Search Results (149)

Lopezia racemosa Cav., commonly known as “cancer herb” in indigenous communities, has long been used for its medicinal properties. The biotechnological production of its bioactive compounds through genetic transformation represents a valuable approach for obtaining pharmacologically relevant substances. The initial focus of this study was to identify compounds previously reported in the species; however, phytochemical analysis by HPLC and NMR led to the isolation and identification of two pentacyclic triterpene esters not previously described in L. racemosa: 3-O-[(E)-feruloyl]-maslinic acid (1) and 3-O-[(E)-feruloyl]-corosolic acid (2), identified as constituents of fraction 33. The LRTC3.1 callus line was obtained from hairy roots generated by infecting L. racemosa leaf explants with Agrobacterium rhizogenes strain ATCC15834/pTDT. The crude extract, specific fractions, and the mixture of these compounds demonstrated significant anti-inflammatory and cytotoxic activities. Anti-inflammatory activity was evaluated using the carrageenan-induced mouse paw edema model, where the crude extract achieved 51.02% inhibition of inflammation compared to meloxicam (30.86%). Cytotoxicity was assessed against three human cancer cell lines: breast carcinoma (MCF7), cervical carcinoma (SiHa), and colon carcinoma (HCT-15). Fractions FD (28–29) and 33 exhibited potent cytotoxic effects, with IC₅₀ values of 0.508 and 1.345 µg/mL against SiHa cells, and 0.053 and 2.693 µg/mL against MCF-7 cells, respectively. These findings suggest that transformed L. racemosa cultures represent a promising source of bioactive compounds for potential therapeutic development. Full article

Agave species are plants with great economic value and multiple possibilities of use as ornamentals, medicinal plants, and fibers, as well as being significant sources of bioethanol. However, their long life cycles hinder their conventional breeding. Therefore, biotechnology tools are the most effective means for clonal propagation and genetic improvement. In vitro micropropagation of A. sisalana via axillary shoot proliferation from bulbil explants was attained using Murashige and Skoog medium (MS) supplemented with cytokinins (CKs), such as 6-benzyladenine (BA), kinetin (KIN), or thidiazuron (TDZ). The optimum significant shoot proliferation (14.67 shoots/explant) was achieved on 1.0 mg L⁻¹ TDZ. The carry-over effect of CKs on subsequent rooting could be detected. Control and KIN treatments could enhance the rooting of shoots on shoot proliferation media. The regenerated plantlets were acclimatized directly with 100% survival. To mitigate this carry-over effect, that causes hindering further root growth and development, and promote healthy growth of roots, subculturing shoots onto a CK-free medium is a recommended practice. The shoots induced on all BA treatments, and TDZ at 0.5 and 1.0 mg L⁻¹ could be rooted after two subcultures on CK-free medium, then they were acclimatized with 100% survival. However, the higher concentrations of TDZ inhibited in vitro rooting even after two subcultures on CK-free medium, and the acclimatization percentage was reduced by increasing the TDZ concentration recorded from 10 to 0%. Full article

Cacao genotypes propagation through plant tissue culture represents a strategic approach for establishing a core collection of elite plants to be used as a donor material source, necessary for increasing new planting areas of cacao. This study aimed to evaluate somatic embryo regeneration in ten native fine-aroma cacao genotypes (INDES-06, INDES-11, INDES-14, INDES-32, INDES-52, INDES-53, INDES-63, INDES-64, INDES-66, INDES-70) from the INDES-CES germplasm collection, under in vitro conditions using culture medium supplemented with different concentrations of Thidiazuron (0, 10, and 20 nM). Our results showed an average of 20 and 100% of callogenesis in all genotypes evaluated, but the callus development did not appear after early stages of its induction; however, primary somatic embryos were observed after 42 days after TDZ treatment in the INDES-52, INDES-53, INDES-64, INDES-66, INDES-70 genotypes. The INDES-52 genotype was more responsive to under 20 nM of TDZ, generating an average of 17 embryos per explant. This study contributes to the adaptation and establishment of a protocol for somatic embryo regeneration of fine-flavor cacao genotypes. Full article

Light influence on shoot regeneration in Prunus salicina is a complex interaction that has been studied for the first time. Japanese plum plants were regenerated from calli and seeds of the scion cultivar ‘Victoria’. The effect of four different light spectra (white, blue, red, and mixed), along with three 6-benzyladenine (BA) concentrations (1, 1.5, and 2 mg L⁻¹), was studied in these two sources of explants. Organogenic calli were derived from the base of stem explants of the scion cultivar ‘Victoria’, whereas cotyledons and embryogenic axis slices were used as seed explants. Calli cultured with 2 mg L⁻¹ of BA and mixed light or 2.5 mg L⁻¹ of BA and control light showed the highest regeneration rates, with no significant differences compared to other treatments. Seed explants exposed to 2.5 mg L⁻¹ of BA and red light exhibited significantly higher organogenesis. In comparison, those in 1.5 mg L⁻¹ of BA with blue light or 2.5 mg L⁻¹ of BA with mixed/control light showed no regeneration. BA concentration did not have a significant effect in the induction of somatic shoots from any explant source. In contrast, a strong interaction between light and BA was noticed. This work presents a protocol that can be applied in transformation and editing research as light spectrum studies continue to advance. Full article

Malania oleifera Chun & S.K. Lee, an endemic monotypic species that belongs to the family Olacaceae, is under continuous pressure of decline owing to several ecological and physiological factors. The present study aimed to establish an efficient in vitro protocol for callus-mediated indirect somatic embryogenesis in M. oleifera by alleviating tissue browning. Internodes and leaves obtained from seedlings were used as explants. Antioxidant pre-treatment (ascorbic acid, AA) followed by different carbon sources (sucrose, maltose, glucose, and fructose) and plant growth regulators in various concentrations and combinations were employed in Woody Plant Medium (WPM) to alleviate explant browning and induce callus formation from the explants. AA pre-treatment and subsequent culture on maltose at a concentration of 116.8 mM were optimal for controlling phenolic exudation on >90% of both explants. The highest responses of 53.77% and 57.43% for embryogenic calli were induced from internode and leaf explants, respectively. The highest responses, 85.22% and 93.80%, were observed for somatic embryos that matured into the globular, heart-shaped and torpedo stages at different percentages on NAA 2.5 mg/L in combination with BA 1.0 mg/L for both explants. The matured somatic embryos were finally germinated at a maximum concentration of GA₃, 2.0 mg/L. All plantlets were successfully hardened and acclimatized under culture room conditions and then transferred to the greenhouse. The current study suggests an efficient protocol for indirect somatic embryogenesis by alleviating phenolic exudation from the explants of M. oleifera. This first successful report of in vitro culture establishment in M. oleifera may offer an effective alternative measure to conserve this species and provide a system for analyzing bioactive chemicals and for use in the oil industry. Full article

Triadica sebifera, an economically and medicinally valuable tree species native to China, was investigated for its in vitro regeneration potential using leaf explants from nodal cuttings of young stems and sprouts. This study evaluated the effects of basal media, plant growth regulators (PGRs), explant sources, and incision methods on adventitious shoot induction, supplemented by histological analysis. The highest shoot regeneration frequency (98.89%) and maximum shoot number (72) were achieved via direct organogenesis using sucker-derived nodal cuttings cultured on MS medium with 2 mg/L 6- benzyladenine (6-BA), 0.3 mg/L kinetin (KT), and 0.2 mg/L α-naphthaleneacetic acid (NAA). Under identical conditions, branch-derived explants showed lower regeneration (84.44%, 64 shoots). Transverse midvein incision proved most effective, with sucker-derived leaves exhibiting superior regeneration. Shoots elongated completely (100%) on Murashige and Skoog (MS) medium containing 0.3 mg/L 6-BA, 0.03 mg/L NAA, and activated charcoal. Rooting was optimal on MS medium with 0.3 mg/L indole-3-butyric acid (IBA), yielding a 98% acclimatization survival rate. Histological analysis revealed de novo meristem formation from parenchyma cells, confirming direct organogenesis without callus intermediation, further validating the enhanced regenerative capacity of sprout-derived explants. This efficient in vitro regeneration system provides a foundation for large-scale propagation and germplasm conservation of T. sebifera, while offering insights for woody plant regeneration studies. Full article

Sophora koreensis Nakai, listed as endangered on the IUCN Red List, is a species native to Korea, specifically found in parts of Gangwon-do. Recent research highlights its potential in hangover relief and as an antioxidant source, sparking interest in enhancing its components through mutation for commercial purposes. Given its limited distribution, micropropagation of S. koreensis is essential for its economic exploitation. This study focuses on in vitro culture to develop an elongation system for micropropagation. The hormonal combination of 6-benzylaminopurine (2 μM), thidiazuron (2 μM), and indole-3-butyric acid (0.5 μM) produced the highest number of shoots (14) with an average length of 0.7 cm compared to the control. Additionally, adjusting photoperiod conditions under specific culture media further increased shoot length to 0.6 cm, which was also higher than that of the corresponding control group under standard light conditions. However, survival rates were generally low across all treatments during subculture. Isolating and individually culturing induced explants resulted in shorter shoots and lower survival rates. Improvements were noted when explants with 10 shoots were subcultured, achieving an 83% survival rate, with an average of 4.93 shoots at 0.95 cm in length. Rooting was most successful with 10 μM IBA, also showing the highest root number, indicating a potential pathway for enhancing micropropagation efficiency. Full article

Agrobacterium-mediated transformation systems using epicotyl explants have been widely used for genetic transformations of citrus. However, their application in lemons is severely constrained by browning of epicotyl tissues, which leads to an extremely low efficiency of transformation. In this study, we developed an optimized Agrobacterium-mediated transformation system using whole cotyledonary node explants of ‘Eureka’ lemon (Citrus limon), which significantly reduced tissue browning and enhanced transformation efficiency up to 14.48%. In addition, preparation of the whole cotyledonary node was simple and rapid, which reduced time and labor. This system facilitated efficient generation of transgenic lemon plantlets and provided a novel explant source for citrus transformation. Full article

Tea plants (Camellia sinensis) are among the world’s most significant economic tree species. Tissue culture serves as a crucial method in commercial breeding by facilitating the rapid propagation of valuable genotypes and the generation of disease-free clones. However, callus browning represents a prevalent challenge in tea plant tissue culture, and may adversely affect explant growth and development. Our research demonstrates that although anti-browning agents can effectively suppress browning, they induce distinct color changes in the callus. These color variations could significantly influence callus induction and subsequent growth patterns. In this study, callus tissues from C. sinensis var. Assamica cv. Mengku were employed as experimental materials and treated with three commonly used anti-browning agents: ascorbic acid (VC), activated carbon (AC), and polyvinylpyrrolidone (PVP). The results demonstrated that while these three reagents effectively inhibited browning, they also induced distinct color changes in the explants, which appeared red, green, and white, respectively. Furthermore, this study investigated the molecular mechanisms underlying callus color changes using transcriptomic and metabolomic approaches. Based on transcriptome analysis, it was revealed that photosynthesis and flavonoid biosynthesis pathways were significantly enriched. Metabolome analysis identified 14 phenolic acids, which exhibited significant variation in accumulation across calluses of different colors. The differential expression of genes involved in flavonoid biosynthesis pathways, coupled with the distinct accumulation patterns of metabolites, can effectively alleviate photooxidative damage and enhance the resistance of callus to browning. AC activates the photosynthesis of callus by regulating carbon source allocation and upregulating the expression of key genes in the psa, psb, and pet families within the photosynthetic system. This process promotes chlorophyll biosynthesis, thereby enabling the callus to grow green, while VC activates the expression of key genes such as CHS, F3H, C4H, CYP75B1, and ANR in the flavonoid pathway, which are involved in the regulation of pigment synthesis in red callus. This study elucidated the molecular mechanisms underlying the effects of anti-browning agents on color variations in C. sinensis callus, thereby providing a robust theoretical foundation for optimization, the establishment of tea plant tissue culture systems, and enhancing cultivar quality. Full article

Araucaria araucana is an emblematic native conifer from Chile and Argentina that has been classified as threatened due to anthropogenic activities. Somatic embryogenesis (SE) is a biotechnological tool used for both the preservation of genetic material and the propagation of valuable genotypes. The present study investigates the effects of explant source and culture medium on SE induction in A. araucana genotypes from three wild plant populations. Immature strobili were collected from different geographical provenances: a coastal area (Villa Araucarias, “VA”), Cordillera de Nahuelbuta (Trongol Alto, “TR”), and the Andes Mountains (Malalcahuello, “MA”). SE induction was observed after 45 days in a basal medium (BM) supplemented with 1-naphthaleneacetic acid (NAA—11 µM), 6-benzylaminopurine (6-BA—2.8 µM), and Kinetin (Kin—2.8 µM). The highest induction rate (75%) was achieved for seeds from VA. Embryogenic cell line (ECL) proliferation requires auxins but is genotype-dependent, as not all genotypes survive. Cytochemical analysis revealed the presence of pro-embryogenic masses (PEMs) in the ECLs, indicating an efficient SE induction protocol. The progression of PEMs to early embryos was observed in the presence of maltose (3% w/v), polyethylene glycol 3350 (PEG—7% w/v), and abscisic acid (ABA—68 µM). Our results establish a baseline for the establishment of in vitro cultures for a diverse range of A. araucana genotypes, enabling the initiation of ex situ preservation programs and further investigation into embryo maturation. Full article

This study aims to establish efficient in vitro propagation protocols for Gymnocalycium cv. Fancy, an ornamental cactus with variegated variants, by evaluating the effects of cytokinin type and explant source on the organogenic response. Plants with different degrees of variegation (0–100%) were classified by size to obtain different explant types (apices, central discs, epicotyls, and hypocotyls). The effects of 6-Benzylaminopurine (BAP, 8 µM), Kinetin (KIN, 4 µM), and Thidiazuron (TDZ, 1 µM) were assessed on shoot production, callus formation, and rhizogenesis. Additionally, we studied the relationship between initial plant variegation and the productivity of the variegated shoots. The best shoot production results were obtained for central discs treated with 1 µM TDZ. Furthermore, a correlation was observed between the activated areole type (green, mixed, or fully colored) and shoot color percentage, enabling precise explant selection. The appearance of differently colored shoots confirms the potential for selecting new lines from this cultivar too. These findings hold significant potential not only for the breeding and propagation of ornamental cacti but also for the cultivation of other edible cacti and their relatives. Full article

Cannabis sativa is used for multiple purposes, notably for its medicinal properties. It produces various secondary metabolites, including cannabinoids, terpenes, and flavonoids, which have therapeutic value and typically produce high amounts in female plants. The growth of the global cannabis market has led to intensive breeding efforts to develop elite cultivars with enhanced secondary metabolite profiles. As a dioecious and anemophilous plant, it produces staminate and pistillate inflorescences on separate plants and relies on wind for pollination, rendering traditional propagation methods challenging owing to high genetic recombination in progeny. Consequently, asexual propagation (micropropagation) is commonly employed to maintain female clones entirely. Micropropagation/direct organogenesis is a tissue culture technique that produces numerous disease-free clone plants in vitro more rapidly than traditional rooted cuttings. Factors such as sterilization, hormonal balance, explant type, nutrient additives, carbon source, pH, and environment influence the success of cultivar-specific micropropagation. In this review, we discussed how these factors affect cannabis micropropagation based on recent findings, emphasizing the importance of optimizing cultivar-specific protocols for long-term germplasm conservation and efficient breeding based on a mechanistic background. Full article

(1) Background: Traditionally, hop is propagated using rhizome fragments or herbaceous stem cuttings. Micropropagation, therefore, offers a viable alternative for the large-scale production of healthy, genetically uniform plants, regardless of the season and within confined spaces. A temporary immersion system (TIS) facilitates plant propagation by alternating immersions of microcuttings in liquid culture medium with dry periods, preventing gas accumulation through forced ventilation. (2) Methods: In this study, the response of hop plantlets, cv. Columbus, cultured in media with and without sucrose, in solid and liquid culture systems (Plantform^TM bioreactor), was evaluated, considering its effect on morpho-physiological parameters, on the total phenolic content, and on antioxidant capacity. Moreover, to make the TIS more efficient, the effect of immersion duration (three and six minutes every twelve hours) was evaluated. (3) Results: The presence of sucrose in the culture medium improved plant proliferation in both culture systems tested: solid and liquid (particularly for explants immersed for three minutes). In the TIS, plantlets with a higher antioxidant capacity were obtained when sucrose-free culture medium was used. (4) Conclusions: This study confirms the efficacy of the TIS as a hop propagation method but also as a valid tool to produce biomass to be used as a source of bioactive compounds. Full article

Fingerroot (Boesenbergia rotunda (L.) Mansf.) is valued for its therapeutic benefits, both in Thailand and internationally. This study optimized in vitro propagation and induced microrhizomes (MRZ) to produce cleaned plantlets to support organic farming using disease-free plantlets, which is crucial for preventing and eradicating diseased plantlets, reducing the use of chemicals, and alternative approaches to enhancing phytochemical diversity. Shoots cultured on ½-strength MS medium with 1 mg L⁻¹ of 6-benzylaminopurine (BAP) showed the highest shoot formation (69%) and shoot multiplication (3.45 ± 0.29 shoots per explant). Plantlets acclimatized in peat moss or a peat moss–coconut coir (1:1) mixture achieved a 100% survival rate. Genetic fidelity was confirmed using SSR markers, showing genetic consistency with the mother plant. The MRZ formation was the highest (98.33%) under white LED light with 30 g L⁻¹ of sucrose. Nuclear magnetic resonance (NMR) analysis in MRZ revealed aspartate, a precursor to pinocembrin and pinostrobin. Additionally, nine unique metabolites not previously identified in fingerroot were detected in the MRZ, suggesting some potential in novel therapeutic applications. These findings support the development of efficient micropropagation methods and highlight MRZ as a source of diverse bioactive compounds, contributing to the medicinal value of B. rotunda in sustainable and large-scale production. Full article

(1) Background: Humulus lupulus L. plants constitute a rich source of bioactive compounds. The synthesis of bioactive compounds in plants is often triggered by the activation of secondary metabolism, which can be induced by biotic or abiotic elicitors. In vitro, the effect of the elicitors can be studied in a controlled environment and in a small space, independently of seasonal variations. Cytokinins are frequently used in plant tissue culture for bud regeneration, branching and shoot elongation due to their role in cell division enhancement. This study aimed to investigate the effects of different cytokinins on the growth parameters, total (poly)phenolic content and antioxidant capacity of in vitro-grown hop plants to evaluate hop vitro-derived biomass as a potential source of bioactive compounds. (2) Methods: unimodal hop (cvs. Cascade and Columbus) explants were cultured on media enriched with four cytokinins (kinetin, 6-benzylaminopurine, meta-topolin and 6-(γ,γ-dimethylallylamino)-purine) at four concentrations. (3) Results: A genotype-dependent response to different cytokinins was encountered. (4) Conclusions Columbus explants could root in culture media auxin-free, providing valuable opportunities for commercial nurseries. Moreover, cytokinins were confirmed to be valuable elicitors to stimulate the bioactive compound biosynthesis in micropropagated hop plants, making them a precious source for various industries. Full article