Lettuce Downy Mildew Research Articles

Revealing real-time 3D in vivo pathogen dynamics in plants by label-free optical coherence tomography

Microscopic imaging for studying plant-pathogen interactions is limited by its reliance on invasive histological techniques, like clearing and staining, or, for in vivo imaging, on complicated generation of transgenic pathogens. We present real-time 3D in vivo visualization of pathogen dynamics with label-free optical coherence tomography. Based on intrinsic signal fluctuations as tissue contrast we image filamentous pathogens and a nematode in vivo in 3D in plant tissue. We analyze 3D images of lettuce downy mildew infection (Bremia lactucae) to obtain hyphal volume and length in three different lettuce genotypes with different resistance levels showing the ability for precise (micro) phenotyping and quantification of the infection level. In addition, we demonstrate in vivo longitudinal imaging of the growth of individual pathogen (sub)structures with functional contrast on the pathogen micro-activity revealing pathogen vitality thereby opening a window on the underlying molecular processes.

A Loop-Mediated Isothermal Amplification (LAMP) Assay for the Detection of Bremia lactucae in the Field.

A real-time loop-mediated isothermal amplification (LAMP) assay for the detection of Bremia lactucae, the causal pathogen of lettuce downy mildew, was developed and validated to aid in-field detection of airborne inoculum. Assay specificity was confirmed against a range of other pathogenic oomycete and fungal spp., and sensitivity of the assay for the detection of DNA extracted from sporangia was evaluated. The B. lactucae LAMP assay reliably detected DNA equivalent to 1 spore/reaction (16.7 pg DNA/reaction). Following extraction of DNA from Rotorod air samplers, to which sporangial suspensions were added, the assay reliably detected 25 sporangia/Rotorod. Detection of airborne inoculum of B. lactucae collected through the season from air samplers deployed in-field in plots infected with B. lactucae and in commercial lettuce fields in Scotland over two growing seasons was assessed. The method can be deployed on samples collected from commercial lettuce production to inform disease management strategies and limit the use of unnecessary prophylactic pesticide applications.

Identification and characterization of lettuce cultivars with high inhibitory activity against the human pathogen Escherichia coli O157:H7: Toward a plant-intrinsic hurdle approach to microbial safety

Foodborne illness linked to fruit and vegetables poses a major challenge to public health and horticulture production. Processed lettuce has been implicated in recurrent outbreaks of pathogenic Shiga toxin-producing Escherichia coli (STEC) infection. We hypothesized that plant defenses elicited by mechanical injury may effect STEC inhibition in cut leaves. Intact and cut leaves of 31 lettuce cultivars (Lactuca spp.) selected from >500 accessions based on their resistance to common insect pests and the necrotrophic lettuce drop pathogen were assessed for STEC serovar O157:H7 (EcO157) survival under cold storage conditions. The cultivars were previously ranked also for phenotypes such as resistance to other phytopathogens and physiological traits. Total leaf phenolic compounds, anthocyanins, and reactive oxygen species (ROS); and phenylalanine lyase (PAL), peroxidase (POD), and polyphenol oxidase (PPO) activity were quantified in each cultivar. Principal Component Analysis (PCA) and K-means cluster analysis were applied to group phenotypes and EcO157 population decline on lettuce. In five cultivars, high resistance to herbivorous insects (aphids, leaf miners and thrips), diseases (lettuce drop, downy mildew, and Impatiens Necrotic Spot Virus), and the disorder tipburn was associated with up to 42-fold EcO157 population decline on cold-stored cut (shredded) leaves, and with up to 11-fold difference in decline on cold-stored intact vs cut leaves (ΔD). The decline on these cold-stored cut cultivars also was up to 24-fold greater than on the cultivar with the lowest decline, and up to 8-fold greater than the mean decline for all 31 cultivars (P < 0.01). Resistance to the hemibiotrophic pathogen causing lettuce bacterial leaf spot, Xanthomonas hortorum pv. vitians, was not indicative of high EcO157 inhibition in this system. High PAL activity, and phenolic and anthocyanin content (PPA); and high PAL and POD activity, were associated with high EcO157 population decline (P < 0.05). One cultivar with high susceptibility to herbivorous insects and phytopathogens, and low levels of defense metabolism nevertheless showed high EcO157 inhibition, suggesting other plant inhibitory characteristics at play. Basal plant immunity and pathways associated with a strong wound response may inform about enteric pathogen survival on minimally processed postharvest lettuce. The related traits identified in this study may serve to breed new genotypes with intrinsically enhanced microbial safety.

Multiplex Quantitative PCR Assay for Detection of Spinach and Lettuce Downy Mildews Using Spore Trapping

Downy mildews are major constraints on spinach and lettuce production globally. Disease management can be achieved with fungicides, but routine applications are costly and can lead to pathogen resistance. Detection of airborne spores could guide sustainable fungicide application, and, considering that spinach and lettuce are often grown in the same cycles, simultaneous detection of both pathogens is practical. Here, a multiplex hydrolysis probe quantitative PCR assay was designed using single copy mitochondrial DNA targets for spinach downy mildew ( Peronospora effusa) and lettuce downy mildew ( Bremia lactucae). To quantify P. effusa and B. lactucae sporangia, a standard curve was developed for each pathogen using the multiplex qPCR to amplify DNA obtained from known dilutions of sporangia. Analysis of these curves revealed a greater sensitivity for B. lactucae, indicating that the sporangia of B. lactucae may harbor more mitochondria than those of P. effusa, providing insight into the biology of these pathogens. The multiplex qPCR assay was partnered with two different spore trap types: a cyclone spore trap and an impaction spore trap. Results from air sampling revealed that the cyclone spore traps collect significantly more sporangia compared with impaction traps. Exposure of P. effusa sporangia to desiccation was performed to assess the environmental impact on the assay, and although detection levels were reduced, they were still apparent. This detection and quantification tool will be useful in efforts to improve the accuracy of downy mildew forecasting, which in return may reduce fungicide usage or improve its efficiency. [Formula: see text] The author(s) have dedicated the work to the public domain under the Creative Commons CC0 “No Rights Reserved” license by waiving all of his or her rights to the work worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law, 2024.

New clades of viruses infecting the obligatory biotroph Bremia lactucae representing distinct evolutionary trajectory for viruses infecting oomycetes.

Recent advances in high throughput sequencing (HTS) approaches allowed a broad exploration of viromes from different fungal hosts, unveiling a great diversity of mycoviruses with interesting evolutionary features. The word mycovirus historically applies also to viruses infecting oomycetes but most studies are on viruses infecting fungi, with less mycoviruses found and characterized in oomycetes, particularly in the obligatory biotrophs. We, here, describe the first virome associated to Bremia lactucae, the causal agent of lettuce downy mildew, which is an important biotrophic pathogen for lettuce production and a model system for the molecular aspects of the plant-oomycetes interactions. Among the identified viruses, we could detect (1) two new negative sense ssRNA viruses related to the yueviruses, (2) the first example of permuted RdRp in a virus infecting fungi/oomycetes, (3) a new group of bipartite dsRNA viruses showing evidence of recent bi-segmentation and concomitantly, a possible duplication event bringing a bipartite genome to tripartite, (4) a first representative of a clade of viruses with evidence of recombination between distantly related viruses, (5) a new open reading frame (ORF)an virus encoding for an RdRp with low homology to known RNA viruses, and (6) a new virus, belonging to riboviria but not conserved enough to provide a conclusive phylogenetic placement that shows evidence of a recombination event between a kitrinoviricota-like and a pisuviricota-like sequence. The results obtained show a great diversity of viruses and evolutionary mechanisms previously unreported for oomycetes-infecting viruses, supporting the existence of a large diversity of oomycetes-specific viral clades ancestral of many fungal and insect virus clades.

EffectorO: Motif-Independent Prediction of Effectors in Oomycete Genomes Using Machine Learning and Lineage Specificity.

Oomycete plant pathogens cause a wide variety of diseases, including late blight of potato, sudden oak death, and downy mildews of plants. These pathogens are major contributors to loss in numerous food crops. Oomycetes secrete effector proteins to manipulate their hosts to the advantage of the pathogen. Plants have evolved to recognize effectors, resulting in an evolutionary cycle of defense and counter-defense in plant-microbe interactions. This selective pressure results in highly diverse effector sequences that can be difficult to computationally identify using only sequence similarity. We developed a novel effector prediction tool, EffectorO, that uses two complementary approaches to predict effectors in oomycete pathogen genomes: i) a machine learning-based pipeline that predicts effector probability based on the biochemical properties of the N-terminal amino-acid sequence of a protein and ii) a pipeline based on lineage specificity to find proteins that are unique to one species or genus, a sign of evolutionary divergence due to adaptation to the host. We tested EffectorO on Bremia lactucae, which causes lettuce downy mildew, and Phytophthora infestans, which causes late blight of potato and tomato, and predicted many novel effector candidates while recovering the majority of known effector candidates. EffectorO will be useful for discovering novel families of oomycete effectors without relying on sequence similarity to known effectors. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

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.

Effectiveness of COPPION 5.2 WV and Rhapynal to control downy mildew of lettuce

The most important lettuce pathogen under Greek agro-ecological conditions is Bremia lactucae, the causal agent of downy mildew. Chemical control methods are widely used to manage this pathogen, with copper fungicides of tensed as the first line of defence, especially for growers who prefer environmentally friendly methods. The main aim of this work was to investigate the effectiveness of the biosurfactant Rhapynal (rhamnolipids) and the commercial copper product COPPION 5.2 WV (an ionic form copper) against downy mildew of lettuce. The experiments were conducted under both greenhouse and field conditions. COPPION 5.2 WV was applied at a rate of 2 mLL−1, while Rhapynal was applied on its own at a rate of 1.5 gL−1 or in a mixture with COPPION 5.2 WV at rates of 0.5, 1.0 and 1.5 gL−1. In greenhouse experiments, COPPION 5.2 WV alone or in a mixture with 0.5, 1.0, or 1.5 gL−1 Rhapynal were the most effective treatments to control downy mildew of lettuce. In field experiments, the percentage of infection in plants treated with COPPION 5.2 WV in a mixture with 0.5, 1, or 1.5 gL−1 Rhapynal was statistically similar. No significant differences were found among treatments with COPPION 5.2 WV, COPPION 5.2 WV in a mixture with 0.5 or 1 gL−1 Rhapynal, or Rhapynal on its own at 1.5 gL−1. Based on the results of this study, a mixture of COPPION 5.2 WV with Rhapynal seems to be a promising alternative method to control downy mildew on lettuce plants.

Genome-wide association mapping reveals genomic regions frequently associated with lettuce field resistance to downy mildew.

GWAS identified 63 QTLs for resistance to downy mildew. Though QTLs were distributed across all chromosomes, the genomic regions frequently associated with resistance were located on chromosomes 4 and 5. Lettuce downy mildew is one of the most economically important diseases of cultivated lettuce worldwide. We have applied the genome-wide association mapping (GWAS) approach to detect QTLs for field resistance to downy mildew in the panel of 496 accessions tested in 21 field experiments. The analysis identified 131 significant marker-trait associations that could be grouped into 63 QTLs. At least 51 QTLs were novel, while remaining 12 QTLs overlapped with previously described QTLs for lettuce field resistance to downy mildew. Unlike race-specific, dominant Dm genes that mostly cluster on three out of nine lettuce chromosomes, QTLs (qDMR loci) for polygenic resistance are randomly distributed across all nine chromosomes. The genomic regions frequently associated with lettuce field resistance to downy mildew are located on chromosomes 4 and 5 and could be used for detailed study of the mechanism of polygenic resistance. The most resistant accessions identified in the current study (cvs. Auburn, Grand Rapids, Romabella, PI 226514, and PI 249536) are being incorporated into our breeding program. Markers closely linked to the resistance QTLs could be potentially used for marker-assisted selection, or in combination with other markers in the genome, for a combined genomic and marker-assisted selection. Up to date this is the most comprehensive study of QTLs for field resistance to downy mildew and the first study that uses GWAS for mapping disease resistance loci in lettuce.

Lactuca georgica, a new wild source of resistance to downy mildew: comparative study to other wild lettuce relatives

Seedlings of 431 accessions representing six and five Armenian natural populations of the wild Lactuca georgica and L. altaica, respectively, and 32, 23, and 20 populations of L. saligna, L. serriola, and L. aculeata, respectively (mostly Israeli natural populations), were screened at seedling stage for resistance to six highly virulent races of Bremia lactucae - the causal of lettuce downy mildew - that were found on several widely grown cultivars and represent the virulence spectrum in the main lettuce production areas in Europe and California. This study is likely the first detailed screening of resistance to B. lactucae races in natural populations of L. georgica and L. altaica. The highest average resistance probability and frequency of highly resistant accessions across races were detected in L. georgica. These were even higher than those shown by the germplasm of known non-host resistance (NHR) species L. saligna, which classified to the secondary lettuce gene pool (LGP-2). Thus, we suggest that L. georgica is a new LGP-2 source of resistance to B. lactucae. Our findings also support previous observations that L. aculeata, a species within the LGP-1, should be prioritized as a source of downy mildew resistance in domesticated lettuce (L. sativa) breeding programs, since some L. aculeata accessions expressed highly resistance across races. For L. serriola, the closest relative of cultivated lettuce, and for L. altaica, also a closer or even a primitive form of L. sativa, probabilities of resistance to each of the investigated B. lactucae races were very low.

Identification of Major Quantitative Trait Loci Controlling Field Resistance to Downy Mildew in Cultivated Lettuce (Lactuca sativa).

Lettuce downy mildew, caused by Bremia lactucae Regel, is the most economically important foliar disease of lettuce (Lactuca sativa L.). The deployment of resistant cultivars carrying dominant resistance genes (Dm genes) plays a crucial role in integrated downy mildew disease management; however, high variability in pathogen populations leads to the defeat of plant resistance conferred by Dm genes. Some lettuce cultivars exhibit field resistance that is only manifested in adult plants. Two populations of recombinant inbred lines (RILs), originating from crosses between the field resistant cultivars Grand Rapids and Iceberg and susceptible cultivars Salinas and PI491224, were evaluated for downy mildew resistance under field conditions. In all, 160 RILs from the Iceberg × PI491224 and 88 RILs from the Grand Rapids × Salinas populations were genotyped using genotyping by sequencing, which generated 906 and 746 high-quality markers, respectively, that were used for quantitative trait locus (QTL) analysis. We found a QTL in chromosome 4 that is present in both Grand Rapids × Salinas and Iceberg × PI491224 populations that has a major effect on field resistance. We also found two additional significant QTLs in chromosomes 2 and 5 in the Iceberg × PI491224 RIL population. Marker-assisted gene pyramiding of multiple Dm genes in combination with QTLs for field resistance provide the opportunity to develop cultivars with more durable resistance to B. lactucae.

Study on the effect of different adjuvants on dimethomorph against lettuce downy mildew

Study on the effect of different adjuvants on dimethomorph against lettuce downy mildew

Downy Mildew of Lettuce in Florida

Lettuce Downy Mildew (LDM), caused by the oomycete &lt;i&gt;Bremia lactucae&lt;/i&gt;, is the most important disease of lettuce worldwide. LDM has a direct effect on both yield quantity and quality because it may infect lettuce at any growth stage, affecting the marketable portion of the crop. This new 6-page publication of the UF/IFAS Horticultural Sciences Department introduces the LDM disease in Florida lettuce and available control methods and strategies. This publication also introduces the work on LDM in the UF/IFAS Lettuce Breeding Program, which was created to release cultivars adapted to Florida conditions. Written by Lis Rodrigues-Porto, Richard N. Raid, and Germán V. Sandoya.https://edis.ifas.ufl.edu/hs1403

Monitoring virulence of Bremia lactucae as a breeding tool against lettuce downy mildew from south and southwest Brazilian regions

Lettuce downy mildew caused by Bremia lactucae is one of the main diseases in high humidity and low temperature conditions. The identification of virulence factors appearing in the pathogen population could help improve breeding programs against this disease. This study aimed to monitor and evaluate virulence dynamic changes in B. lactucae virulence among two Brazilian regions to recommend resistance genes for breeding programs. Isolates from Sao Paulo, Rio Grande do Sul, and Parana states were assessed using the sextet code EU-C composed by 16 Lactuca spp. genotypes, between 2015 and 2016. To understand the population dynamics, the frequencies of virulence phenotypes and factors and the virulence complexity per isolate (Ci), phenotype (Cp), and Gleason (Ig), indexes were calculated. B. lactucae virulence from Parana, Rio Grande do Sul and Sao Paulo were similar and shared nine out of the 15 evaluated virulence factors. In total, 90 isolates were analyzed, and 27 virulence phenotypes were found. The most frequent sextet codes were 31–00-00, 31–00-02, 31–01-00, and 31–01-02. The genes or resistance factors present in ‘Argeles’ (Dm38), ‘Balesta’, and ‘Bartoli’ could be used as sources of resistance by Brazilian lettuce breeders.

Identification and mapping of new genes for resistance to downy mildew in lettuce

Key messageEleven new major resistance genes for lettuce downy mildew were introgressed from wild Lactuca species and mapped to small regions in the lettuce genome.Downy mildew, caused by the oomycete pathogen Bremia lactucae Regel, is the most important disease of lettuce (Lactuca sativa L.). The most effective method to control this disease is by using resistant cultivars expressing dominant resistance genes (Dm genes). In order to counter changes in pathogen virulence, multiple resistance genes have been introgressed from wild species by repeated backcrosses to cultivated lettuce, resulting in numerous near-isogenic lines (NILs) only differing for small chromosome regions that are associated with resistance. Low-pass, whole genome sequencing of 11 NILs was used to identify the chromosome segments introgressed from the wild donor species. This located the candidate chromosomal positions for resistance genes as well as additional segments. F2 segregating populations derived from these NILs were used to genetically map the resistance genes to one or two loci in the lettuce reference genome. Precise knowledge of the location of new Dm genes provides the foundation for marker-assisted selection to breed cultivars with multiple genes for resistance to downy mildew.

Effector prediction and characterization in the oomycete pathogen Bremia lactucae reveal host-recognized WY domain proteins that lack the canonical RXLR motif.

Pathogens that infect plants and animals use a diverse arsenal of effector proteins to suppress the host immune system and promote infection. Identification of effectors in pathogen genomes is foundational to understanding mechanisms of pathogenesis, for monitoring field pathogen populations, and for breeding disease resistance. We identified candidate effectors from the lettuce downy mildew pathogen Bremia lactucae by searching the predicted proteome for the WY domain, a structural fold found in effectors that has been implicated in immune suppression as well as effector recognition by host resistance proteins. We predicted 55 WY domain containing proteins in the genome of B. lactucae and found substantial variation in both sequence and domain architecture. These candidate effectors exhibit several characteristics of pathogen effectors, including an N-terminal signal peptide, lineage specificity, and expression during infection. Unexpectedly, only a minority of B. lactucae WY effectors contain the canonical N-terminal RXLR motif, which is a conserved feature in the majority of cytoplasmic effectors reported in Phytophthora spp. Functional analysis of 21 effectors containing WY domains revealed 11 that elicited cell death on wild accessions and domesticated lettuce lines containing resistance genes, indicative of recognition of these effectors by the host immune system. Only two of the 11 recognized effectors contained the canonical RXLR motif, suggesting that there has been an evolutionary divergence in sequence motifs between genera; this has major consequences for robust effector prediction in oomycete pathogens.

Essential Tea Tree Oil Activity against Bremia lactucae in Lettuce

Downy mildew caused by the oomycete Bremia lactucae Regel is a serious disease of lettuce in field and greenhouse production. Here we report on the antifungal activity of essential Australian tea tree oil (TTO) derived from Melaleuca alternifolia against downy mildew in lettuce. Preventive treatments with Timorex Gold (STK Ltd., Petach Tikva, Israel), fungicide containing TTO, suppressed the development of B. lactucae on cotyledons and young lettuce plants. Epifluorescence microscopy showed that TTO had a moderate inhibitory effect on spore germination of the pathogen but a strong inhibitory effect on sporophore emergence and hence on sporulation. Timorex Gold (22.3 EC W/V) was as effective as copper hydroxide in controlling the disease in organic field plots. It was as effective as the fungicides Bellis (Boscalid + Pyraclostrobin; BASF, Germany) and Commet (pyraclostrobin; BASF, Germany) in conventional production. The results demonstrate that Timorex Gold effectively suppresses the development of downy mildew in lettuce in growth chambers as well as in the field, and thus suitable biopesticide for its control.

Host interactors of effector proteins of the lettuce downy mildew Bremia lactucae obtained by yeast two-hybrid screening.

Plant pathogenic bacteria, fungi and oomycetes secrete effector proteins to manipulate host cell processes to establish a successful infection. Over the last decade the genomes and transcriptomes of many agriculturally important plant pathogens have been sequenced and vast candidate effector repertoires were identified using bioinformatic analyses. Elucidating the contribution of individual effectors to pathogenicity is the next major hurdle. To advance our understanding of the molecular mechanisms underlying lettuce susceptibility to the downy mildew Bremia lactucae, we mapped physical interactions between B. lactucae effectors and lettuce candidate target proteins. Using a lettuce cDNA library-based yeast-two-hybrid system, 61 protein-protein interactions were identified, involving 21 B. lactucae effectors and 46 unique lettuce proteins. The top ten interactors based on the number of independent colonies identified in the Y2H and two interactors that belong to gene families involved in plant immunity, were further characterized. We determined the subcellular localization of the fluorescently tagged lettuce proteins and their interacting effectors. Importantly, relocalization of effectors or their interactors to the nucleus was observed for four protein-pairs upon their co-expression, supporting their interaction in planta.

Distribution of races and virulence factors of Bremia lactucae in the main lettuce production area in Brazil

Lettuce downy mildew, caused by Bremia lactucae, occurs throughout Sao Paulo, the main lettuce production state in Brazil, during winter due to low temperatures and high humidity. Thus, the aim of this work was to monitor the races and virulence factors in Sao Paulo’s B. lactucae population from 2003 to 2015, and to compare the results with similar studies in other countries. Data from 582 isolates collected from Lactuca sativa cultivars in 33 municipalities of Sao Paulo were analyzed. Seventeen virulence patterns (races) were identified. Only few races have been stable over the years. The virulence factors v1, v2, v3, v4, v5/8, v7, v10, v11, v12 and v13 have always been present since the first screening in 2003 while the factors v6, v14, v15, v16, v36 and v37 were found in varying frequencies. The population diversity in Sao Paulo increased over time with five new races emerging in 2014 and 2015. However, the virulence factors identified are the same as previously described in other areas. The races of Sao Paulo state are similar to the new races from Australia, California, Europe and Japan, grouping in the same clade by analysis the virulence patterns. The genes Dm17, Dm18, R38 and the resistance factors of the cultivars Balesta and Bellisimo can be recommended as source of resistance to the pathotypes characterized to date in Sao Paulo.

Measurements of Aerial Spore Load by qPCR Facilitates Lettuce Downy Mildew Risk Advisement.

The lettuce downy mildew pathogen, Bremia lactucae, is an obligate oomycete that causes extensive produce losses. Initial chlorotic symptoms that severely reduce the market value of the produce are followed by the appearance of white, downy sporulation on the abaxial side of the leaves. These spores become airborne and disseminate the pathogen. Controlling lettuce downy mildew has relied on repeated fungicide applications to prevent outbreaks. However, in addition to direct economic costs, heterogeneity and rapid adaptation of this pathogen to repeatedly applied fungicides has led to the development of fungicide-insensitivity in the pathogen. We deployed a quantitative PCR assay-based detection method using a species-specific DNA target for B. lactucae coupled with a spore trap system to measure airborne B. lactucae spore loads within three commercial fields that each contained experimental plots, designated EXP1 to EXP3. Based upon these measurements, when the spore load in the air reached a critical level (8.548 sporangia per m3 air), we advised whether or not to apply fungicides on a weekly basis within EXP1 to EXP3. This approach saved three sprays in EXP1, and one spray each in EXP2 and EXP3 without a significant increase in disease incidence. The reduction in fungicide applications to manage downy mildew can decrease lettuce production costs while slowing the development of fungicide resistance in B. lactucae by eliminating unnecessary fungicide applications.