Search Results (130)

To overcome the seasonal constraints of explant availability and facilitate genetic improvement in Catalpa ovata, this study established a dual-pathway in vitro regeneration system (encompassing adventitious shoot organogenesis and somatic embryogenesis) using mature zygotic embryos. We systematically evaluated the synergistic effects of maternal genotypes, plant growth regulators (PGRs), basal media, and the histone deacetylase inhibitor Trichostatin A (TSA). Genotype screening revealed significant divergence in regenerative potential, with the half-sib family 32F17 exhibiting superior responsiveness (84.7% callus induction). A high cytokinin-to-auxin ratio (ZA3 medium) optimally drove direct shoot organogenesis. For adventitious shoot proliferation, the addition of TDZ significantly improved the multiplication coefficient (up to 2.99 on ZB4 medium), although a physiological trade-off with shoot elongation was observed. In parallel, the application of 10 µM TSA significantly enhanced somatic embryogenesis from embryogenic calli, effectively alleviating the inhibitory constraints of exogenous PGRs. For rhizogenesis, the DKW basal medium proved superior to half-strength MS, with the ZE3 treatment (0.1 mg·L⁻¹ NAA + 0.1 mg·L⁻¹ IBA) yielding the highest rooting frequency (69.6%) and robust root architecture. Notably, while somatic embryo conversion remained recalcitrant, plantlets derived exclusively from the adventitious shoot organogenesis pathway were successfully acclimatized ex vitro. These transplanted plantlets exhibited consistently high survival rates (83.1–84.4%) across all tested genotypes, effectively overcoming the initial genotype-dependent recalcitrance. Collectively, this optimized protocol provides a reliable technical platform for the large-scale clonal propagation and biotechnological breeding of C. ovata. Full article

Garlic (Allium sativum L.) is an important vegetable with high nutritional and medicinal value. Its reliance on asexual reproduction causes variety degradation and low propagation efficiency, severely limiting the garlic industry. This study established an efficient shoot organogenesis system for the garlic cultivar Gailiangsuan through optimizing tissue culture protocols. Various explants, media, and hormone combinations were tested to determine the optimal conditions for improving in vitro propagation efficiency. The results demonstrated that for garlic inflorescence explants, immature inflorescences protruding 0–5 cm from the leaf sheath or not protruding were the optimal explants, exhibiting the highest shoot number. The Gamborg B5 (B₅) medium supplemented with a hormone combination of zeatin (ZT) 2 mg/L + indole-3-acetic acid (IAA) 0.05–0.2 mg/L at the first stage and ZT 0.2 mg/L + IAA 0.05 mg/L at the second stage was the most effective for improving in vitro propagation efficiency. For in vitro stem disc culture, the B₅ medium containing 6-benzylaminopurine (6–BA) 2 mg/L + 1-naphthaleneacetic acid (NAA) 0.2 mg/L was optimal. Moreover, a sucrose concentration of 7% was identified as optimal for microbulb development, resulting in significantly larger microbulbs than those grown in a medium with 3% sucrose. These results provide a technical basis for large-scale production of high-quality garlic seedlings. Full article

Background/Objectives: Hylocereus undatus is a high-value crop whose conventional propagation is inefficient for commercial scaling. This study aimed to develop an optimized protocol for in vitro establishment and to define optimal plant growth regulator (PGR) formulations for shoot multiplication. Methods: methods involved testing six surface sterilization protocols using combinations of a surfactant, a systemic fungicide, ethanol, and sodium hypochlorite. Subsequently, nodal explants were cultured on Murashige and Skoog (MS) medium supplemented with ten different concentrations of benzylaminopurine (BAP) and indole-3-butyric acid (IBA), with morphogenic responses evaluated over 60 days. Results: We identified a sterilization treatment that achieved contamination-free cultures with high explant survival percentages. Shoot multiplication was strictly dependent on cytokinin supplementation, with the highest BAP concentration inducing maximal shoot proliferation, while lower concentrations favored shoot elongation. The inclusion of IBA demonstrated a synergistic effect; a balanced BAP–IBA combination optimized shoot proliferation and vigor, whereas a high auxin-to-cytokinin ratio severely repressed organogenesis. Conclusion: this research establishes a reproducible, two-phase protocol that integrates rigorous aseptic establishment with tailored PGR application, effectively balancing high multiplication with superior shoot morphology for the commercial micropropagation of pitahaya. Full article

P. peruviana is a species of agronomic and biotechnological interest; however, the relationship between in vitro regeneration and phenolic compound production remains poorly explored. This study evaluated the combined effects of thidiazuron (TDZ), explant type (cotyledon and hypocotyl), auxin type (naphthaleneacetic acid, NAA, or indole-3-butyric acid, IBA), and auxin concentration on shoot organogenesis, photosynthetic pigment content, and phenolic accumulation. An initial screening identified 4.54 µM TDZ as the optimal concentration for shoot induction. Subsequent experiments showed that morphogenic and physiological responses were strongly dependent on the interaction among explant type, auxin type, and auxin dose. Cotyledon explants consistently exhibited higher shoot regeneration, vigor, biomass accumulation, and photosynthetic pigment content than hypocotyl explants, which showed reduced physiological performance and a higher tendency for callus formation. NAA-based treatments primarily enhanced morphogenic traits, whereas IBA-based treatments were associated with increased photosynthetic pigment content and phenolic accumulation. Multivariate analysis integrating morphogenic, physiological, and biochemical variables identified cotyledon explants cultured with 0.5 µM IBA in the presence of 4.54 µM TDZ as the treatment achieving the most favorable balance between shoot regeneration, physiological stability, and controlled phenolic accumulation. These findings provide a robust basis for optimizing in vitro culture systems of P. peruviana that balance growth, physiological integrity, and secondary metabolism. Full article

Adventitious shoot regeneration in woody species is regulated by interactions between plant growth regulators, endogenous hormone metabolism, and environmental cues such as light quality. Here, we investigated the effects of thidiazuron (TDZ) and the cytokinin oxidase/dehydrogenase (CKX) inhibitors INCYDE and phenyladenine (PA), in combination with different light spectra, on morphogenesis in Melia volkensii leaf explants. TDZ induced the highest frequencies of callus formation and adventitious shoot regeneration, particularly under white light. INCYDE promoted localized regeneration responses, including activation of dormant meristematic regions in secondary leaf axils, whereas PA showed limited regeneration efficiency. Light quality significantly influenced morphogenesis, with white and blue light favoring organized shoot development, while red and far-red light suppressed shoot regeneration and promoted callus formation. Cytokinin profiling revealed treatment-dependent shifts in endogenous cytokinin composition, most notably in isopentenyladenine (iP)-type cytokinins, which is consistent with altered cytokinin degradation dynamics. Cis-zeatin-type cytokinins were abundant across treatments, likely reflecting regulation associated with in vitro culture conditions. These findings indicate that cytokinin metabolism and light quality jointly influence organogenic competence in Melia volkensii Gürke, providing a physiological basis for optimizing regeneration strategies in woody plants. This study provides the first integrated analysis of cytokinin-modulating compounds and light spectra on adventitious shoot regeneration in Melia volkensii. The findings establish a physiological basis for improving regeneration protocols in recalcitrant woody species and support future biotechnological applications, including genetic improvement and advanced propagation strategies. Full article

This study presents the first integrated analysis of genotype–medium interactions and temporal morphogenesis profiling in sunflower regeneration. It aims to characterize genotype-specific responses, identify predictive morphological markers, and develop a scalable framework for breeding and transformation. Eighteen sunflower genotypes were evaluated to assess organogenic performance. The model genotype Ha-26-PR was used for a complementary experiment, testing varying sucrose concentrations to examine their influence on morphogenic outcomes. Hierarchical Cluster Analysis (HCA), guided by the Elbow method, identified four optimal clusters (K = 4). These aligned with three biologically meaningful categories: High Regenerators (Cluster 1), Moderate/Specific Regenerators (Clusters 2 and 3), and Non-Regenerators (Cluster 4). On S1 medium, NO-SU-12 and AS-1-PR showed superior shoot regeneration, while on R4 medium, HA-26-PR-SU and NO-SU-12 performed best. Genotypes such as NO-SU-12 and AS-1-PR consistently excelled across both media, whereas AB-OR-8 and FE-7 remained non-regenerators. Medium R4 supported superior regeneration, primarily through root formation, while S1 failed to induce roots in any genotype, highlighting the importance of hormonal composition. Although sucrose promoted callus induction, it did not trigger organogenesis. Callus was consistently present across media and time points, but its correlations with shoot and root formation were weak and temporally unstable, limiting its predictive value. Root formation at 14 days (Root 14D) emerged as a robust early predictor of organogenic success. This integration of morphological, temporal, and statistical analyses offers a genotype-tailored regeneration framework with direct applications in molecular breeding and CRISPR/Cas-based genome editing. Full article

Ficus carica L. is a fruit crop of notable nutritional and economic value. The ‘Xinjiang Zaohuang’ cultivar, rich in flavonoids, also holds considerable medicinal potential. To address the constraints of conventional propagation for mass production, this study developed an efficient and genetically stable generation protocol using healthy sprouted branches. MS medium was identified as the most effective basal medium for shoot growth. The highest adventitious bud induction rate (89.67%) and the greatest mean bud number (6.29) were achieved when explants were cultured on MS medium supplemented with 1 mg/L 6-BA and 0.1 mg/L IBA. In the organogenesis process, indole-3-butyric acid (IBA) promoted direct shoot formation with minimal callus intervention compared to naphthaleneacetic acid (NAA). The optimal combination for shoot elongation was 0.1 mg/L 6-BA and 0.01 mg/L IBA, which produced morphologically uniform shoots. For rooting, an IBA concentration of 1 mg/L was optimal, achieving a 96.7% success rate. Inter-simple sequence repeat (ISSR) analysis confirmed the genetic stability of all regenerated plants. These findings establish a reliable technical framework for the large-scale propagation of this valuable fig cultivar. Full article

Iris setosa is a characteristic perennial wild herbaceous flower in the Changbai Mountain region of China, boasting significant ornamental and medicinal values. Given the increasing scarcity of its wild resources, this study developed an efficient in vitro regeneration system using shoot tips as explants via the direct organogenesis pathway. The optimal surface sterilization protocol was achieved with a treatment of 0.1% HgCl₂ for 8 min, resulting in an explant survival rate of 57.78%. The highest multiple shoot induction rate (88.89%) of shoot tips was achieved on MS medium supplemented with 2.0 mg·L⁻¹ 6-benzylaminopurine (6-BA), 0.5 mg·L⁻¹ naphthalene acetic acid (NAA), and 1.0 mg·L⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D). The optimal shoot differentiation and proliferation medium was MS + 2.0 mg·L⁻¹ 6-BA + 0.3 mg·L⁻¹ NAA, achieving a proliferation coefficient of 3.37. The optimal medium for rooting was confirmed to be 1/2 MS + 0.5 mg·L⁻¹ indole-3-butyric acid (IBA), exhibiting a high rooting rate reached 98.33%. During transplantation, plantlets exhibited high survival rates (over 90%) and vigorous growth across all three tested substrates, with no significant differences in survival rates among substrates. The key advance of this study lies in the development of a highly efficient and stable regeneration protocol for I. setosa derived from shoot tip explants, providing critical technical backing for the conservation and sustainable exploitation of its wild-type germplasm. Full article

The spice Ferula assa-foetida L., also known as asafoetida, is widely recognized for its medicinal and culinary applications. The non-native status of the plant and the prolonged dormancy of its seeds pose significant challenges for large-scale cultivation in India. In vitro organogenesis offers an effective solution to these obstacles. Establishing reliable in vitro regeneration protocols requires standardized statistical methods to evaluate univariate and multivariate data for optimizing specific traits. However, these methods have limitations when handling complex, nonlinear inputs, often producing large prediction errors that reduce the reliability of trait optimization. This study developed an in vitro regeneration system for F. assa-foetida L. and identified optimal PGRs for somatic embryogenesis and shoot organogenesis through image-based morphological analysis. Predictive models were created using DL and ML algorithms. Calli induced from leaf explants was cultured on the Murashige and Skoog medium supplemented with various combinations and concentrations of thidiazuron (TDZ), 6-benzylaminopurine (BAP), and α-naphthaleneacetic acid (NAA), as experimental variables. Seven ML approaches, namely random forest (RF), support vector machine (SVM), k-nearest neighbours (kNN), decision tree (DT), extreme gradient boosting (XG Boost), naïve bayes, and logistic regression, alongside five DL models—convolutional neural network (CNN), MobileNet, region-based convolutional neural network (RCNN), residual neural network (ResNet), and visual geometry group (VGG19)—were employed to predict the best PGRs for somatic embryogenesis and shoot organogenesis. Among them, the convolutional neural network (CNN) achieved the highest accuracy (87%), outperforming baseline ML models such as logistic regression and decision tree (82%). This pioneering study in F. assa-foetida L. presents an AI-driven, image-based framework for predicting optimal PGRs, offering a scalable approach to enhance micropropagation in endangered medicinal plants. Full article

The genus Corydalis, belonging to the Papaveraceae family, is widely distributed across the Northern Hemisphere, primarily in Asia. This study aimed to investigate the effect of plant growth regulators (PGRs) on callus induction, and of light quality and intensity on indirect shoot organogenesis, dl-Tetrahydropalmatine (dl-THP) accumulation, and activities of antioxidant enzymes in Corydalis turtschaninovii Besser. Calli were successfully induced from the leaf, tuber, and petiole explants with different PGR combinations. The best callus induction from leaf, tuber, and petiole explants were obtained in the medium supplemented with 3 mg·L⁻¹ kinetin (Kn) combined with 0.8 mg·L⁻¹ naphthalene acetic acid (NAA), 3 mg·L⁻¹ benzyl adenine (BA) combined with 0.8 mg·L⁻¹ NAA, and 2 mg·L⁻¹ BA combined with 0.5 mg·L⁻¹ NAA, respectively. For indirect shoot organogenesis, calli were cultured on the Murashige and Skoog (MS) medium under dark (D), white (W), red (R), blue (B), or 1:1 mixture of red and blue (RB) light-emitting diodes (LEDs) at an intensity of 25 or 50 µmol·m⁻²·s⁻¹ photosynthetic photon flux density (PPFD) for six weeks. The RB treatment increased biomass accumulation of the callus, and promoted the induction of the shoot from the callus, whereas the R treatment promoted the dl-THP accumulation, especially with the higher light intensity. Light quality and intensity significantly influenced the activities of antioxidant enzymes and the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging capacity in calli, with the most pronounced effects observed under B or RB light treatments. Taken together, the application of monochromatic LED or combinations of red and blue LEDs could be used for the callus culture for different purposes in vitro. Full article

Agave species possess substantial cultural, ecological, and economic significance, particularly in Mexico, where they are traditionally utilized for food, fiber, and beverages. Their industrial relevance has expanded to include bioenergy, nutraceuticals, and sustainable agriculture. However, conventional propagation methods are constrained by long life cycles, low seed germination rates, and susceptibility to phytopathogens. In vitro culture has emerged as a pivotal biotechnological strategy for clonal propagation, germplasm conservation, and physiological enhancement. This review presents a critical synthesis of plant growth regulators (PGRs) employed in agave micropropagation, emphasizing their roles in organogenesis, somatic embryogenesis, shoot proliferation, and rooting. Classical PGRs such as 6-benzylaminopurine (BAP), benzyladenine (BA), 2,4-dichlorophenoxyacetic acid (2,4-D), indole-3-acetic acid (IAA), and indole-3-butyric acid (IBA) are widely utilized, with BA + 2,4-D and BA + IAA combinations demonstrating high efficiency in embryogenic callus induction and shoot multiplication. Additionally, non-traditional regulators such as abscisic acid (ABA) and putrescine (Put) have been shown to affect embryo maturation. This review synthesizes recent studies on agave in vitro culture protocols, identifies trends in PGR use, and highlights key research gaps. These insights reveal opportunities for innovation and underscore the need for species-specific optimization and molecular validation to improve reproducibility and scalability. Full article

Pothos (Epipremnum aureum G.S. Bunting), which belongs to the Arum family (Araceae Juss.), can be used for medicinal, ornamental, and pollutant-purifying purposes. Due to the usefulness of pothos, the market demand for this species is increasing. Our study attempts to fill in the shortcomings of previous studies on the effect of activated carbon and plant growth regulators on the ability of shoots to take root in vitro, as well as the effect of inexpensive and readily available materials on the transition of seedlings from in vitro to the greenhouse stage. To evaluate the shooting results, Murashige and Skoog medium (MS) was used, which included 6-benzylaminopurine (BA), kinetin (Kn), α-naphthaleneacetic acid (α-NAA), coconut water, activated carbon, and indole-3-butyric acid (IBA) in various concentrations and combinations. Our results showed that the MS medium with the addition of 2.5 mg/L BA and 1.0 mg/L Kn was optimal for propagation by shoots. In this variant, 2.86 shoots per explant, 1.87 cm of shoot length, and 1.59 leaves per shoot were obtained. Despite the fact that this treatment provided the highest total cytokinin concentration, it was significantly more effective than only BA (2.5 mg/L) and all combinations of BA+α-NAA or Kn+α-NAA. For rooting, the micro shoots obtained on the above medium were transferred to MS + 0.25 mg/L α-NAA + 0.5 g/L AC, which allowed for rooting by 93.33%, 1.93 roots per explant, and root lengths by 2.37 cm. This is higher than with the IBA-based treatment, which led to a shortening of the roots and a reduction in their branching. Acclimatization in a 1:1 mixture (by volume) of loamy garden soil (pH 6.2, 2.1% organic matter) and coconut coir (particle size 0.5–2 mm) gave 75% survival after 40 days. These results have opened up the prospect of developing an effective method for reproducing pothos species in vitro by organogenesis at the lowest cost. Full article

Generation of state-of-the-art highly productive cabbage genotypes (Brassica oleracea convar. capitata (L.) Alef.) with improved agronomic traits is attainable using modern biotechnological approaches. However, capitata cabbage is relatively recalcitrant to de novo shoot organogenesis from callus tissue, especially with loss of somatic cell totipotency during genetic transformation. An effective and rapid protocol for in vitro indirect shoot organogenesis from hypocotyl and cotyledon explants derived from 6-day-old aseptic donor seedlings of Russian cabbage genotypes (the DH line as well as cvs. Podarok and Parus) has been developed. In order to obtain standardized donor explants, aseptic cabbage seeds were germinated under dim light conditions (30–40 µmol m⁻² s⁻¹) with a 16 h light/8 h dark photoperiod. Multiple indirect shoot organogenesis (1.47–4.93 shoots per explant) from both cotyledonary leaves and hypocotyl segments with a frequency of 55.2–89.1% was achieved through 45 days of culture on the 0.7% agar-solidified (w/v) Murashige and Skoog (MS) basal medium containing 2 mg/L 6-benzylaminopurine (6-BAP), 0.02 mg/L 1-naphthalene acetic acid (NAA), and 5 mg/L AgNO₃. The regenerants were successfully rooted on an MS basal medium (69.2%) without plant growth regulators (PGRs), as well as supplemented with 0.5 mg/L NAA (86.8%). Subsequently, in vitro rooted cabbage plantlets were adapted to soil conditions with an efficiency of 85%. This rapid protocol, allowing for the performance of a full cycle from in vitro seed germination to growing adapted plantlets under ex vitro conditions over 95 days, can be successfully applied to induce an indirect shoot formation in various cabbage genotypes, and it is recommended to produce transgenic plants with improved quality traits and productivity. Full article

Introduction: Stevia rebaudiana Bertoni has recently gained significant attention due to the presence of intensely sweet yet low-calorie steviol glycosides (SGs) in its leaves, making it a promising natural sugar alternative with applications in the food, pharmaceutical, and cosmetics industries. The primary goal of this study was to determine whether generating somaclonal variation from plant material obtained by indirect regeneration results in further genetic changes identifiable using the SCoT marker (Start Codon Targeted). Methods: In the first stage, callus tissue was initiated from first-generation somaclones on MS medium supplemented with 4.0 mg/L 6-benzylaminopurine (BAP), 2.0 mg/L 1-naphthaleneacetic acid (NAA), and 2.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D). Their morphogenetic potential was analyzed on four media with different BAP and Kinetin concentrations. Donor plants, first and second generation somaclones, were also analyzed for genetic diversity using SCoT markers. Results: All first-generation somaclones demonstrated a very high callus initiation capacity, ranging from 95 to 100%. It was found that for most of the studied somaclones, the greatest number of shoots were developed by explants grown in a medium supplemented with 0.5 mg/L BAP and 0.25 mg/L Kin. The studied group of somaclones exhibits a high degree of polymorphism (55.2%). The analysis of genetic similarity of somaclones presented in the form of individual dendrograms indicates that in most cases, greater genetic diversity was revealed as a result of indirect regeneration in the first generation of somaclones compared to the second. Indirect organogenesis allows for the production of subsequent generations of genetically unstable somaclones, creating the potential for obtaining new phenotypic variants useful in plant breeding. Full article

Coconut (Cocos nucifera L.) is one of the most widely cultivated crops, with increasing popularity and demand for its products, which necessitates increased production. However, the lack of high-quality planting materials is a major limitation in replanting the senile palms worldwide. This study examined the possibility of using a direct shoot organogenesis pathway as an alternative to somatic embryogenesis to produce clonal coconut plantlets using immature inflorescence explants obtained from Indonesia and Australia, through investigation of the explant types, exogenous plant growth regulators, and additives. Histological analysis showed suitable stages of immature inflorescence explants to be used, which led to the formation of shoot-like structures resembling true vegetative shoots, in all treatments consisting of exogenous plant growth regulators except for those without activated charcoal. The culture medium supplemented with thidiazuron (100 μM) alone or those supplemented with various combinations of other plant growth regulators showed similar shoot induction percentages (ca. 63 to 80%) or shoot-like structures per explant (ca. 6 to 8). The addition of adenine sulphate (217 μM) was found to significantly improve shoot induction (ca. 50%) from immature inflorescence explants compared to the control (ca. 5%), whereas phloroglucinol was found to negatively impact shoot induction, and L-glutamine showed a positive influence. The current study showed several improvements, which warrant further studies to develop commercial protocol for mass production of clonal coconut plantlets through direct organogenesis. Full article