Leptosphaeria Maculans Research Articles

Strategic genetic insights and integrated approaches for successful management of blackleg in canola/rapeseed farming

AbstractThis review describes a triumphant narrative in the battle against the devastating plant pathogen complex, Leptosphaeria maculans and L. biglobosa, and the success of the world's second‐largest oilseed crop, canola/oilseed rape. Emphasizing global collaborations, the article explores successfully mitigating this destructive disease in canola/rapeseed production across Australia, Canada and Europe. It highlights how strategies may vary between continents to adapt to specific contexts. While initial resistance (R) genes proved effective, the evolution of the pathogen under crop‐induced disease pressure led to the breakdown of these genes. Now, growers in these regions have been equipped with new tools, allowing them to make informed decisions that help to keep the disease at generally low levels. A pivotal factor in this success has been a deepened understanding of the intricate science underlying the host–pathogen interaction. Concerted efforts of individual laboratories and collaborative initiatives have played an essential role in this success, including novel methods for disease control based on extensive research that has translated into developing highly resistant varieties, enhanced pathogen monitoring, improved cultivar recommendation and integrated management strategies. The review showcases many milestone advancements, including the cloning of numerous avirulence genes within the pathogen, characterization of specific R genes, the development of various molecular tools for monitoring both pathogen and host, the introduction of groundbreaking disease management strategies such as R gene labelling, rotation and stacking, and establishment of a universal pathogen isolate collection that facilitates the exchange of information among multiple laboratories and adds a new dimension to this triumph.

Host Genetic Resistance in Brassica napus: A Valuable Tool for the Integrated Management of the Fungal Pathogen Leptosphaeria maculans.

Management of plant disease in agro-ecosystems ideally relies on a combination of host genetic resistance, chemical control, and cultural practices. Growers increasingly rely on chemical and genetic options, but their relative benefits in disease control, yield, and economic outcomes are rarely quantified. We explore this relationship for blackleg crown canker disease (caused by Leptosphaeria maculans), a major biotic constraint limiting canola production globally. Data from 20 field trials conducted from 2013 to 2015 in canola-growing regions of Australia were used to assess the effects of host resistance and fungicide treatment on blackleg severity, grain yield, and gross margin. In the absence of fungicide, blackleg disease was 88% lower in the most resistant compared with the most susceptible blackleg resistance category. In the most susceptible resistance category, the most effective fungicide treatment significantly reduced blackleg severity (from 50 to 6%) and increased grain yield (478 kg/ha, 41%) and gross margin (AU$120/ha, 17%). However, the mean benefits of fungicide tended to decrease with increasing levels of genetic resistance, to the point that yield, disease, and gross margin benefits were close to zero in the most resistant cultivars. Overall, these findings suggest that fungicides can reduce blackleg severity, but the benefits of application strongly depend on associated levels of genetic resistance. Canola cultivars with higher genetic resistance reliably reduced blackleg disease and maintained grain yield without the associated cost of fungicide application. The intensification of canola production to meet increasing global demand will require strategies to sustainably manage and protect finite genetic resistance resources to control blackleg disease.

Interactions between Weeds, Pathogen Symptoms and Winter Rapeseed Stand Structure

Rapeseed, weeds, and pathogens interact with each other. However, these interactions are not well understood. The aim of our work was to describe the relationships between weed vegetation and pathogen manifestations in rapeseed stands. Results from the four seasons show that different rapeseed stand structures produce different weed and pathogen responses. Eighteen weed species were identified in the rapeseed stands. The selected characteristics of rapeseed stands, pathogens, and weed manifestations were evaluated using redundancy analysis. Rapeseed stands with the highest levels of pathogens present (Alternaria brassiceae (Berk.) Sacc., Botrytis cinerea (De Bary) Whetzel, Sclerotinia sclerotiorum (Lib.) de Bary, Verticilium longisporum (C.Stark) Karapapa, Bainbr & Heale) had the lowest seed yield. There, the weeds Cirsium arvense (L.) Scop., Tripleurospermum inodorum (L.) Sch. Bip., Sonchus arvensis L. were more abundant in dense stands, and Phoma lingam (telomorph: Leptosphaeria maculans Ces. & De Not.) was more common. Mutual positive interactions may also include the relationship between weed species of the Asteraceae family and increased manifestations of Phoma lingam. A similar relationship can be expected for the weeds Capsella bursa-pastoris (L.) Medik., Descurainia sophia (L.) Prantl and Sclerotinia sclerotiorum symptoms.

Inter- and Intra-Pathogen Interactions Emanating from Coinfection with Different Fungal and Viral strains in Canola Cultivars with Differing Host Resistances.

Few recent investigations examine coinfection interactions between fungal and viral plant pathogens. Here, we investigated coinfections between Leptosphaeria maculans and turnip mosaic virus (TuMV) in canola (Brassica napus). Different combinations of L. maculans isolate P11 and resistance breaking isolates L. maculans UWA192 and TuMV 12.1, were inoculated to three cultivars with differing pathogen resistances/susceptibilities. They were inoculated first to entire or half cotyledons 10-12 days after emergence, and second to opposite entire or half cotyledons on the same day (day 0) or 3 or 7 days afterwards. The parameters measured were L. maculans cotyledon disease index (%CDI), and TuMV systemically infected leaf symptom intensity (SI) and virus concentration (VC). Except when both day 0 inoculations were with isolate UWA192, %CDI values were supressed strongly or only weakly when isolates P11 and/or UWA192 were inoculated to plants with L. maculans single gene resistance (SGR) or polygenic resistance, respectively. However, except when isolate P11 was inoculated first and UWA192 second, these values declined after inoculation day 0 when SGR was absent. TuMV infection suppressed %CDI values, although this decrease was usually smaller following day 0 half cotyledon inoculations. When TuMV temperature sensitive extreme resistance was present and both inoculations were with TuMV, SI and VC values diminished greatly. However, the extent of this decrease was reduced when second inoculations were with L. maculans. SI and VC values were also smaller when SGR was present and second inoculations were with L. maculans. When L. maculans resistance was lacking, SI and VC values were smaller when second inoculations to entire cotyledons were with L. maculans rather than TuMV. This also occurred after second half cotyledon inoculations with isolate P11 but not isolate UWA192. Therefore, diverse inter- or intra-pathogen interactions developed depending upon host resistance, isolate combination, cotyledon inoculation approach and second inoculation timing.

Studying temperature’s impact on Brassica napus resistance to identify key regulatory mechanisms using comparative metabolomics

To investigate the effects of temperature on Brassica napus (canola) resistance to Leptosphaeria maculans (LM), the causal agent of blackleg disease, metabolic profiles of LM infected resistant (R) and susceptible (S) canola cultivars at 21 °C and 28 °C were analyzed. Metabolites were detected in cotyledons of R and S plants at 48- and 120-h post-inoculation with LM using UPLC-QTOF/MS. The mock-inoculated plants were used as controls. Some of the resistance-related specific pathways, including lipid metabolism, amino acid metabolism, carbohydrate metabolism, and aminoacyl-tRNA biosynthesis, were down-regulated in S plants but up-regulated in R plants at 21 °C. However, some of these pathways were down-regulated in R plants at 28 °C. Amino acid metabolism, lipid metabolism, alkaloid biosynthesis, phenylpropanoid biosynthesis, and flavonoid biosynthesis were the pathways linked to combined heat and pathogen stresses. By using network analysis and enrichment analysis, these pathways were identified as important. The pathways of carotenoid biosynthesis, pyrimidine metabolism, and lysine biosynthesis were identified as unique mechanisms related to heat stress and may be associated with the breakdown of resistance against the pathogen. The increased susceptibility of R plants at 28 °C resulted in the down-regulation of signal transduction pathway components and compromised signaling, particularly during the later stages of infection. Deactivating LM-specific signaling networks in R plants under heat stress may result in compatible responses and deduction in signaling metabolites, highlighting global warming challenges in crop disease control.

Frequency of avirulence genes and race structure in a Leptosphaeria maculans population in Tunisia

Frequency of avirulence genes and race structure in a <i>Leptosphaeria maculans</i> population in Tunisia

Molecular Characterization and Expression Analysis of a Gene Encoding 3-Hydroxy-3-Methylglutaryl-CoA Reductase (HMGR) from Bipolaris eleusines, an Ophiobolin A-Producing Fungus.

Ophibolin A, a fungal sesterterpene, exerts a pivotal influence in a diverse array of biological processes, encompassing herbicidal, bactericidal, fungicidal, and cytotoxic activities. Sixty genes associated with sesterterpene compound biosynthesis were obtained from Bipolaris eleusines via transcriptome sequencing, and those closely linked to ophiobolin A biosynthesis were subsequently filtered. A gene encoding 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) that catalyzes the first committed step of ophiobolin biosynthesis in the mevalonic acid (MVA) pathway was isolated and characterized using RACE (Rapid Amplification of cDNA Ends) technology from ophiobolin A-producing fungus, B. eleusines. The full-length cDNA of the B. eleusines HMGR gene (BeHMGR) was 3906 bp and contained a 3474 bp open reading frame (ORF) encoding 1157 amino acids. Sequence analysis revealed that deduced BeHMGR had high homology to the known HMGRs from Pyrenophora tritici-repentis and Leptosphaeria maculans. It had a calculated molecular mass of about 124.65 kDa and an isoelectric point (pI) of 6.90. It contained two putative HMG-CoA-binding motifs and two NADP(H)-binding motifs. Induced expression analysis of the BeHMGR gene by methyl jasmonate treatment using quantitative fluorescence PCR showed that it significantly elevated after 3 h of methyl jasmonate treatment, peaked at 6 h, and then gradually decreased. This demonstrates that BeHMGR gene expression is induced by methyl jasmonate.

The ilv2 gene, encoding acetolactate synthase for branched chain amino acid biosynthesis, is required for plant pathogenicity by Leptosphaeria maculans

BackgroundControl of blackleg disease of canola caused by the fungus Leptosphaeria maculans relies on strategies such as the inhibition of growth with fungicides. However, other chemicals are used during canola cultivation, including fertilizers and herbicides. There is widespread use of herbicides that target the acetolactate synthase (ALS) enzyme involved in branched chain amino acid synthesis and low levels of these amino acids within leaves of Brassica species. In L. maculans the ilv2 gene encodes ALS and thus ALS-inhibiting herbicides may inadvertently impact the fungus.Methods and resultsHere, the impact of a commercial herbicide targeting ALS and mutation of the homologous ilv2 gene in L. maculans was explored. Exposure to herbicide had limited impact on growth in vitro but reduced lesion sizes in plant disease experiments. Furthermore, the mutation of the ilv2 gene via CRISPR-Cas9 gene editing rendered the fungus non-pathogenic.ConclusionHerbicide applications can influence disease outcome, but likely to a minor extent.

A Specific Resistance Gene to Leptosphaeria maculans, Rlm11, Has a Limited Impact on Brassica napus Mycobiota Compared with Plant Compartment or Cropping Season Effects

One important ecological question regarding the use of plant resistance genes against fungal pathogens concerns whether and how such resistance genes may modify pathogenic or beneficial members of the plant-associated microbiota. We studied the impact of a plant resistance gene by analyzing the mycobiota associated with Brassica napus organs over two cropping seasons. Sampling dates coincided with key stages of the life cycle of the B. napus pathogen Leptosphaeria maculans. Leaf samples were collected at three time points in autumn and spring, and stem base samples were collected at two time points a few weeks before and at harvest. Stem residues, where L. maculans survives in the intercropping season and develops sexual reproduction, were also analyzed at four time points between the two cropping seasons. The sampling was performed on two plant genotypes, Darmor and Darmor- Rlm11, only differing by the effective resistance gene against L. maculans, Rlm11. Altogether, 419 samples were analyzed using two barcode: internal transcribed spacer (ITS) and Actin. The plant organ was shown to be the main mycobiota structuring factor, as clear-cut alternation of the species suggested that each plant organ represented a specific ecological niche. The cropping season and plant genotype also significantly influenced the community structure in lower proportions. The resistance gene contributed differently to the community structure depending on the year and the organ concerned. A significant but low impact of Rlm11 on other B. napus fungal pathogens was detected. The ITS and Actin barcodes showed similar results, but the species assignation was limited for the latter.

Identification of new lines resistance to blackleg in an Argentinian rapeseed-breeding program

Rapeseed (Brassica napus L.) is the third most important oil crop worldwide and it has a high demand for its industrial characteristics that make it suitable for human consumption and production of biofuels. The main yield losses of the crop are produce by blackleg, caused by Leptosphaeria maculans (Desmaz.) Ces. and of Not. [Anamorph Phoma lingam (Tode ex Fr.) Desmaz]. In this context, 469 inbred lines (ILs) of the INTA-EEA Paraná breeding program were phenotypic characterized for its resistance to Blackleg by artificial inoculations with a local isolation. From the initial 469 ILs, a total of 59 (13%) were identified as phenotypically resistant and were characterized by molecular makers associated to resistant genes (Rlm1, Lepr3, Rpg3Dun, and Rlm4) with the aim to identify lines carrying different resistance genes to be selected as parental lines in the development of new resistant genotypes. Moreover, the resistant ILs were characterized for its agronomics performance, including yield; percentage of oil; and content of fatty acids, over two crop seasons (2018 and 2019). A Generalized Procrustes Analysis was conducted to correlate all the traits evaluated and identify elite lines. From the 59 resistants ILs, four lines were selected based on their high performances and multiple resistant gene combinations, as started lines to develop a pyramiding gene strategy. The selection of these elite lines to combine traditional breeding and gene-pyramiding based on MAS in our breeding program is a useful strategy to develop superior rapeseed genotypes.

Evolution of decreased sensitivity to azole fungicides in western European populations of Plenodomus lingam (Phoma stem canker on oilseed rape)

AbstractPlenodomus lingam (Leptosphaeria maculans) and P. biglobosus (L. biglobosa) are fungi causing Phoma leaf spot/stem canker, an international damaging disease of oilseed rape (Brassica napus) and other brassicas. In Europe, fungicides used for disease management are mainly sterol 14α‐demethylase (CYP51) inhibitors (DMIs/azoles); quinone‐outside inhibitors (QoIs) and succinate dehydrogenase inhibitors (SDHIs) are also used. Decreased DMI sensitivity has emerged in Australian and eastern European P. lingam populations and is mediated by CYP51 promoter inserts resulting in target site overexpression. In this study using in vitro sensitivity testing, we report decreased DMI (prothioconazole‐desthio, mefentrifluconazole) sensitivity in modern western European P. lingam isolates (collected 2022–2023) compared to older baseline (1992–2005) isolates. Around 85% of modern western European P. lingam isolates collected, for which the CYP51 promoter region was sequenced, carried a promoter insert, but target site alterations were not detected. Six different CYP51 promoter inserts were identified, most commonly a 237 bp fragment of the Sahana transposable element. Inserts were associated with an approximately 3‐ to 10‐fold decrease in sensitivity to the DMIs tested. In contrast to P. lingam, PCR screening revealed CYP51 promoter inserts were absent in modern western European P. biglobosus isolates (2021–2023). Combined data indicate P. lingam isolates lacking an insert were similarly (or slightly more) sensitive to the DMIs tested for P. biglobosus, whereas those carrying an insert were slightly less sensitive than P. biglobosus. No evidence for substantive sensitivity shifts to the QoI (pyraclostrobin) or SDHI (boscalid) fungicides tested was obtained for either Plenodomus species.

No safe haven: Loss of avirulence in the plant pathogen Leptosphaeria maculans by DNA duplication and repeat‐induced point mutation

AbstractMicrobes can overcome the ability of plant resistance genes to confer protection against disease by mutating their corresponding avirulence genes. The fungus Leptosphaeria maculans causes blackleg disease on canola crops and subverts Brassica napus resistance genes through several DNA mutation mechanisms. One of these is repeat‐induced point (RIP) mutation, which can ‘leak’ into the avirulence genes from the adjacent repetitive sequences that the mutation process is targeting. Here, we identified populations of L. maculans in Australia that have extensive RIP mutations in the avirulence gene AvrLm2 and show that this has been triggered by a duplication of the gene and surrounding DNA that includes the distant (&gt;55 kb in total) AvrLm6 gene. This finding provides another mechanism of mutation by which pathogens can overcome host resistance, and more broadly contributes to understanding the complex balance between gene duplication and genome defence.

Does fungal infection increase the palatability of oilseed rape to insects?

Multiple and simultaneous attacks by pathogens and insect pests frequently occur in nature. Plants respond to biotic stresses by activating distinct defense mechanisms, but little is known about how plants cope with multiple stresses. The focus of this study was the combined interaction of fungal infection caused by Leptosphaeria maculans (synonym Plenodomus lingam) and arthropod infestation by the diamondback moth (Plutella xylostella) in oilseed rape (Brassica napus). We hypothesized that infection by the fungal pathogen L. maculans could alter oilseed rape palatability to P. xylostella-chewing caterpillars. Feeding preference tests were complemented with analyses of defense gene transcription, and levels of glucosinolates (GLSs) and volatile organic compounds (VOCs) in L. maculans-inoculated and non-inoculated (control) leaves to determine possible causes of larval choice. Caterpillars preferred true leaves to cotyledons, hence true leaves were used for further experiments. True leaves inoculated with L. maculans were more palatable to caterpillars over control leaves during the early stage of infection at 3 days post inoculation (dpi), but this preference disappeared in the later stages of infection at 7 dpi. In parallel, genes involved in the salicylic acid and ethylene pathways were up-regulated in L. maculans-inoculated leaves at 3 and 7 dpi; L. maculans increased the level of total aliphatic GLSs, specifically glucobrassicanapin, and decreased the level of glucoiberin at 3 dpi and altered the content of specific VOCs. A group of 55 VOCs with the highest variability between treatments was identified. We suggest that the P. xylostella preference for L. maculans-inoculated leaves in the early stage of disease development could be caused by the underlying mechanisms leading to changes in metabolic composition. Further research should pinpoint the compounds responsible for driving larval preference and evaluate whether the behavior of the adult moths, i.e. the stage that makes the first choice regarding host plant selection in field conditions, correlates with our results on larval host acceptance. © 2024 Society of Chemical Industry.

Facilitation of Symplastic Effector Protein Mobility by Paired Effectors Is Conserved in Different Classes of Fungal Pathogens.

It has been discovered that plant pathogens produce effectors that spread via plasmodesmata (PD) to allow modulation of host processes in distal uninfected cells. Fusarium oxysporum f. sp. lycopersici (Fol) facilitates effector translocation by expansion of the size-exclusion limit of PD using the Six5/Avr2 effector pair. How other fungal pathogens manipulate PD is unknown. We recently reported that many fungal pathogens belonging to different families carry effector pairs that resemble the SIX5/AVR2 gene pair from Fol. Here, we performed structural predictions of three of these effector pairs from Leptosphaeria maculans (Lm) and tested their ability to manipulate PD and to complement the virulence defect of a Fol SIX5 knockout mutant. We show that the AvrLm10A homologs are structurally related to FolSix5 and localize at PD when they are expressed with their paired effectors. Furthermore, these effectors were found to complement FolSix5 function in cell-to-cell mobility assays and in fungal virulence. We conclude that distantly related fungal species rely on structurally related paired effector proteins to manipulate PD and facilitate effector mobility. The wide distribution of these effector pairs implies Six5-mediated effector translocation to be a conserved propensity among fungal plant pathogens. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Transcriptomics of temperature-sensitive R gene-mediated resistance identifies a WAKL10 protein interaction network

Understanding temperature-sensitivity of R gene-mediated resistance against apoplastic pathogens is important for sustainable food production in the face of global warming. Here, we show that resistance of Brassica napus cotyledons against Leptosphaeria maculans was temperature-sensitive in introgression line Topas-Rlm7 but temperature-resilient in Topas-Rlm4. A set of 1,646 host genes was differentially expressed in Topas-Rlm4 and Topas-Rlm7 in response to temperature. Amongst these were three WAKL10 genes, including BnaA07g20220D, representing the temperature-sensitive Rlm7-1 allele and Rlm4. Network analysis identified a WAKL10 protein interaction cluster specifically for Topas-Rlm7 at 25 °C. Diffusion analysis of the Topas-Rlm4 network identified WRKY22 as a putative regulatory target of the ESCRT-III complex-associated protein VPS60.1, which belongs to the WAKL10 protein interaction community. Combined enrichment analysis of gene ontology terms considering gene expression and network data linked vesicle-mediated transport to defence. Thus, dysregulation of effector-triggered defence in Topas-Rlm7 disrupts vesicle-associated resistance against the apoplastic pathogen L. maculans.

Regulation of effector gene expression as concerted waves in Leptosphaeria maculans: a two-player game.

Effector genes, encoding molecules involved in disease establishment, are concertedly expressed throughout the lifecycle of plant-pathogenic fungi. However, little is known about how effector gene expression is regulated. Since many effector genes are located in repeat-rich regions, the role of chromatin remodeling in their regulation was recently investigated, notably establishing that the repressive histone modification H3K9me3, deposited by KMT1, was involved in several fungal species including Leptosphaeria maculans. Nevertheless, previous data suggest that a second regulatory layer, probably involving a specific transcription factor (TF), might be required. In L. maculans, a Dothideomycete causing stem canker of oilseed rape, we identified the ortholog of Pf2, a TF belonging to the Zn2Cys6 fungal-specific family, and described as essential for pathogenicity and effector gene expression. We investigated its role together with KMT1, by inactivating and over-expressing LmPf2 in a wild-type strain and a ∆kmt1 mutant. Functional analyses of the corresponding transformants highlighted an essential role of LmPf2 in the establishment of pathogenesis and we found a major effect of LmPf2 on the induction of effector gene expression once KMT1 repression is lifted. Our results show, for the first time, a dual control of effector gene expression.

Synthesis and biological evaluation of burnettiene A derivatives enabling discovery of novel fungicide candidates.

An antifungal polyene-decalin polyketide natural product, burnettiene A (1) has been re-discovered from the culture broth of Lecanicillium primulinum (current name: Flavocillium primulinum) FKI-6715 strain utilizing our original multidrug-sensitive yeast system. This polyene-decalin polyketide natural product was originally isolated from Aspergillus burnettii. The antifungal activity of 1 against Candida albicans has been reported. However, only one fungal species for the antifungal activity of 1 has been revealed, and details of the antifungal activity against other pathogenic fungus remain unknown. After extensive screening for antifungal activity, we found that 1 exhibits broad antifungal activity against pathogenic plant fungi, including Colletotrichum gloeosporioides, Botrytis cinerea, Pyricularia oryzae, Leptosphaeria maculans, and Rhizoctonia solani. Furthermore, we synthesized 12 derivatives from 1 and evaluated their antifungal activity to reveal the detailed structure-activity relationship. The methyl ester derivative showed antifungal activity against Saccharomyces cerevisiae 12geneΔ0HSR-iERG6 100-fold more potent than that of 1. Our research indicates that 1 would be a promising natural product as a new fungicidal candidate and the methyl ester derivative especially has great potential.

Evaluation of the effect of magnetic field on rapeseed growth and the causal agent of blackleg disease, Phoma lingam.

In recent years, with the increased production of oilseed rape, there has been a simultaneous enhancement in reports on pathogens causing diseases. Magnetic technology has been recognized as a new agricultural method aimed at improving health and crop production. In this work, the effect of magnetic fields was studied on the mycelial growth and conidia formation of Leptosphaeria maculans Gol125 and Leptosphaeria biglobosa KH36, the causal agents of Phoma stem cancer (blackleg) disease in rapeseed. In addition, seeds exposed to eight direct frequencies of magnetic fields were impregnated with pathogen suspension and grown under greenhouse conditions. The growth speed of both pathogen isolates decreased by 1-28% in GOL125 and 6-46% in KH36 over time in cultures exposed to magnetic fields. However, the number of conidia increased significantly under magnetic field exposure, reaching 5.4 × 107 and 7.7 × 107 SFU/ml in KH36 and GOL125 isolates, respectively. Furthermore, in greenhouse conditions, an increase in photosynthetic pigment levels was observed in almost all of the magnetic field-treated plants. In addition, disease incidence decreased by around 6% in the magnetic field-treated plants. This study represents the first evaluation of magnetic technology in controlling plant diseases. The use of magnetic fields may present a viable strategy for a sustainable production system; however, it requires further advanced studies to improve plant health and productivity.

Artificial intelligence analysis of contributive factors in determining blackleg disease severity in canola farmlands

Canola (Brassica napus L.) production is threatened by blackleg disease caused by Leptosphaeria maculans. Disease outcome is determined by interactions among pathogens, plants, farming practices, and environmental factors. Although the gene-for-gene interactions between the pathogen and its plant host are relatively clear, how precisely the pathogen interacts with the environment and farming practices is still poorly understood, making disease forecasting challenging for commercial farmlands. In recent years, artificial intelligence (AI) has been successful in forecasting disease risks based on environmental factors. In this study, we evaluated two AI methods and a data augmentation method to forecast disease risk using a dataset collected from 116 farmlands in Alberta in 2021 and 2022. We first assessed a machine learning model (support vector machine or SVM) and a deep-learning model (convolutional neural network or CNN) to predict blackleg severity based on five weather variables, flea beetle damage, root maggot damage, and crop-rotation variables. Both SVM and CNN predicted the disease risk with an accuracy of over 66%. The data augmentation method did not improve model performance. Flea beetle feeding and maggot damage contribute little to the model’s performance, and omitting these data did not appear to affect the results. In contrast, crop rotation contributes substantially to model performance. The five weather variables contribute roughly equally to the model’s performance, and removing any of the individual weather variables did not impact prediction ability for both models.

Genetic Diversity and Population Structure of Leptosphaeria biglobosa from the Winter Oilseed Rape Region in China.

Phoma stem canker (blackleg), caused by the fungi Leptosphaeria maculans (anamorph Phoma lingam) and L. biglobosa, is one of the most devastating diseases in oilseed rape (Brassica napus L.) production worldwide. However, the population structure and genetic variation of L. biglobosa populations in China have rarely been investigated. Here, a collection of 214 fungal strains of blackleg from China (including Shaanxi, Jiangxi, Hubei, Jiangsu, Chongqing, Sichuan, Guangxi, Guizhou, Hunan, and Henan) and Europe (France and Ukraine) was identified as L. biglobosa. Three simple sequence repeat (SSR) markers were developed to characterize their population structure. The results showed that the Nei's average gene diversity ranged from 0.6771 for the population from Jiangsu to 0.3009 for the population from Hunan. In addition, most of the genetic variability (96%) occurred within groups and there were only relatively small amounts among groups (4.0%) (FST = 0.043, p = 0.042 < 0.05). Pairwise population differentiation (FST) suggested that significant genetic differentiation was observed between different L. biglobosa populations. Bayesian and unweighted average method analysis revealed that these L. biglobosa strains were clustered into three branches, and three European strains were similar to those from eastern China. The pathogenicity assay showed that those in Group III were significantly more virulent than those in Group I (t = 2.69, p = 0.016). The study also showed that Group III was dominant in Chinese L. biglobosa populations, which provides new insights for the further study of population evolution and the management of this pathogen.