2019
Authors
Raghuram Badmi Yupeng Zhang Torstein Tengs May Bente Brurberg Paal Krokene Carl Gunnar Fossdal Timo Hytönen Tage ThorstensenAbstract
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.
Authors
Raghuram Badmi Yupeng Zhang Torstein Tengs May Bente Brurberg Paal Krokene Carl Gunnar Fossdal Timo Hytönen Tage ThorstensenAbstract
No abstract has been registered
Authors
Raghuram Badmi Yupeng Zhang Torstein Tengs May Bente Brurberg Paal Krokene Carl Gunnar Fossdal Timo Hytönen Tage ThorstensenAbstract
No abstract has been registered
Authors
Andre van Eerde Aniko Varnai John-Kristian Jameson Lisa Paruch Anders Moen Jan Haug Anonsen Piotr Chylenski Hege Særvold Steen Inger Heldal Ralph Bock Vincent Eijsink Jihong Liu ClarkeAbstract
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.
Abstract
No abstract has been registered
Authors
Igor A. Yakovlev Daniel Gackowski Abdulkadir Abakir Marcos Viejo Somoano Alexey Ruzov Ryszard Olinski Marta Starczak Carl Gunnar Fossdal Konstantin V. KrutovskyAbstract
5-Methylcytosine (5mC) is an epigenetic modification involved in regulation of gene expression in metazoans and plants. Iron-(II)/α-ketoglutarate-dependent dioxygenases can oxidize 5mC to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC). Although these oxidized forms of 5mC may serve as demethylation intermediates or contribute to transcriptional regulation in animals and fungi, experimental evidence for their presence in plant genomes is ambiguous. Here, employing reversed-phase HPLC coupled with sensitive mass spectrometry, we demonstrated that, unlike 5caC, both 5hmC and 5fC are detectable in non-negligible quantities in the DNA of a conifer, Norway spruce. Remarkably, whereas 5hmC content of spruce DNA is approximately 100-fold lower relative to human colorectal carcinoma cells, the levels of both - 5fC and a thymine base modification, 5-hydroxymethyluracil, are comparable in these systems. We confirmed the presence of modified DNA bases by immunohistochemistry in Norway spruce buds based on peroxidase-conjugated antibodies and tyramide signal amplification. Our results reveal the presence of specific range of noncanonical DNA bases in conifer genomes implying potential roles for these modifications in plant development and homeostasis.
Authors
Marcos Viejo Somoano Torstein Tengs Igor A. Yakovlev Jorunn Elisabeth Olsen Carl Gunnar FossdalAbstract
No abstract has been registered
Authors
Simeon Rossmann Merete Wiken Dees Torfinn Torp Vinh Hong Le Monica Skogen Borghild Glorvigen Jan van der Wolf May Bente BrurbergAbstract
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.
Authors
Melissa Magerøy Erik Christiansen Bo Långström Anna-Karin Borg-Karlson Halvor Solheim Niklas Björklund Tao Zhao Axel Schmidt Carl Gunnar Fossdal Paal KrokeneAbstract
No abstract has been registered
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.