Importance of Leptosphaeria biglobosa as a cause of phoma stem canker on winter oilseed rape in the UK

Phoma stem canker is a major disease of oilseed rape in the UK, leading to annual yield losses worth more than £100M. The disease is caused by two closely related species, Leptosphaeria maculans and L. biglobosa. L. maculans is generally considered more damaging, causing stem base canker; L. biglobosa is generally less damaging, causing upper stem lesions. Therefore, previous work has mainly focused on L. maculans and there has been little work on L. biglobosa. This work investigated the contribution of L. biglobosa to stem canker epidemics by assessing the amounts of DNA of L. maculans and L. biglobosa in upper stem lesions or stem base cankers on winter oilseed rape cultivars with different types of resistance against L. mac ulans. Diseased upper stem and stem base samples were collected from nine oilseed rape cultivars in a 2011/2012 field experiment at Rothamsted. The presence of L. maculans or L. biglobosa in each stem sample was detected by speciesspecific PCR. The abundance of L. maculans or L. biglobosa in each stem sample was measured by quantification of L. maculans DNA and L. biglobosa DNA using quantitative PCR (qPCR). The amounts of L. biglobosa DNA were greater than those of L. maculans DNA in both upper stem and stem base samples. These results suggest that the severe upper stem lesions and stem base cankers in the 2011/2012 cropping season were mainly caused by L. biglobosa, suggesting that L. biglobosa can sometimes cause considerable yield loss in the UK. There were differences between cultivars in the amounts of L. maculans DNA and L. biglobosa DNA, with the susceptible cultivar Drakkar having more L. maculans DNA than L. biglobosa DNA while resistant cultivars had less L. maculans DNA than L. biglobosa DNA. These results suggest that L. biglobosa can be an important cause of phoma stem canker on oilseed rape in the UK.Peer reviewedFinal Published versio

Disease management in oilseed rape: insights into the Brassica napus-Pyrenopeziza brassicae pathosystem

Final Published versio

Effects of plant age and inoculum concentration on light leaf spot disease phenotypes on oilseed rape

© The Author(s). All rights reserved.Light leaf spot is caused by the fungal pathogen Pyrenopeziza brassicae and is the mosteconomically damaging disease of oilseed rape (Brassica napus) in the UK. Current controlrelies on repeated fungicide applications; however, pathogen fungicide-insensitivitydevelopment highlights the need for non-chemical controls like host resistance. A study wasdone to assess light leaf spot disease phenotype on the susceptible B. napus cultivar Charger indifferent treatment conditions; factors studied included plant age and inoculum concentration.Results showed that older plants grown in a controlled-environment cabinet produced the mostvisible symptoms. Plants that received a greater inoculum concentration (105spores/ml) weresignificantly shorter by 5 cm than those inoculated with a smaller inoculum concentration (104spores/ml), suggesting possible correlations between fungal inoculum concentration and plantgrowth. Additionally, > 20 P. brassicae field isolates were collected from leaf samples acrossEngland through single-spore isolation and will be screened for virulence

Improved understanding of novel sources of resistance against the light leaf spot pathogen, Pyrenopeziza brassicae

Chinthani Shanika Karandeni Dewage, Kavithra Jayani Wijerathna, Henrik U. Stotz, and Bruce D. L. Fitt, 'Improved understanding of novel sources of resistance against the light leaf spot pathogen, Pyrenopeziza brassicae', paper presented at the Association of Applied Biologists Conference Crop Production in Southern Britain 2017, 15 - 16 February 2017, Peterborough, UK. Proceedings available online at: http://www.aab.org.uk/contentok.php?id=501.In this work, the endophytic growth phase of the light leaf spot pathogen Pyrenopeziza brassicae in selected lines from a doubled haploid (DH) population of oilseed rape, which is known to segregate for resistance against P. brassicae, was characterised using controlled environment (CE) experiments. Fungal staining techniques and pathogen-specific quantitative polymerase chain reactions (qPCR) were used to observe and quantify the pathogen biomass, respectively. The qPCR results showed that the resistant lines contained little P. brassicae DNA and there seemed to be little to no change in the amount of DNA over time. In contrast, there was a considerable increase in pathogen DNA in susceptible lines from 0 to 24 days post inoculation (dpi). These results were also reflected in observations made by a fungal staining method. In addition, leaf samples of these DH lines, collected at three different times from winter oilseed rape field experiments, were analysed using qPCR. The resistant lines had a considerably smaller amount of P. brassicae DNA in leaf samples collected later in the cropping season than that in susceptible lines

Effects of cultivar resistance and fungicide application on stem canker of oilseed rape (Brassica napus) and potential interseasonal transmission of Leptosphaeria spp. inoculum

© 2021 The Authors. Plant Pathology published by John Wiley & Sons Ltd on behalf of British Society for Plant Pathology. This is an open access article under the terms of the Creative Commons Attribution License, https://creativecommons.org/licenses/by/4.0/Phoma stem canker is a damaging disease of oilseed rape (Brassica napus) that causesannual yield losses to UK oilseed rape growers worth approximately £100 million,despite the use of fungicides. In the UK, oilseed rape is sown in August/Septemberand harvested in the following July. The disease epidemics are initiated by ascosporesreleased from Leptosphaeria spp. pseudothecia (ascocarps) on stem stubble in theautumn/winter. Control of this disease is reliant on the use of cultivars with “fieldresistance” and azole fungicides. This study investigated the effects of cultivar resistanceand application of the fungicide prothioconazole on the severity of stem cankerbefore harvest and the subsequent production of pseudothecia on the infected stubbleunder natural conditions in the 2017/2018, 2018/2019, and 2019/2020 croppingseasons. The application of prothioconazole and cultivar resistance decreased theseverity of phoma stem canker before harvest, and the subsequent production ofLeptosphaeria spp. pseudothecia on stubble in terms of pseudothecial density. Resultsshowed that stems with less severe stem cankers produced fewer mature pseudotheciaof Leptosphaeria spp. on the infected stubble. This investigation suggests that themost sustainable and effective integrated control strategy for phoma stem canker inseasons with low quantities of inoculum is to use cultivars with medium or good fieldresistance and apply only one spray of prothioconazole when required.Peer reviewe

Chemical warfare: the fungal quest to conquer oilseed rape

Final Accepted Versio

Co‐inoculation timing affects the interspecific interactions between phoma stem canker pathogens Leptosphaeria maculans and Leptosphaeria biglobosa

© 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY), https://creativecommons.org/licenses/by/4.0BACKGROUND: Phoma stem canker is an economically important disease of oilseed rape, caused by two co‐existing fungal pathogen species, Leptosphaeria maculans (Plenodomus lingam) and Leptosphaeria biglobosa (Plenodomus biglobosus). Leptosphaeria maculans produces a phytotoxin called sirodesmin PL. Our previous work showed that L. biglobosa has an antagonistic effect on the production of sirodesmin PL if it is simultaneously co‐inoculated with L. maculans. However, the effects of sequential co‐inoculation on interspecific interactions between the two pathogens are not understood. RESULTS: The interactions between L. maculans and L. biglobosa were investigated in liquid culture by inoculation with L. maculans first, followed by L. biglobosa sequentially at 1, 3, 5 or 7 days later and vice versa; the controls were inoculated with L. maculans only, L. biglobosa only, or L. maculans and L. biglobosa simultaneously. The results showed that L. biglobosa inhibited the growth of L. maculans, the production of both sirodesmin PL and its precursors if L. biglobosa was inoculated before, or simultaneously with, L. maculans. However, the antagonistic effects of L. biglobosa were lost if it was co‐inoculated 5 or 7 days after L. maculans. CONCLUSION: For the first time, the results of this study provided evidence that the timing when L. maculans and L. biglobosa meet significantly influences the outcome of the interspecific competition between them. Leptosphaeria biglobosa can inhibit the production of sirodesmin PL and the growth of L. maculans if it is inoculated before L. maculans or less than 3 days after L. maculans in liquid culture. There is a need to further investigate the timing of co‐inoculation on interactions between L. maculans and L. biglobosa in their host plants for improving the control of phoma stem canker. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.Peer reviewe

Leptosphaeria biglobosa inhibits the production of sirodesmin PL by L. maculans

© 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY), https://creativecommons.org/licenses/by/4.0/BACKGROUND: Phoma stem canker is caused by two coexisting pathogens, Leptosphaeria maculans and L. biglobosa. They coexist because of their temporal and spatial separations, which are associated with the differences in timing of their ascospore release. L. maculans produces sirodesmin PL, while L. biglobosa does not. However, their interaction/coexistence in terms of secondary metabolite production is not understood. RESULTS: Secondary metabolites were extracted from liquid cultures, L. maculans only (Lm only), L. biglobosa only (Lb only), L. maculans and L. biglobosa simultaneously (Lm&Lb) or sequentially 7 days later (Lm+Lb). Sirodesmin PL or its precursors were identified in extracts from ‘Lm only’ and ‘Lm+Lb’, but not from ‘Lm&Lb’. Metabolites from ‘Lb only’, ‘Lm&Lb’ or ‘Lm+Lb’ caused significant reductions in L. maculans colony area. However, only the metabolites containing sirodesmin PL caused a significant reduction to L. biglobosa colony area. When oilseed rape cotyledons were inoculated with conidia of ‘Lm only’, ‘Lb only’ or ‘Lm&Lb’, ‘Lm only’ produced large gray lesions, while ‘Lm&Lb’ produced small dark lesions similar to lesions caused by ‘Lb only’. Sirodesmin PL was found only in the plant extracts from ‘Lm only’. These results suggest that L. biglobosa prevents the production of sirodesmin PL and its precursors by L. maculans when they grow simultaneously in vitro or in planta. CONCLUSION: For the first time, L. biglobosa has been shown to inhibit the production of sirodesmin PL by L. maculans when interacting simultaneously with L. maculans either in vitro or in planta. This antagonistic effect of interspecific interaction may affect their coexistence and subsequent disease progression and management.Peer reviewe

Host pathogen interactions in relation to management of light leaf spot disease (caused by Pyrenopeziza brassicae) on Brassica species

Light leaf spot, caused by Pyrenopeziza brassicae, is currently the most damaging disease problem in oilseed rape in the UK. According to recent survey data, the severity of epidemics has increased progressively across the UK, with current yield losses of up to £160M per annum in England and more severe epidemics in Scotland. Light leaf spot is a polycyclic disease with primary inoculum consisting of air-borne ascospores produced on diseased debris from the previous cropping season. Splash-dispersed conidia produced on diseased leaves are the main component of the secondary inoculum. P. brassicae is also able to infect and cause considerable yield losses on vegetable brassicas, especially Brussels sprouts. There may be spread of light leaf spot among different brassica species. Since they have a wide host range, Pyrenopeziza brassicae populations are likely to have considerable genetic diversity and there is evidence suggesting population variations between different regions, which need further study. Available disease-management tools are not sufficient to provide adequate control of the disease. There is a need to identify new sources of resistance, which can be integrated with fungicide applications to achieve sustainable management of light leaf spot. Several major resistance genes and quantitative trait loci have been identified in previous studies, but rapid improvements in the understanding of molecular mechanisms underpinning B. napus – P. brassicae interactions can be expected through exploitation of novel genetic and genomic information for brassicas and extracellular fungal pathogens.Peer reviewe

Current understanding of phoma stem canker and light leaf spot on oilseed rape in the UK

© The Author(s). All rights reserved.Oilseed rape is the third most important arable crop in the UK. Phoma stem cankerand light leaf spot are two economically important diseases of this crop. These twodiseases cause annual yield losses of winter oilseed rape worth > £100M, despite theuse of fungicides. Phoma stem canker is caused by two closely related fungalpathogens Leptosphaeria maculans and L. biglobosa, whereas light leaf spot is causedby the fungal pathogen Pyrenopeziza brassicae. Epidemics of both diseases areinitiated in autumn by ascospores released from crop debris from the previous croppingseason. However, phoma stem canker is a monocyclic disease, while light leaf spot isa polycyclic disease. Understanding the pathogen biology, disease epidemiology andhost resistance are essential for effective control of these two diseases. This minireview summarises current understanding of these two diseases in relation to pathogenbiology, disease epidemiology and host resistance