 2, and 3, respectively, and ampicillin-resistant Salmonella spp. of 101.93, 101.31, and 102.07 CFU/g lettuce at sites 1, 2, and 3, respectively. No colonies were obtained from lettuce samples collected from site 4. Notably, we detected no isolates resistant to ciprofloxacin at any of the sites. Salmonella spp. concentrations varied greatly among sampling sites. Salmonella spp. concentrations were used to predict the daily probability of illness, with a probability of 0.59 (0.33 to 0.78, CI 95%) for Salmonella spp. and 0.3 (0.03 to 0.53, CI 95%) for ampicillin-resistant Salmonella spp.

Influence of brassinosteroid and silicon on growth, antioxidant enzymes, and metal uptake of leafy vegetables under wastewater irrigation.

Vegetable cultivation under sewage irrigation is a common practice mostly in developing countries due to a lack of freshwater. Long-term usage provokes heavy metals accumulation in soil and ultimately hinders the growth and physiology of crop plants and deteriorates the quality of food. A study was performed to investigate the role of brassinosteroid (BRs) and silicon (Si) on lettuce, spinach, and cabbage under lead (Pb) and cadmium (Cd) contaminated sewage water. The experiment comprises three treatments (control, BRs, and Si) applied under a completely randomized design (CRD) in a growth chamber. BRs and Si application resulted in the highest increase of growth, physiology, and antioxidant enzyme activities when applied under canal water followed by distilled water and sewage water. However, BRs and Si increased the above-determined attributes under the sewage water by reducing the Pb and Cd uptake as compared to the control. It's concluded that sewerage water adversely affected the growth and development of vegetables by increasing Pb and Cd, and foliar spray of Si and BRs could have great potential to mitigate the adverse effects of heavy metals and improve the growth. The long-term alleviating effect of BRs and Si will be evaluated in the field conditions at different ecological zones.