May Bente Brurberg

Research Professor

(+47) 926 09 364
may.brurberg@nibio.no

Place
Ås H7

Visiting address
Høgskoleveien 7, 1433 Ås

Biography

Research area: molecular plant pathology.

My research includes basic studies of pathogen biology, pesticide resistance, molecular diagnostics, barcoding and metabarcoding, as well as plant-pathogen interactions.

I have a 50 % position as professor in plant pathology at the Norwegian University of Life Sciences (1994).

Education: Cand. Scient. (MSc) in biotechnology, University of Oslo (1989), and Dr. Scient. (PhD) in molecular microbiology, Agricultural University of Norway – currently Norwegian University of Life Sciences (1994).

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Abstract

The oomycete pathogen Phytophthora cactorum causes crown rot, a major disease of cultivated strawberry. We report the draft genome of P. cactorum isolate 10300, isolated from symptomatic Fragaria x ananassa tissue. Our analysis revealed that there are a large number of genes encoding putative secreted effectors in the genome, including nearly 200 RxLR domain containing effectors, 77 Crinklers (CRN) grouped into 38 families, and numerous apoplastic effectors, such as phytotoxins (PcF proteins) and necrosis inducing proteins. As in other Phytophthora species, the genomic environment of many RxLR and CRN genes differed from core eukaryotic genes, a hallmark of the two-speed genome. We found genes homologous to known Phytophthora infestans avirulence genes including Avr1, Avr3b, Avr4, Avrblb1 and AvrSmira2 indicating effector sequence conservation between Phytophthora species of clade 1a and clade 1c. The reported P. cactorum genome sequence and associated annotations represent a comprehensive resource for avirulence gene discovery in other Phytophthora species from clade 1 and, will facilitate effector informed breeding strategies in other crops.

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Abstract

The oomycete Phytophthora infestans, the cause of late blight, is one of the most important potato pathogens. During infection, it secretes effector proteins that manipulate host cell function, thus contributing to pathogenicity. This study examines sequence differentiation of two P. infestans effectors from 91 isolates collected in Poland and Norway and five reference isolates. A gene encoding the Avr-vnt1 effector, recognized by the potato Rpi-phu1 resistance gene product, is conserved. In contrast, the second effector, AvrSmira1 recognized by Rpi-Smira1, is highly diverse. Both effectors contain positively selected amino acids. A majority of the polymorphisms and all selected sites are located in the effector C-terminal region, which is responsible for their function inside host cells. Hence it is concluded that they are associated with a response to diversified target protein or recognition avoidance. Diversification of the AvrSmira1 effector sequences, which existed prior to the large-scale cultivation of plants containing the Rpi-Smira1 gene, may reduce the predicted durability of resistance provided by this gene. Although no isolates virulent to plants with the Rpi-phu1 gene were found, the corresponding Avr-vnt1 effector has undergone selection, providing evidence for an ongoing ‘arms race’ between the host and pathogen. Both genes remain valuable components for resistance gene pyramiding.

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Abstract

Pathogenic soft rot Enterobacteriaceae (SRE) belonging to the genera Pectobacterium and Dickeya cause diseases in potato and numerous other crops. Seed potatoes are the most important source of infection, but how pathogen-free tubers initially become infected remains an enigma. Since the 1920s, insects have been hypothesized to contribute to SRE transmission. To validate this hypothesis and to map the insect species potentially involved in SRE dispersal, we have analyzed the occurrence of SRE in insects recovered from potato fields over a period of 2 years. Twenty-eight yellow sticky traps were set up in 10 potato fields throughout Norway to attract and trap insects. Total DNA recovered from over 2,000 randomly chosen trapped insects was tested for SRE, using a specific quantitative PCR (qPCR) TaqMan assay, and insects that tested positive were identified by DNA barcoding. Although the occurrence of SRE-carrying insects varied, they were found in all the tested fields. While Delia species were dominant among the insects that carried the largest amount of SRE, more than 80 other SRE-carrying insect species were identified, and they had different levels of abundance. Additionally, the occurrence of SRE in three laboratory-reared insect species was analyzed, and this suggested that SRE are natural members of some insect microbiomes, with herbivorous Delia floralis carrying more SRE than the cabbage moth (Plutella xylostella) and carnivorous green lacewing larvae (Chrysoperla carnea). In summary, the high proportion, variety, and ubiquity of insects that carried SRE show the need to address this source of the pathogens to reduce the initial infection of seed material.

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Abstract

Helminthosporium solani causes silver scurf, which affects the quality of potato. The biocontrol agent Clonostachys rosea greatly limited the severity of silver scurf symptoms and amount of H. solani genomic DNA in laboratory experiments. Transcriptomic analysis during interaction showed that H. solani gene expression was highly reduced when coinoculated with the biocontrol agent C. rosea, whereas gene expression of C. rosea was clearly boosted as a response to the pathogen. The most notable upregulated C. rosea genes were those encoding proteins involved in cellular response to oxidative stress, proteases, G-protein signaling, and the methyltransferase LaeA. The most notable potato response to both fungi was downregulation of defense-related genes and mitogen-activated protein kinase kinase kinases. At a later stage, this shifted, and most potato defense genes were turned on, especially those involved in terpenoid biosynthesis when H. solani was present. Some biocontrol-activated defense-related genes in potato were upregulated during early interaction with C. rosea alone that were not triggered by H. solani alone. Our results indicate that the reductions of silver scurf using C. rosea are probably due to a combination of mechanisms, including mycoparasitism, biocontrol-activated stimulation of plant defense mechanisms, microbial competition for nutrients, space, and antibiosis.

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Abstract

Soft rot and blackleg of potato caused by pectinolytic bacteria lead to severe economic losses in potato production worldwide. To investigate the species composition of bacteria causing soft rot and black leg of potato in Norway and Poland, bacteria were isolated from potato tubers and stems. Forty-one Norwegian strains and 42 Polish strains that formed cavities on pectate medium were selected for potato tuber maceration assays and sequencing of three housekeeping genes (dnaX, icdA and mdh) for species identification and phylogenetic analysis. The distribution of the species causing soft rot and blackleg in Norway and Poland differed: we have demonstrated that mainly P. atrosepticum and P. c. subsp. carotovorum are the causal agents of soft rot and blackleg of potatoes in Norway, while P. wasabiae was identified as one of the most important soft rot pathogens in Poland. In contrast to the other European countries, D. solani seem not to be a major pathogen of potato in Norway and Poland. The Norwegian and Polish P. c. subsp. carotovorum and P. wasabiae strains did not cluster with type strains of the respective species in the phylogenetic analysis, which underlines the taxonomic complexity of the genus Pectobacterium. No correlation between the country of origin and clustering of the strains was observed. All strains tested in this study were able to macerate potato tissue. The ability to macerate potato tissue was significantly greater for the P. c. subsp. carotovorum and Dickeya spp., compared to P. atrosepticum and P. wasabiae.

Abstract

The genus Pectobacterium, which belongs to the bacterial family Enterobacteriaceae, contains numerous species that cause soft rot diseases in a wide range of plants. The species Pectobacterium carotovorum is highly heterogeneous, indicating a need for re-evaluation and a better classification of the species. PacBio was used for sequencing of two soft-rot-causing bacterial strains (NIBIO1006T and NIBIO1392), initially identified as P. carotovorum strains by fatty acid analysis and sequencing of three housekeeping genes (dnaX, icdA and mdh). Their taxonomic relationship to other Pectobacterium species was determined and the distance from any described species within the genus Pectobacterium was less than 94% average nucleotide identity (ANI). Based on ANI, phylogenetic data and genome-to-genome distance, strains NIBIO1006T, NIBIO1392 and NCPPB3395 are suggested to represent a novel species of the genus Pectobacterium, for which the name Pectobacterium polaris sp. nov. is proposed. The type strain is NIBIO1006T (=DSM 105255T=NCPPB 4611T).

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Abstract

Non-native pests, climate change, and their interactions are likely to alter relationships between trees and tree-associated organisms with consequences for forest health. To understand and predict such changes, factors structuring tree-associated communities need to be determined. Here, we analysed the data consisting of records of insects and fungi collected from dormant twigs from 155 tree species at 51 botanical gardens or arboreta in 32 countries. Generalized dissimilarity models revealed similar relative importance of studied climatic, host-related and geographic factors on differences in tree-associated communities. Mean annual temperature, phylogenetic distance between hosts and geographic distance between locations were the major drivers of dissimilarities. The increasing importance of high temperatures on differences in studied communities indicate that climate change could affect tree-associated organisms directly and indirectly through host range shifts. Insect and fungal communities were more similar between closely related vs. distant hosts suggesting that host range shifts may facilitate the emergence of new pests. Moreover, dissimilarities among tree-associated communities increased with geographic distance indicating that human-mediated transport may serve as a pathway of the introductions of new pests. The results of this study highlight the need to limit the establishment of tree pests and increase the resilience of forest ecosystems to changes in climate.

Abstract

Crown rot, caused by Phytophthora cactorum, is a devastating disease of strawberry. While most commercial octoploid strawberry cultivars (Fragaria × ananassa Duch) are generally susceptible, the diploid species Fragaria vesca is a potential source of resistance genes to P. cactorum. We previously reported several F. vesca genotypes with varying degrees of resistance to P. cactorum. To gain insights into the strawberry defence mechanisms, comparative transcriptome profiles of two resistant genotypes (NCGR1603 and Bukammen) and a susceptible genotype (NCGR1218) of F. vesca were analysed by RNA-Seq after wounding and subsequent inoculation with P. cactorum. Differential gene expression analysis identified several defence-related genes that are highly expressed in the resistant genotypes relative to the susceptible genotype in response to P. cactorum after wounding. These included putative disease resistance (R) genes encoding receptor-like proteins, receptor-like kinases, nucleotide-binding sites, leucine-rich repeat proteins, RPW8-type disease resistance proteins, and ‘pathogenesis-related protein 1’. Seven of these R-genes were expressed only in the resistant genotypes and not in the susceptible genotype, and these appeared to be present only in the genomes of the resistant genotypes, as confirmed by PCR analysis. We previously reported a single major gene locus RPc-1 (Resistance to Phytophthora cactorum 1) in F. vesca that contributed resistance to P. cactorum. Here, we report that 4–5% of the genes (35–38 of ca 800 genes) in the RPc-1 locus are differentially expressed in the resistant genotypes compared to the susceptible genotype after inoculation with P. cactorum. In particular, we identified three defence-related genes encoding wall-associated receptor-like kinase 3, receptor-like protein 12, and non-specific lipid-transfer protein 1-like that were highly expressed in the resistant genotypes compared to the susceptible one. The present study reports several novel candidate disease resistance genes that warrant further investigation for their role in plant defence against P. cactorum.

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Abstract

Fusarium verticillioides is the most common fungal pathogen of maize in Ethiopia. Many strains of this pathogen produce fumonisin myotoxins that are harmful to human and animal health. This study was conducted to determine the fumonisin-producing ability of isolates of F. verticillioides isolated from maize kernels collected from different maize- growing areas of the country. Eighty F. verticillioides isolates were grown on autoclaved maize cultures for one month, and the fumonisin content was quantified using Enzyme Linked Immunosorbent Assay (ELISA). All the 80 isolates evaluated were able to produce detectable levels of total fumonisins in the maize culture with values ranging from 0.25 to 38.01 mg of the toxin per kg of culture material (fungal biomass and maize kernels). The mean levels of total fumonisins produced by the F. verticillioides isolates were not significantly (p>0.05) different among maize growing areas, however, the total fumonisins levels produced by isolates obtained from the same area as well as agroecological zones were wide-ranging. The results indicate that the majority (57.5%) of the F. verticillioides isolates associated with maize grains in Ethiopia produced total fumonisins >4 mg/kg, while 35% of the isolates produced total fumonisins <2 mg/kg. The widespread occurrence of higher fumonisin-producing strains across all maize-growing areas in Ethiopia indicates a possible food safety risk. Thus, efforts should be made to prevent the spread of this fungus with good agronomic practices and to implore all possible ways to avoid maize contamination with fumonisin both in the field and in storage.

Abstract

Phytophthora cactorum has two distinct pathotypes that cause crown rot and leather rot in strawberry (Fragaria × ananassa). Strains of the crown rot pathotype can infect both the rhizome (crown) and fruit tissues, while strains of the leather rot pathotype can only infect the fruits of strawberry. The genome of a highly virulent crown rot strain, a low virulent crown rot strain, and three leather rot strains were sequenced using PacBio high fidelity (HiFi) long read sequencing. The reads were de novo assembled to 66.4–67.6 megabases genomes in 178–204 contigs, with N50 values ranging from 892 to 1,036 kilobases. The total number of predicted complete genes in the five P. cactorum genomes ranged from 17,286 to 17,398. Orthology analysis identified a core secretome of 8,238 genes. Comparative genomic analysis revealed differences in the composition of potential virulence effectors, such as putative RxLR and Crinklers, between the crown rot and the leather rot pathotypes. Insertions, deletions, and amino acid substitutions were detected in genes encoding putative elicitors such as beta elicitin and cellulose-binding domain proteins from the leather rot strains compared to the highly virulent crown rot strain, suggesting a potential mechanism for the crown rot strain to escape host recognition during compatible interaction with strawberry. The results presented here highlight several effectors that may facilitate the tissue-specific colonization of P. cactorum in strawberry.

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Abstract

Control of grey mould, caused by Botrytis spp., is a major challenge in open field strawberry production. Botrytis was isolated from plant parts collected from 19 perennial strawberry fields with suspected fungicide resistance in the Agder region of Norway in 2016. Resistance to boscalid, pyraclostrobin and fenhexamid was high and found in 89.1%, 86.0% and 65.4% of conidia samples, respectively. Multiple fungicide resistance was common; 69.6% of conidia samples exhibited resistance to three or more fungicides. Botrytis group S and B. cinerea sensu stricto isolates were obtained from 19 and 16 fields, respectively. The sdhB, cytb, erg27 and mrr1 genes of a selection of isolates were examined for the presence of mutations known to confer fungicide resistance to boscalid, pyraclostrobin, fenhexamid and pyrimethanil plus fludioxonil, respectively. Allele-specific PCR assays were developed for efficient detection of resistance-conferring mutations in cytb. Among B. cinerea isolates, 84.7%, 86.3% and 61.3% had resistance-conferring mutations in sdhB, cytb and erg27, respectively. A triplet deletion in mrr1, resulting in ΔL497, commonly associated with the multidrug resistance phenotype MDR1h, was detected in 29.2% of Botrytis group S isolates. High frequencies of resistance to several fungicides were also detected in Botrytis from both imported and domestically produced strawberry transplants. Fungicide resistance frequencies were not different among fields grouped by level of grey mould problem assessed by growers, indicating factors other than fungicide resistance contributed to control failure, a fact that has important implications for future management of grey mould.

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Abstract

Bacteria isolated from onion bulbs suffering from bacterial decay in the United States and Norway were previously shown to belong to the genus Rahnella based on partial housekeeping gene sequences and/or fatty acid analysis. However, many strains could not be assigned to any existing Rahnella species. Additionally, strains isolated from creek water and oak as well as a strain with bioremediation properties were assigned to Rahnella based on partial housekeeping gene sequences. The taxonomic status of these 21 strains was investigated using multilocus sequence analysis, whole genome analyses, phenotypic assays and fatty acid analysis. Phylogenetic and phylogenomic analyses separated the strains into five clusters, one of which corresponded to Rahnella aceris . The remaining four clusters could be differentiated both genotypically and phenotypically from each other and existing Rahnella species. Based on these results, we propose the description of four novel species: Rahnella perminowiae sp. nov. (type strain SL6T=LMG 32257T=DSM 112609T), Rahnella bonaserana sp. nov. (H11bT=LMG 32256T=DSM 112610T), Rahnella rivi sp. nov. (FC061912-KT=LMG 32259T=DSM 112611T) and Rahnella ecdela sp. nov. (FRB 231T=LMG 32255T=DSM 112612T).

Abstract

Grey mold caused by the necrotrophic fungal pathogen Botrytis cinerea can affect leaves, flowers, and berries of strawberry, causing severe pre- and postharvest damage. The defense elicitor β-aminobutyric acid (BABA) is reported to induce resistance against B. cinerea and many other pathogens in several crop plants. Surprisingly, BABA soil drench of woodland strawberry (Fragaria vesca) plants two days before B. cinerea inoculation caused increased infection in leaf tissues, suggesting that BABA induce systemic susceptibility in F. vesca. To understand the molecular mechanisms involved in B. cinerea susceptibility in leaves of F. vesca plants soil drenched with BABA, we used RNA sequencing to characterize the transcriptional reprogramming 24 h post-inoculation. The number of differentially expressed genes (DEGs) in infected vs. uninfected leaf tissue in BABA-treated plants was 5205 (2237 upregulated and 2968 downregulated). Upregulated genes were involved in pathogen recognition, defense response signaling, and biosynthesis of secondary metabolites (terpenoid and phenylpropanoid pathways), while downregulated genes were involved in photosynthesis and response to auxin. In control plants not treated with BABA, we found a total of 5300 DEGs (2461 upregulated and 2839 downregulated) after infection. Most of these corresponded to those in infected leaves of BABA-treated plants but a small subset of DEGs, including genes involved in ‘response to biologic stimulus‘, ‘photosynthesis‘ and ‘chlorophyll biosynthesis and metabolism’, differed significantly between treatments and could play a role in the induced susceptibility of BABA-treated plants.

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Abstract

Soft rot Pectobacteriaceae (SRP) are ubiquitous on earth as there are records of findings from all continents where host plants are grown. This chapter describes information on soft rot diseases on these continents. For some countries, detailed information is provided by local experts on the SRP present, their economic damage, and the management strategies applied for their control. The focus of the chapter is mainly on SRP as causative agents of potato blackleg, although in specific cases details are provided on SRP in other host plants. In Europe, the SRP cause important economic losses mainly on potato, with most species described in the literature being found. In Latin America significant losses are also reported due to potato diseases caused by various Dickeya and Pectobacterium species, while in Australia and Oceania, recent outbreaks of D. dianthicola in potato have resulted in high economic losses. In Asia, however, SRP cause economic losses mainly in vegetable crops other than potato, while in North America SRP cause diseases on a wide range of crops (including potato and ornamental plants) in both field and storage. In Africa SRP are only known to occur in 17 of the 54 African countries but where it is known, potato is the most affected crop.

Abstract

Plants with roots and soil clumps transported over long distances in plant trading can harbor plant pathogenic oomycetes, facilitating disease outbreaks that threaten ecosystems, biodiversity, and food security. Tools to detect the presence of such oomycetes with a sufficiently high throughput and broad scope are currently not part of international phytosanitary testing regimes. In this work, DNA metabarcoding targeting the internal transcribed spacer (ITS) region was employed to broadly detect and identify oomycetes present in soil from internationally shipped plants. This method was compared to traditional isolation-based detection and identification after an enrichment step. DNA metabarcoding showed widespread presence of potentially plant pathogenic Phytophthora and Pythium species in internationally transported rhizospheric soil with Pythium being the overall most abundant genus observed. Baiting, a commonly employed enrichment method for Phytophthora species, led to an increase of golden-brown algae in the soil samples, but did not increase the relative or absolute abundance of potentially plant pathogenic oomycetes. Metabarcoding of rhizospheric soil yielded DNA sequences corresponding to oomycete isolates obtained after enrichment and identified them correctly but did not always detect the isolated oomycetes in the same samples. This work provides a proof of concept and outlines necessary improvements for the use of environmental DNA (eDNA) and metabarcoding as a standalone phytosanitary assessment tool for broad detection and identification of plant pathogenic oomycetes.

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Abstract

Dollar spot, caused by at least five Clarireedia species (formerly Sclerotinia homoeocarpa F. T. Benn.), is one of the economically most important turfgrass diseases worldwide. The disease was detected for the first time in Scandinavia in 2013. There is no available information from Scandinavian variety trials on resistance to dollar spot in turfgrass species and cultivars (http://www.scanturf.org/). Our in vitro screening (in glass vials) of nine turfgrass species comprising a total of 20 cultivars showed that on average for ten Clarireedia isolates of different origin, the ranking for dollar spot resistance in turfgrass species commonly found on Scandinavian golf courses was as follows: perennial ryegrass = slender creeping red fescue > strong creeping red fescue > Kentucky bluegrass = velvet bentgrass > colonial bentgrass = Chewings fescue ≥ creeping bentgrass = annual bluegrass. Significant differences in aggressiveness among Clarireedia isolates of different origin were found in all turfgrass species except annual bluegrass (cv. Two Putt). The U.S. C. jacksonii isolate MB-01 and Canadian isolate SH44 were more aggressive than C. jacksonii isolates from Denmark and Sweden (14.10.DK, 14.15.SE, and 14.16.SE) in velvet bentgrass and creeping bentgrass. The Swedish isolate 14.112.SE was generally more aggressive than 14.12.NO despite the fact that they most likely belong to the same Clarireedia sp. The U.S. C. monteithiana isolate RB-19 had similar aggressiveness as the Scandinavian C. jacksonii isolates, but was less aggressive than two U.S. C. jacksonii isolates MB-01 and SH44. Thus, aggressiveness of Clarireedia isolates was more impacted by their geographic origin and less by species of the isolate and/or the host turfgrass species.

Abstract

The soft rot Pectobacteriaceae (SRP) infect a wide range of plants worldwide and cause economic damage to crops and ornamentals but can also colonize other plants as part of their natural life cycle. They are found in a variety of environmental niches, including water, soil and insects, where they may spread to susceptible plants and cause disease. In this chapter, we look in detail at the plants colonized and infected by these pathogens and at the diseases and symptoms they cause. We also focus on where in the environment these organisms are found and their ability to survive and thrive there. Finally, we present evidence that SRP may assist the colonization of human enteric pathogens on plants, potentially implicating them in aspects of human/animal as well as plant health.

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Abstract

Oomycetes are spore-forming eukaryotic microbes responsible for infections in animal and plant species worldwide, posing a threat to natural ecosystems, biodiversity and food security. Genomics and transcriptomics approaches, together with host interaction studies, give promising results towards better understanding of the infection mechanisms in oomycetes and their general biology. Significant development and progress in oomycetes genomic studies have been achieved over the past decades but further understanding of molecular processes, gene regulations and infection mechanisms are still needed. The use of molecular tools such as CRISPR/Cas and RNAi helped elucidate some of the molecular processes involved in host invasion and infection both in plant and animal pathogenic oomycetes. These methods provide an opportunity for accurate and detailed functional analysis involving various fields of studies such as genomics, epigenomics, proteomics, and interactomics. Functional gene characterisation is essential for filling the knowledge gaps in dynamic biological processes. However, every method has both advantages and limitations that should be considered before choosing the best method for investigating a particular research question. Here we review transformation systems, gene silencing and gene editing techniques in oomycetes, how they function, in which species and what are their main advantages and disadvantages.

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Abstract

The increasing use of seaweeds in European cuisine led to cultivation initiatives funded by the European Union. lactuca, commonly known as sea lettuce, is a fast growing seaweed in the North Atlantic that chefs are bringing into the local cuisine. Here, different strains of Arctic U. lactuca were mass-cultivated under controlled conditions for up to 10 months. We quantified various chemical constituents associated with both health benefits (carbohydrates, protein, fatty acids, minerals) and health risks (heavy metals). Chemical analyses showed that long-term cultivation provided biomass of consistently high food quality and nutritional value. Concentrations of macroelements (C, N, P, Ca, Na, K, Mg) and micronutrients (Fe, Zn, Co, Mn, I) were sufficient to contribute to daily dietary mineral intake. Heavy metals (As, Cd, Hg and Pb) were found at low levels to pose health risk. The nutritional value of Ulva in terms of carbohydrates, protein and fatty acids is comparable to some selected fruits, vegetables, nuts and grains.

Abstract

Pythium species are ubiquitous organisms known to be pathogens to terrestrial plants and marine algae. While several Pythium species (hereafter, Pythium) are described as pathogens to marine red algae, little is known about the pathogenicity of Pythium on marine green algae. A strain of a Pythium was isolated from a taxonomically unresolved filamentous Ulva collected in an intertidal area of Oslo fjord. Its pathogenicity to a euryhaline Ulva intestinalis collected in the same area was subsequently tested under salinities of 0, 15, and 30 parts per thousand (ppt). The Pythium isolate readily infected U. intestinalis and decimated the filaments at 0 ppt. Mycelium survived on U. intestinalis filaments for at least 2 weeks at 15 and 30 ppt, but the infection did not progress. Sporulation was not observed in the infected algal filaments at any salinity. Conversely, Pythium sporulated on infected grass pieces at 0, 15, and 30 ppt. High salinity retarded sporulation, but did not prevent it. Our Pythium isolate produced filamentous non-inflated sporangia. The sexual stage was never observed and phylogenetic analysis using internal transcribed spacer suggest this isolate belongs to the clade B2. We conclude that the Pythium found in the Oslo fjord was a pathogen of U. intestinalis under low salinity.

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Abstract

Microdochium majus and Microdochium nivale cause serious disease problems in grasses and cereal crops in the temperate regions. Both fungi can infect the plants during winter (causing pink snow mould) as well as under cool humid conditions during spring and fall. We conducted a pathogenicity test of 15 M. nivale isolates and two M. majus isolates from Norway at low temperature on four different grass cultivars of Lolium perenne and Festulolium hybrids. Significant differences between M. nivale isolates in the ability to cause pink snow mould were detected. The M. nivale strains originally isolated from grasses were more pathogenic than isolates from cereals. The genetic diversity of M. nivale and M. majus isolates was studied by sequencing four genetic regions; Elongation factor-1 alpha (EF-1α), β-tubulin, RNA polymerase II (RPB2) and the Internal Transcribed Spacer (ITS). Phylogenetic trees based on the sequences of these four genetic regions resolved M. nivale and M. majus isolates into separate clades. Higher genetic diversity was found among M. nivale isolates than among M. majus isolates. M. nivale isolates revealed genetic differences related to different host plants (grasses vs. cereals) and different geographic regions (Norway and UK vs. North America). Sequence results from the RPB2 and β-tubulin genes were more informative than those from ITS and EF-1α. The genetic and phenotypic differences detected between Norwegian M. nivale isolates from cereals and grasses support the assumption that host specialization exist within M. nivale isolates.

Abstract

Invasive alien species and new plant pests are introduced into new regions at an accelerating rate, due to increasing international trade with soil, plants and plant products. Exotic, plant pathogenic oomycetes in soil from the root zone of imported plants pose a great threat to endemic ecosystems and horticultural production. Detecting them via baiting and isolation, with subsequent identification of the isolated cultures by Sanger sequencing, is labour intensive and may introduce bias due to the selective baiting process. We used metabarcoding to detect and identify oomycetes present in soil samples from imported plants from six different countries. We compared metabarcoding directly from soil both before and after baiting to a traditional approach using Sanger-based barcoding of cultures after baiting. For this, we developed a standardized analysis workflow for Illumina paired-end oomycete ITS metabarcodes that is applicable to future surveillance efforts. In total, 73 soil samples from the rhizosphere of woody plants from 33 genera, in addition to three samples from transport debris, were analysed by metabarcoding the ITS1 region with primers optimized for oomycetes. We detected various Phytophthora and Pythium species, with Pythium spp. being highly abundant in all samples. We also found that the baiting procedure, which included submerging the soil samples in water, resulted in the enrichment of organisms other than oomycetes, compared to non-baited soil samples.

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Abstract

Potato soft rot Pectobacteriaceae (SRP) cause large yield losses and are persistent in seed lots once established. In Norway, different Pectobacterium species are the predominant cause of soft rot and blackleg disease. This work aimed to evaluate the potential of real-time PCR for quantification of SRP in seed tubers, as well as investigating the status of potato seed health with respect to SRP in Norway. A total of 34 seed potato lots, including certified seeds, was grown and monitored over three consecutive years. All seed lots contained a quantifiable amount of SRP after enrichment, with very few subsamples being free of the pathogens. A high SRP prevalence based on a qPCR assay, as well as a high symptom incidence in certified seeds were observed, suggesting that current criteria for seed certification are insufficient to determine tuber health and predict field outcomes. Pectobacterium atrosepticum was the most abundant species in the examined seed lots and present in all lots. Consistently good performance of first generation seed lots with respect to blackleg and soft rot incidence, as well as low quantity of SRP in these seed lots demonstrated the importance of clean seed potatoes. Weather conditions during the growing season seemed to govern disease incidence and SRP prevalence more than seed grade. The impact of temperature, potato cultivar and Pectobacterium species on tuber soft rot development were further examined in tuber infection experiments, which showed that temperature was the most important factor in nearly all cultivars. Large-scale quantification of latent infection and predictive models that include contributing factors like weather, infecting bacterial species and cultivar are needed to reduce soft rot and blackleg.

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Abstract

Strawberry powdery mildew (Podosphaera aphanis Wallr.) is a pathogen which infects the leaves, fruit, stolon and flowers of the cultivated strawberry (Fragaria ×ananassa), causing major yield losses, primarily through unmarketable fruit. The primary commercial control of the disease is the application of fungicidal sprays. However, as the use of key active ingredients of commercial fungicides is becoming increasingly restricted, interest in developing novel strawberry cultivars exhibiting resistance to the pathogen is growing rapidly. In this study, a mapping population derived from a cross between two commercial strawberry cultivars (‘Sonata’ and ‘Babette’) was genotyped with single nucleotide polymorphism (SNP) markers from the Axiom iStraw90k genotyping array and phenotyped for powdery mildew susceptibility in both glasshouse and field environments. Three distinct, significant QTLs for powdery mildew resistance were identified across the two experiments. Through comparison with previous studies and scrutiny of the F. vesca genome sequence, candidate genes underlying the genetic control of this trait were identified.

Abstract

Plants are exposed to various pathogens in their environment and have developed immune systems with multiple defense layers to prevent infections. However, often pathogens overcome these resistance barriers, infect plants and cause disease. Pathogens that cause diseases on economically important crop plants incur huge losses to the agriculture industry. For example, the 2016 outbreak of strawberry grey mold (Botrytis cinerea) in Norway caused up to 95% crop losses. Such outbreaks underline the importance of developing novel and sustainable tools to combat plant diseases, for example by increasing the plants’ natural disease resistance. Priming plant defenses using chemical elicitors may enhance resistance against multiple pathogens. Such an approach may reduce the use of chemical fungicides and pesticides that often select for resistant strains of pests and pathogens. My presentation will focus on the effectiveness of different chemical agents to prime woodland strawberry (Fragaria vesca) defenses against the necrotroph B. cinerea. We have identified several genes that seem to play a role in disease resistance in strawberry and associated epigenetic memory mechanisms. Our results point out new management avenues for more sustainable crop protection schemes.

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Abstract

Fusarium is one of the most diverse fungal genera affecting several crops around the world. This study describes the phylogeny of Fusarium species associated with grains of sorghum and finger millet from different parts of Ethiopia. Forty-two sorghum and 34 finger millet grain samples were mycologically analysed. All of the sorghum and more than 40% of the finger millet grain samples were contaminated by the Fusarium species. The Fusarium load was higher in sorghum grains than that in finger millet grains. In addition, 67 test isolates were phylogenetically analysed using EF-1α and β-tubulin gene primers. Results revealed the presence of eight phylogenetic placements within the genus Fusarium, where 22 of the isolates showed a close phylogenetic relation to the F. incarnatum–equiseti species complex. Nevertheless, they possess a distinct shape of apical cells of macroconidia, justifying the presence of new species within the Fusarium genus. The new species was the most dominant, represented by 33% of the test isolates. The current work can be seen as an important addition to the knowledge of the biodiversity of fungal species that exists within the Fusarium genus. It also reports a previously unknown Fusarium species that needs to be investigated further for toxin production potential.

Abstract

Aims Bacterial decays of onion bulbs have serious economic consequences for growers, but the aetiologies of these diseases are often unclear. We aimed to determine the role of Rahnella, which we commonly isolated from bulbs in the United States and Norway, in onion disease. Methods and Results Isolated bacteria were identified by sequencing of housekeeping genes and/or fatty acid methyl ester analysis. A subset of Rahnella spp. strains was also assessed by multilocus sequence analysis (MLSA); most onion strains belonged to two clades that appear closely related to R. aquatilis. All tested strains from both countries caused mild symptoms in onion bulbs but not leaves. Polymerase chain reaction primers were designed and tested against strains from known species of Rahnella. Amplicons were produced from strains of R. aquatilis, R. victoriana, R. variigena, R. inusitata and R. bruchi, and from one of the two strains of R. woolbedingensis. Conclusions Based on binational testing, strains of Rahnella are commonly associated with onions, and they are capable of causing mild symptoms in bulbs. Significance and Impact of the Study While Rahnella strains are commonly found within field‐grown onions and they are able to cause mild symptoms, the economic impact of Rahnella‐associated symptoms remains unclear.

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Abstract

The oomycete pathogen Phytophthora cactorum causes crown rot, a major disease of cultivated strawberry. We report the draft genome of P. cactorum isolate 10300, isolated from symptomatic Fragaria x ananassa tissue. Our analysis revealed that there are a large number of genes encoding putative secreted effectors in the genome, including nearly 200 RxLR domain containing effectors, 77 Crinklers (CRN) grouped into 38 families, and numerous apoplastic effectors, such as phytotoxins (PcF proteins) and necrosis inducing proteins. As in other Phytophthora species, the genomic environment of many RxLR and CRN genes differed from core eukaryotic genes, a hallmark of the two-speed genome. We found genes homologous to known Phytophthora infestans avirulence genes including Avr1, Avr3b, Avr4, Avrblb1 and AvrSmira2 indicating effector sequence conservation between Phytophthora species of clade 1a and clade 1c. The reported P. cactorum genome sequence and associated annotations represent a comprehensive resource for avirulence gene discovery in other Phytophthora species from clade 1 and, will facilitate effector informed breeding strategies in other crops.

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Abstract

The oomycete Phytophthora infestans, the cause of late blight, is one of the most important potato pathogens. During infection, it secretes effector proteins that manipulate host cell function, thus contributing to pathogenicity. This study examines sequence differentiation of two P. infestans effectors from 91 isolates collected in Poland and Norway and five reference isolates. A gene encoding the Avr-vnt1 effector, recognized by the potato Rpi-phu1 resistance gene product, is conserved. In contrast, the second effector, AvrSmira1 recognized by Rpi-Smira1, is highly diverse. Both effectors contain positively selected amino acids. A majority of the polymorphisms and all selected sites are located in the effector C-terminal region, which is responsible for their function inside host cells. Hence it is concluded that they are associated with a response to diversified target protein or recognition avoidance. Diversification of the AvrSmira1 effector sequences, which existed prior to the large-scale cultivation of plants containing the Rpi-Smira1 gene, may reduce the predicted durability of resistance provided by this gene. Although no isolates virulent to plants with the Rpi-phu1 gene were found, the corresponding Avr-vnt1 effector has undergone selection, providing evidence for an ongoing ‘arms race’ between the host and pathogen. Both genes remain valuable components for resistance gene pyramiding.

Abstract

Total forfattarliste: Franić, I., Prospero, S., Adamson, K., Allan, A., Auger-Rozenberg, A-M, Augustin, S., Avtzis, D., Barta, M., Boroń, P., Bragança, H., Brestovanská, T., Brurberg, M. B., Burgess, B., Burokienė, D., Černý, K., Cleary, M., Corley, J., Coyle, D. R., Csóka, G., Davydenko, K., Elsafy, M. A. O., Eötvös, C., de Groot, M., Diez, J. J., Lehtijärvi, H. T. D., Drenkhan, R., Fan, J., Grabowski, M., Grad, B., Havrdova, L., Hrabetova, M., Iede, E. T., Kacprzyk, M., Kenis, M., Kirichenko25,45, N., Lacković26,N., Lazarević, J., Leskiv, M., Li, H., Madsen, C.L., Matošević, D., Matsiakh, I., Meffert, J., Migliorini, D., Mikó, Á., Nikolov, C., O'Hanlon, R., Oskay, F., Paap, T., Parpan, T., Petrakis, P.V., Piškur, B., Ravn, H.P., Ronse, A., Roques, A., Schühli, G.S., Sivickis, K., Talgø, V., Tomoshevich, M., Uimari, A., Ulyshen, M., Vettraino, A.M., Villari, C., Wang, Y., Witzell, J., Zlatković, M., Eschen, R.

Abstract

Total forfattarliste: Franić, I., Prospero, S., Adamson, K., Allan, A., Auger-Rozenberg, A-M, Augustin, S., Avtzis, D., Barta, M., Boroń, P., Bragança, H., Brestovanská, T., Brurberg, M. B., Burgess, B., Burokienė, D., Černý, K., Cleary, M., Corley, J., Coyle, D. R., Csóka, G., Davydenko, K., Elsafy, M. A. O., Eötvös, C., de Groot, M., Diez, J. J., Lehtijärvi, H. T. D., Drenkhan, R., Fan, J., Grabowski, M., Grad, B., Havrdova, L., Hrabetova, M., Iede, E. T., Kacprzyk, M., Kenis, M., Kirichenko25,45, N., Lacković26,N., Lazarević, J., Leskiv, M., Li, H., Madsen, C.L., Matošević, D., Matsiakh, I., Meffert, J., Migliorini, D., Mikó, Á., Nikolov, C., O'Hanlon, R., Oskay, F., Paap, T., Parpan, T., Petrakis, P.V., Piškur, B., Ravn, H.P., Ronse, A., Roques, A., Schühli, G.S., Sivickis, K., Talgø, V., Tomoshevich, M., Uimari, A., Ulyshen, M., Vettraino, A.M., Villari, C., Wang, Y., Witzell, J., Zlatković, M., Eschen, R.

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Abstract

Phytophthora cryptogea, P. gonapodyides, P. lacustris, P. megasperma, P. plurivora, P. taxon paludosa and an unknown Phytophthora species were isolated from waterways and soil samples in Christmas tree fields in southern Sweden. In addition, P. megasperma was isolated from a diseased Norway spruce (Picea abies) plant from one of the fields in Svalöv. Inoculation tests were sequentially carried out with one isolate from each of the three species P. cryptogea, P. megasperma, and P. plurivora, all known pathogens on conifers. The same three isolates were used to study a few morphological features to confirm the identification, and temperature-growth relationships were carried out to see how well the organisms fit into Swedish climatic conditions. Seedlings of Norway spruce and Nordmann fir (Abies nordmanniana) were inoculated in the roots and the stems. None of the isolates caused extensive root rot under the experimental conditions, but all three species could be re-isolated from both Norway spruce and Nordmann fir. Phytophthora root rot is currently of minor concern for Christmas tree growers in Sweden. However, the Phytophthora isolations from soil and water indicate the presence of this damaging agent, which may lead to future problems.

Abstract

Plants are exposed to various pathogens in their environment and have developed immune systems with multiple layers of defence to fight-back. However, often pathogens overcome the resistance barriers, infect the plants to cause the disease. Pathogens that cause diseases on economically important crop plants like strawberry incur huge losses to the agriculture industry. For example, The 2016 outbreak of strawberry grey mould (Botrytis cinerea) in Norway caused up to 95% crop losses. Outbreaks like this underline the importance of developing novel and sustainable tools to combat plant diseases, for example by increasing the plants’ natural disease resistance. Priming plant defences using chemical elicitors may be effective in providing the enhanced resistance against multiple pathogens. We have used β-aminobutyric acid (BABA) as a chemical priming agent to induce resistance in Fragaria vesca against Botrytis cinerea. Effects of BABA on disease progression and defence responses of Fragaria are being characterized using molecular tools like RNAseq, RT-PCR and ChIP. As priming chemicals may induce an epigenetic memory in treated plants, we also plan to study the histone methylation patterns in primed plants and the genes that are regulated. Our long-term aim is to understand the duration of the epigenetic memory and its cross-generational transmission to the progeny in Fragaria. Our results will help guide various crop protection strategies in addition to providing new insights to develop novel tools for plant disease management.

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Abstract

Pathogenic soft rot Enterobacteriaceae (SRE) belonging to the genera Pectobacterium and Dickeya cause diseases in potato and numerous other crops. Seed potatoes are the most important source of infection, but how pathogen-free tubers initially become infected remains an enigma. Since the 1920s, insects have been hypothesized to contribute to SRE transmission. To validate this hypothesis and to map the insect species potentially involved in SRE dispersal, we have analyzed the occurrence of SRE in insects recovered from potato fields over a period of 2 years. Twenty-eight yellow sticky traps were set up in 10 potato fields throughout Norway to attract and trap insects. Total DNA recovered from over 2,000 randomly chosen trapped insects was tested for SRE, using a specific quantitative PCR (qPCR) TaqMan assay, and insects that tested positive were identified by DNA barcoding. Although the occurrence of SRE-carrying insects varied, they were found in all the tested fields. While Delia species were dominant among the insects that carried the largest amount of SRE, more than 80 other SRE-carrying insect species were identified, and they had different levels of abundance. Additionally, the occurrence of SRE in three laboratory-reared insect species was analyzed, and this suggested that SRE are natural members of some insect microbiomes, with herbivorous Delia floralis carrying more SRE than the cabbage moth (Plutella xylostella) and carnivorous green lacewing larvae (Chrysoperla carnea). In summary, the high proportion, variety, and ubiquity of insects that carried SRE show the need to address this source of the pathogens to reduce the initial infection of seed material.

Abstract

The plant pathogenic fungus Fusarium langsethiae produces the highly potent mycotoxins HT-2 and T-2. Since these toxins are frequently detected at high levels in oat grain lots, they pose a considerable risk for food and feed safety in Norway, as well as in other north European countries. To reduce the risk of HT-2/T- 2-contaminated grain lots to enter the food and feed chain, it is important to identify factors that influence F. langsethiae infection and mycotoxin development in oats. However, the epidemiology of F. langsethiae is unclear. A three-year survey was performed to reveal more of the life cycle of F. langsethiae and its interactions with oats, other Fusarium species, as well as insects, mites and weeds. We searched for inoculum sources by quantifying the amount of F. langsethiae DNA in crop residues, weeds, and soil sampled from a selection of oat-fields. To be able to define the onset of infection, we analysed the amount of F. langsethiae DNA in oat plant material sampled at selected growth stages (between booting and maturation), as well as the amount of F. langsethiae DNA and HT-2 and T-2 toxins in the mature grain. We also studied the presence of possible insect- and mite vectors sampled at the selected growth stages using Berlese funnel traps. The different types of materials were also analysed for the presence F. graminearum DNA, the most important deoxynivalenol producer observed in Norwegian cereals, and which presence has shown a striking lack of correlation with the presence of F. langsethiae in oat. Results show that F. langsethiae DNA may occur in the oat plant already before heading and flowering. Some F. langsethiae DNA was observed in crop residues and weeds, though at relatively low levels. No Fusarium DNA was detected in soil samples. Of the arthropods that were associated with the collected oat plants, aphids and thrips species were dominating. Further details will be given at the meeting.

Abstract

Sclerotinia stem rot (SSR) is the most important disease of oilseed Brassica crops in Norway. Fungicide applications should be aligned with the actual need for control, but the SSR prediction models used lack accuracy. We have studied the importance of precipitation, and the role of petal and leaf infection for SSR incidence by using data from Norwegian field and trap plant trials over several years. In the trials, SSR incidence ranged from 0 to 65%. Given an infection threshold of 25% SSR, regression and Receiver Operating Characteristics (ROC) analysis were used to evaluate different precipitation thresholds. The sum of precipitation two weeks before and during flowering appeared to be a poor predictor for SSR infection in our field and trap plant trials (P = 0.24, P = 0.11, respectively). Leaves from three levels (leaf one, three, five), and petals were collected at three to four different times during flowering from nine field sites over two years and tested for SSR infection with real-time PCR. Percentage total leaf and petal infection explained 57 and 45% of variation in SSR incidence, respectively. Examining the different leaves and petals separately, infection of leaf three sampled at full flowering showed the highest explanation of variation in later SSR incidence (R2 = 65%, P < 0.001). ROC analysis showed that given an infection threshold of 45%, both petal and leaf infection recommended spraying when spraying was actually needed. Combining information on petal and leaf infection during flowering with relevant microclimate factors in the canopy, instead of the sum of precipitation might improve prediction accuracy for SSR.

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Abstract

Helminthosporium solani causes silver scurf, which affects the quality of potato. The biocontrol agent Clonostachys rosea greatly limited the severity of silver scurf symptoms and amount of H. solani genomic DNA in laboratory experiments. Transcriptomic analysis during interaction showed that H. solani gene expression was highly reduced when coinoculated with the biocontrol agent C. rosea, whereas gene expression of C. rosea was clearly boosted as a response to the pathogen. The most notable upregulated C. rosea genes were those encoding proteins involved in cellular response to oxidative stress, proteases, G-protein signaling, and the methyltransferase LaeA. The most notable potato response to both fungi was downregulation of defense-related genes and mitogen-activated protein kinase kinase kinases. At a later stage, this shifted, and most potato defense genes were turned on, especially those involved in terpenoid biosynthesis when H. solani was present. Some biocontrol-activated defense-related genes in potato were upregulated during early interaction with C. rosea alone that were not triggered by H. solani alone. Our results indicate that the reductions of silver scurf using C. rosea are probably due to a combination of mechanisms, including mycoparasitism, biocontrol-activated stimulation of plant defense mechanisms, microbial competition for nutrients, space, and antibiosis.

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Abstract

Soft rot and blackleg of potato caused by pectinolytic bacteria lead to severe economic losses in potato production worldwide. To investigate the species composition of bacteria causing soft rot and black leg of potato in Norway and Poland, bacteria were isolated from potato tubers and stems. Forty-one Norwegian strains and 42 Polish strains that formed cavities on pectate medium were selected for potato tuber maceration assays and sequencing of three housekeeping genes (dnaX, icdA and mdh) for species identification and phylogenetic analysis. The distribution of the species causing soft rot and blackleg in Norway and Poland differed: we have demonstrated that mainly P. atrosepticum and P. c. subsp. carotovorum are the causal agents of soft rot and blackleg of potatoes in Norway, while P. wasabiae was identified as one of the most important soft rot pathogens in Poland. In contrast to the other European countries, D. solani seem not to be a major pathogen of potato in Norway and Poland. The Norwegian and Polish P. c. subsp. carotovorum and P. wasabiae strains did not cluster with type strains of the respective species in the phylogenetic analysis, which underlines the taxonomic complexity of the genus Pectobacterium. No correlation between the country of origin and clustering of the strains was observed. All strains tested in this study were able to macerate potato tissue. The ability to macerate potato tissue was significantly greater for the P. c. subsp. carotovorum and Dickeya spp., compared to P. atrosepticum and P. wasabiae.

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Abstract

The genus Microbacterium contains bacteria that are ubiquitously distributed in various environments and includes plant-associated bacteria that are able to colonize tissue of agricultural crop plants. Here, we report the 3,508,491 bp complete genome sequence of Microbacterium sp. strain BH-3-3-3, isolated from conventionally grown lettuce (Lactuca sativa) from a field in Vestfold, Norway. The nucleotide sequence of this genome was deposited into NCBI GenBank under the accession CP017674.

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Abstract

Dollar spot is a destructive and widespread disease affecting most turfgrass species, but until recently it has been absent from the Scandinavian countries of northern Europe. In the fall of 2014, disease symptoms consistent with dollar spot were observed on a golf course fairway in Sweden. A fungus was isolated from symptomatic turf and identified as Sclerotinia homoeocarpa on the basis of ribosomal deoxyribonucleic acid (rDNA) internal transcribed spacer (ITS) sequences, morphology, and culture characteristics. The ITS sequence was identical to isolates of S. homoeocarpa from the eastern and midwestern United States. Koch’s postulates were fulfilled, confirming the S. homoeocarpa isolate as the causal agent. This is the first report of turfgrass dollar spot in Sweden and only the second report of the disease from Scandinavia. Because pesticides are rarely used in the cultivation of Scandinavian turfgrass, dollar spot disease may prove difficult to control through conventional means and potentially represents a major threat to the industry.

Abstract

Knowledge about the reproduction strategies of invasive species is fundamental for effective control. The invasive Fallopia taxa (Japanese knotweed s.l.) reproduce mainly clonally in Europe, and preventing spread of vegetative fragments is the most important control measure. However, high levels of genetic variation within the hybrid F. × bohemica indicate that hybridization and seed dispersal could be important. In Norway in northern Europe, it is assumed that these taxa do not reproduce sexually due to low temperatures in the autumn when the plants are flowering. The main objective of this study was to examine the genetic variation of invasive Fallopia taxa in selected areas in Norway in order to evaluate whether the taxa may reproduce by seeds in their most northerly distribution range in Europe. Fallopia stands from different localities in Norway were analyzed with respect to prevalence of taxa, and genetic variation within and between taxa was studied using amplified fragment length polymorphism (AFLP). Taxonomic identification based on morphology corresponded with identification based on simple sequence repeats (SSR) and DNA ploidy levels (8× F. japonica, 6× F. × bohemica and 4× F. sachalinensis). No genetic variation within F. japonica was detected. All F. × bohemica samples belonged to a single AFLP genotype, but one sample had a different SSR genotype. Two SSR genotypes of F. sachalinensis were also detected. Extremely low genetic variation within the invasive Fallopia taxa indicates that these taxa do not reproduce sexually in the region, suggesting that control efforts can be focused on preventing clonal spread. Climate warming may increase sexual reproduction of invasive Fallopia taxa in northern regions. The hermaphrodite F. × bohemica is a potential pollen source for the male-sterile parental species. Targeted eradication of the hybrid can therefore reduce the risk of increased sexual reproduction under future warmer climate.

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Abstract

The natural occurrence of fungi, mycotoxins and fungal metabolites was investigated in 100 samples of maize grains collected from south and southwestern Ethiopia in 2015. The maize samples were contaminated by Fusarium, Aspergillus and Penicillium species. Using liquid chromatography tandem mass spectrometry 127 secondary metabolites were analysed. Zearalenone was the most prevalent mycotoxin, occurring in about 96% of the samples. Zearalenone sulfate was the second most prevalent, present in 81% of the samples. Fumonisin B1 was detected in 70% of the samples with a mean level of 606 μg kg−1 in positive samples, while FB2, FB3 and FB4 were detected in 62%, 51% and 60% of the maize samples with mean levels of 202, 136 and 85 μg kg−1, respectively. Up to 8% of the samples were contaminated with aflatoxins, with a maximum level of aflatoxin B1 of 513 μg kg−1. Results were higher than earlier reports for maize from Ethiopia.

Abstract

The genus Pectobacterium, which belongs to the bacterial family Enterobacteriaceae, contains numerous species that cause soft rot diseases in a wide range of plants. The species Pectobacterium carotovorum is highly heterogeneous, indicating a need for re-evaluation and a better classification of the species. PacBio was used for sequencing of two soft-rot-causing bacterial strains (NIBIO1006T and NIBIO1392), initially identified as P. carotovorum strains by fatty acid analysis and sequencing of three housekeeping genes (dnaX, icdA and mdh). Their taxonomic relationship to other Pectobacterium species was determined and the distance from any described species within the genus Pectobacterium was less than 94% average nucleotide identity (ANI). Based on ANI, phylogenetic data and genome-to-genome distance, strains NIBIO1006T, NIBIO1392 and NCPPB3395 are suggested to represent a novel species of the genus Pectobacterium, for which the name Pectobacterium polaris sp. nov. is proposed. The type strain is NIBIO1006T (=DSM 105255T=NCPPB 4611T).

Abstract

Important losses in strawberry production are often caused by the oomycete Phytophthora cactorum, the causal agent of crown rot. However, very limited studies at molecular levels exist of the mechanisms related to strawberry resistance against this pathogen. To begin to rectify this situation, a PCR-based approach (NBS profiling) was used to isolate strawberry resistance gene analogs (RGAs) with altered expression in response to P. cactorum during a time course (2, 4, 6, 24, 48, 96 and 192 h post-infection). Twenty-three distinct RGA fragments of the NB-LRR type were identified from a resistance genotype (Bukammen) of the wild species Fragaria vesca. The gene transcriptional profiles after infection showed that the response of most RGAs was quicker and stronger in the resistance genotype (Bukammen) than in the susceptible one (FDP821) during the early infection stage. The transcriptional patterns of one RGA (RGA109) were further monitored and compared during the P. cactorum infection of two pairs of resistant and susceptible genotype combinations (Bukammen/FDP821 and FDR1218/1603). The 5′ end sequence was cloned, and its putative protein was characteristic of NBS-LRR R protein. Our results yielded a first insight into the strawberry RGAs responding to P. cactorum infection at molecular level.

Abstract

The prevalence of Fusarium dry rot in potatoes produced in Norway was investigated in a survey for three consecutive years in the period 2010 to 2012. A total of 238 samples (comprising 23,800 tubers) were collected, representing different cultivars and production regions in Norway. Fusarium spp. were detected in 47% of the samples, with one to three species per sample. In total, 718 isolates of Fusarium spp. were recovered and identified to seven species. The most commonly isolated species was Fusarium coeruleum, comprising 59.6% of the total Fusarium isolates and found in 17.2% of the collected samples, followed by Fusarium avenaceum (27.2% of the isolates and found in 27.7% of the samples). Fusarium sambucinum was the third most prevalent species (6.4% in 8.8% of the samples) and Fusarium culmorum the fourth (5.2% in 6.3% of the samples). Less prevalent species included Fusarium cerealis, Fusarium graminearum, and Fusarium equiseti (<1% in 0.4 to 1.3% of the samples). F. coeruleum was the most prevalent species in northern and southwestern Norway, whereas F. avenaceum was dominating in eastern Norway. The potato cultivars Berber and Rutt were susceptible to all Fusarium spp. A new TaqMan real-time PCR assay specific for F. coeruleum was developed, which successfully identified Norwegian isolates. This and other previously developed real-time PCR assays targeting different Fusarium species were evaluated for their ability to detect latent infections in potatoes at harvest. This study provides new information on the current occurrence of different Fusarium species causing Fusarium dry rot in potatoes in Europe including areas far into the arctic in the north of Norway.

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Abstract

Amplified fragment length polymorphism (AFLP) was used to study the genetic variation among 80 F. verticillioides isolates from kernels of Ethiopian maize, collected from 20 different maize growing areas in four geographic regions. A total of 213 polymorphic fragments were obtained using six EcoRI/MseI primer combinations. Analysis of the data based on all 213 polymorphic AFLP fragments revealed high level of genetic variation in the F. verticillioides entities in Ethiopia. About 58% of the fragments generated were polymorphic. The genetic similarity among F. verticillioides isolates varied from 46% to 94% with a mean Dice similarity of 73%. Unweighted Pair Group Method with Arithmetic Average (UPGMA) analysis revealed two main groups and four subgroups. The principal coordinate analysis (PCO) also displayed two main groups that agreed with the results of UPGMA analysis, and there was no clear pattern of clustering of isolates according to geographic origin. Analysis of molecular variance: (AMOVA) showed that only 1.5% of the total genetic variation was between geographic regions, while 98.5% was among isolates from the same geographic regions of Ethiopia. Eighty distinct haplotypes were recognized among the 80 isolates analyzed. Hence, breeding efforts should concentrate on quantitative resistance that is effective against all genotypes of the pathogen.

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Abstract

Lack of resistance to pink snow mould (Microdochium nivale) is a major constraint for adaptation of perennial ryegrass (Lolium perenne L.) to continental regions with long-lasting snow cover at higher latitudes. Almost all investigations of genetic variation in resistance have been performed using cold acclimated plants. However, there may be variation in resistance mechanisms that are functioning independently of cold acclimation. In this study our aim was to identify candidate genes involved in such resistance mechanisms. We first characterized variation in resistance to M. nivale among non-acclimated genotypes from the Norwegian cultivar ‘Fagerlin’ based on relative regrowth and fungal quantification by real-time qPCR. One resistant and one susceptible genotype were selected for transcriptome analysis using paired-end sequencing by Illumina Hiseq 2000. Transcriptome profiles, GO enrichment and KEGG pathway analysis indicate that defense response related genes are differentially expressed between the resistant and the susceptible genotype. A significant up-regulation of defense related genes, as well as genes involved in cell wall cellulose metabolic processes and aryl-alcohol dehydrogenase (NADP+) activity, was observed in the resistant genotype. The candidate genes identified in this study might be potential molecular marker resources for breeding perennial ryegrass cultivars with improved resistance to pink snow mould.

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Abstract

Fusarium species causing maize kernel rot are major threats to maize production, due to reduction in yield as well as contamination of kernels by mycotoxins that poses a health risk to humans and animals. Two-hundred maize kernel samples, collected from 20 major maize growing areas in Ethiopia were analyzed for the identity, species composition and prevalence of Fusarium species and fumonisin contamination. On average, 38 % (range: 16 to 68 %) of maize kernels were found to be contaminated by different fungal species. Total of eleven Fusarium spp. were identified based on morphological characteristics and by sequencing the partial region of translation elongation factor 1-alpha (EF-1α) gene. Fusarium verticillioides was the dominant species associated with maize kernels (42 %), followed by F. graminearum species complex (22.5 %) and F. pseudoanthophilium (13.4 %). The species composition and prevalence of Fusarium species differed among the areas investigated. Fusarium species composition was as many as eight and as few as four in some growing area. The majority of the maize samples (77 %) were found positive for fumonisin, with concentrations ranging from 25 μg kg−1 to 4500 μg kg−1 (mean: 348 μg kg−1 and median: 258 μg kg−1). Slight variation in fumonisin concentration was also observed among areas. Overall results indicate widespread occurrence of several Fusarium species and contamination by fumonisin mycotoxins. These findings are useful for intervention measures to reduce the impact of the main fungal species and their associated mycotoxins, by creating awareness and implementation of good agricultural practices.

Abstract

The plant pathogenic fungus Fusarium langsethiae produces the highly potent mycotoxins HT-2 and T-2. Since these toxins are frequently detected at high levels in oat grain lots, they pose a considerable risk for food and feed safety in Norway, as well as in other north European countries. To reduce the risk of HT-2/T- 2-contaminated grain lots to enter the food and feed chain, it is important to identify factors that influence F. langsethiae infection and mycotoxin development in oats. However, the epidemiology of F. langsethiae is unclear. A three-year survey was performed to reveal more of the life cycle of F. langsethiae and its interactions with oats, other Fusarium species, as well as insects, mites and weeds. We searched for inoculum sources by quantifying the amount of F. langsethiae DNA in weeds, crop residues, and soil, sampled from a predetermined selection of oat-fields. To be able to define the onset of infection, we analysed the amount of F. langsethiae DNA in oat plant material sampled at selected growth stages (between booting and maturation), as well as the amount of F. langsethiae DNA and HT-2 and T-2 toxins in the mature grain. We also studied the presence of possible insect- and mite vectors sampled at the selected growth stages using Berlese funnel traps. All the different types of materials were also analysed for the presence F. graminearum DNA, the most important deoxynivalenol producer observed in Norwegian cereals, and which presence has shown a striking lack of correlation with the presence F. langsethiae in oat. Preliminary results show that F. langsethiae DNA may occur in the oat plant before heading and flowering. Some F. langsethiae DNA was observed in crop residues and weeds, though at relatively low levels. More results from this work will be presented at the meeting.

Abstract

The plant pathogenic fungus Fusarium langsethiae produces the highly potent mycotoxins HT-2 and T-2. Since these toxins are frequently detected at high levels in oat grain lots, they pose a considerable risk for food and feed safety in Norway, as well as in other north European countries. To reduce the risk of HT-2/T- 2-contaminated grain lots to enter the food and feed chain, it is important to identify factors that influence F. langsethiae infection and mycotoxin development in oats. However, the epidemiology of F. langsethiae is unclear. A three-year survey was performed to reveal more of the life cycle of F. langsethiae and its interactions with oats, other Fusarium species, as well as insects, mites and weeds. We searched for inoculum sources by quantifying the amount of F. langsethiae DNA in weeds, crop residues, and soil, sampled from a predetermined selection of oat-fields. To be able to define the onset of infection, we analysed the amount of F. langsethiae DNA in oat plant material sampled at selected growth stages (between booting and maturation), as well as the amount of F. langsethiae DNA and HT-2 and T-2 toxins in the mature grain. We also studied the presence of possible insect- and mite vectors sampled at the selected growth stages using Berlese funnel traps. All the different types of materials were also analysed for the presence F. graminearum DNA, the most important deoxynivalenol producer observed in Norwegian cereals, and which presence has shown a striking lack of correlation with the presence F. langsethiae in oat. Preliminary results show that F. langsethiae DNA may occur in the oat plant before heading and flowering. Some F. langsethiae DNA was observed in crop residues and weeds, though at relatively low levels. More results from this work will be presented at the meeting.

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Abstract

Delphinella shoot blight (Delphinella abietis) attacks true firs (Abies spp.) in Europe and North America. Especially subalpine fir (A. lasiocarpa), one of the main Christmas tree species in Norway, is prone to the disease. The fungus kills current year needles, and in severe cases entire shoots. Dead needles become covered with black fruiting bodies, both pycnidia and pseudothecia. Delphinella shoot blight has mainly been a problem in humid, coastal regions in the northwestern part of Southern Norway, but, probably due to higher precipitation in inland regions during recent years, heavy attacks were found in 2011 in a field trial with 76 provenances of subalpine fir in Southeastern Norway. However, the amount of precipitation seemed less important once the disease had established in the field. Significant differences in susceptibility between provenances were observed. In general, the more bluish the foliage was, the healthier the trees appeared. The analysis of provenance means indicated that, at least for the southern range, the disease ratings were correlated with foliage color. This study also includes isolation, identification, a pathogenicity test, a seed test and electron microscopy of the wax layer on the needles. The fungus was identified based on the morphology of spores and by sequencing the Internal Transcribed Spacer (ITS) regions of the ribosomal DNA. Koch’s postulates were fulfilled. The fungus was found present on newly harvested seeds and may therefore spread via international seed trade. When comparing the wax layers on green and blue needles, those of the latter were significantly thicker, a factor that may be involved in disease resistance.

Abstract

The woodland strawberry (Fragaria vesca) has become the model plant for the economically important, but genetically complex, octoploid F. × ananassa. Crown rot caused by the oomycete Phytophthora cactorum is a major problem for the strawberry industry and the identification and incorporation of efficient resistance genes into superior cultivars are important for breeding. In the present study, two experimental populations were used in inoculation experiments under controlled greenhouse condition. Studies of a sparse diallel cross between resistant and susceptible F. vesca genotypes concluded that resistance to crown rot is inherited as a dominant trait under nuclear control. Subsequently, an F2 population derived from the grandparents Bukammen (resistant) and Haugastøl 3 (susceptible) collected in Norway, were phenotyped in infection experiments and genotyped using genotyping-by-sequencing. A 416.2-cM linkage map was constructed, and a single major gene locus was identified on linkage group 6 that we attributed to resistance to Phytopthora infection. We propose to name the resistance locus RPc-1 (Resistance to Phytophthora cactorum 1). Gene prediction of the 3.3 Mb QTL recovered 801 genes of which 69 had a potential role in plant disease resistance.

Abstract

Pythium species are fungal-like organisms distributed all over the world. Most Pythium spp. live as saprophytes, but some of them are pathogenic. Here we report on disease incidence in Norway spruce (Picea abies) seedlings caused by Pythium undulatum, and pathogenicity in vitro of Norwegian isolates of P. undulatum and P. anandrum.

Abstract

A survey of nematodes associated with terrestrial slugs was conducted for the first time in Norway. A total of 611 terrestrial slugs were collected from 32 sample sites. Slugs were identified by means of morphological examination, dissection of genitalia and molecular analysis using mitochondrial DNA. Twelve slug species were identified, representing four different slug families. Internal nematodes were identified by means of morphological analysis and the sequencing of the 18S rRNA gene. Of the sample sites studied, 62.5% were found to be positive for nematode parasites, with 18.7% of all slugs discovered being infected. Five nematode species were identified in this study: Alloionema appendiculatum, Agfa flexilis, Angiostoma limacis, Angiostoma sp. and Phasmarhabditis hermaphrodita. Of these species, only one nematode was previously undescribed (Angiostoma sp.). This is the first record of the presence of A. appendiculatum, A. flexilis and A. limacis in Norway.

Abstract

Initial sources of inoculum of Phytophthora infestans were investigated in ten potato fields with early outbreaks of potato late blight. Infected plant samples and isolates from these fields were examined with respect to mating type prevalence, fungicide resistance and genotypes based on microsatellites A high proportion (91 %) of the isolates recovered were of mating type A1. However, both mating types were found in 3 of 9 fields with more than one isolate recovered, and sometimes both mating types were found on the same plant. Most of the isolates recovered from fields treated with metalaxyl-M prior to sampling had reduced sensitivity or were resistant to metalaxyl-M, and most of the isolates recovered form fields without metalaxyl treatment were sensitive. The isolates recovered from fields treated with propamocarb prior to sampling had a higher frequency of reduced sensitivity to propamocarb than isolates from fields without propamocarb treatment. We found that most plants contained more than one P. infestans SSR-genotype. Clustering analysis of the infected samples revealed that most samples clustered together according to fields. By combining information from P. infestans isolates and DNA extracts from the leaf lesions we found examples of both mating type A1 and A2 having the same multilocus genotype. This result indicates that both of these genotypes have a common ancestor, hence the inoculum originates from oospores. Although this a minor study of only 10 fields with a limited amount of isolates and plant samples, the results indicate oospores in the soil is an inoculum source. Hence the forecasting model to predict outbreaks of potato late blight should be modified to include this.

Abstract

In 2008, an epidemic caused by a new Neonectria sp. was discovered on white fir (Abies concolor) in several counties in southern Norway [1]. Later the pathogen was also found on other fir species in Norway and Denmark [2]. Typical symptoms and signs were dead shoots, flagging (dead branches), canker wounds, heavy resin flow, and occasionally red fruiting bodies (perithecia). Pathogenicity tests on several Abies spp. proved the fungus to be very aggressive, which corresponds well with observations of mortality of white fir and subalpine fir (A. lasiocarpa) from different age classes under field conditions. Sequencing of the internal transcribed regions (ITS) of the ribosomal DNA showed that this Neonectria sp. was most similar to N. ditissima (only 5 bp different from isolates in the GenBank), a common pathogen worldwide on broad leaf trees. The ITS sequences were very different (> 20 bp) from N. fuckeliana, a well-known fungus on Norway spruce in Scandinavia and other parts of the world, especially in the northern hemisphere. In 2011, the new Neonectria species was found on diseased trees in a Danish nordmann fir (Abies nordmanniana) seed orchard. Resin flow was seen from mature cones, and tests revealed that the seeds were infected by the Neonectria sp.