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15 pages, 8189 KB

Open AccessArticle

Comparative Effects of Cytokinins and Spermidine on In Vitro Organogenesis of Ruscus hypoglossum L. and Rooting of Regenerated Shoots

by İbrahim Halil Hatipoğlu and Ümmü Özgül Karagüzel

Plants 2026, 15(17), 2615; https://doi.org/10.3390/plants15172615 - 27 Aug 2026

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Abstract

Ruscus hypoglossum L. is an evergreen ornamental species valued as cut foliage, but its commercial production is constrained by slow vegetative propagation and limited availability of uniform planting material. This study aimed to develop an efficient in vitro propagation protocol for R. hypoglossum [...] Read more.

Ruscus hypoglossum L. is an evergreen ornamental species valued as cut foliage, but its commercial production is constrained by slow vegetative propagation and limited availability of uniform planting material. This study aimed to develop an efficient in vitro propagation protocol for R. hypoglossum by evaluating the effects of different cytokinins and spermidine on shoot organogenesis and subsequent rooting. Thidiazuron (TDZ), zeatin, kinetin, and spermidine were applied at 0.5 and 1.0 mg L⁻¹ to rhizome explants derived from in vitro-grown seedlings. Explant length, shoot and cladode number, fresh weight, shoot diameter, chlorophyll a and b, total carotenoid content, SPAD value, and rooting performance were assessed. Significant treatment effects were observed for most traits. Kinetin at 1.0 mg L⁻¹ produced the highest shoot proliferation (4.66 shoots explant⁻¹), cladode number (12.00 explant⁻¹), fresh weight (0.31 g), chlorophyll a (0.72 mg g⁻¹ FW), chlorophyll b (0.29 mg g⁻¹ FW), and SPAD value (40.25). Spermidine at 1.0 mg L⁻¹ resulted in the highest carotenoid content (0.59 mg g⁻¹ FW) and promoted greater explant elongation, whereas TDZ at 1.0 mg L⁻¹ reduced cladode formation and pigment accumulation. Multivariate analyses consistently identified kinetin as the most favorable treatment during multiplication. Shoots regenerated with 1.0 mg L⁻¹ kinetin also showed the highest rooting response following a 10 s pulse treatment with 2.0 mg L⁻¹ IBA and transfer to hormone-free half-strength MS medium. Overall, kinetin was the most effective treatment under the conditions tested, providing a practical basis for efficient clonal propagation and ex situ conservation of R. hypoglossum. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Plant Regeneration—2nd Edition)

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21 pages, 3433 KB

Open AccessArticle

Microbial Inoculation Enhances Growth and Physiological Traits of Tissue Cultured Panicum turgidum Forssk. Plantlets During Acclimatization

by Muhammad M. Habib, Yaser Hassan Dewir, Thobayet S. Alshahrani, Jahangir A. Malik, Basharat A. Dar and Abdulaziz A. Al-Qarawi

Plants 2026, 15(15), 2252; https://doi.org/10.3390/plants15152252 - 23 Jul 2026

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Abstract

Panicum turgidum is an important keystone forage grass crucial for ecosystem stability in arid and semi-arid rangelands of Asia and Africa. This study aimed to investigate the use of beneficial bioinoculants for enhancing the growth and acclimatization of micropropagated desert grass under controlled [...] Read more.

Panicum turgidum is an important keystone forage grass crucial for ecosystem stability in arid and semi-arid rangelands of Asia and Africa. This study aimed to investigate the use of beneficial bioinoculants for enhancing the growth and acclimatization of micropropagated desert grass under controlled environmental conditions. Tissue-cultured plants were transferred to pots with a sand–soil mixture (1:1, v/v) and kept in a temperature and light-controlled environment (25 ± 2 °C, 100 µmol m⁻²·s⁻¹, and 16/8 light/dark photoperiod) for 10 weeks. We used a factorial approach to investigate the effects of the arbuscular mycorrhizal fungus (AMF) Rhizophagus fasciculatus, Trichoderma harzianum, and Bacillus subtilis on transplanted desert grass plants. Two AMF levels control (AMF, 5%; w/w) or without AMF (NAMF); and three microbial inoculation treatments (T. harzianum, B. subtilis, and a combination of T. harzianum + B. subtilis) were employed. Microscopic investigation indicated the extent of AMF colonization in the roots of micropropagated P. turgidum plantlets during acclimatization. Growth parameters with shoots and roots, chlorophyll and carotenoid content, and nitrogen uptake were all improved in bioinoculants-treated plants. Under non-AMF conditions, co-inoculation with T. harzianum and B. subtilis resulted in the highest biomass and root traits, whereas under AMF conditions, T. harzianum alone or with AMF was generally most effective. In the plants treated with the combination of inoculants, heatmap correlation, principal component analysis, and hierarchical clustering demonstrated positive association between growth and physiological traits and the absorption of mineral nutrients, although these associations varied among treatment combinations. These findings highlight the potential of beneficial bioinoculants to improve the growth and acclimatization of micropropagated desert grass plants, providing a foundation for tailored bioinoculant compositions towards the rehabilitation of degraded grazing lands. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Plant Regeneration—2nd Edition)

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21 pages, 2643 KB

Open AccessArticle

ROSEA1-Based Visual Selection Reduces Plant Regeneration and Alters Developmental Regulator Expression

by Tao Jiang, Sameena Ejaz Tanwir, Fangchen Liu, Wisnu Handoyo Ardi, Yeyen Novitasari, Sandy Zammar, Fida Hussain and Heqiang Huo

Plants 2026, 15(13), 2004; https://doi.org/10.3390/plants15132004 - 28 Jun 2026

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Abstract

Anthocyanin-based visual reporters enable rapid, non-destructive identification of transgenic tissues, but their pigment output may not be physiologically neutral during organogenesis, raising concerns about their suitability as selection markers in transformation pipelines. To address this, we examined the regeneration performance of constitutive ROSEA1 [...] Read more.

Anthocyanin-based visual reporters enable rapid, non-destructive identification of transgenic tissues, but their pigment output may not be physiologically neutral during organogenesis, raising concerns about their suitability as selection markers in transformation pipelines. To address this, we examined the regeneration performance of constitutive ROSEA1 expression across four eudicot species from three families: tomato, petunia, begonia, and marigold. Stable ROSEA1-overexpressing lines were evaluated in tomato and petunia, whereas transformation-stage assays were performed in begonia and marigold. Regeneration frequencies were quantified relative to controls, and transcript-level changes were assessed for anthocyanin biosynthetic genes and regeneration regulators; complementation assays with PLT5 co-expression and hormonal manipulations were also performed. In tomato and petunia, ROSEA1-overexpressing lines exhibited reduced regeneration frequencies and elevated anthocyanin accumulation, with two independent petunia lines showing that stronger pigment activation corresponded to more severe regeneration defects. In tomato, ROSEA1 coordinately upregulated CHI, F3H, F3′5′H, and DFR and was associated with reduced expression of PLT5, WUS, and LBD16 during early regeneration. PLT5 co-expression partially restored regeneration with modestly reduced pigmentation, indicating that pigment output alone does not fully explain the phenotype. The penalty extended to begonia and marigold, with genotype- and hormone-dependent severity in marigold. These findings indicate a context-dependent regeneration penalty associated with constitutive ROSEA1 expression and suggest that hormone optimization, genotype selection, and developmental support may help mitigate this limitation in some contexts. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Plant Regeneration—2nd Edition)

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23 pages, 18655 KB

Open AccessArticle

Synthetic Small Molecules as Regulators of In Vitro Multiplication in Selenicereus Hybrids

by Malen Escánez, Alejandro Miralles-Rodríguez, Sandra Gil, Francisco Bermúdez, Santiago Vilanova, Elena Carneros, Ana Martinez, Carmen Gil, Pilar S. Testillano and Edgar García-Fortea

Plants 2026, 15(13), 1931; https://doi.org/10.3390/plants15131931 - 23 Jun 2026

Viewed by 498

Abstract

Micropropagation of Selenicereus hybrids is a key tool for breeding and conservation; however, further refining the balance between high multiplication rates and morphological quality remains a complex challenge within conventional protocols. This study explores targeted signaling modulation using nine bioactive small molecules—including three [...] Read more.

Micropropagation of Selenicereus hybrids is a key tool for breeding and conservation; however, further refining the balance between high multiplication rates and morphological quality remains a complex challenge within conventional protocols. This study explores targeted signaling modulation using nine bioactive small molecules—including three mammalian glycogen synthase kinase 3 (GSK3) inhibitors (TDZD-9, VP3.15 and VP0.7), three leucine rich repeat kinase 2 (LRRK2) inhibitors (JZ1.24, JZ1.3 and IGS4.75), and three phosphodiesterase (PDE) inhibitors—to complement traditional micropropagation. Explants were evaluated in two distinct contexts: a hormone-free basal medium (BM) and a plant growth regulator-supplemented medium (PIT2) and the response rates, yield, and quality were measured and integrated using a Global Efficiency Index (GEI). Results demonstrate that inhibitor efficacy is strictly context-dependent; while most molecules repressed budding in BM, they acted as response modulators by determining the specific type of morphogenic pathway in PIT2. Notably, the GSK3 inhibitor TDZD-9 reached the highest GEI (0.85) by maximizing productivity, whereas LRRK2 inhibitors effectively preserved architectural integrity. Flow cytometry confirmed cytogenetic stability across all treatments, with a 98.5% plantlet survival rate during acclimatization. In conclusion, the strategic integration of targeted signaling modulators and multi-parametric indices offers a refined and objective framework to enhance the efficiency of mass propagation protocols in pitahaya and other recalcitrant species. Furthermore, our findings provide new evidence of the strong potential of these small molecules as novel tools to improve plant micropropagation beyond traditional plant growth regulators. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Plant Regeneration—2nd Edition)

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12 pages, 13440 KB

Open AccessArticle

Ectomycorrhizal Symbiosis as a Bio-Enhancement Strategy for Transplantation of Somatic Embryo-Derived Pinus elliottii

by Zhen-Xing Tian, Xin Ke, Xin-Yi Ji, Xi-Yuan Chen and Li-Hua Zhu

Plants 2026, 15(11), 1701; https://doi.org/10.3390/plants15111701 - 30 May 2026

Viewed by 519

Abstract

Somatic embryo-derived plantlets of pines often fail to survive acclimatization, which limits commercial micropropagation. Conventional hardening methods do not correct the physiological weaknesses of in vitro plantlets, especially the lack of beneficial microbes. Here we developed a practical protocol for resistant Pinus elliottii [...] Read more.

Somatic embryo-derived plantlets of pines often fail to survive acclimatization, which limits commercial micropropagation. Conventional hardening methods do not correct the physiological weaknesses of in vitro plantlets, especially the lack of beneficial microbes. Here we developed a practical protocol for resistant Pinus elliottii. First, we used an optimized maturation protocol (three sequential ABA pre-treatments) and glucose for germination. Substrate screening showed that a peat:vermiculite:perlite mixture (3:1:1) gave the highest survival (98.9%). Then, before transplantation, we introduced a key bio-enhancement step: in vitro inoculation with the ectomycorrhizal fungus Pisolithus orientalis cfcc7668. This treatment achieved a mycorrhization rate of 97.7% and transformed root morphology from thin, sparsely branched roots to a coralloid, dichotomously branched system with a well-developed Hartig net. As a result, mycorrhizal plantlets had 100% transplant survival at 30 days and remained above 94% over 360 days, whereas non-inoculated controls dropped to 95.6% at 30 days and further declined to about 73% after three months. Pre-establishing ectomycorrhizal symbiosis effectively restores a key root function missing in in vitro plantlets. Our integrated procedure provides a practical method for clonal propagation of conifers. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Plant Regeneration—2nd Edition)

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21 pages, 3413 KB

Open AccessArticle

In Vitro Regeneration Strategies in Woody Citrus of Northeast India (Citrus jambhiri and Citrus aurantifolia)

by Priyanka Sharma, Bidhan Roy, Gopal Shukla, Monish Roy, Gadge Sushant Sundarrao and Vijay Dunna

Plants 2026, 15(11), 1677; https://doi.org/10.3390/plants15111677 - 29 May 2026

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Abstract

Since the status of C. jambhiri is “rare” and C. aurantifolia is endowed with folk medicinal properties, our study aimed at producing true-to-type seedlings for further conservation by using PGRs, ferrous sulfate, and casein hydrolysate in MS medium to induce in vitro germination [...] Read more.

Since the status of C. jambhiri is “rare” and C. aurantifolia is endowed with folk medicinal properties, our study aimed at producing true-to-type seedlings for further conservation by using PGRs, ferrous sulfate, and casein hydrolysate in MS medium to induce in vitro germination and “callus” formation from seeds of C. jambhiri Lush. It also focused on evaluation of suitable species and the best type of explant for organogenesis. The present study was undertaken to develop an efficient micropropagation protocol for C. jambhiri Lush. and C. aurantifolia. The frequency of callus induction increased to its maximum when 2.0 mg/L 2,4-D and 1.0 mg/L picloram were added individually. The calli derived from 2,4-D exhibited maximum regeneration potentiality. In addition, sucrose (30 g/L), dextrose (60 g/L), and coconut water (10 mL/L and 15 mL/L) also enhanced callus induction. Regarding heavy metals, 100 ppm of Fe₂SO₄ exhibited maximum germination percentage (84.33%) from seeds of C. jambhiri Lush. However, maximum callus induction (50.00%) was induced from the seeds of C. aurantifolia incubated in Fe₂SO₄ (400 ppm). The maximum number of shoots per callus was produced (5.13) with the addition of 200 mg/L casein hydrolysate in the callus-induction medium. Pearson correlation analysis revealed a positive and significant association of the number of plantlets/calluses with shoot length and regeneration percentage, respectively. It was phenotypically observed that the tissue culture traits performed better from explants derived from C. jambhiri Lush. than C. aurantifolia when subjected to varying concentrations of PGRs, carbon sources, organic adjuvants, and heavy metals, respectively. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Plant Regeneration—2nd Edition)

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17 pages, 19300 KB

Open AccessArticle

Effects of Explant Source and Orientation on Secondary Somatic Embryogenesis in Hevea brasiliensis

by Xiaochuan Gu, Jingyu Ao, Lisheng Kong, Xuemei Dai, Huasun Huang, Huabo Du, Xiaoyi Wang and Tiandai Huang

Plants 2025, 14(21), 3274; https://doi.org/10.3390/plants14213274 - 27 Oct 2025

Viewed by 1185

Abstract

Propagation of rubber tree (Hevea brasiliensis) via secondary somatic embryogenesis (SSEis) is a reliable method. However, its efficiency is relatively low. The aim of this study was to understand more about the factors related to SSEis in rubber trees, trying to [...] Read more.

Propagation of rubber tree (Hevea brasiliensis) via secondary somatic embryogenesis (SSEis) is a reliable method. However, its efficiency is relatively low. The aim of this study was to understand more about the factors related to SSEis in rubber trees, trying to improve the efficiency of somatic embryo (SE) yield. Our study showed that the orientations of explants, i.e., the fragments of primary SE (PSE), on the medium affected secondary SE (SSE) yield significantly. Among five experimental tests, the highest yield was 2.6 ± 0.9 secondary somatic embryos (SSEs) per explant, which was achieved by orienting the abaxial side of the explant in contact with the medium and then the adaxial side after a period of culture time. Based on histological evidence, SSEis was induced from the epidermal cells and adjacent cells on the adaxial side of the explants. A remarkable difference in embryogenic capacity difference existed among individual PSE. The concentrations of soluble proteins, starch, soluble sugars, and the superoxide dismutase activity (SOD) levels in the explants were measured during a 25-day long SSEis induction treatment and compared between explants of high and low embryogenic capacity. This study proves that the explant orientation toward the culture medium plays a crucial role in SSEis, while the concentration changes of these biochemical compounds correlate to morphological changes in the explants during induction, as do the changes in SOD activity. Furthermore, the trend of the dynamic changes in the explants reflected a process of de-differentiation and re-differentiation, which started from mature SE tissues during SSE induction. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Plant Regeneration—2nd Edition)

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22 pages, 5266 KB

Open AccessArticle

Integrated Multi-Omics Reveals Mechanism of Adventitious Buds Regeneration in In Vitro Cultures of Cinnamomum parthenoxylon

by Chenglin Luo, Xin Qiao, Xiaoying Dai, Yuntong Zhang, Xinliang Liu and Yanfang Wu

Plants 2025, 14(19), 2945; https://doi.org/10.3390/plants14192945 - 23 Sep 2025

Cited by 1 | Viewed by 1682

Abstract

A pluripotent callus is central to genetic transformation in Cinnamomum parthenoxylon; however, the molecular and cellular mechanisms regulating callus formation and subsequent differentiation remain unelucidated, hindering progress in its genetic improvement. This study systematically investigated the dynamic changes during the in vitro [...] Read more.

A pluripotent callus is central to genetic transformation in Cinnamomum parthenoxylon; however, the molecular and cellular mechanisms regulating callus formation and subsequent differentiation remain unelucidated, hindering progress in its genetic improvement. This study systematically investigated the dynamic changes during the in vitro regeneration of C. parthenoxylon through morphological observations, physiological assays, and transcriptomic analyses, while comparing differences in callus formation under varying induction conditions to elucidate the mechanism of its high-efficiency regeneration. The results showed that the formation of a pluripotent callus is a critical step in C. parthenoxylon regeneration, characterized by the presence of highly proliferative cell zones. Compared to an ordinary callus (P3C), a pluripotent callus (P3) exhibited higher activities of polyphenol oxidase (PPO) and indole-3-acetic acid oxidase (IAAO), as well as elevated levels of zeatin riboside (ZR) and abscisic acid (ABA). In contrast, P3 showed lower levels of soluble sugars, soluble proteins, malondialdehyde (MDA), indole-3-acetic acid (IAA), and gibberellins (GA), a reduced IAA/ZR ratio, and diminished peroxidase (POD) activity. Weighted gene co-expression network analysis (WGCNA) identified 27 hub transcription factors (TFs) strongly associated with IAA/ZR, primarily from the ERF, bHLH, MYB, WRKY, and C3H families. Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses revealed the significant enrichment of differentially expressed genes (DEGs) related to plant hormone signal transduction and cell wall metabolism during pluripotent callus acquisition. Further investigations revealed that five genes encoding a putative indole-3-acetic acid-amido synthetase GH3.1, protein TIFY 10A, a two-component response regulator ARR2-like isoform X2, and xyloglucan endotransglucosylase/hydrolase, likely promoting callus pluripotency by modulating plant hormone signaling and cell wall metabolism, thereby enhancing in vitro regeneration in C. parthenoxylon. In summary, this study provides critical insights into the molecular mechanisms of C. parthenoxylon regeneration and offers valuable germplasm resources for establishing an efficient and stable genetic transformation system via tissue culture. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Plant Regeneration—2nd Edition)

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16 pages, 801 KB

Open AccessArticle

Superior In Vitro Responses of a Native Rose Genotype to Driver Kuniyuki Walnut (DKW) Medium in a Comparative Study Using Natural and Synthetic Plant Growth Regulators

by Mahboubeh Davoudi Pahnekolayi, Zahra Parchianloo, Majid Babouyehdarabi and Meysam Ghasemi

Plants 2025, 14(16), 2606; https://doi.org/10.3390/plants14162606 - 21 Aug 2025

Cited by 2 | Viewed by 2139

Abstract

Rosa canina is one of the precious native rose rootstocks with a high reputation among plant producers, which has potential horticultural and pharmacological properties related to the cosmetic values and the production of secondary metabolites. Due to high horticultural consumption, applying the plant [...] Read more.

Rosa canina is one of the precious native rose rootstocks with a high reputation among plant producers, which has potential horticultural and pharmacological properties related to the cosmetic values and the production of secondary metabolites. Due to high horticultural consumption, applying the plant tissue culture technique as a major tool for healthy and massive-scale production of R. canina plants is not unexpected. However, the response of R. canina in vitro plantlets to various plant tissue culture ingredients is not well understood to tender an efficient applied protocol for qualitative and quantitative in vitro propagation. In this regard, the main objective of this study is to investigate the influence of several abiotic in vitro variants including six plant tissue culture media formulations (McCown’s Woody Plant Medium (WPM), Murashige and Skoog (MS), Van der Salm (VS), Schenk and Hildebrant (SH), Driver Kuniyuki Walnut (DKW), and Gamburg B5 (B5)) in combination with four concentrations (0, 1.5, 3, 4 mgL⁻¹) of two types of cytokinins (6-Benzyaminopurine (BAP) and Kinetin (Kin)) simultaneously. Notably, it is perceived that DKW culture medium containing 1.5 mgL⁻¹ BAP and 0.1 mgL⁻¹ NAA is the best treatment for both in vitro morphological and flowering properties. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Plant Regeneration—2nd Edition)

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18 pages, 1967 KB

Open AccessEditor’s ChoiceArticle

Optimizing Growth Regulator Concentrations for Cannabis sativa L. Micropropagation

by Gabrielle A. Johnson, Carissa L. Jackson, Antonio Timoteo, Jr., Papaiah Sardaru, Michael H. Foland, Purushothaman Natarajan and Sadanand A. Dhekney

Plants 2025, 14(16), 2586; https://doi.org/10.3390/plants14162586 - 20 Aug 2025

Cited by 3 | Viewed by 3462

Abstract

In this study, the effect of growth regulators on shoot proliferation and rooting were evaluated to develop an efficient micropropagation protocol for the Cannabis sativa L. cultivars ‘Cherry Soda’ and ‘Purple’. Apical meristems were isolated from actively growing shoots of stock plants and [...] Read more.

In this study, the effect of growth regulators on shoot proliferation and rooting were evaluated to develop an efficient micropropagation protocol for the Cannabis sativa L. cultivars ‘Cherry Soda’ and ‘Purple’. Apical meristems were isolated from actively growing shoots of stock plants and transferred to Driver and Kuniyuki Walnut (DKW) culture medium containing either 0.0, 0.5, 1.0, 2.0, or 5.0 μM meta-Topolin to study their shoot proliferation response. Resulting shoot cultures were transferred to medium containing varying levels of Indole Acetic Acid (IAA), Indole Butyric Acid (IBA), or Naphthalene Acetic Acid (NAA), solely or in combination, and were subjected to a 10-day dark incubation followed by a 16 h/8 h light/dark period to identify the best treatment for root production. Among the different shoot proliferation treatments studied, the maximum number of shoots was produced on the control medium that was devoid of any meta-Topolin. Cultures grown on medium containing 5.0 μM meta-Topolin exhibited hyperhydricity, where shoots appeared translucent and pale green in color; were characterized by water-soaked lesions; and leaves appeared curled and brittle in contrast to healthy looking cultures. Among the various rooting treatments studied, shoots grown in the dark for 10 days exhibited the highest frequency of rooting on medium containing 4.0 μM NAA or 6.0 μM IBA + 1.0 μM NAA. Full developed plants with a robust shoot and root system were transferred to soil, acclimatized under conditions for high humidity, and then transferred to ambient conditions in 4 weeks. The micropropagation protocol developed here allows for rapid multiplication of disease-free plants in C. sativa cultivars. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Plant Regeneration—2nd Edition)

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Review

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57 pages, 3492 KB

Open AccessReview

Application of Nanoparticles in Plant In Vitro Culture for Micropropagation and Secondary Metabolite Production: A Review

by Natalia A. Semenova, Dmitry A. Zakharov, Dmitry A. Serov, Sergey V. Gudkov, Alexey S. Dorokhov and Andrey Yu. Izmailov

Plants 2026, 15(13), 2071; https://doi.org/10.3390/plants15132071 - 3 Jul 2026

Viewed by 903

Abstract

This review summarizes recent studies on the use of nanoparticles (NPs) in plant in vitro clonal micropropagation and secondary metabolite production. The analyzed applications include culture initiation, shoot multiplication, rooting, acclimatization, callus culture, and hairy root culture. Because NPs’ effects are strongly endpoint-dependent, [...] Read more.

This review summarizes recent studies on the use of nanoparticles (NPs) in plant in vitro clonal micropropagation and secondary metabolite production. The analyzed applications include culture initiation, shoot multiplication, rooting, acclimatization, callus culture, and hairy root culture. Because NPs’ effects are strongly endpoint-dependent, their effectiveness is evaluated separately for micropropagation endpoints, including contamination control, explant survival, multiplication rate, shoot and root development, and plantlet quality, and for metabolite-production endpoints, including biomass accumulation, target metabolite concentration, and total metabolite yield. Based on the quantitative analysis of published data, the most frequently beneficial NP size range was 20–60 nm, whereas effective concentrations, combining a positive effect on plant growth and the synthesis of secondary metabolites, were mainly within 1–120 mg L⁻¹. For the most extensively studied NPs, the corresponding indicative concentration ranges were 15–45 mg L⁻¹ for 25–40 nm Ag-NPs, 75–200 mg L⁻¹ for 15–45 nm ZnO-NPs, 120–150 mg L⁻¹ for 50–80 nm Se-NPs, and 90–145 mg L⁻¹ for 54–55 nm Si-based NPs. Ag- and carbon-based nanomaterials showed relatively strong overall responses in micropropagation datasets, whereas Ag- and Se-NPs were often associated with enhanced target metabolite accumulation. NP responses depend on particle composition, synthesis method, surface properties, dose, culture system, species, genotype, and may involve phytotoxic or residue-related risks. Full article

(This article belongs to the Special Issue Plant Tissue Culture and Plant Regeneration—2nd Edition)

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