Mushrooms have long served as both food and medicine, providing polysaccharides, terpenoids, phenolics, and peptides with diverse health benefits. Extensive studies have begun to clarify the molecular mechanisms underlying these therapeutic effects, which include anti-aging, immunomodulatory, anticancer, and anti-inflammatory activities. Translational research is progressing from preclinical models to clinical trials, reinforcing the biomedical potential of mushroom-derived compounds. Advances in fungal genetic modification and gene editing have further positioned edible mushrooms as promising platforms for recombinant biopharmaceutical production. Their eukaryotic protein-processing capacity, natural bioencapsulation, and GRAS (Generally Recognized as Safe) status make them well-suited for sustainable and orally deliverable therapeutics. Engineered mushrooms show strong potential as platforms for oral vaccines and recombinant protein production, bridging traditional medicinal use with modern molecular farming.

Traditional stable genetic transformation in plant biotechnology remains largely inaccessible at many Primarily Undergraduate Institution (PUIs) due to high costs, long timelines, and specialized facility demands. Viral vector-based transient expression systems offer an efficient and accessible alternative method that enables meaningful undergraduate research within a single academic term. These systems utilize plant virus-derived vectors (e.g., TMV or Geminivirus) to transiently express target genes, producing detectable recombinant proteins within 3–7 days. Requiring only basic lab tools, they align well with Course-based Undergraduate Research Experiences (CUREs), lab courses, and capstone projects. Students gain practical experience in gene cloning, agroinfiltration, protein or metabolite chemical analysis, while faculty benefit from increased research capacity and funding potential. This mini-review highlights the advantages, implementation strategies, and funding opportunities of viral vector-based transient expression systems at PUIs, underscoring their value in expanding access to synthetic biology, plant-based biomanufacturing, and interdisciplinary STEM education.

Several studies across this Research Topic highlight the continual refinement of N. benthamiana as a host for improved recombinant protein production. Suppressing the defense-associated genes Nb-SABP2 and Nb-COI1 markedly increased transient expression efficiency by reducing immune responses that normally limit heterologous protein accumulation, offering a practical strategy for enhancing yield without affecting plant growth (Kopertekh, 2024). Yang et al. (2025) also characterized the papain-like cysteine protease (PLCP) gene family and identified specific proteases that degrade recombinant proteins, revealing targets whose suppression can further improve protein stability in planta. Additional refinement comes from Kaur et al. (2025) employed multiplex CRISPR/Cas9 editing to disrupt all seven core glycosyltransferase genes responsible for plant-specific N-glycan motifs, generating stable, Cas9-free N. benthamiana lines completely lacking α-1,3-fucosyltransferase and β-1,2-xylosyltransferase activities. These glycoengineered plants grew normally and formed a robust platform for producing biotherapeutics with human-compatible glycosylation profiles.Glyco-optimization remains central to enabling therapeutic-grade protein production in plants. Göritzer et al. (2025) demonstrated that modifying amino acids adjacent to the conserved Asn297 site, particularly through the Y300L variant, markedly increases N-glycan occupancy in plantproduced IgG1 and enhances Fc receptor binding and thermal stability, thereby reducing functional differences between plant-and mammalian-derived antibodies. Moss systems offer an additional avenue for controlled glycan engineering. Jonner et al. (2025) showed that introducing Spodoptera frugiperda mannosidase III and hexosaminidase into Physcomitrium patens, together with promoter tuning and targeted subcellular localization, enables production of recombinant human lysosomal acid α-glucosidase (Repleva GAA, RPV-002) enriched with paucimannosidic (MM) glycans. The engineered moss lines retained normal growth and achieved up to 43.5% of MM glycans, offering a practical strategy for improving mannose receptor-mediated uptake of therapeutic proteins.Several articles in this Topic highlight the production of functional therapeutic molecules in plant systems. Yu et al. (2025) demonstrated that expression of an epidermal growth factor (EGF)transdermal peptide fusion in A. thaliana yields a recombinant protein capable of penetrating the skin, and topical application markedly improves skin barrier repair in mice, illustrating the potential of plants for dermatological biologics. Melendez et al. (2024) reported that N. benthamiana can rapidly produce a monoclonal antibody targeting the extracellular enveloped virion (EV) form of Monkeypox virus; the plant-derived antibody bound the MPXV A35 antigen and neutralized EV particles, demonstrating that plant-based systems can generate antibodies with functional antiviral activity against this clinically important virion form. Wang et al. ( 2025) also compared the expression of full-length and mature nattokinase, revealing that the full-length precursor induces rapid leaf necrosis, whereas the mature enzyme accumulates efficiently and retains strong fibrinolytic activities, supporting its feasibility as a plant-produced therapeutic enzyme.Viral nanoparticle technologies represent another major thematic area in this Research Topic. Ljumović et al. (2025) developed an innovative foliar spray infection method for producing Tomato Bushy Stunt Virus (TBSV)-derived viral nanoparticles in N. benthamiana, offering a simple and scalable alternative to syringe or vacuum infiltration and demonstrating compatibility with vertical farming systems. This advance is paired with a detailed description of a GMPcompliant manufacturing facility for producing clinical-grade plant-made nanomaterials, which outlines upstream cultivation, downstream purification, and regulatory considerations and provides a practical blueprint for industrial-scale nanoparticle production (Pivotto et al., 2025). Vater et al. (2025) also demonstrated that N. benthamiana can produce functional human Galectin-1 (hGAL1) with correct folding, glycan binding, and immunomodulatory activities, reinforcing the suitability of plant systems for generating active immunotherapeutic proteins.The Research Topic features significant contributions in metabolic engineering and agricultural biotechnology. Gerasymenko et al. (2025) demonstrated that co-expression of specialized isoprenyl diphosphate synthases (IDSs) and substrate-producing enzymes in N. benthamiana enables efficient biosynthesis of irregular monoterpene malonyl glucosides that are otherwise difficult to obtain, illustrating the versatility of plant systems for generating structurally diverse metabolites with pharmaceutical potential. Qi et al. (2024) reviewed advances in RNA interference as a precise and environmentally sustainable strategy for crop protection, highlighting developments in delivery platforms, regulatory considerations, and pathways for integration into large-scale agricultural practice. Schubert et al. (2025) utilized multilayered zeinbased protein bodies in N. benthamiana to encapsulate both wild-type and hypoallergenic parvalbumin, enhancing resistance to gastrointestinal digestion and enabling controlled release. These engineered protein bodies were efficiently internalized by intestinal epithelial cells, establishing a promising plant-derived bioencapsulation platform for oral administration.Together, these studies reflect a field moving rapidly toward technical maturity, translational relevance, and manufacturing readiness. We thank all contributing authors and reviewers for advancing the scientific and practical foundations of this evolving field, and we hope this collection will inspire continued innovation across molecular farming applications.

Nattokinase is a potent fibrinolytic enzyme widely used in the treatment of cardiovascular diseases for its ability to directly degrade fibrin and plasmin substrates, effectively dissolving blood clots. In this study, both full-length and mature forms of the nattokinase coding sequence were transiently expressed in Nicotiana benthamiana using a modified Bean Yellow Dwarf Virus (BeYDV) replicon system. Overexpression of the full-length (pre-pro) construct resulted in severe leaf necrosis within 2.5 days post-infiltration (dpi), with electrolyte leakage analysis indicating an 83.5% loss of membrane integrity by 3 dpi. In contrast, the mature form of nattokinase was successfully expressed without early cytotoxicity and exhibited strong caseinolytic and fibrinolytic activity, reaching 22,500 FU/g—comparable to commercial standards. These findings demonstrate the feasibility of producing biologically active nattokinase in plants and highlight the potential of plant-based expression systems as scalable, cost-effective platforms for therapeutic enzyme production.

Many primarily undergraduate institutions (PUIs) offer coursework in plant biology, agriculture, and environmental sciences but lack dedicated plant growth facilities and controlled-environment infrastructure. Hydroponic systems provide an indoor platform for year-round plant cultivation, hypothesis-driven experimentation, and integration with course-based undergraduate research experiences (CUREs). At our institution, commercial and custom-built hydroponic systems were integrated into existing teaching laboratories and maintained by faculty, laboratory staff, and undergraduate work-study students. This structure supported consistent growth conditions, repeated student-led experimentation, and practical training in plant cultivation and biotechnology. Based primarily on this institutional experience, this mini-review synthesizes pedagogical outcomes, implementation onsiderations, and funding pathways, highlighting hydroponic platforms as effective infrastructure for experiential, research-centered STEM education at PUIs.

A dual-platform mushroom cultivation system integrating artificial intelligence (AI)-assisted environmental monitoring and controlled-environment agriculture (CEA) was developed to support experiential STEM education across K–12 and undergraduate settings. Hands-on instruction with multicellular fungi is often limited by reliance on microbial models and by constraints associated with field-based activities. To address this gap, we implemented an indoor instructional platform that combines a commercial AI-assisted automated cultivation unit with a tent-based chamber for hands-on environmental control. Representative cultivated species included oyster mushrooms (Pleurotus spp.) and lion’s mane (Hericium erinaceus). The AI-assisted system provided sensor/camera-based monitoring, app-based feedback, and software-assisted regulation of humidity, light, and airflow, whereas the tent-based system enabled direct student manipulation of cultivation conditions. Together, the systems allowed students to observe fungal development, manage environmental parameters, and collect quantitative and qualitative data within a single academic term. Post-harvest activities, including mushroom-based food preparation and tasting, further connected fungal biology with food and sustainability. A matched pre- and post-course survey (n = 30) showed increases in students’ self-reported perceived understanding, cultivation confidence, and engagement, with mean scores increasing from approximately 2–4 to 6–8. Because the survey instrument was not formally validated and no control group was included, these results are interpreted as preliminary self-reported perceptions rather than objective evidence of learning gains. The platform provides a practical model for integrating fungal biology, AI-assisted environmental monitoring, and CEA into STEM education.