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Publications

NIBIOs employees contribute to several hundred scientific articles and research reports every year. You can browse or search in our collection which contains references and links to these publications as well as other research and dissemination activities. The collection is continously updated with new and historical material.

2019

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Abstract

Shallot (Allium cepa var. aggregatum) is an important vegetable crop belonging to the genus Allium. The present study attempted to develop an efficient droplet-vitrification cryopreservation method for shallot ‘10603’ shoot tips. In vitro stock shoots were maintained on Murashige and Skoog (1962) medium (MS) supplemented with 30 g L-1 sucrose, 0.5 mg L-1 BAP, 0.1 mg L-1 NAA and 8 g L-1 agar (pH=5.8). Shoot tips (2.0-3.0 mm in length) were excised from 4-week-old stock shoots and stepwise precultured with increased sucrose concentrations from 0.3 to 0.5 M, each concentration for 1 day. The precultured shoot tips were then loaded for 20 min with a solution composed of 2 M glycerol and 0.5 M sucrose, before exposure to PVS3 for 3 h at room temperature. Dehydrated shoot tips were transferred onto aluminum foils (2×0.8 cm), prior to direct immersion into liquid nitrogen (LN) for cryostorage. For thawing, frozen aluminum foils were moved from LN and immediately transferred into unloading solution composed of liquid MS containing 1.2 M sucrose. After incubation at room temperature for 20 min, shoot tips were post-cultured on solidified MS medium containing 0.3 M sucrose for 2 days and then transferred onto a recovery medium for shoot regrowth. With this procedure, 94% shoot tips survived, and 58% shoot tips regenerated into shoots following cryopreservation.

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Abstract

We present the results of an inventory and status assessment of alien species in Norway. The inventory covered all known multicellular neobiota, 2496 in total, 1039 of which were classified as naturalised. The latter constitute c. 3% of all species known to be stably reproducing in Norway. These figures are higher than expected from Norway’s latitude, which may be due a combination of climatic and historical factors, as well as sampling effort. Most of the naturalised neobiota were plants (71%),followed by animals (21%) and fungi (8%). The main habitat types colonised were open lowlands (79%), urban environments (52%) and woodlands (42%). The main areas of origin were Europe (67%), North America (15%) and Asia (13%). For most taxa, the rate of novel introductions seems to have been increasing during recent decades. Within Norway, the number of alien species recorded per county was negatively correlated with latitude and positively correlated with human population density. In the high-Arctic territories under Norwegian sovereignty, i.e. Svalbard and Jan Mayen, 104 alien species were recorded, of which 5 were naturalised.

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.

Abstract

Complex communities of microorganisms influence plant and agroecosystem health and productivity. Bacteria and fungi constitute a major part of the wheat head microbiome. A microorganism’s ability to colonize or infect a wheat seed is influenced by interacting microbiome. In Norway, wheat seed lots are routinely analysed for the infestation by Fusarium head blight and seedling blight diseases, such as Fusarium and Microdochium spp., and glume blotch caused by Parastagonospora nodorum using traditional methods (plating grain on PDA, recording presence or absence of fungal colonies) The purpose is to decide if the seed quality is suitable for sowing and whether seed treatment is needed. This method is time consuming, require knowledge within fungal morphology, and do not facilitate identification to species level in all cases. Molecular methods such as sequencing could allow detection and quantification of “all” microbial DNA, only limited by the specificity of the primers. Microbial profiling (metabarcoding) can be very time and cost-effective, since a mixture of many samples can be analysed simultaneously for both fungi and bacteria, and other microbes if required. In our project “Phytobiome” we used metabarcoding to analyse microbial communities in wheat heads and verify this information with results from qPCR and plate studies for a more complete study. Around 150 spring wheat seed lots from the years 2016-2017 (including two cultivars) were selected for analysis. One of the main objectives was to find microorganisms associated with seed germination. We will present findings from this work, but also some challenges when using PCR-based sequencing methods, especially regarding Fusarium head blight fungi.

Abstract

Occasionally, high mycotoxin levels are observed in Norwegian oat grain lots. The development of moderate resistant oat cultivars is therefore highly valued in order to increase the share of high quality grain into the food and feed industry. The Norwegian SafeOats project (2016-2020) aims to develop resistance screening methods to facilitate the phase-out of Fusarium-susceptible oat germplasm. Furthermore, SafeOats will give new insight into the biology of F. langsethiae and HT2+T2 accumulation in oats. The relative ranking of oat varieties according to F. graminearum/DON versus F. langsethiae/HT2+T2 content has been explored in naturally infested as well as in inoculated field trials. Routine testing of the resistance to F. graminearum in oat cultivars and breeding lines has been conducted in Norway since 2007. We are currently working on ways to scale up the inoculum production and fine tune the methodology of F. langsethiae inoculation of field trials to be routinely applied in breeding programs. Through greenhouse studies, we have analysed the content of Fusarium DNA and mycotoxins in grains of selected oat varieties inoculated at different development stages. Furthermore, we are studying the transcriptome during F. langsethiae and F. graminearum infestation of oats. The project also focus on the occurrence of F. langsethiae in oat seeds and possible influence of the fungus on seedling development in a selection of oat varieties. On average, the fungus was observed on 5% of the kernels in 168 seed lots tested during 2016-2018. No indication of transmission of F. langsethiae from germinating seed to seedlings was found in a study with germination of naturally infected seeds. So far, the studies have shown that the ranking of oat varieties according to HT2+T2 content in non-inoculated field trials resembles the ranking observed in inoculated field trials. The ranking of oat varieties according to DON content is similar in non-inoculated and F. graminearum inoculated field trials. However, the ranking of oat varieties according to DON content does not resemble the ranking for HT2+T2. The results from SafeOats will benefit consumers nationally and internationally by providing tools to increase the share of high quality grain into the food and feed industry. The project is financed by The Foundation for Research Levy on Agricultural Products/Agricultural Agreement Research Fund/Research Council of Norway with support from the industry partners Graminor, Lantmännen, Felleskjøpet Agri, Felleskjøpet Rogaland & Agder, Fiskå Mølle Moss, Norgesmøllene, Strand Unikorn/Norgesfôr and Kimen Seed Laboratory.

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

Sustainable production of biofuels from lignocellulose feedstocks depends on cheap enzymes for degradation of such biomass. Plants offer a safe and cost‐effective production platform for biopharmaceuticals, vaccines and industrial enzymes boosting biomass conversion to biofuels. Production of intact and functional protein is a prerequisite for large‐scale protein production, and extensive host‐specific post‐translational modifications (PTMs) often affect the catalytic properties and stability of recombinant enzymes. Here we investigated the impact of plant PTMs on enzyme performance and stability of the major cellobiohydrolase TrCel7A from Trichoderma reesei, an industrially relevant enzyme. TrCel7A was produced in Nicotiana benthamiana using a vacuum‐based transient expression technology, and this recombinant enzyme (TrCel7Arec) was compared with the native fungal enzyme (TrCel7Anat) in terms of PTMs and catalytic activity on commercial and industrial substrates. We show that the N‐terminal glutamate of TrCel7Arec was correctly processed by N. benthamiana to a pyroglutamate, critical for protein structure, while the linker region of TrCel7Arec was vulnerable to proteolytic digestion during protein production due to the absence of O‐mannosylation in the plant host as compared with the native protein. In general, the purified full‐length TrCel7Arec had 25% lower catalytic activity than TrCel7Anat and impaired substrate‐binding properties, which can be attributed to larger N‐glycans and lack of O‐glycans in TrCel7Arec. All in all, our study reveals that the glycosylation machinery of N. benthamiana needs tailoring to optimize the production of efficient cellulases.