Plant Engineering Research Articles

Enhancing drought stress tolerance in horticultural plants through melatonin-mediated phytohormonal crosstalk.

Melatonin and melatonin-mediated phytohormonal crosstalk play a multifaceted role in improving drought stress tolerance via molecular mechanisms and biochemical interactions in horticultural plants. The physical, physiological, biochemical, and molecular characteristics of plants are all affected by drought stress. Crop yield and quality eventually decline precipitously as a result. A phytohormone, melatonin, controls several plant functions during drought stress. However, the interactions between melatonin and other phytohormones, particularly how they control plant responses to drought stress, have not been clearly explored. This review explores the effects of melatonin and particular phytohormones on improving plant tolerance to drought stress. Specifically, the key melatonin roles in improved photosynthetic performance, better antioxidant activities, up-regulated gene expression, increased plant growth, and yield, etc., during drought stress have been elucidated in this review. Furthermore, this review explains how the intricate networks of melatonin-mediated crosstalk phytohormones, such as IAA, BR, ABA, GA, JA, CK, ET, SA, etc., enable horticultural plants to tolerate drought stress. Thus, this research provides a better understanding of the role of phytohormones, mainly melatonin, elucidates phytohormonal cross-talks in drought stress response, and future perspectives of phytohormonal contributions in plant improvements including engineering plants for better drought stress tolerance via targeting melatonin interactions.

A Constructivist Leader Avoids the Bankruptcy of an Emblematic Company and the Deepening of Deindustrialization in Europe: Lessons From a Case Study

Confronting a complex reality always causes a mixture of shock that continues, even after acceptance. The moment Bertrand Martin takes over C.M.M. Sulzer-France in 1984, the company is in the disaster situation. With the Naval Construction crisis, the order book quickly ran out, the stock warehouses were full, and people's morale was at a level close to exhaustion. Everyone expected either closure or, at the very least, a massive collective dismissal. B. Martin's position surprises everyone and everything, equivalent to a real electroshock, according to his own expression. In a staff meeting, called by him to “say what he came for”, B. Martin (or a member of his management team) holds back and does not even formulate a word that evokes “solutions”. Faced with the dramatic events that the company was experiencing, B. Martin says he is at this staff meeting to hear… solutions, as an alternative to closure or massive layoffs. Rarely have words like these been heard in companies in a country accustomed to a hierarchical organizational culture and job cuts in response to crises. Voluntary problem-solving groups, involving more than 300 people from all hierarchical levels, developed a plan with 450 measures. The workers' assembly votes on the plan unanimously. In a short time, the company changes radically and comes out of the red, accepting a difficult challenge, associated with an order for two large power plant engines, for China, previously considered unrealizable. The dynamics introduced into the company become proverbial with unusual repercussions throughout France. The case seems to fit more into the order of counter factuality than a plausible situation. Four orders of reasons seem to have contributed to the novelty of the director general's communication: (1) A general director who, instead of starting by presenting a speech of hope (to create a climate of positive expectation), as is customary to do, exposes the naked reality for everyone to observe. (2) A leader who says he has no solutions but was available to listen to the solutions to be found within the organization. (3) An entrepreneur who knew how to listen to himself and listen to what he would call a “crazy dream” that mobilized the talents of each subject. (4) A people engineer who knew how to intuit, as an organizational reference, the thirst for collectively shared change and which would allow 70% of all four hundred proposals from the working groups to be carried out in six months. The constructivist transformational leadership of this leader, unlikely given the context, Bertrand Martin, made the difference, in a situation where “hope seemed to have disappeared”, in the words of Dante, in the “Divine Comedy” – sequel to Inferno.

A promiscuous mechanism to phase separate eukaryotic carbon fixation in the green lineage.

CO2 fixation is commonly limited by inefficiency of the CO2-fixing enzyme Rubisco. Eukaryotic algae concentrate and fix CO2 in phase-separated condensates called pyrenoids, which complete up to one-third of global CO2 fixation. Condensation of Rubisco in pyrenoids is dependent on interaction with disordered linker proteins that show little conservation between species. We developed a sequence-independent bioinformatic pipeline to identify linker proteins in green algae. We report the linker from Chlorella and demonstrate that it binds a conserved site on the Rubisco large subunit. We show that the Chlorella linker phase separates Chlamydomonas Rubisco and that despite their separation by ~800 million years of evolution, the Chlorella linker can support the formation of a functional pyrenoid in Chlamydomonas. This cross-species reactivity extends to plants, with the Chlorella linker able to drive condensation of some native plant Rubiscos in vitro and in planta. Our results represent an exciting frontier for pyrenoid engineering in plants, which is modelled to increase crop yields.

Engineering Plant Metabolism for Synthesizing Amino Acid Derivatives of Animal Origin Using a Synthetic Modular Approach.

The biosynthesis of amino acid derivatives of animal origin in plants represents a promising frontier in synthetic biology, offering a sustainable and eco-friendly approach to enhancing the nutritional value of plant-based diets. This study leverages the versatile capabilities of Nicotiana benthamiana as a transient expression system to test a synthetic modular framework for the production of creatine, carnosine, and taurine-compounds typically absent in plants but essential for human health. By designing and stacking specialized synthetic modules, we successfully redirected the plant metabolic flux toward the synthesis of these amino acid derivatives of animal origin. Our results revealed the expression of a standalone creatine module resulted in the production of 2.3 μg/g fresh weight of creatine in N. benthamiana leaves. Integrating two modules significantly carnosine yield increased by 3.8-fold and minimized the impact on plant amino acid metabolism compared to individual module application. Unexpectedly, introducing the taurine module caused a feedback-like inhibition of plant cysteine biosynthesis, revealing complex metabolic adjustments that can occur when introducing foreign pathways. Our findings underline the potential for employing plants as biofactories for the sustainable production of essential nutrients of animal origin.

Polymeric Nanocarriers Autonomously Cross the Plant Cell Wall and Enable Protein Delivery for Stress Sensing.

Delivery of proteins in plant cells can facilitate the design of desired functions by modulation of biological processes and plant traits but is currently limited by narrow host range, tissue damage, and poor scalability. Physical barriers in plants, including cell walls and membranes, limit protein delivery to desired plant tissues. Herein, a cationic high aspect ratio polymeric nanocarriers (PNCs) platform is developed to enable efficient protein delivery to plants. The cationic nature of PNCs binds proteins through electrostatic. The ability to precisely design PNCs' size and aspect ratio allowed us to find a cutoff of ≈14nm in the cell wall, below which cationic PNCs can autonomously overcome the barrier and carry their cargo into plant cells. To exploit these findings, a reduction-oxidation sensitive green fluorescent protein (roGFP) is deployed as a stress sensor protein cargo in a model plant Nicotiana benthamiana and common crop plants, including tomato and maize. In vivo imaging of PNC-roGFP enabled optical monitoring of plant response to wounding, biotic, and heat stressors. These results show that PNCs can be precisely designed below the size exclusion limit of cell walls to overcome current limitations in protein delivery to plants and facilitate species-independent plant engineering.

Direct protein delivery into intact Arabidopsis cells for genome engineering

Intracellular delivery of biomolecules is a prerequisite for genetic techniques such as recombinant engineering and genome editing. Realizing the full potential of this technology requires the development of safe and effective methods for delivering protein tools into cells. In this study, we demonstrated the spontaneous internalization of exogenous proteins into intact cells and root tissue of whole plants of Arabidopsis thaliana. We termed this internalization phenomenon as protein Delivery Independent of Vehicles or Equipment (DIVE), which efficiently delivered genome engineering proteins including Cre recombinase and zinc-finger nucleases (ZFN) into plant cells. Using protein DIVE, we achieved less toxic protein delivery than electroporation with up to 94% efficiency in Arabidopsis cell culture and 19% genome modification in Arabidopsis plants that was maintained in regenerated tissue. This work illustrates the potential of protein DIVE for a wide range of applications, including genome engineering in plants.

Technical and economic aspects of a nuclear power engineering plant for hydrogen production

Technical and economic aspects of a nuclear power engineering plant for hydrogen production

Double Transformation in Mechanical and Plant Engineering - Digitalization and sustainability for one-of-a-kind and small-batch manufacturers

Double Transformation in Mechanical and Plant Engineering - Digitalization and sustainability for one-of-a-kind and small-batch manufacturers

Enhancing the Catalytic Activity of Geranylgeranyl Diphosphate Synthase through Ancestral Sequence Reconstruction and Semirational Design.

Geranylgeranyl diphosphate synthase (GGPPS) is the crucial bottleneck in carotenoid biosynthesis. However, low activity limits the broad application of GGPPS. In this study, OsGGPPS1 in rice was engineered based on ancestral sequence reconstruction (ASR) and semirational design to improve the catalytic performances of existing GGPPS. The better mutant of A22R/A26P with improved enzyme activity was generated based on ASR. Additionally, the improved enzyme activity of mutants as V162A/M218S/F227Y was designed using a semirational design. The combinatorial assembly of the d-OsGGPPS1 mutant (A22R/A26P/V162A/M218S/F227Y) exhibited higher conversion of IPP and each cosubstrate of DMAPP for 9.8-fold in GPP production, GPP for 6.4-fold in FPP production, and FPP for 1.4-fold in GGPP production relative to wild-type OsGGPPS1 at 25 °C, which showed higher conversion than wild-type OsGGPPS1 at temperatures as high as 50 °C. The successful design of OsGGPPS1 was representative of protein engineering, which will shed new light on GGPPS engineering and active plant pigment resource utilization.

Conquering Limitations: Exploring the Factors that Drive Successful Agrobacterium-Mediated Genetic Transformation of Recalcitrant Plant Species.

Agrobacterium-mediated transformation is a preferred method for genetic engineering and genome editing of plants due to its numerous advantages, although not all species exhibit transformability. Genetic engineering and plant genome editing methods are technically challenging in recalcitrant crop plants. Factors affecting the poor rate of transformation in such species include host genotype, Agrobacterium genotype, type of explant, physiological condition of the explant, vector, selectable marker, inoculation method, chemical additives, antioxidative compounds, transformation-enhancing compounds, medium formulation, optimization of culture conditions, and pre-treatments. This review provides novel insights into the key factors involved in gene transfer facilitated by Agrobacterium and proposes potential solutions to overcome existing barriers to transformation in recalcitrant species, thereby contributing to improvement programs for these species. This review introduces the key factors that impact the effectiveness of a molecular breeding program using Agrobacterium-mediated transformation, specifically focusing on recalcitrant plant species.

Expression of a mammalian RNA demethylase increases flower number and floral stem branching in Arabidopsis thaliana

AbstractRNA methylation plays a central regulatory role in plant biology and is a relatively new target for plant improvement efforts. In nearly all cases, perturbation of the RNA methylation machinery results in deleterious phenotypes. However, a recent landmark paper reported that transcriptome‐wide use of the human RNA demethylase FTO substantially increased the yield of rice and potatoes. Here, we have performed the first independent replication of those results and demonstrated broader transferability of the trait, finding increased flower and fruit count in the model species Arabidopsis thaliana. We also performed RNA‐seq of our FTO‐transgenic plants, which we analyzed in conjunction with previously published datasets to detect several previously unrecognized patterns in the functional and structural classification of the upregulated and downregulated genes. From these, we present mechanistic hypotheses to explain these surprising results with the goal of spurring more widespread interest in this promising new approach to plant engineering.

Predictive modeling of combined cycle power plant performance using a digital twin-based neural ODE approach

This study presents a novel digital twin-based predictive modeling approach to enhance the operation, maintenance, and management of combined cycle power plants. The research aimed to accurately forecast the combined cycle power (CCP) output, a critical performance indicator, to support intelligent decision-making for plant operators and engineers. The proposed framework leverages a neural ordinary differential equation (NODE) model integrated within a digital twin architecture to capture the complex, nonlinear dynamics of combined cycle gas turbine unit (CCGU) system. The predictive model was trained and validated using multivariate time-series data collected from a CCGU, achieving high accuracy with an R2_score of 0.993, a mean absolute percentage error of 0.325 %, and a root mean squared error of 1.518. Importantly, the NODE model demonstrated superior forecasting capabilities compared to traditional machine learning techniques. The digital twin (DT) concept enables seamless real-time data integration, physics-based models, and advanced data analytics to facilitate comprehensive CCGU lifecycle management. This novel approach provides operators with reliable, real-time insights to optimize plant performance, reduce maintenance costs, and improve overall sustainability. The generalization of the NODE model to an independent CCGU unit validated its applicability across diverse power plant configurations, highlighting the originality and practical value of the research. The key contribution of this work is the development of a digital twin-based predictive modeling framework centered on the innovative use of neural ordinary differential equations to enhance the operation and maintenance of critical energy infrastructure. This research advances the state-of-the-art in intelligent asset management for the built environment.

(Invited) The “Duckweed Dip”: DNA-Wrapped Carbon Nanotubes for Transient Gene Expression

This study reports on usage of functionalized carbon nanotubes as a transgene delivery cargo into higher plants. We show that aquatic Spirodela polyrhiza (Duckweed) plant, when incubated in a solution containing carbon nanotubes wrapped with a plasmid containing the gene for reporter protein β-glucuronidase (GUS), can directly uptake the DNA-CNTs with high efficiency. The mechanism of the DNA-CNT delivery, directly from growth media, and plasmid release is discussed. We show that transgenes encoded within the plasmids are expressed by the plants. This method allows to avoid using large doses of nanomaterials or direct infiltration of agrobacterium into plant tissue. We expect this new approach for transgene delivery will allow for more efficient plant engineering and can serve as a useful tool in synthetic biology.ACKNOWLEDGMENTThe work was supported by the National Institutes of Health/National Institute of General Medical Sciences [1R35GM133483] and Department of Defense [Contract #W911QY2220006]. This work was performed in part at the Joint School of Nanoscience and Nanoengineering, a member of the Southeastern Nanotechnology Infrastructure Corridor (SENIC) and National Nanotechnology Coordinated Infrastructure (NNCI), which is supported by the National Science Foundation [ECCS-1542174].

Processing of crack-free Nickel- and Cobalt-based wear protection coatings and defined surfaces by subsequent milling processes

In the area of plant engineering, steel components are provided with a wear protection coating for efficient use to protect them against corrosive, tribological, thermal and mechanical stresses. The use of innovative ultrasound-assisted milling processes and plasma-welded nickel- and cobalt-based wear protection coatings are being investigated to determine how more favourable machinability can be achieved while retaining the same wear protection potential. The focus is on the NiCrSiFeB alloy, which is intended to replace CoCr alloys in the area of screw machines. The utilization of ultrasonic-assisted milling for the machining of coating materials is a novel approach. The modification of hard facing layers in terms of microstructure and precipitation morphology as well as suitability for machining is investigated and compared with the CoCr alloy. The alloy modifications are generated by a PTA process by systematically adjusting the preheating and interpass temperatures, a crack-free wear-resistant layer can be generated, which is subsequently machined by a milling process. In addition to the crack-free properties, the microstructure, the bonding as well as the mixing between the NiCrSiFeB alloy and a 1.8550 as well as between the CoCr alloy and a 1.4828 are analysed and compared in the joining areas. In addition, heating and cooling rates are determined and a chemical analysis of the weld metals is performed. Furthermore, it was found that the build-up layers of NiCrSiFeB alloy are more difficult to machine using the milling process than the CoCr alloy, as higher milling forces are required.

CRISPR/Cas-Mediated Genome Engineering in Plants: Application and Prospectives.

Genetic engineering has become an essential element in developing climate-resilient crops and environmentally sustainable solutions to respond to the increasing need for global food security. Genome editing using CRISPR/Cas [Clustered regulatory interspaced short palindromic repeat (CRISPR)-associated protein (Cas)] technology is being applied to a variety of organisms, including plants. This technique has become popular because of its high specificity, effectiveness, and low production cost. Therefore, this technology has the potential to revolutionize agriculture and contribute to global food security. Over the past few years, increasing efforts have been seen in its application in developing higher-yielding, nutrition-rich, disease-resistant, and stress-tolerant "crops", fruits, and vegetables. Cas proteins such as Cas9, Cas12, Cas13, and Cas14, among others, have distinct architectures and have been used to create new genetic tools that improve features that are important for agriculture. The versatility of Cas has accelerated genomic analysis and facilitated the use of CRISPR/Cas to manipulate and alter nucleic acid sequences in cells of different organisms. This review provides the evolution of CRISPR technology exploring its mechanisms and contrasting it with traditional breeding and transgenic approaches to improve different aspects of stress tolerance. We have also discussed the CRISPR/Cas system and explored three Cas proteins that are currently known to exist: Cas12, Cas13, and Cas14 and their potential to generate foreign-DNA-free or non-transgenic crops that could be easily regulated for commercialization in most countries.

Deep mutational scanning reveals sequence to function constraints for SWEET family transporters.

Protein science is entering a transformative phase enabled by deep mutational scans that provide an unbiased view of the residue level interactions that mediate function. However, it has yet to be extensively used to characterize the mutational and evolutionary landscapes of plant proteins. Here, we apply the method to explore sequence-function relationships within the sugar transporter AtSWEET13. DMS results describe how mutational interrogation throughout different regions of the protein affects AtSWEET13 abundance and transport function. Our results identify novel transport-enhancing mutations that are validated using the FRET sensor assays. Extending DMS results to phylogenetic analyses reveal the role of transmembrane helix 4 (TM4) which makes the SWEET family transporters distinct from prokaryotic SemiSWEETs. We show that transmembrane helix 4 is intolerant to motif swapping with other clade-specific SWEET TM4 compositions, despite accommodating single point-mutations towards aromatic and charged polar amino acids. We further show that the transfer learning approaches based on physics and ML based In silico variant prediction tools have limited utility for engineering plant proteins as they were unable to reproduce our experimental results. We conclude that DMS can produce datasets which, when combined with the right predictive computational frameworks, can direct plant engineering efforts through derivative phenotype selection and evolutionary insights.

Evolving Roles in Systems Engineering —‐ Insights from Germany's Mechanical and Plant Engineering Sector

AbstractSystems Engineering is developing differently in each sector and region. In German industry, especially in mechanical and plant engineering, Systems Engineering is of major importance. The introduction of Systems Engineering raises the question of which roles and competencies are required. This article examines the evolution of roles in Systems Engineering from a German perspective. Based on a literature review, the evolution of the identified Systems Engineering roles over time is shown, starting with the seminal publication by Sheard in 1996. It points out that only minimal adjustments and occasional role renaming have occurred. However, the review shows a common understanding of essential areas of responsibility within the SE and changes over time. The next step is to examine the current understanding of Systems Engineering roles in the industry. A quantitative analysis of job postings in Germany reveals a diverse interpretation of the term 'Systems Engineer; more than half of the positions cannot be categorized according to the INCOSE definitions. The job postings are used to determine which tasks are associated with the job, how often they occur, and in what combination. The primary responsibilities of systems engineers include creating and managing requirements, architecture processes, validation, and verification processes, and coordinating with customers and stakeholders. Finally, three representative companies from the mechanical and plant engineering sector were selected to analyze existing roles and tasks. From this, a common understanding of tasks and responsibilities is combined and organized into clusters. These are used by the companies to locate and thus derive their roles. The result is in an integrative approach that enables companies, especially in the midsize and medium sectors, to design the introduction in line with stakeholder demands. In summary, the industry's ongoing adaptation necessitates the evolution of Systems Engineering roles and competencies for successful and sustainable development and implementation of systems.

Role of IIoT as New Business Model Innovation in the Manufacturing Industry

The manufacturing Industry in Europe plays a significant role in economic growth and development. Given the increased role of technology in this sector, the Industrial Internet of Things (IIoT) is one of the technological advancements that improve the Business Model (BM) efficacy in various manufacturing industries. However, research into the role of IIoT as a BM in the manufacturing industry is limited despite its contribution to the success and performance of the firms. Therefore, this study explores the role that IIoT plays as a BM in various manufacturing industries. A survey was conducted utilizing open-ended questionnaires across 60 manufacturing firms in Europe including automotive suppliers, information and communication, machine and plant engineering, electrical engineering, and medical engineering. The findings indicate that IIoT as a BM has different levels of contribution across the manufacturing industry with glaring variations in terms of performance between information and communication firms as well as automotive. This study can act as a guideline for further research on the complex relationship between IIoT as a BM and firms’ performance to influence decision-making processes within the manufacturing industry. Given that this is a pre-study, the limitations, and recommendations for future research are highlighted.

Transcriptional response of Arabidopsis thaliana's root-tip to spaceflight.

Plants are expected to play a critical role in the biological life support systems of crewed spaceflight missions, including in the context of upcoming missions targeting the Moon and Mars. Therefore, understanding the response of plants to spaceflight is essential for improving the selection and engineering of plants and spaceflight conditions. In particular, understanding the root-tip's response to spaceflight is of importance as it is the center of orchestrating the development of the root, the primary organ for the absorption of nutrients and anchorage. GLDS-120 is a pioneering study by Paul et al. that used transcriptomics to evaluate the spaceflight response of the root-tip of the model plant Arabidopsis thaliana in dark and light through separate analyses of three genotype groups (Wassilewskija, Columbia-0, and Columbia-0 PhyD) and comparison of genotype responses. Here, we provide a complementary analysis of this dataset through a combined analysis of all samples while controlling for the genotypes in a paired analysis. We identified a robust transcriptional response to spaceflight with 622 DEGs in light and 200 DEGs in dark conditions. Gene enrichment analysis identified 37 and 13 significantly enriched terms from biological processes in light and dark conditions, respectively. Prominent enrichment for hypoxia-related terms in both conditions suggests hypoxia is a key stressor for root development during spaceflight. Additional enriched terms in light conditions include the circadian cycle, light response, and terms for the metabolism of flavonoid and indole-containing compounds. These results further our understanding of plants' responses to the spaceflight environment.

DNA Delivery by Virus-Like Nanocarriers in Plant Cells.

Tobacco mild green mosaic virus (TMGMV)-like nanocarriers were designed for gene delivery to plant cells. High aspect ratio TMGMVs were coated with a polycationic biopolymer, poly(allylamine) hydrochloride (PAH), to generate highly charged nanomaterials (TMGMV-PAH; 56.20 ± 4.7 mV) that efficiently load (1:6 TMGMV:DNA mass ratio) and deliver single-stranded and plasmid DNA to plant cells. The TMGMV-PAH were taken up through energy-independent mechanisms in Arabidopsis protoplasts. TMGMV-PAH delivered a plasmid DNA encoding a green fluorescent protein (GFP) to the protoplast nucleus (70% viability), as evidenced by GFP expression using confocal microscopy and Western blot analysis. TMGMV-PAH were inactivated (iTMGMV-PAH) using UV cross-linking to prevent systemic infection in intact plants. Inactivated iTMGMV-PAH-mediated pDNA delivery and gene expression of GFP in vivo was determined using confocal microscopy and RT-qPCR. Virus-like nanocarrier-mediated gene delivery can act as a facile and biocompatible tool for advancing genetic engineering in plants.