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Academic – The “humanized” N-glycosylation pathway in CRISPR/Cas9-edited Nicotiana benthamiana significantly enhances the immunogenicity of a S/preS1 Hepatitis B Virus antigen and the virus-neutralizing antibody response in vaccinated mice
Ana-Maria Madalina Pantazica, André van Eerde, Mihaela-Olivia Dobrica, ...
AuthorsAna-Maria Madalina Pantazica André van Eerde Mihaela-Olivia Dobrica Iuliana Caras I Ionescu Adriana Costache Catalin Tucureanu Hege Særvold Steen Catalin Lazar Inger Heldal Sissel Haugslien Adrian Onu Crina Stavaru Norica Branza-Nichita Jihong Liu Clarke
No abstract has been registered
Academic – Patterns and roles of lignan and terpenoid accumulation in the reaction zone compartmentalizing pathogen-infected heartwood of Norway spruce
Nina Elisabeth Nagy, Hans Ragnar Norli, Monica Fongen, ...
AuthorsNina Elisabeth Nagy Hans Ragnar Norli Monica Fongen Runa Berg Østby Inger Heldal Jahn Davik Ari Hietala
Tree defense against xylem pathogens involves both constitutive and induced phenylpropanoids and terpenoids. The induced defenses include compartmentalization of compromised wood with a reaction zone (RZ) characterized by polyphenol deposition, whereas the role of terpenoids has remained poorly understood. To further elucidate the tree–pathogen interaction, we profiled spatial patterns in lignan (low-molecular-weight polyphenols) and terpenoid content in Norway spruce (Picea abies) trees showing heartwood colonization by the pathogenic white-rot fungus Heterobasidion parviporum. There was pronounced variation in the amount and composition of lignans between different xylem tissue zones of diseased and healthy trees. Intact RZ at basal stem regions, where colonization is the oldest, showed the highest level and diversity of these compounds. The antioxidant properties of lignans obviously hinder oxidative degradation of wood: RZ with lignans removed by extraction showed significantly higher mass loss than unextracted RZ when subjected to Fenton degradation. The reduced diversity and amount of lignans in pathogen-compromised RZ and decaying heartwood in comparison to intact RZ and healthy heartwood suggest that α-conindendrin isomer is an intermediate metabolite in lignan decomposition by H. parviporum. Diterpenes and diterpene alcohols constituted above 90% of the terpenes detected in sapwood of healthy and diseased trees. A significant finding was that traumatic resin canals, predominated by monoterpenes, were commonly associated with RZ. The findings clarify the roles and fate of lignan during wood decay and raise questions about the potential roles of terpenoids in signal transduction, synthesis, and translocation of defense compounds upon wood compartmentalization against decay fungi.
Academic – Establishment of a piscine myocarditis virus (PMCV) challenge model and testing of a plant-produced subunit vaccine candidate against cardiomyopathy syndrome (CMS) in Atlantic salmon Salmo salar
Hang Su, Andre van Eerde, Hege Særvold Steen, ...
AuthorsHang Su Andre van Eerde Hege Særvold Steen Inger Heldal Sissel Haugslien Irene Ørpetveit Stefanie Caroline Wüstner Makoto Inami Marie Løvoll Espen Rimstad Jihong Liu Clarke
Cardiomyopathy syndrome (CMS) is a severe cardiac disease occurring in the grow-out sea phase of farmed Atlantic salmon with approximately 100 outbreaks annually in Norway. Piscine myocarditis virus (PMCV) is believed to be the causative agent of CMS. There is no vaccine available to control CMS, partially because PMCV withstands propagation in known cell cultures. In the present study, we selected the putative capsid protein of PMCV as the candidate antigen for immunization experiments and produced it in the plant Nicotiana benthamiana by transient expression. The recombinant PMCV antigen formed virus-like particles (VLPs). To evaluate the efficacy of the plant made VLP vaccine, a PMCV infection model was established. In an experimental salmon vaccination trial, the VLP vaccine triggered innate immunity, and indicative but not significant inhibition of viral replication in heart, spleen and kidney tissues was observed. Similarly, a reduction of inflammatory lesions in cardiomyocytes and subendocardial infiltration by mononuclear leukocytes were observed. Therefore, there was no difference in efficacy or immune response observed post the plant made PMCV VLP antigen vaccination. Taken together, this study has demonstrated that plant made VLP antigens should be investigated further as a possible platform for the development of PMCV antigens for a CMS vaccine.
Academic – Transcriptome profiling and in silico detection of the antimicrobial peptides of red king crab Paralithodes camtschaticus
Igor A. Yakovlev, Erik Lysøe, Inger Heldal, ...
Endogenous antimicrobial peptides (AMPs) are evolutionarily ancient factors of innate immunity, which are produced by all multicellular organisms and play a key role in their protection against infection. Red king crab (Paralithodes camtschaticus), also called Kamchatka crab, is widely distributed and the best known species of all king crabs belonging to the family Lithodidae. Despite their economic importance, the genetic resources of king crabs are scarcely known and no fullgenome sequences are available to date. Therefore, analysis of the red king crab transcriptome and identifcation and characterization of its AMPs could potentially contribute to the development of novel antimicrobial drug candidates when antibiotic resistance has become a global health threat. In this study, we sequenced the P. camtschaticus transcriptomes from carapace, tail fap and leg tissues using an Illumina NGS platform. Libraries were systematically analyzed for gene expression profles along with AMP prediction. By an in silico approach using public databases we defned 49 cDNAs encoding for AMP candidates belonging to diverse families and functional classes, including buforins, crustins, paralithocins, and ALFs (anti-lipopolysaccharide factors). We analyzed expression patterns of 27 AMP genes. The highest expression was found for Paralithocin 1 and Crustin 3, with more than 8,000 reads. Other paralithocins, ALFs, crustins and ubiquicidins were among medium expressed genes. This transcriptome data set and AMPs provide a solid baseline for further functional analysis in P. camtschaticus. Results from the current study contribute also to the future application of red king crab as a bio-resource in addition to its being a known seafood delicacy.
Academic – In-depth characterization of Trichoderma reesei cellobiohydrolase TrCel7A produced in Nicotiana benthamiana reveals limitations of cellulase production in plants by host-specific post-translational modifications
Andre van Eerde, Aniko Varnai, John-Kristian Jameson, ...
AuthorsAndre 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 Clarke
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.
Lecture – Plant biotechnology for low cost production of fish vaccines
Jihong Liu Clarke, Andre van Eerde, Lisa Paruch, ...
AuthorsJihong Liu Clarke Andre van Eerde Lisa Paruch Inger Heldal Hege Særvold Steen Yanliang Wang Astrid Sivertsen Sissel Haugslien
No abstract has been registered
Lecture – Plant-derived antigens for fish vaccines
Jihong Liu Clarke, Lisa Paruch, Hege Særvold Steen, ...
AuthorsJihong Liu Clarke Lisa Paruch Hege Særvold Steen Andre van Eerde Yanliang Wang Astrid Sivertsen Sissel Haugslien Inger Heldal
No abstract has been registered
Academic – Quantifying Neodiprion sertifer nucleopolyhedrovirus DNA from insects, foliage and forest litter using the quantitative real-time polymerase chain reaction
Paal Krokene, Inger Heldal, Carl Gunnar Fossdal
1. Neodiprion sertifer nucleopolyhedrovirus (NeseNPV) is widely used as a viral bio-insecticide against larvae of the European pine sawfly N. sertifer (Geoff.) (Hymenoptera: Diprionidae), which is one of the most harmful defoliators of pines in Northern Europe. A major obstacle to studying this pathogenic virus in nature is the difficulty of confirming and quantifying the presence of NeseNPV. 2. In the present study, we developed real-time polymerase chain reaction (PCR) primers, based on the caspid gene 39 sequence, for the specific and quantitative detection of NeseNPV. The quantitative real-time PCR (qPCR) assay can detect virus from any substrate tested, including different insect life stages (egg, larval, adult), pine foliage, and litter or ground vegetation. The reproducible detection limit for the real-time assay is 0.013 pg of viral DNA (0.013 × 10−12 g), corresponding to 136 viral genomes or approximately one to seven virus occlusion bodies per sample. 3. qPCR is a specific, quantitative, sensitive, reliable and flexible procedure, and is a good supplement to conventional microscopy- or bioassay-based methods for detection of the virus. We have used qPCR to quantify the level of NeseNPV in samples collected in the field after aerial application of the virus, and demonstrated significantly higher virus levels in sawfly larvae from sprayed areas compared with unsprayed control areas 4 weeks after spraying. 4. This qPCR assay can be used to determine important aspects of the biology of NeseNPV (e.g. virus levels in different insect life stages and in their microhabitats on pine foliage and in forest litter).
Academic – Effects of Rhizoctonia infection and drought on peroxidase and chitinase activity in Norway spruce (Picea abies)
Nina Elisabeth Nagy, Carl Gunnar Fossdal, Lars Sandved Dalen, ...
AuthorsNina Elisabeth Nagy Carl Gunnar Fossdal Lars Sandved Dalen Anders Lønneborg Inger Heldal Øystein Johnsen
No abstract has been registered
Division of Biotechnology and Plant Health
Healthy feed to healthy aquatic food via Sino-Norwegian cooperation- Feed2Food
The project is at the forefront of scientific research in utilizing molecular, physiological and high advanced methodology to quantify the challenges with feed additives in combination with high fat diets (HFD).
SmartVac - Next Generation Viral Hepatitis B and C vaccine development in plantsand algae using advanced biotechnological tools
Hepatitis B (HBV) and C viruses (HCV) infect the human liver, triggering persistent inflammation and eventually cirrhosis and hepatocellular carcinoma (HCC), the second leading cause of cancer-related mortality worldwide. Currently, more than 500 million people are chronically infected with HBV or HCV and at high risk of developing end stage liver disease and HCC.
Scientists from NIBIO and CAAS are working together using innovative technologies in order to improve productivity, food safety and sustainability in Chinese agriculture.