Search Results (498)

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

The Earth’s environment is deteriorating, and biodiversity is declining. The use of plant-based cellulose nanofibers (CNFs) as structural materials can reduce environmental impact, and further technological developments are anticipated. This review article introduces types of cellulose derived from wood, weeds, bamboo, and fruits, and examines the potential technological applications of CNFs. It is hypothesized that maintaining an appropriate content of Mn, Ca, and O—including the interactions of Ca within carbon-based structures—could contribute to plant health, while the exclusion of elements such as V, Cd, and Sn (regardless of the effectiveness of partial sequestration) could promote cell activity. Calcium deposition can influence wood growth depending on the elemental composition in the bark, and a hypothesis regarding pH adjustment for shoot formation is proposed (see textbook on inorganic chemistry). Furthermore, manufacturing processes for CNFs and their mechanical properties—including evaluation methods—are summarized. This overview focuses on nanostructures that exhibit heterogeneous functional and mechanical properties and offer potential benefits in reducing environmental impact through processes such as 3D printing and coating. CNFs derived from fruit peels can yield lightweight and durable materials. Furthermore, the roles of proteins and fruit-derived components in neutralizing acidic environments and reducing oxides are discussed. A concept is proposed that links the processing of fruit-peel-based materials with environmental applications such as forest restoration and combating desertification. The hypothesis is put forward that cytoplasmic activity and cell wall strengthening could be enhanced through chlorophyll-related processes and water transport mechanisms. Optimizing pH conditions could promote shoot formation in plants such as conifers. Sustainable greening can be achieved through the use of cellulose-based materials in combination with water-retaining components such as bamboo-derived resources. The interaction between CNFs, plant bark, and water-bound proteins can contribute to forest regeneration and the curbing of slash-and-burn practices. Overall, this approach can contribute to environmental remediation, the reduction of environmental impact, and urban greening in degraded regions. Full article

Macadamia (Macadamia spp.) is an economically important nut crop whose severe recalcitrance to genetic transformation substantially hinders progress in functional genomics and molecular breeding. To overcome this critical technical bottleneck, this study established a highly efficient and broadly applicable in planta hairy root genetic transformation system with integrated visual screening. This system utilizes an Agrobacterium rhizogenes-mediated transformation method, employing multiple visual reporter gene systems (DsRed2, eGFP, RUBY, and AtPAP2) to achieve antibiotic-independent and non-destructive screening of transgenic roots. Notably, the system innovatively incorporates the air layering (marcotting) technique to extend in planta genetic transformation to branches of mature trees in the field. By circumventing the stringent sterile conditions required for conventional in vitro tissue culture, this approach achieves a largely genotype-independent transformation across open-pollinated seedlings with diverse genetic backgrounds (A4, GR1, HAES900, and O.C.). The transgenic hairy root induction frequencies ranged from 39.25% to 47.38%, although the GR1 genotype exhibited a notable developmental stage-dependent decline in transformation efficiency. Furthermore, transgenic hairy roots were successfully induced on mature tree branches, with a maximum induction rate of 28.2%. Gene expression analyses confirmed the stable, high-level expression of the target transgenes in all the transgenic hairy root lines. This in planta transformation system provides a reliable in vivo experimental platform for the rapid functional validation of candidate genes and the investigation of root biology in Macadamia. Moreover, it establishes a novel strategy for plant regeneration via root-to-shoot organogenesis, offering a promising avenue for the genetic improvement of recalcitrant woody plants. Full article

Chamaecyparis obtusa is a valuable conifer species, prized for its high-quality timber and bioactive essential oils. However, commercial micropropagation is challenging due to its resistance to vegetative propagation. This study aimed to develop an efficient in vitro plant regeneration system using leaf explants and to profile the metabolic changes during organogenesis. Leaf explants from one-year-old in vitro-grown plantlets were cultured on media with various plant growth regulators (PGRs) to optimize callus induction, shoot multiplication, and rooting. Secondary metabolites were systematically analyzed throughout the developmental stages: explant, callus, regenerated shoot, and rooted plantlet. The most effective shoot regeneration, leading to whole plantlets, was achieved on a medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid and 2.0 mg/L thidiazuron. Metabolic profiling revealed significant stage-specific biochemical transitions. High-Performance Liquid Chromatography (HPLC) precisely quantified individual phenolics, resolving cross-reactivity issues seen in total flavonoid colorimetric assays. Gas Chromatography-Mass Spectrometry (GC-MS) also identified substantial shifts in volatile terpenoid biosynthesis during shoot morphogenesis. This integrated protocol provides a reliable platform for mass propagation of C. obtusa and offers fundamental insights into the metabolic dynamics of in vitro development, with significant potential for future horticultural and biotechnological applications. Full article

In vitro regeneration and flowering systems provide valuable platforms for rose breeding and biotechnological applications. Although the effects of plant growth regulators (PGRs) on Rose regeneration have been extensively studied, the contribution of growth additives, either alone or in combination with PGRs, remains insufficiently understood in Rosa chinensis. This study investigated the effects of PGRs (6-Benzylaminopurine (BAP), 1-Naphthaleneacetic acid (NAA), and indole-butyric acid (IBA)) combined with three growth additives, putrescine (Put), phloroglucinol (PG), and casein hydrolysate (CH), at different concentrations on the vegetative and reproductive responses of R. chinensis in an in vitro system. No flowering occurred in media without PGRs, regardless of the growth additive treatment, highlighting the essential role of PGRs in floral induction. However, the integration of growth additives with PGRs significantly enhanced regeneration and flowering performance. Among the tested additives, Put showed the strongest stimulatory effect, increasing in vitro flower production and improving floral characteristics. PG promoted several morphological traits, including shoot regeneration and flower stem elongation, but was less effective than Put. CH exhibited limited effects on both vegetative growth and reproductive development. These findings indicate that Put supplementation in PGR-based media is an effective approach for improving regeneration and flowering efficiency in R. chinensis, with potential applications in rose biotechnology and breeding programs. Full article

Shallot (Allium cepa var. aggregatum) is an annual herbaceous plant of the Amaryllidaceae family cultivated worldwide for its gastronomic importance, unique flavour, and nutritional properties. In Ecuador, shallot is considered an emerging crop; however, the lack of effective detection and treatment options for phytoviruses remains a challenge for farmers. These viruses negatively impact crop yield, leading to substantial economic losses and declining crop quality. Simultaneous infections can synergistically exacerbate crop damage. In this study, the molecular detection of the five most common shallot viruses—Shallot latent virus (SLV), Onion yellow dwarf virus (OYDV), Shallot virus X (ShVX), Leek yellow stripe virus (LYSV), and Iris yellow spot virus (IYSV)—were investigated in shallots obtained from different market suppliers in Quito, Ecuador. We identified two viruses using RT-PCR: SLV (latent) and IYSV (responsible for severe disease symptoms). Shoot tip culture produced shallot plants completely free of IYSV, while SLV persisted in the regenerated plants. This study reports IYSV for the first time in shallots in Ecuador and shows that shoot tip culture can serve as a partial cleaning method to improve crop quality and support more effective pathogen management and control strategies. Full article

Ajania achilleoides, a wild species with strong environmental adaptability, is a valuable germplasm resource for genetic improvement and functional gene studies. However, efficient in vitro regeneration and genetic transformation systems for this species remain undeveloped, limiting functional gene characterization and genetic improvement of this valuable germplasm resource. In this study, leaf explants were used to establish an efficient regeneration system by evaluating different combinations of plant growth regulators and to develop an Agrobacterium tumefaciens-mediated genetic transformation system for A. achilleoides. MS medium supplemented with 2.0 mg/L 6-benzyladenine (6-BA) and 1.0 mg/L α-naphthaleneacetic acid (NAA) achieved the highest adventitious shoot regeneration rate (99%), while 1/2 MS medium containing 0.5 mg/L indole-3-butyric acid (IBA) was optimal for rooting, resulting in a 100% rooting rate. Critical hygromycin concentrations for shoot regeneration and rooting selection were determined to be 25 mg/L and 10 mg/L, respectively. For transformation, the best results were obtained with a 3-day preculture, bacterial suspension at OD₆₀₀ = 0.2, 30 min infection, and 2-day co-cultivation. PCR and sequencing confirmed the successful integration of the target genes into the genome, yielding three positive transgenic lines with a molecular confirmation rate of 30%. This study establishes a stable and efficient regeneration and transformation system for A. achilleoides, providing a platform for functional gene analysis and molecular breeding of wild chrysanthemum species. Full article

Bamboo shoots, as the most nutritionally valuable food source for giant pandas during key reproductive seasons, are critical for conservation because their availability and timing directly influence panda foraging and habitat use. However, the molecular mechanisms through which altitudinal temperature variation governs rhizome shooting in staple food bamboos remain largely unknown. Here, we performed integrated metabolomic and transcriptomic analyses of Fargesia rufa rhizomes collected along an elevational gradient (1000 m, 1500 m, and 2000 m), with a critical paired comparison at 2000 m between a non-shooting cold gully-edge site (16.4 °C) and a shooting warm gully-center site (21.1 °C), where soil temperature is elevated by approximately 4 °C due to prolonged solar exposure. Our results demonstrate that soil temperature, rather than elevation per se, acts as the primary driver of rhizome shooting, with an apparent threshold near 20 °C. A core shooting metabolome (CSM) comprising 843 metabolites was consistently accumulated across all shooting conditions, which featured gibberellin/auxin precursors, TCA cycle intermediates, and phenylpropanoid compounds. Correspondingly, a core shooting transcriptome (Rh_shooting) of 10,970 genes was identified, which resolved into three functionally distinct temporal clusters: “shooting-on” (activated upon threshold crossing, enriched in hormone signaling and cell wall metabolism), “temperature-dose” (progressively upregulated with rising temperature, enriched in energy metabolism and defense), and “microenvironment-enhanced” (specifically upregulated in the high-elevation warm gully, enriched in photosynthesis and antioxidant pathways). Integrative network analysis further revealed zeatin riboside and multiple hub genes as central coordinators linking hormone signaling, energy metabolism, and cell wall remodeling. Collectively, these findings establish a molecular framework linking altitudinal temperature variation to bamboo rhizome regeneration—a process that directly determines the spatiotemporal availability of bamboo shoots for giant pandas. This work provides mechanistic insights into giant panda foraging ecology and has direct implications for predicting habitat quality under climate change and informing evidence-based conservation strategies for this flagship species and its critical food resource. Full article

Physalis peruviana L. is an Andean crop of high nutritional and commercial value; however, the limited availability of uniform planting material restricts its large-scale propagation. Indole-3-butyric acid (IBA) is widely used to promote adventitious rooting, although its concentration and application methods influence the response observed throughout the different stages of root development. This study evaluated how IBA concentration and explant origin influenced rooting-related traits and early vegetative growth of in vitro-regenerated P. peruviana shoots during a 30-day ex vitro acclimatization phase. The evaluated variables included rooting percentage, root number, longest root length, root fresh and dry mass, shoot length, leaf and node number, stem diameter, shoot fresh and dry mass, leaf area, and photosynthetic pigment contents. The experiment followed a completely randomized design with a 2 × 4 factorial arrangement consisting of two explant origins (cotyledon and hypocotyl) and four IBA concentrations (0, 400, 800, and 1600 mg L⁻¹), with five biological replicates per treatment combination. All shoots formed at least one visible root, resulting in 100% rooting across all treatment combinations. IBA concentration significantly affected root fresh and dry mass and several shoot-growth traits, whereas root number and longest root length were not significantly affected. Among the concentrations tested, 800 mg L⁻¹ produced the highest root biomass and favorable responses in selected shoot-growth traits, whereas 1600 mg L⁻¹ was associated with lower values for some growth variables. Hypocotyl-derived shoots had more leaves and nodes, greater stem diameter, and higher shoot dry mass than cotyledon-derived shoots. These results indicate a concentration- and trait-dependent response to IBA and identify 800 mg L⁻¹ as the most favorable concentration among those tested for increasing root biomass and selected shoot-growth traits under the evaluated acclimatization conditions. Full article

Handroanthus chrysanthus (Jacq.) S.O. Grose is a Neotropical tree species highly valued for its ornamental beauty, durable timber, and medicinal properties. However, overexploitation and the recalcitrant nature of its seeds have constrained propagation and conservation efforts, and the species has been listed as Vulnerable on the IUCN Red List since 2020. In this study, a direct adventitious shoot regeneration system was developed for H. chrysanthus by systematically evaluating explant type, basal medium, plant growth regulator combination, and light quality. Hypocotyls were identified as the most responsive explants for shoot induction, and the highest adventitious shoot induction frequency, 51.79%, was obtained on Murashige and Skoog medium (MS) supplemented with 5 mg·L⁻¹ 6-benzylaminopurine, 0.5 mg·L⁻¹ indole-3-butyric acid, and 0.2 mg·L⁻¹ thidiazuron under white fluorescent light. The highest shoot multiplication coefficient, 3.27, was obtained on MS medium containing 4 mg·L⁻¹ 6-benzylaminopurine and 0.3 mg·L⁻¹ gibberellic acid. The maximum rooting frequency, 80%, was obtained on R14 medium after 30 days of culture. After acclimatization, 95% of the regenerated plantlets survived and grew vigorously under greenhouse conditions. To our knowledge, no direct adventitious shoot regeneration system has been reported for the Tabebuia alliance. This efficient regeneration protocol provides a practical platform for clonal propagation, germplasm conservation, and genetic improvement of H. chrysanthus and may also support regeneration and conservation in related endangered taxa within the Tabebuia alliance. Full article

This study aimed to develop an efficient micropropagation protocol for Plectranthus amboinicus (Lour.) Spreng, evaluate the genetic fidelity of in vitro-regenerated plants using RAPD molecular markers, and perform a biochemical assessment of micropropagated material compared with acclimatized plants. For the initiation of in vitro cultures, explants (uninodal fragments) were inoculated on Murashige and Skoog (MS) medium without plant growth regulators (PGRs). Various plant growth regulator treatments were tested to evaluate the in vitro regenerative potential of the explants. The highest multiplication rate (3.43 ± 0.21 shoots per explant) was obtained on MS basal medium supplemented with 3 mg/L 6-benzylaminopurine (BAP) and 3 mg/L gibberellic acid (GA₃), while the greatest shoot length (3.22 ± 0.22 cm) was achieved on MS medium containing 1 mg/L BAP and 3 mg/L GA₃. Indole-3-butyric acid (IBA) (0.5 mg/L), applied alone or in combination with GA₃ (3 mg/L), was the most effective treatment for inducing a well-developed root system, facilitating successful acclimatization of regenerated plants (100% survival rate). In vivo rooting of shoots simultaneously with acclimatization was also tested. The best results (100% rooting rate) were obtained using perlite or Jiffy peat pellets as the rooting substrate, and the shoots were treated with Radi Stim. Molecular analyses confirmed the genetic fidelity of the micropropagated plants. Biochemical analyses revealed higher levels of phenolic compounds and flavonoids in in vitro-cultured plants than in acclimatized plants. These results validate the micropropagation protocol morphologically, genetically, and biochemically, and highlight in vitro culture as an efficient approach for producing high-quality P. amboinicus plant material and for the sustainable production of valuable bioactive compounds. Full article

Due to Ludwigia octovalvis’ aquatic habitat’s vulnerability to climate change, this study developed an in vitro regeneration system using indirect organogenesis to ensure sustainable production of biomass and secondary metabolites. Treatment T16 (0.1 mg/L BAP and 1.0 mg/L NAA) was identified as the optimal hormonal regimen for callus induction and shoot differentiation. Phytochemical analysis by GC-MS revealed that seedlings regenerated under treatment T16 exhibited a diverse profile of 18 phytoconstituents, enhancing the accumulation of phytosterols, terpenes, and tocopherols. In vitro biological evaluation demonstrated that T16 extract possesses significant antibacterial activity (MIC < 62.5 µg/mL) against methicillin-resistant Staphylococcus aureus, and moderate antioxidant capacity. T16 extract showed anti-inflammatory effects superior to indomethacin at a low quantity (0.5 mg/ear) in adult CD1 mice of both sexes. In conclusion, the indirect organogenesis of L. octovalvis not only conserves the species but also optimizes its pharmacological potential, consolidating it as an efficient biotechnological platform for the development of advanced phytopharmaceuticals. Full article

Madhuca indica J.F.Gmel. holds significant economic and industrial value due to its applications in traditional and modern medicine. Its oil is especially important for biodiesel production, owing to its high acid value and suitability as a non-edible feedstock. However, propagation is difficult due to low seed germination, seed recalcitrance, and poor rooting of stem cuttings, limiting large-scale multiplication through conventional methods. To address these limitations, a regeneration protocol using nodal explants was developed. Murashige and Skoog (MS) medium augmented with BA (5.0 µM) and NAA (0.5 µM) produced a maximum of 7.10 ± 0.11 shoots per explant with an average shoot length of 4.53 ± 0.22 cm after six weeks. Rooting was achieved on half-strength medium supplemented with IBA (1.0 µM), resulting in 4.83 ± 0.17 roots per shoot and a root length of 4.50 ± 0.20 cm. In vitro-derived plants were successfully acclimatised in Soilrite with an 82.3% survival rate. The explants were derived from aseptic seedling material, representing juvenile rather than mature elite donor sources. Direct shoot bud development was verified by histological examination. Within the resolution of the employed marker systems, no polymorphism was found utilising RAPD and ISSR markers. SEM showed similar leaf surface characteristics, and physiological and biochemical studies were carried out throughout acclimatisation. A partial overlap in metabolite composition with qualitative and relative quantitative differences between mother and in vitro-derived plants was shown by GC–MS-based profiling. Overall, the study establishes a reproducible regeneration system for M. indica, providing a basis for further optimisation and conservation-oriented applications. Full article

Baikal skullcap (Scutellaria baicalensis) is one of the most important herbs used for centuries in traditional Chinese medicine. The main objective of the research was to micropropagate and introduce the most vigorous lines into garden cultivation. Eleven lines representing individual genotypes were selected and propagated using nodal segments. 6-Benzylaminopurine (BAP) at 1.0 mg L⁻¹, alone or with indole-3-acetic acid (IAA) at 0.1 mg L⁻¹, was the most effective for shoot regeneration. Significant variability in multiplication rate (2.5–6.4 shoots per explant) was observed among the lines. Shoots were successfully rooted (97.9%) and then acclimatized. After six months of cultivation, 113 well-developed plants representing all genotypes were obtained. Depending on the genotype, the final survival rates ranged from 26.9 to 80.0%. Flow cytometry (FCM-DAPI) analysis confirmed the ploidy stability of the micropropagated plants and ISSR markers revealed notable variability among the lines. Moreover, a polymorphism level of 36.4% was detected within line SB_6, whereas the other two analyzed lines (SB_3 and SB_7) maintained in vitro for four years showed no somaclonal variation. Therefore, genetic stability must be monitored, particularly in long-term cultures. In subsequent studies, the acclimatized lines will be evaluated for root yield and phytochemical content under garden conditions. Full article

Pinus massoniana Lamb. is an important native economic and ecological tree species in southern China. However, adventitious root regeneration severely restricts the asexual propagation of elite traits. The WOX gene family, a plant-specific transcription factor family, plays critical roles in various aspects of plant growth and development, particularly in root development, including the maintenance of the root apical meristem, root elongation, and the initiation and formation of lateral roots. In this study, a total of 21 WOX family proteins were identified from the genome of P. massoniana and designated as PmWOX1 to PmWOX21 based on their chromosomal locations. These 21 members were unevenly distributed across nine chromosomes, with a clustered distribution observed on chr7. Collinearity analysis suggested that gene duplication and purifying selection may serve as key driving forces in the evolution of WOX genes. Conserved motif analysis revealed divergence among different PmWOX clades, implying distinct functions in growth and development. Cis-element analysis indicated that PmWOX genes may be involved in the regulation of stress responses, hormone signaling, and developmental processes. Expression pattern analysis based on transcriptome data from root tips and shoot tips identified nine PmWOX genes with significantly higher expression in root tips. RT-qPCR further screened PmWOX4, which exhibited the highest expression in root primordia. Functional experiments verified that PmWOX4 genes encoded nuclear-localized proteins with transcriptional activity. The PPI results indicate that PmWOX4 may interact with CLE41/44, LBD, ATHB8 and CLV3/CLV1 to regulate adventitious root formation. Collectively, our findings suggest that PmWOX4 possesses functional potential in adventitious rooting, which could be further exploited to improve the efficiency of asexual propagation in P. massoniana. Full article