Opportunities for Improved Potato Late Blight Management in the Republic of Ireland: Field Evaluation of the Modified Irish Rules Crop Disease Risk Prediction Model.

Potato late blight remains the most significant disease threat of potato cultivation globally, often requiring expensive, time-consuming, and environmentally unfriendly approaches to disease management. The goal of this research was to evaluate whether an estimation of potato late blight risk based on environmental factors can be reliably used to adjust the standard potato late blight management practices and the role of cultivar resistance under growing conditions and contemporary Phytophthora infestans populations in the Republic of Ireland. The modified Irish Rules model made it possible to reduce fungicide usage by 58.7% on average, compared with current standard practices used by growers and without adversely compromising disease control and yield, with similar results achieved by the half-dose program. Host resistance levels were found to be correlated with a delay in the initiation of the epidemics, final foliar disease levels, and reduction of fungicide usage. Disease levels on the highly resistant cultivars remained low, and a clear selection pattern toward the P. infestans genotypes EU_13_A2 and EU_6_A1 was observed. An increase in the frequency of strains belonging to genotypes EU_13_A2 and EU_6_A1 was also observed to occur in the latter part of the trial growing seasons. Because of the increasingly dynamic nature of the population structure, associated with the continued evolution of the P. infestans population and the arrival of EU_36_A2 in the Republic of Ireland, routine population monitoring is necessary to ensure that potato late blight control strategies remain effective.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Isolate-Specific Responses of the Nonhost Grass Brachypodium distachyon to the Fungal Pathogen Zymoseptoria tritici Compared with Wheat.

Septoria tritici blotch (STB) is an important foliar disease of wheat that is caused by the fungal pathogen Zymoseptoria tritici. The grass Brachypodium distachyon has been used previously as a model system for cereal-pathogen interactions. In this study, we examined the nonhost resistance (NHR) response of B. distachyon to two different Z. tritici isolates in comparison with wheat. These isolates vary in aggressiveness on wheat cultivar Remus, displaying significant differences in disease and pycnidia coverage. Using microscopy, we found that similar isolate-specific responses were observed for hydrogen peroxide accumulation and cell death in both wheat and B. distachyon. Despite this, induction of isolate-specific patterns of defense gene expression by Z. tritici did differ between B. distachyon and wheat. Our results suggest that expression of the phenylalanine ammonia lyase PAL gene may be important for NHR in B. distachyon, while pathogenesis-related PR genes and expression of genes regulating reactive oxygen species may be important to limit disease in wheat. Future studies of the B. distachyon-Z. tritici interaction may allow identification of conserved plant immunity targets that are responsible for the isolate-specific responses observed in both plant species.

Mefentrifluconazole sensitivity amongst European Zymoseptoria tritici populations and potential implications for its field efficacy.

BACKGROUND: Septoria tritici blotch caused by Zymoseptoria tritici continues to be one of the most economically destructive diseases of winter wheat in north-western Europe. Control is heavily reliant on the application of fungicides, in particular those belonging to the azole group. Here we describe the sensitivity of European Z. tritici populations to the novel azole mefentrifluconazole and the analysis of associated mechanisms of resistance. RESULTS: A wide range of sensitivity to mefentrifluconazole was observed amongst the Z. tritici collections examined, with strong cross-resistances also observed between mefentrifluconazole, difenoconazole and tebuconazole. Overall, the Irish population displayed the lowest sensitivity to all azoles tested. Further detailed analysis of the Irish population in 2021 demonstrated differences in sensitivity occurred between sampling sites, with these differences associated with the frequencies of key resistance mechanisms (CYP51 alterations and MFS1 promoter inserts linked to overexpression). Under glasshouse conditions reductions in the efficacy of mefentrifluconazole were observed towards those strains exhibiting the lowest in vitro sensitivities. CONCLUSIONS: This study demonstrates that a large range of sensitivity to mefentrifluconazole exists in European Z. tritici populations. Those strains exhibiting the lowest sensitivity to the azoles tested had the most complex CYP51 haplotypes in combination with the 519 bp insert, associated with enhanced activity of MFS1. The future use of mefentrifluconazole should take these findings into consideration to minimise the selection of these strains. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Baseline sensitivity of European Zymoseptoria tritici populations to the complex III respiration inhibitor fenpicoxamid.

BACKGROUND: Fenpicoxamid is a recently developed fungicide belonging to the quinone inside inhibitor (QiI) group. This is the first fungicide within this group to be active against the Zymoseptoria tritici, which causes Septoria tritici blotch on wheat. The occurrence of pre-existing resistance mechanisms was monitored, using sensitivity assays and Illumina sequencing, in Z. tritici populations sampled in multiple European countries before the introduction of fenpicoxamid. RESULTS: Although differences in sensitivity to all three fungicides tested (fenpicoxamid, fentin chloride and pyraclostrobin) existed between the isolate collections, no alterations associated with QiI resistance were detected. Among the isolates, a range in sensitivity to fenpicoxamid was observed (ratio between most sensitive/least sensitive = 53.1), with differences between the most extreme isolates when tested in planta following limited fenpicoxamid treatment. Sensitivity assays using fentin chloride suggest some of the observed differences in fenpicoxamid sensitivity are associated with multi-drug resistance. Detailed monitoring of the wider European population using Illumina-based partial sequencing of the Z. tritici also only detected the presence of G143A, with differences in frequencies of this alteration observed across the region. CONCLUSIONS: This study provides a baseline sensitivity for European Z. tritici populations to fenpicoxamid. Target-site resistance appears to be limited or non-existing in European Z. tritici populations prior to the introduction of fenpicoxamid. Non-target site resistance mechanisms exist, but their impact in the field is predicted to be limited. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Azole fungicide sensitivity and molecular mechanisms of reduced sensitivity in Irish Pyrenopeziza brassicae populations.

BACKGROUND: Light leaf spot, caused by Pyrenopeziza brassicae, is amongst the most damaging diseases of winter oilseed rape (Brassica napus), and currently the sterol 14α-demethylase (CYP51) inhibitors (azoles) represent the main class of fungicides used to control light leaf spot development. However, a shift in sensitivity to azole fungicides in P. brassicae populations has been observed in different European countries, including Ireland. RESULTS: To assess the sensitivity status of Irish P. brassicae populations to azole fungicides, three collections of P. brassicae from 2018-2020 were tested in vitro against tebuconazole and prothioconazole-desthio, and the PbCYP51 gene targeted by this class of fungicides was genotyped in different isolates. A change in sensitivity to azole fungicides was observed and differences in sensitivity to tebuconazole between Irish populations were present. There were two substitutions within PbCYP51 (G460S and S508T) and inserts of different sizes in its promoter region. The presence of the G460S/S508T double mutant was reported for the first time, and the diversity in insert size was greater than previously known. Compared to wild type isolates, those carrying G460S or S508T were less sensitive to both fungicides and, where inserts were also identified, they further reduced sensitivity to azole fungicides. CONCLUSIONS: The results of this study suggest that azole fungicides are still very effective in controlling light leaf spot in Ireland. However, using azole fungicides in mixtures of fungicides with different modes of action is recommended. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Shifting sensitivity of septoria tritici blotch compromises field performance and yield of main fungicides in Europe.

Septoria tritici blotch (STB; Zymoseptoria tritici) is a severe leaf disease on wheat in Northern Europe. Fungicide resistance in the populations of Z. tritici is increasingly challenging future control options. Twenty-five field trials were carried out in nine countries across Europe from 2019 to 2021 to investigate the efficacy of specific DMI and SDHI fungicides against STB. During the test period, two single DMIs (prothioconazole and mefentrifluconazole) and four different SDHIs (fluxapyroxad, bixafen, benzovindiflupyr and fluopyram) along with different co-formulations of DMIs and SDHIs applied at flag leaf emergence were tested. Across all countries, significant differences in azole performances against STB were seen; prothioconazole was outperformed in all countries by mefentrifluconazole. The effects also varied substantially between the SDHIs, with fluxapyroxad providing the best efficacy overall, while the performance of fluopyram was inferior to other SDHIs. In Ireland and the UK, the efficacy of SDHIs was significantly lower compared with results from continental Europe. This reduction in performances from both DMIs and SDHIs was reflected in yield responses and also linked to decreased sensitivity of Z. tritici isolates measured as EC50 values. A clear and significant gradient in EC50 values was seen across Europe. The lower sensitivity to SDHIs in Ireland and the UK was coincident with the prevalence of SDH-C-alterations T79N, N86S, and sporadically of H152R. The isolates' sensitivity to SDHIs showed a clear cross-resistance between fluxapyroxad, bixafen, benzovindiflupyr and fluopyram, although the links with the latter were less apparent. Co-formulations of DMIs + SDHIs performed well in all trials conducted in 2021. Only minor differences were seen between fluxapyroxad + mefentrifluconazole and bixafen + fluopyram + prothioconazole; the combination of benzovindiflupyr + prothioconazole gave an inferior performance at some sites. Fenpicoxamid performed in line with the most effective co-formulations. This investigation shows a clear link between reduced field efficacy by solo SDHIs as a result of increasing problems with sensitivity shifting and the selection of several SDH-C mutations. The presented data stress the need to practice anti-resistance strategies to delay further erosion of fungicide efficacy.

Natural Co-Occurrence of Multiple Mycotoxins in Unprocessed Oats Grown in Ireland with Various Production Systems.

The natural co-occurrence of 42 mycotoxins was investigated in unprocessed oat grains grown in Ireland. The sample set included a total of 208 oat crops harvested during 2015-2016 and produced using conventional, organic, or gluten free farming systems. A range of different toxins was identified, including the major type A (neosolaniol, HT-2 and T-2 toxins, T-2 triol, and T-2-glucoside, co-occurring in 21 samples) and B trichothecenes (deoxynivalenol, nivalenol, and deoxynivalenol-3-glucoside), enniatins (B1, B, and A1, co-occurring in 12 samples), as well as beauvericin, alternariol, mycophenolic acid, and sterigmatocystin. The influences of sowing season, year, and production system were investigated, eventually indicating that the latter factor may have a higher impact than others on the production of certain mycotoxins in oats. The most frequently quantified compounds were HT-2 (51%) and T-2 (41%) toxins, with gluten free oats containing significantly lower concentrations of HT-2 compared to conventionally produced oats. Although the prevalence and concentrations of mycotoxin found in oat samples in this study should be substantially reduced by processing. However, as mycotoxin occurrence is clearly influenced by multiple factors, controlled field trials should be carried out to define optimal agronomic practices and mitigate mycotoxin production. Furthermore, this work highlights the need for regularly testing cereal-based foods with multi-residue analytical methods with wider specificities than the traditionally screened and regulated toxins, to generate knowledge on the occurrence of several mycotoxins that are, to date, rarely investigated.

Development of a PacBio Long-Read Sequencing Assay for High Throughput Detection of Fungicide Resistance in Zymoseptoria tritici.

Fungicide resistance has become a challenging problem in management of Septoria tritici blotch (STB), caused by Zymoseptoria tritici, the most destructive disease of winter wheat throughout western and northern Europe. To ensure the continued effectiveness of those fungicides currently used, it is essential to monitor the development and spread of such resistance in field populations of the pathogen. Since resistance to the key families of fungicides used for STB control (demethyalation inhibitors or azoles, succinate dehydrogenase inhibitors or SDHIs and Quinone outside Inhibitors or QoIs) is conferred through target-site mutations, the potential exists to monitor resistance through the molecular detection of alterations in the target site genes. As more efficient fungicides were developed and applied, the pathogen has continuously adapted through accumulating multiple target-site alterations. In order to accurately monitor these changes in field populations, it is therefore becoming increasingly important to completely sequence the targeted genes. Here we report the development of a PacBio assay that facilitates the multiplex amplification and long-read sequencing of the target gene(s) for the azole (CYP51), SDHI (Sdh B, C, and D), and QoI (cytochrome b) fungicides. The assay was developed and optimised using three Irish Z. tritici collections established in spring 2017, which capture the range of fungicide resistance present in modern European populations of Z. tritici. The sequences obtained through the PacBio assay were validated using traditional Sanger sequencing and in vitro sensitivity screenings. To further exploit the long-read and high throughput potential of PacBio sequencing, an additional nine housekeeping genes (act, BTUB, cal, cyp, EF1, GAPDH, hsp80-1, PKC, TFC1) were sequenced and used to provide comprehensive Z. tritici strain genotyping.

Identifying the drivers and constraints to adoption of IPM among arable farmers in the UK and Ireland.

BACKGROUND: Arable crops in temperate climatic regions such as the UK and Ireland are subject to a multitude of pests (weeds, diseases and vertebrate/invertebrate pests) that can negatively impact productivity if not properly managed. Integrated pest management (IPM) is widely promoted as a sustainable approach to pest management, yet there are few recent studies assessing adoption levels and factors influencing this in arable cropping systems in the UK and Ireland. This study used an extensive farmer survey to address both these issues. RESULTS: Adoption levels of various IPM practices varied across the sample depending on a range of factors relating to both farm and farmer characteristics. Positive relationships were observed between IPM adoption and farmed area, and familiarity with IPM. Choice of pest control information sources was also found to be influential on farmer familiarity with IPM, with those who were proactive in seeking information from impartial sources being more engaged and reporting higher levels of adoption. CONCLUSION: Policies that encourage farmers to greater levels of engagement with their pest management issues and more proactive information seeking, such as through advisory professionals, more experienced peers through crop walks, open days and discussion groups should be strongly encouraged.

Spatio-temporal distribution of DMI and SDHI fungicide resistance of Zymoseptoria tritici throughout Europe based on frequencies of key target-site alterations.

BACKGROUND: Over the past decade, demethylation inhibitor (DMI) and succinate dehydrogenase inhibitor (SDHI) fungicides have been extensively used to control to septoria tritici blotch, caused by Zymoseptoria tritici on wheat. This has led to the development and selection of alterations in the target-site enzymes (CYP51 and SDH, respectively). RESULTS: Taking advantage of newly and previously developed qPCR assays, the frequency of key alterations associated with DMI (CYP51-S524T) and SDHI (SDHC-T79N/I, C-N86S and C-H152R) resistance was assessed in Z. tritici-infected wheat leaf samples collected from commercial crops (n = 140) across 14 European countries prior to fungicide application in the spring of 2019. This revealed the presence of a West to East gradient in the frequencies of the most common key alterations conferring azole (S524T) and SDHI resistance (T79N and N86S), with the highest frequencies measured in Ireland and Great Britain. These observations were corroborated by sequencing (CYP51 and SDH subunits) and sensitivity phenotyping (prothioconazole-desthio and fluxapyroxad) of Z. tritici isolates collected from a selection of field samples. Additional sampling made at the end of the 2019 season confirmed the continued increase in frequency of the targeted alterations. Investigations on historical leaf DNA samples originating from different European countries revealed that the frequency of all key alterations (except C-T79I) has been gradually increasing over the past decade. CONCLUSION: Whilst these alterations are quickly becoming dominant in Ireland and Great Britain, scope still exists to delay their selection throughout the wider European population, emphasizing the need for the implementation of fungicide antiresistance measures. © 2021 Society of Chemical Industry.

Measuring the unmeasurable? A method to quantify adoption of integrated pest management practices in temperate arable farming systems.

BACKGROUND: The impetus to adopt integrated pest management (IPM) practices has re-emerged in the last decade, mainly as a result of legislative and environmental drivers. However, a significant deficit exists in the ability to practically monitor and measure IPM adoption across arable farms; therefore, the aim of the project reported here was to establish a universal metric for quantifying adoption of IPM in temperate arable farming. This was achieved by: (i) identifying a set of key activities that contribute to IPM; (ii) weighting these in terms of their importance to the achievement of IPM using panels of expert stakeholders to create the metric (scoring system from 0 to 100 indicating level of IPM practised); (iii) surveying arable farmers in the UK and Ireland about their pest management practices; and (iv) measuring level of farmer adoption of IPM using the new metric. RESULTS: This new metric was found to be based on a consistent conception of IPM between countries and professional groups. The survey results showed that, although level of adoption of IPM practices varied over the sample, all farmers had adopted IPM to some extent (minimum 32.6 [corrected] points, mean score of 67.1), [corrected] but only 15 [corrected] of 225 farmers (5.8%) had adopted more than 67.1% [corrected] of what is theoretically possible, as measured by the new metric. CONCLUSION: We believe that this new metric would be a viable and cost-effective system to facilitate the benchmarking and monitoring of national IPM programmes in temperate zone countries with large-scale arable farming systems. © 2019 Society of Chemical Industry.

Changes in field dose-response curves for demethylation inhibitor (DMI) and quinone outside inhibitor (QoI) fungicides against Zymoseptoria tritici, related to laboratory sensitivity phenotyping and genotyping assays.

BACKGROUND: Insensitivity of Zymoseptoria tritici to demethylation inhibitor (DMI) and quinone outside inhibitor (QoI) fungicides has been widely reported from laboratory studies, but the relationships between laboratory sensitivity phenotype or target site genotype and field efficacy remain uncertain. This article reports field experiments quantifying dose-response curves, and investigates the relationships between field performance and in vitro half maximal effective concentration (EC50 ) values for DMIs, and the frequency of the G143A substitution conferring QoI resistance. RESULTS: Data were analysed from 83 field experiments over 21 years. Response curves were fitted, expressed as percentage control, rising towards an asymptote with increasing dose. Decline in DMI efficacy over years was associated with a decrease in the asymptote, and reduced curvature. Field ED50 values were positively related to in vitro EC50 values for isolates of Z. tritici collected over a 14-year period. Loss of QoI efficacy was expressed through a change in asymptote. Increasing frequency of G143A was associated with changes in field dose-response asymptotes. CONCLUSION: New resistant strains are often detected by resistance monitoring and laboratory phenotyped/genotyped before changes in field performance are detected. The relationships demonstrated here between laboratory tests and field performance could aid translation between laboratory and field for other fungicide groups. © 2017 Society of Chemical Industry.

Detection of the cytochrome b mutation G143A in Irish Rhynchosporium commune populations using targeted 454 sequencing.

BACKGROUND: Rhynchosporium commune is a major fungal pathogen of barley crops, and the application of fungicides, such as quinone outside inhibitors (QoIs), plays an important role in crop disease control. The genetic mechanisms linked to QoI resistance have been identified in the cytochrome b gene, with QoI resistance conferred by the G143A substitution. The objective of this study was to develop a high-throughput molecular assay to detect and identify mutations associated with QoI resistance within the Irish R. commune population. RESULTS: Leaf lesions of R. commune sampled from 74 sites during 2009-2014 and isolates from 2006 and 2007 were screened for non-synonymous mutations of the cytochrome b gene using 454 targeted sequencing. The presence of the G143A substitution was confirmed in R. commune samples at one site in 2013 and at four sites in 2014; however, the frequency of the substitution in these samples was low (2-18%). The 454 sequencing results were confirmed by PCR-RFLP and Sanger sequencing. CONCLUSION: The molecular assay that has been applied to this monitoring programme has shown that the application of 454 next-generation sequencing offers the potential for high throughput and accurate characterisation of non-synonymous mutations associated with fungicide resistance in a crop pathogen. © 2016 Society of Chemical Industry.

The effect of succinate dehydrogenase inhibitor/azole mixtures on selection of Zymoseptoria tritici isolates with reduced sensitivity.

BACKGROUND: Combining fungicides with different modes of action is regarded as one of the most effective means of slowing the selection of resistance. Field trials were used to study the effects of such mixtures on selection for Zymoseptoria tritici with reduced sensitivity to the succinate dehydrogenase inhibitors (SDHIs) and azole fungicides. The SDHI isopyrazam and the azole epoxiconazole were applied individually as solo products, and together in a preformulated mixture. All fungicide treatments were included at both full and half the recommended doses. RESULTS: Compared with using epoxiconazole alone, mixing epoxiconazole with isopyrazam led to an increase in epoxiconazole-sensitive isolates. In contrast, all treatments containing isopyrazam reduced the sensitivity of Z. tritici to isopyrazam compared with those without. Reducing doses to half the recommended rate had no effect on sensitivity of isolates to either active ingredient. In a subgroup of isolates least sensitive to isopyrazam, non-synonymous mutations were found in the SdhC and SdhD subunits, but their presence was unrelated to sensitivity. CONCLUSION: Mixing an azole and SDHI was clearly beneficial for the azole, but not for the SDHI component. This dynamic might change if strains conferring reduced sensitivity to the SDHIs were to arise. © 2015 Society of Chemical Industry.

Detection of Zymoseptoria tritici SDHI-insensitive field isolates carrying the SdhC-H152R and SdhD-R47W substitutions.

BACKGROUND: Succinate dehydrogenase inhibitor (SDHI) fungicides are important in the management of Zymoseptoria tritici in wheat. New active ingredients from this group of fungicides have been introduced recently and are widely used. Because the fungicides act at a single enzyme site, resistance development in Z. tritici is classified as medium-to-high risk. RESULTS: Isolates from Irish experimental plots in 2015 were tested against the SDHI penthiopyrad during routine monitoring. The median of the population was approximately 2 times less sensitive than the median of the baseline population. Two of the 93 isolates were much less sensitive to penthiopyrad than the least sensitive of the baseline isolates. These isolates were also insensitive to most commercially available SDHIs. Analysis of the succinate dehydrogenase coding genes confirmed the presence of the substitutions SdhC-H152R and SdhD-R47W in the very insensitive isolates. CONCLUSION: This is the first report showing that the SdhC-H152R mutation detected in laboratory mutagenesis studies also exists in the field. The function and relevance of this mutation, combined with SdhD-R47W, still needs to be determined. © 2016 Society of Chemical Industry.

The wheat-Septoria conflict: a new front opening up?

In the utopic absence of abiotic and/or biotic stressors, attaining the predicted increase (up to 70%) in wheat demand by 2050 in response to global population trends is a challenge. This objective becomes daunting, however, when one factors in the continuous constraint on global wheat production posed by Septoria tritici blotch (STB) disease. This is because, despite resistant loci being identified, a deficit of commercially relevant STB-resistant wheat germplasm remains. The issue is further compounded for growers by the emergence and prevalence of fungicide-resistant/insensitive strains of the causative pathogen Zymoseptoria tritici (formerly known as Mycosphaerella graminicola/Septoria tritici). However, biotechnology-based research is providing new opportunities in this struggle. As the exome response of wheat to STB attack begins to be deciphered, genes intrinsic to resistant and susceptible phenotypes will be identified. Combined with the application of genome-editing techniques and a growing appreciation of the complexity of wheat's and the dynamism of Z. tritici's genome, the generation of resulting STB-resistant wheat varieties will counter the prevalent threat of STB disease in wheat-production systems.

European Union policy on pesticides: implications for agriculture in Ireland.

European Community (EC) legislation has limited the availability of pesticide active substances used in effective plant protection products. The Pesticide Authorisation Directive 91/414/EEC introduced the principle of risk assessment for approval of pesticide active substances. This principle was modified by the introduction of Regulation (EC) 1107/2009, which applies hazard, the intrinsic toxicity of the active substance, rather than risk, the potential for hazard to occur, as the approval criterion. Potential impacts of EC pesticide legislation on agriculture in Ireland are summarised. While these will significantly impact on pesticide availability in the medium to long term, regulations associated with water quality (Water Framework Directive 2000/60/EC and Drinking Water Directive 1998/83/EC) have the potential to restrict pesticide use more immediately, as concerns regarding public health and economic costs associated with removing pesticides from water increase. This rationale will further reduce the availability of effective pesticide active substances, directly affecting crop protection and increasing pesticide resistance within pest and disease populations. In addition, water quality requirements may also impact on important active substances used in plant protection in Ireland. The future challenge for agriculture in Ireland is to sustain production and profitability using reduced pesticide inputs within a framework of integrated pest management.