Lignin engineering in forest trees

Wood is a renewable resource that is mainly composed of lignin and cell wall polysaccharides. The polysaccharide fraction is valuable as it can be converted into pulp and paper, or into fermentable sugars. On the other hand, the lignin fraction is increasingly being considered a valuable source of aromatic building blocks for the chemical industry. The presence of lignin in wood is one of the major recalcitrance factors in woody biomass processing, necessitating the need for harsh chemical treatments to degrade and extract it prior to the valorization of the cell wall polysaccharides, cellulose and hemicellulose. Over the past years, large research efforts have been devoted to engineering lignin amount and composition to reduce biomass recalcitrance toward chemical processing. We review the efforts made in forest trees, and compare results from greenhouse and field trials. Furthermore, we address the value and potential of CRISPR-based gene editing in lignin engineering and its integration in tree breeding programs

Stacking of a low-lignin trait with an increased guaiacyl and 5-hydroxyguaiacyl unit trait leads to additive and synergistic effects on saccharification efficiency in Arabidopsis thaliana

Background: Lignocellulosic biomass, such as wood and straw, is an interesting feedstock for the production of fer-mentable sugars. However, mainly due to the presence of lignin, this type of biomass is recalcitrant to saccharification. In Arabidopsis, lignocellulosic biomass with a lower lignin content or with lignin with an increased fraction of guaiacyl (G) and 5-hydroxyguaiacyl (5H) units shows an increased saccharification efficiency. Here, we stacked these two traits and studied the effect on the saccharification efficiency and biomass yield, by combining either transaldolase (tra2), cinnamate 4-hydroxylase (c4h-3), or 4-coumarate:CoA ligase (4cl1-1) with caffeic acid O-methyltransferase (comt-7 or comt-4) mutants. Results: The three double mutants (tra2 comt-1, c4h-3 comt-4, and 4cl1-7 comt-4) had a decreased lignin amount and an increase in G and 5H units in the lignin polymer compared to wild-type (WT) plants. The tra2 comt-1 double mutant had a better saccharification efficiency compared to the parental lines when an acid or alkaline pretreatment was used. For the double mutants, c4h-3 comt-4 and 4cl1-1 comt-4, the saccharification efficiency was significantly higher compared to WT and its parental lines, independent of the pretreatment used. When no pretreatment was used, the saccharification efficiency increased even synergistically for these mutants. Conclusion: Our results show that saccharification efficiency can be improved by combining two different mutant lignin traits, leading to plants with an even higher saccharification efficiency, without having a yield reduction of the primary inflorescence stem. This approach can help improve saccharification efficiency in bio-energy crops

Vessel-specific reintroduction of CINNAMOYL-COA REDUCTASE1 (CCR1) in dwarfed ccr1 mutants restores vessel and xylary fiber integrity and increases biomass

Lignocellulosic biomass is recalcitrant toward deconstruction into simple sugars due to the presence of lignin. To render lignocellulosic biomass a suitable feedstock for the bio-based economy, plants can be engineered to have decreased amounts of lignin. However, engineered plants with the lowest amounts of lignin exhibit collapsed vessels and yield penalties. Previous efforts were not able to fully overcome this phenotype without settling in sugar yield upon saccharification. Here, we reintroduced CINNAMOYL-COENZYME A REDUCTASE1 (CCR1) expression specifically in the protoxylem and metaxylem vessel cells of Arabidopsis (Arabidopsis thaliana) ccr1 mutants. The resulting ccr1 ProSNBE: CCR1 lines had overcome the vascular collapse and had a total stem biomass yield that was increased up to 59% as compared with the wild type. Raman analysis showed that monolignols synthesized in the vessels also contribute to the lignification of neighboring xylary fibers. The cell wall composition and metabolome of ccr1 ProSNBE: CCR1 still exhibited many similarities to those of ccr1 mutants, regardless of their yield increase. In contrast to a recent report, the yield penalty of ccr1 mutants was not caused by ferulic acid accumulation but was (largely) the consequence of collapsed vessels. Finally, ccr1 ProSNBE: CCR1 plants had a 4-fold increase in total sugar yield when compared with wild-type plants

Modelling of annual sand transports at the Dutch lower shoreface

Dutch coastal policy aims for a safe, economically strong and attractive coast. This is achieved by maintaining the part of the coast that support these functions; the coastal foundation. The coastal foundation is maintained by means of sand nourishments. Up to now, it has been assumed that net transports across the coastal foundation's offshore boundary at the 20 m depth contour are negligibly small. In the framework of the Coastal Genesis 2.0 program we investigate sand transports across this boundary and across other depth contours at the lower shoreface. The purpose of this paper is to provide knowledge for a well-founded choice of the seaward boundary of the coastal foundation. The lower shoreface is the zone where the mixed action of shoreface currents (tide-, wind- and density gradient driven) and shoaling and refracting waves is predominant. Transport rates are relatively small and hence the bed levels in the lower shoreface undergo relatively slow changes

Tailoring poplar lignin without yield penalty by combining a null and haploinsufficient CINNAMOYL-CoA REDUCTASE2 allele

Lignin causes lignocellulosic biomass recalcitrance to enzymatic hydrolysis. Engineered low-lignin plants have reduced recalcitrance but often exhibit yield penalties, offsetting their gains in fermentable sugar yield. Here, CRISPR/Cas9-generated CCR2(−/*) line 12 poplars have one knockout CCR2 allele while the other contains a 3-bp deletion, resulting in a 114I115A-to-114T conversion in the corresponding protein. Despite having 10% less lignin, CCR2(−/*) line 12 grows normally. On a plant basis, the saccharification efficiency of CCR2(−/*) line 12 is increased by 25–41%, depending on the pretreatment. Analysis of monoallelic CCR2 knockout lines shows that the reduced lignin amount in CCR2(−/*) line 12 is due to the combination of a null and the specific haploinsufficient CCR2 allele. Analysis of another CCR2(−/*) line shows that depending on the specific CCR2 amino-acid change, lignin amount and growth can be affected to different extents. Our findings open up new possibilities for stably fine-tuning residual gene function in planta

Perioperative SARS-CoV-2 infections increase mortality, pulmonary complications, and thromboembolic events : a Dutch, multicenter, matched-cohort clinical study

Background: A direct comparison of severe acute respiratory syndrome coronavirus 2 positive patients with a severe acute respiratory syndrome coronavirus 2 negative control group undergoing an operative intervention during the current pandemic is lacking, and a reliable estimate of the assumed difference in morbidity and mortality between both patient categories remains unknown. Methods: We included all consecutive patients with a confirmed pre- or postoperative severe acute respiratory syndrome coronavirus 2 positive status (operated in 27 hospitals) and negative control patients (operated in 4 hospitals) undergoing emergency or elective operations. A propensity score-matched comparison of clinical outcomes was performed between severe acute respiratory syndrome coronavirus 2 positive and negative tested patients (control group). Primary outcome was overall 30-day mortality rate between both groups. Main secondary outcomes were overall, pulmonary, and thromboembolic complications. Results: In total, 161 severe acute respiratory syndrome coronavirus 2 positive and 342 control severe acute respiratory syndrome coronavirus 2 negative patients were included in this study. The 30-day overall postoperative mortality rate was greater in the severe acute respiratory syndrome coronavirus 2 positive cohort compared with the negative control group (16% vs 4% respectively; P = .007). After propensity score matching, the severe acute respiratory syndrome coronavirus 2 positive group consisted of 123 patients (median 70 years of age [interquartile range 59-77] and 55% male) were compared with 196 patients in the matched control group (median 69 years (interquartile range 58-75] and 53% male). The 30-day mortality rate and risk were greater in the severe acute respiratory syndrome coronavirus 2 positive group compared with the matched control group (12% vs 4%; P = .009 and odds ratio 3.4 [95% confidence interval 1.5-8.5]; P = .005, respectively). Overall, pulmonary and thromboembolic complications occurred more often in severe acute respiratory syndrome coronavirus 2 positive patients (P < .01). Conclusion: Patients diagnosed with perioperative severe acute respiratory syndrome coronavirus 2 have an increased risk of 30-day mortality, pulmonary complications, and thromboembolic events. These findings serve as an evidence-based argument to postpone elective surgery and selected emergency cases. (C) 2020 The Author(s). Published by Elsevier Inc

Robotically driven construction of buildings: Exploring on-demand building components production

Robotically Driven Construction of Buildings (RDCB) is an exploration into design to production solutions for robotically driven construction of buildings initiated by the faculties of Civil Engineering and Architecture, TU Delft and Architecture, TU Eindhoven and implemented 2014 within the 3TU Lighthouse framework. The aim of was to involve the disciplines of architecture, robotics, materials science, and structural design in order to integrate knowledge from the individual disciplines and develop new numerically controlled manufacturing techniques and building-design optimisation methods for adding creative value to buildings in a cost-effective and sustainable way.RDCB builds up on expertise developed at Hyperbody with respect to applications of robotics in architecture and this paper presents the contribution of the Robotic Building team from Hyperbody, Faculty of Architecture, TU Delft to the RDCB project. The contribution is in line with Europe’s aim to improve material and energy efficiency of buildings and efficiency of construction processes. Robotically driven construction and customised building materials have the potential to realise this in a cost-effective way and at the same time reduce accidents and health hazards for workers in the building sector. In order to achieve this RDCB is distributing materials as needed and where needed. This requires exploration of a variety of techniques and implies working with customised materials and techniques while finding the best methods of applying materials in the logic of specific force flows or thermal dissipation patterns.RDCB advances multi- and trans-disciplinary knowledge in robotically driven construction by designing and engineering new building systems for the on-demand production of customisable building components (Bier, 2014). The main consideration is that in architecture and building construction the factory of the future employs building materials and components that can be on site robotically processed and assembled

Assessing and facilitating warehouse safety

Purpose – The purpose of this paper is to investigate how warehouse safety can be assessed and facilitated. Methodology – Through a literature study, we build a theoretical framework to provide insights in how safety in Logistics Service Providers (LSPs) can be assessed and facilitated. We perform a case study at a large Dutch LSP using interviews and questionnaires to determine the relevance of the sub-dimensions to assess warehouse safety. Findings – Using literature, we identify people, procedures and technology related sub-dimensions of safety culture and safety behavior and factors that may affect how safety culture translates to safety behavior. Using a case study our findings indicate which sub-dimensions and influencing factors LSP employees find important and why. We found differences in the importance assigned to safety, which may point to the existence of sub-cultures across warehouses. Research limitations/implications – This paper contributes to the limited existing warehouse safety literature in which the factors that influence safety are not well explored. Although the case study investigates one LSP and as such does not generalize across LSPs, it provides valuable insights in important aspects of safety and how they can be influenced. Practical implications – This paper offers safety managers insights in how to assess and facilitate safety within their warehouses. Originality – Although warehouse safety is important, there is scarce academic research that explores this issue

Probabilistic Flood Defence Assessment Tools

The WTI2017 project is responsible for the development of flood defence assessment tools for the 3600 km of Dutch primary flood defences, dikes/levees, dunes and hydraulic structures. These tools are necessary, as per January 1st 2017, the new flood risk management policy for the Netherlands will be implemented. Then, the seven decades old design practice (maximum water level methodology of 1958) and two decades old safety standards (and maximum hydraulic load methodology of 1996) will formally be replaced by a more risked based approach for the national policy in flood risk management. The formal flood defence assessment is an important part of this new policy, especially for flood defence managers, since national and regional funding for reinforcement is based on this assessment. This new flood defence policy is based on a maximum allowable probability of flooding. For this, a maximum acceptable individual risk was determined at 1/100 000 per year, this is the probability of life loss of for every protected area in the Netherlands. Safety standards of flood defences were then determined based on this acceptable individual risk. The results were adjusted based on information from cost -benefit analysis, societal risk and large scale societal disruption due to the failure of critical infrastructure e.g. power stations. The resulting riskbased flood defence safety standards range from a 300 to a 100 000 year return period for failure. Two policy studies, WV21 (Safety from floods in the 21st century) and VNK-2 (the National Flood Risk in 2010) provided the essential information to determine the new risk based safety standards for flood defences. The WTI2017 project will provide the safety assessment tools based on these new standards and is thus an essential element for the implementation of this policy change. A major issue to be tackled was the development of user-friendly tools, as the new assessment is to be carried out by personnel of the institutions managing flood the defences, and not by just a small number of experts in probabilistic assessment. Therefore, data management and use of software are main issues that have been covered in courses and training in 2016 and 2017. All in all, this is the largest change in the assessment of Dutch flood defences since 1996. In 1996 probabilistic techniques were first introduced to determine hydraulic boundary conditions (water levels and waves (wave height, wave period and direction for different return periods). To simplify the process, the assessment continues to consist of a three-step approach, moving from simple decision rules, to the methods for semi-probabilistic assessment, and finally to a fully probabilistic analysis to compare the strength of flood defences with the hydraulic loads. The formal assessment results are thus mainly based on the fully probabilistic analysis and the ultimate limit state of the strength of a flood defence. For complex flood defences, additional models and software were developed. The current Hydra software suite (for policy analysis, formal flood defence assessment and design) will be replaced by the model Ringtoets. New stand-alone software has been developed for revetments, geotechnical analysis and slope stability of the foreshore. Design software and policy analysis software, including the Delta model, will be updated in 2018. A fully probabilistic method results in more precise assessments and more transparency in the process of assessment and reconstruction of flood defences. This is of increasing importance, as large-scale infrastructural projects in a highly urbanized environment are increasingly subject to political and societal pressure to add additional features. For this reason, it is of increasing importance to be able to determine which new feature really adds to flood protection, to quantify how much its adds to the level of flood protection and to evaluate if it is really worthwhile. Please note: The Netherlands government policy is to develop open source models