Academic – Low phosphorus availability increases shoot boron concentration in canola and potato but not in wheat
Yanliang Wang, Nicholas Clarke, Anne Falk Øgaard
A large proportion of global agricultural soils contain suboptimal available phosphorus (P) for the growth of many plant species. Boron (B) plays important roles in plant growth and development, but limited research has been conducted to study B uptake under low P availability. This study comprised a hydroponic and a mini-rhizobox experiment with canola (Brassica napus L.), potato (Solanum tuberosum L.) and wheat (Triticum aestivum L.) under P sufficient and deficient conditions. Boron concentrations, rhizosphere soil pH, and gene expression of BnBOR1 in canola were determined. Shoot B concentrations were found significantly increased (11–149%) by low P availability in potato and canola but not in wheat. Reverse transcription polymerase chain reaction (RT-PCR) indicated that BnBOR1;2a, BnBOR1;2c, and BnBOR1;3c were up-regulated after seven days of low P treatment in canola roots. Our results indicate that plant shoot B concentration was dramatically influenced by P availability, and dicots and monocots showed a contrasting B concentration response to low P availability.
Academic – Transcriptome and metabolome analyses provide insights into root and root-released organic anion responses to phosphorus deficiency in oat
Yanliang Wang, Erik Lysøe, Tegan Armarego-Marriott, ...
Academic – Impact of phosphorus on rhizosphere organic anions of wheat at different growth stages under field conditions
Yanliang Wang, Tore Krogstad, Nicholas Clarke, ...
Phosphorus (P) is one of the main limiting factors for crop productivity while rhizosphere organic anions have been hypothesized to play an important role in P acquisition. Sampling in a long-term field experiment was carried out in order to understand the impact of long-term differences in P fertilization on secretion of organic anions under field conditions. Rhizosphere organic anions were extracted and analyzed every week from three leaves stage to completed flowering stage of wheat (Triticum aestivum) grown on plots that have received 0 (P0) or 48 (P48) kg P ha−1 year−1 since 1966. The study showed that it is possible to extract and quantify rhizosphere organic anions from field plots. In P48 plots, root P concentrations decreased around 40 % at the early stages (before heading), compared with the first sampling, and then increased slightly, while plants grown in P0 plots showed the opposite trend. Malate was the main organic anion secreted throughout all the wheat growth stages. Rhizosphere citrate and malate showed negative and positive correlations (P < 0.05) respectively with root P concentrations at 29 and 42 days after emergence (DAE). Rhizosphere organic anion concentrations were quite low until 29 DAE and then increased up to 4–10-fold until 42 DAE; these concentrations declined at later stages, indicating that root-released organic anions may have been affected by developmental stage and root P concentration. The present study provides valuable information about the relationship between rhizosphere organic anions and various P concentrations of wheat grown in the field.
Academic – Lettuce-produced hepatitis C virus E1E2 heterodimer triggers immune responses in mice and antibody production after oral vaccination
Jihong Liu Clarke, Lisa Paruch, Mihaela-Olivia Dobrica, ...
The hepatitis C virus (HCV) is a major etiologic agent for severe liver diseases ( e.g . cirrhosis, fibrosis and hepatocellular carcinoma). Approximately 140 million people have chronic HCV infections and about 500 000 die yearly from HCV-related liver pathologies. To date, there is no licensed vaccine available to prevent HCV infection and production of a HCV vaccine remains a major challenge. Here, we report the successful production of the HCV E1E2 heterodimer, an important vaccine candidate, in an edible crop (lettuce, Lactuca s ativa ) using Agrobacterium - mediated transient expression technology. The wild-type dimer (E1E2) and a variant without an N-glycosylation site in the E2 polypeptide (E1E2 Δ N6) were expressed, and appropriate N-glycosylation pattern and functionality of the E1E2 dimers were demonstrated. The humoral immune response induced by the HCV proteins was investigated in mice following oral administration of lettuce antigens with or without previous intramuscular prime with the mammalian HEK293T cell-expressed HCV dimer. Immunization by oral feeding only resulted in development of weak serum levels of anti-HCV IgM for both antigens; however, the E1E2 Δ N6 proteins produced higher amounts of secretory IgA, suggesting improved immunogenic properties of the N-glycosylation mutant. The mice group receiving the intramuscular injection followed by two oral boosts with the lettuce E1E2 dimer developed a systemic but also a mucosal immune response, as demonstrated by the presence of anti-HCV secretory IgA in faeces extracts. In summary, our study demonstrates the feasibility of producing complex viral antigens in lettuce, using plant transient expression technology, with great potential for future low-cost oral vaccine development.
Lecture – Plant biotechnology for low cost production of fish vaccines
Jihong Liu Clarke, Andre Eerde, Lisa Paruch, ...
No abstract has been registered
Poster – production of cell-wall degrading enzymes in N. benthamiana plants
Andre van Eerde, Yanliang Wang, John-Kristian Jameson, ...
No abstract has been registered
Lecture – Plant-derived antigens for fish vaccines
Jihong Liu Clarke, Lisa Paruch, Hege Særvold Steen, ...
No abstract has been registered
Academic – Production of dengue virus envelope protein domain III-based antigens in tobacco chloroplasts using inducible and constitutive expression systems
Johanna Gottschamel, Andreas G. Lössl, Stephanie Ruf, ...
Abstract Dengue fever is a disease in many parts of the tropics and subtropics and about half the world’s popula- tion is at risk of infection according to the World Health Organization. Dengue is caused by any of the four related dengue virus serotypes DEN-1, -2, -3 and -4, which are transmitted to people by Aedes aegypti mosquitoes. Cur- rently there is only one vaccine (DengvaxiaÒ) available (limited to a few countries) on the market since 2015 after half a century’s intensive efforts. Affordable and accessible vaccines against dengue are hence still urgently needed. The dengue envelop protein domain III (EDIII), which is capable of eliciting serotype-specific neutralizing antibod- ies, has become the focus for subunit vaccine development. To contribute to the development of an accessible and affordable dengue vaccine, in the current study we have used plant-based vaccine production systems to generate a dengue subunit vaccine candidate in tobacco. Chloroplast genome engineering was applied to express serotype- specific recombinant EDIII proteins in tobacco chloroplasts using both constitutive and ethanol-inducible expression systems. Expression of a tetravalent antigen fusion con- struct combining EDIII polypeptides from all four ser- otypes was also attempted. Transplastomic EDIII- expressing tobacco lines were obtained and homoplasmy was verified by Southern blot analysis. Northern blot analyses showed expression of EDIII antigen-encoding genes. EDIII protein accumulation levels varied for the different recombinant EDIII proteins and the different expression systems, and reached between 0.8 and 1.6 % of total cellular protein. Our study demonstrates the suitability of the chloroplast compartment as a production site for an EDIII-based vaccine candidate against dengue fever and presents a GatewayÒ plastid transformation vector for inducible transgene expression.
Academic – Rhizosphere Organic Anions Play a Minor Role in Improving Crop Species' Ability to Take Up Residual Phosphorus (P) in Agricultural Soils Low in P Availability
Yanliang Wang, Tore Krogstad, Jihong Liu Clarke, ...
Many arable lands have accumulated large reserves of residual phosphorus (P) and a relatively large proportion of soil P is less available for uptake by plants. Root released organic anions are widely documented as a key physiological strategy to enhance P availability, while limited information has been generated on the contribution of rhizosphere organic anions to P utilization by crops grown in agricultural soils that are low in available P and high in extractable Ca, Al, and Fe. We studied the role of rhizosphere organic anions in P uptake from residual P in four common crops Triticum aestivum, Avena sativa, Solanum tuberosum, and Brassica napus in low- and high-P availability agricultural soils from long-term fertilization field trials in a mini-rhizotron experiment with four replications. Malate was generally the dominant organic anion. More rhizosphere citrate was detected in low P soils than in high P soil. B. napus showed 74–103% increase of malate in low P loam, compared with clay loam. A. sativa had the greatest rhizosphere citrate concentration in all soils (5.3–15.2 μmol g−1 root DW). A. sativa also showed the highest level of root colonization by arbuscular mycorrhizal fungi (AMF; 36 and 40%), the greatest root mass ratio (0.51 and 0.66) in the low-P clay loam and loam respectively, and the greatest total P uptake (5.92 mg P/mini-rhizotron) in the low-P loam. B. napus had 15–44% more rhizosphere acid phosphatase (APase) activity, ~0.1–0.4 units lower rhizosphere pH than other species, the greatest increase in rhizosphere water-soluble P in the low-P soils, and the greatest total P uptake in the low-P clay loam. Shoot P content was mainly explained by rhizosphere APase activity, water-soluble P and pH within low P soils across species. Within species, P uptake was mainly linked to rhizosphere water soluble P, APase, and pH in low P soils. The effects of rhizosphere organic anions varied among species and they appeared to play minor roles in improving P availability and uptake.
Academic – Contrasting responses of root morphology and root-exuded organic acids to low phosphorus availability in three important food crops with divergent root traits
Yanliang Wang, Marit Almvik, Nicholas Clarke, ...
Phosphorus (P) is an important element for crop productivity and is widely applied in fertilizers. Most P fertilizers applied to land are sorbed onto soil particles, so research on improving plant uptake of less easily available P is important. In the current study, we investigated the responses in root morphology and root-exuded organic acids (OAs) to low available P (1 mM P) and sufficient P (50 mM P) in barley, canola and micropropagated seedlings of potato— three important food crops with divergent root traits, using a hydroponic plant growth system.We hypothesized that the dicots canola and tuber-producing potato and the monocot barley would respond differently under various P availabilities. WinRHIZO and liquid chromatography triple quadrupole mass spectrometry results suggested that under low P availability, canola developed longer roots and exhibited the fastest root exudation rate for citric acid. Barley showed a reduction in root length and root surface area and an increase in root-exudedmalic acid under low-P conditions. Potato exuded relativelysmall amounts of OAs under low P, while therewas a marked increase in root tips. Based on the results, we conclude that different crops show divergent morphological and physiological responses to low P availability, having evolved specific traits of root morphology and root exudation that enhance their P-uptake capacity under low-P conditions. These results could underpin future efforts to improve P uptake of the three crops that are of importance for future sustainable crop production.