Showing papers on "Kinetin published in 2022"

Efficient micropropagation of Thunbergia coccinea Wall. and genetic homogeneity assessment through RAPD and ISSR markers

Abstract: Thunbergia coccinea Wall. ex D. Don being a rare, ornamental and medicinal plant of India, is needed to propagate for conserving the germplasm and analyzing its phytochemical compounds in the future. A reliable protocol for direct in vitro propagation using nodal shoot meristem of T. coccinea as explant was standardized. The highest number of shoots per explant (22.17 ± 0.54) with maximum shoot length (2.36 ± 0.28) in cm was obtained in Murashige and Skoog (MS) medium supplemented with 9.70 µM of 6-furfurylaminopurine (Kinetin) and 0.053 µM of α-naphthaleneacetic acid (NAA) combination, among all the different plant growth regulators (PGR's) and concentrations tested. The aforesaid PGR's combination was optimum for axillary shoot bud induction and multiplication in T. coccinea. The best rooting was observed on the half-strength MS medium fortified with 2.68 µM NAA with the highest number of roots per shoot (3.75 ± 0.12) and maximum length (5.22 ± 0.32) in cm. All the in vitro raised plantlets were acclimatized in sterile sand and soil mixture (1:1) with a survival rate of 70% on earthen pots under greenhouse conditions. PCR-based RAPD (Random Amplified Polymorphic DNA) and ISSR (Inter-Simple Sequence Repeat) molecular markers were employed to determine the genetic homogeneity amongst the plantlets. Twelve (12) RAPD and nine (9) ISSR primers developed a total of 104 and 91 scorable bands, respectively. The band profiles of micropropagated plantlets were monomorphic to the mother, donor in vivo plant, and similarity values varied from 0.9542-1.000. The dendrogram generated through UPGMA (unweighted pair group method with arithmetic mean) showed 99% similarities amongst all tested plants confirming the genetic uniformity of in vitro raised plants.

Cisplatin-Induced Reproductive Toxicity and Oxidative Stress: Ameliorative Effect of Kinetin

Abstract: Cisplatin is a commonly used chemotherapeutic agent; however, its potential side effects, including gonadotoxicity and infertility, are a critical problem. Oxidative stress has been implicated in the pathogenesis of cisplatin-induced testicular dysfunction. We investigated whether kinetin use at different concentrations could alleviate gonadal injury associated with cisplatin treatment, with an exploration of the involvement of its antioxidant capacity. Kinetin was administered in different doses of 0.25, 0.5, and 1 mg/kg, alone or along with cisplatin for 10 days. Cisplatin toxicity was induced via a single IP dose of 7 mg/kg on day four. In a dose-dependent manner, concomitant administration of kinetin with cisplatin significantly restored testicular oxidative stress parameters, corrected the distorted sperm quality parameters and histopathological changes, enhanced levels of serum testosterone and testicular StAR protein expression, as well as reduced the up-regulation of testicular TNF-α, IL-1β, Il-6, and caspase-3, caused by cisplatin. It is worth noting that the testicular protective effect of the highest kinetin dose was comparable/more potent and significantly higher than the effects of vitamin C and the lowest kinetin dose, respectively. Overall, these data indicate that kinetin may offer a promising approach for alleviating cisplatin-induced reproductive toxicity and organ damage, via ameliorating oxidative stress and reducing inflammation and apoptosis.

Flow cytometry and start codon targeted (SCoT) genetic fidelity assessment of regenerated plantlets in Tylophora indica (Burm.f.) Merrill

Abstract: Tylophora indica (Burm.f.) Merrill. is a pharmacologically important plant, popular for alkaloidal and non-alkaloidal richness. Large scale propagation of T. indica is difficult in the wild as the seeds are small and the frequency of germination is very poor. In the present study, the genome size estimation of in vitro regenerated (indirect, direct and somatic embryo mediated) T. indica was made by flow cytometric method. Clonal fidelity of the regenerants was assessed using a start codon targeted (SCoT) molecular marker. Initially, the explants were inoculated on Murashige and Skoog basal medium supplemented with various concentrations of plant growth regulators like 2,4-dichlorophenoxy acetic acid (2,4-D), Kinetin, 6-benzyl amino purine (BAP) and 1-naphthalene acetic acid either singly or in combinations. The highest callus induction frequency (87.75%) was obtained in 6.7 µM 2,4-D added MS medium which metamorphosed into progressive stages (globular, heart, torpedo, and cotyledonary) of embryos. Mature and healthy somatic embryos efficiently germinated into plantlets on 8.8 µM BAP + 1.4 µM GA3 enriched MS medium. Histological and scanning electron microscopic study confirmed the above developing stages. The regenerated shoots were rooted best in 2.45 µM Indole-3-butyric acid supplemented solid MS medium. The plants were hardened and acclimatized with 90% survivability. The flow cytometric 2C DNA content of indirect, direct and somatic embryo derived plants was 1.896 pg, 1.940 pg and 1.926 pg respectively, very similar to the mother plant (1.928 pg). SCoT marker generated a high percentage of monomorphic bands (94%) revealing similarity with the mother plant, thus ensuring genetic fidelity. To the best of our knowledge, this is perhaps the first ever report of 2C DNA content estimation and SCoT marker based genetic homogeneity study in T. indica. An efficient regeneration method has been developed in T. indica. Histology and SEM indicated somatic embryogenesis based morphogenesis. 2C genome size and SCoT marker revealed genetic homogeneity of the regenerated population.

Sterilization protocols and the effect of plant growth regulators on callus induction and secondary metabolites production in in vitro cultures Melia azedarach L.

Abstract: Melia azedarach L. is a valuable source of antioxidants and secondary metabolites. This study is a first extensive report about the effect of different serialization protocols and plant growth regulators (PGRs) on explant disinfection efficiency, callus induction and secondary metabolites production and accumulation in callus cultures of M. azedarach L. In this regard, the effect of plant growth regulators on callus induction and secondary metabolites production were examined. In addition, different sterilization agents were evaluated for disinfection of chinaberry leaf explants. The results showed that the lowest percentage of explant contamination and browning with the highest percentage of callus induction and callus growth obtained with explants pretreated with benomyl (2 g/L) for 2 h and sterilized with 7% H2O2 for 10 min and NaOCl 2% (without pH adjustment) for 12 min. Although adjusting the pH of NaOCl to pH = 7 and 10 significantly reduced the microbial contamination and increased the percentage of contamination-free cultures of M. azedarach L., adversely influenced the explant viability and callus induction and growth. The highest percentage of callus induction obtained on the MS medium containing 3 mg/L NAA/2,4-D and 1 or 3 mg/L Kin/BAP, and the highest callus yield (1804.833 mg/explant) belonged to the MS medium supplemented with 5 mg/L 2,4-D and 5 mg/L Kin. The callus cultures grown on the MS medium supplemented with 3 mg/L NAA and 1 mg/L Kin produced the highest amount of Quercetin (2.06 mg/g fresh weight), Rutin (5.56 mg/g fresh weight) and Kaempferol (1.84 mg/g fresh weight).

Foliar application of thiourea, salicylic acid, and kinetin alleviate salinity stress in maize grown under etiolated and de-etiolated conditions

Abstract: Abstract Salinity stress and the absence of light negatively impact growth and development of the plants. Morpho-physiological and biochemical attributes of maize ( Zea mays L.) get severely affected by salt stress and subdue light. Therefore, a pot experiment was conducted under the prevailing environmental conditions of Turbat, Balochistan, to explore etiolation and the de-etiolation response of maize hybrid (SP-17S23) to salinity stress under exogenous application of plant growth regulators (PGRs). Maize seedlings in three sets, i.e., non-etiolated, etiolated, de-etiolated, subjected to salinity stress (120 mM NaCl) after 15 days of seed germination. After a week, the seedlings were sprayed with optimized levels of different PGRs, including thiourea (TU; 10 mM), salicylic acid (SA; 250 µM), and kinetin (KIN; 3 µM). Salinity stress hampered plant growth and affected morpho-physiological attributes. However, PGRs foliar treatment proved effective, thus ameliorating the impact of salinity and etiolation on maize seedlings. Growth attributes (root/shoot length, leaf area, root/shoot fresh and dry weight), photosynthetic pigments (Chl a, b and carotenoids) were significantly enhanced under the foliar treatment of PGRs, especially under TU and KIN treatments. However, the oxidative damage parameters, i.e., malondialdehyde (MDA) and hydrogen peroxide (H 2 O 2 ), decreased under the treatment of PGRs, thereby protecting seedlings under salinity and etiolated conditions. Overall, PGRs enhanced tolerance potential of plants under salinity stress with the consideration of light variations remain the key concern for developing healthy and vigor seedling strands.

6-Benzylaminopurine and kinetin modulations during in vitro propagation of Quercus robur (L.): an assessment of anatomical, biochemical, and physiological profiling of shoots

Abstract: For many woody species, such as Quercus robur, cytokinins in the culture medium are required to maintain in vitro plant material. Among synthetic cytokinins, 6-benzylaminopurine (BAP) and kinetin (KIN) are the most frequently used. In addition to inducing shoots, cytokinins can cause morphophysiological disorders. Therefore, we aimed to investigate the anatomical, biochemical, and physiological alterations and profiles of Q. robur shoots exposed to two cytokinins, applied alone and in combination. Shoots previously established in vitro were transferred to WPM culture media supplemented with BAP at concentrations of 0, 1.25, and 3.50 µM combined with KIN at concentrations of 0, 0.62, and 1.25 µM totaling 9 treatments. Anatomical, physiological, and biochemical analyses were performed after 40 days of culture. BAP induced the formation of new buds with anatomically underdeveloped leaves; induced shoot-tip necrosis, which is considered a response to the inefficient transport of water and nutrients; reduced the thickness of the cell walls of phloem fibers; and decreased the content of phenolic compounds and photosynthetic pigments. These responses were less pronounced with co-exposure to KIN. In contrast, KIN alone stimulated a larger area of secondary xylem and more lignified cell walls. BAP can induce shoots with underdeveloped anatomical and biochemical characteristics. Shoots that grew with KIN alone had stem and leaf anatomical characteristics, indicating greater commitment to cellular differentiation than proliferation. When both cytokinins are combined, KIN can partially mitigate the deleterious effects of BAP on in vitro growth. BAP generated shoots with underdeveloped anatomical and biochemical characteristics. KIN can partially mitigate the deleterious effects of BAP on in vitro growth.

New approaches for micropropagation and cryopreservation of Agave peacockii, an endangered species

Abstract: More than 50% out of 129 of Agave species are endemic to Mexico. Among them, Agave peacockii is among the list of threatened species that require special protection. In this work, we aimed at developing new supplementary strategies to achieve micropropagation and perform cryopreservation of in vitro grown shoot-tips of A. peacockii. For multiplication, the addition of two cytokinins, 6-benzylaminopurine (26.6 μM) and kinetin (27.84 μM) to MS semisolid medium significantly favoured the morphogenetic response and produced the highest (87.00 ± 17.18) shoot generation number after 60 days of culture. This interaction was more effective than using the same growth regulators separately. Propagated and rooted plantlets were acclimatized with 100% survival and normal morphological development. For cryopreservation, an optimized protocol following droplet-vitrification allowed obtaining 98% and 96% regrowth before and after cryopreservation, respectively. Shoot-tips (1 mm in length × 1 mm wide) were excised of in vitro-propagated plants, subjected to preculture on MS semisolid medium with 0.3 M sucrose for 1d, loaded in solution with 0.4 M sucrose and 1.6 M glycerol for 20 min, exposed to Plant Vitrification Solution 2 for 15 min, and then, immersed in liquid nitrogen in droplets of PVS2 placed on aluminium foil strips. The vegetative growth of cryo-derived plants and of the in vitro propagated plants was compared under greenhouse conditions. No significant differences were detected in most assessed characteristics after 120 days of culture. The results presented here constitute new viable biotechnological approaches for the in vitro propagation and long-term conservation of endangered Agave germplasm. Agave peacockii shoot micropropagation was induced combining 6-benzylaminopurine (BAP) and 6-furfuryl-aminopurine (kinetin). A droplet-vitrification protocol was optimized to cryopreserve shoot-tips. Greenhouse performance of in vitro and cryo-derived plants was similar.

In Vitro Propagation of Aconitum violaceum Jacq. ex Stapf through Seed Culture and Somatic Embryogenesis

Abstract: Aconitum violaceum Jacq. ex Stapf is a threatened medicinal plant with restricted global distribution. The highest frequency of seed germination was recorded on Murashige and Skoog’s (MS) basal medium, supplemented with 0.5 mg L−1 kinetin with a germination rate of 77.32% and mean germination time of 27 days. Among the various plant growth regulators examined, 0.1 mg L−1 kinetin (Kn) + 0.5 mg L−1 indole-3-acetic acid (IAA) proved to be effective for maximum embryogenic callus production (51.0%) within 31 days of inoculation. The conversion rate of somatic embryos into complete plantlets was highest in the MS medium augmented with 0.1 mg L−1 Kn + 0.5 mg L−1 IAA (68.00%), with an average root initiation time of 25 days. The rooted plantlets were subsequently hardened into jiffy pots with a combination of loamy soil, coco-peat, and vermicompost (1:1:1 v/v), and then transplanted into a greenhouse with a 60% survival rate. To our knowledge, this is the first study on direct in vitro propagation and embryogenic callus induction from seeds. The established regeneration protocol could be employed to propagate A. violaceum on a large scale in a short time. This would contribute significantly to its rapid propagation and germplasm conservation, and establish a framework for the domestication of this highly valued threatened medicinal plant.

In Vitro Propagation of the Mount Parnitha Endangered Species Sideritis raeseri subsp. Attica

Abstract: Over the past few decades, both wildfires and human-sparked fires have ravaged Mount Parnitha, destroying the mountain’s unique natural ecosystem, applying pressure to its flora, and subjecting the vulnerable populations of Sideritis raeseri subsp. attica to excessive stress. The present study aims to establish an efficient micropropagation method starting from in vitro-grown seedlings. The in vitro germination study carried out during the production of seedlings revealed a higher germination rate (34.0% and 37.0%, respectively) at 20.0 °C and 25.0 °C. The in vitro-derived seedlings studied were used as the starting material for the establishment of various media. Murashige and Skoog (MS) media, hormone-free and containing 0.5 mg L−1 6-benzyladenine (BA), led to the satisfactory (84.0–89.0%) establishment of plantlets. During the multiplication phase, the study used BA in conjunction with α-naphthaleneacetic acetic acid and four different cytocinins (BA; kinetin (KIN); 6-(γ-γ-dimethylallylamino) purine; zeatin) at low concentrations (0.5 mg L−1). During the second subculture, a high multiplication index (7.3 and 6.4, respectively) was found for the hormone-free MS medium and the MS medium containing KIN at 0.5 mg L−1. Hyperhydricity took place on the media supplemented with hormones. Rooting occurred on the half-strength MS medium (51.0%). After two months, the plants’ survival rate stood at 100.0%, as did their ex vitro acclimatisation rate, which also registered at 100.0%. The present results could encourage not only the introduction of S. raeseri subsp. attica into the industry of floriculture as a new ornamental plant but also its ex vitro conservation.

Establishment of In Vitro Regeneration Protocol for Sabah’s Jewel Orchid, Macodes limii J.J. Wood & A.L. Lamb

Abstract: Habitat disturbance and excessive collection of wild orchids from their natural habitat have threatened many orchids species at risk of extinction. In this study, the in vitro regeneration protocol for Macodes limii, a jewel orchid endemic to Sabah was established. The effects of explant source and plant growth regulators (PGRs) including naphthaleneacetic acid, picloram, 2,4-dichlorophenoxyacetic acid, 6-benzylaminopurine, kinetin, and thidiazuron on the in vitro regeneration capacity of M. limii plantlets were examined. Both factors showed a significant interaction in promoting axillary shoot formation. Nodal explants from the third and fourth positions cultured with 1.0 mg/L TDZ, induced 95% of shoot regeneration, with an average of three shoots/explant (1.6–1.8 cm of shoot length) after 90 days of culture. The well-developed plantlets went through an acclimatization phase for 60 days with a 60% of survival rate. An inter simple sequence repeat (ISSR) marker analysis confirmed the genetic stability of the in vitro regenerated plants to the mother plant. The successfully acclimatized plantlets were finally transferred to Poring Orchid Conservation Centre for reintroduction. The established protocol provides the means for large-scale production of this endemic jewel orchid, as well as a basis for further research aimed at the conservation and genetic improvement of this plant.

In Vitro Propagation of the Dendrobium anosmum Lindl. Collected in Vietnam

Abstract: Hoa Binh province is one of the best places for orchids in Vietnam. The climate and environment of Hoa Binh province are favorable for the development of orchids, especially rare indigenous ones. Dendrobium anosmum Lindl., which stands out because of the unique fragrance and colors, is one of the most popular varieties in Hoa Binh province. To meet the increasing demands of the industrial market as well as to contribute to the preservation and development of genetic resources of Dendrobium sp. in Hoa Binh province, propagating D. anosmum Lindl. is a crucial step. Plant tissue culture, which has been applied to improve reproducibility of orchids for many years, is still an effective method, especially for large-scale propagation. Studies on in vitro propagation of D. anosmum Lindl. from Hoa Binh province showed that growth regulators (BA, kinetin, α-NAA) did not have a significant effect on protocorm initiation because D. anosmum Lind. from Hoa Binh province already has a high rate of regeneration. However, MS medium + 1.0 mg/L kinetin + 0.5 mg/L α-NAA + 30 g sucrose + 8.0 g agar per liter, pH 5.7–5.8 was the optimal medium to increase shoot length. The MS medium + 1.0 g activated charcoal + 30 g sucrose + 8.0 g agar per liter, pH 5.7–5.8 was the most suitable medium for shoot growth—after 6 weeks of culture, the average shoot length was 1.09 cm, the average number of leaves was 6.13, the average number of roots was 3.17, and the average root length was 1.11 cm—about 3.3, 4.17, 3.41, and 1.67 times higher, respectively, than in the control (without activated charcoal).

Mathematical modeling and optimizing the in vitro shoot proliferation of wallflower using multilayer perceptron non-dominated sorting genetic algorithm-II (MLP-NSGAII)

Abstract: Novel computational methods such as artificial neural networks (ANNs) can facilitate modeling and predicting results of tissue culture experiments and thereby decrease the number of experimental treatments and combinations. The objective of the current study is modeling and predicting in vitro shoot proliferation of Erysimum cheiri (L.) Crantz, which is an important bedding flower and medicinal plant. Its micropropagation has not been investigated before and as a case study multilayer perceptron- non-dominated sorting genetic algorithm-II (MLP-NSGAII) can be applied. MLP was used for modeling three outputs including shoots number (SN), shoots length (SL), and callus weight (CW) based on four variables including 6-benzylaminopurine (BAP), kinetin (Kin), 1-naphthalene acetic acid (NAA) and gibberellic acid (GA3). The R2 correlation values of 0.84, 0.99 and 0.93 between experimental and predicted data were obtained for SN, SL, and CW, respectively. These results proved the high accuracy of MLP model. Afterwards the model connected to Non-dominated Sorting Genetic Algorithm-II (NSGA-II) was used to optimize input variables for obtaining the best predicted outputs. The results of sensitivity analysis indicated that SN and CW were more sensitive to BA, followed by Kin, NAA and GA. For SL, more sensitivity was obtained for GA3 than NAA. The validation experiment indicated that the difference between the validation data and MLP-NSGAII predicted data were negligible. Generally, MLP-NSGAII can be considered as a powerful method for modeling and optimizing in vitro studies.

Micropropagation of Feverfew (Tanacetum parthenium) and Quantification of Parthenolide Content in Its Micropropagated and Conventionally Grown Plants

Abstract: Feverfew (Tanacetum parthenium) is a well-known multi-functional plant with anti-inflammatory, cardiotonic, antiangiogenic, and anticancer effects. The therapeutic value of this plant is due to its phytochemical constitutes, especially parthenolide. Tissue culture techniques have been applied to improve the bioactive components of many herbal plants. Hence, this study, was carried out to establish a protocol for micropropagation of the feverfew plant and to quantify parthenolide content in its micropropagated and conventionally grown plants. To establish an aseptic culture, different concentrations of sodium hypochlorite (NaOCl) were investigated for seed surface sterilization. Besides, the effects of plant growth regulators (PGRs) on the callus induction, shoot organogenesis from callus and in vitro rooting were evaluated. Additionally, the parthenolide yield of the micropropagated and conventionally grown plants was determined by using high-performance liquid chromatography (HPLC). The results showed that surface sterilization of feverfew seeds with 6% NaOCl for 15 min obtained 65.00 ± 2.69% aseptic seeds. Murashige and Skoog (MS) medium supplemented with 0.4 mg/L thidiazuron (TDZ) and 2 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D) resulted in 86.00 ± 1.72% callus induction. The highest number of shoots (5.00 ± 0.15) per explant was obtained in the treatment of MS medium supplemented with 5 mg/L zeatin. MS medium fortified with 3 mg/L indole-3-butyric acid (IBA) produced the maximum number of roots per plantlet (8.90 ± 0.35). A total of 90% of the micropropagated plantlets survived when planted in perlite + peat moss (1:1 v/v); the micropropagated plantlets were successfully established in the ex vitro conditions. According to parthenolide analysis, its level was significantly higher in the micropropagated plants than conventionally grown plants. Among different solvents, ethanolic extraction obtained the highest parthenolide content of the feverfew plant. Hence, it can be concluded that micropropagation of feverfew could be applied to produce disease-free planting materials and to improve the parthenolide content of the feverfew plant.

Establishment of an Efficient In Vitro Propagation Method for a Sustainable Supply of Plectranthus amboinicus (Lour.) and Genetic Homogeneity Using Flow Cytometry and SPAR Markers

Abstract: Plectranthus amboinicus (Lour.) Spreng is a medicinally important aromatic perennial herb used for the treatment of skin diseases, constipation, asthma, flu, fever, cough, and headache as well as a flavoring ingredient in traditional drinks, food, and meat stuffing. In this study, a high-performance in vitro propagation system of P. amboinicus through direct shoot organogenesis was developed using axillary node explants cultured on MS (Murashige and Skoog) medium augmented with 0.5, 2.5, 5.0, 7.5, and 10.0 µM of 6-benzyladenine (BA) or kinetin (Kin), alone or with 0.1, 0.5, 2.5, and 5.0 µM of indole-3-acetic acid (IAA) or α-naphthalene acetic acid (NAA). To optimize the regeneration potential of node explants, the effects of basal media strength and pH were also investigated. After 8 weeks of culture, explants cultured in full strength MS basal medium (pH 5.7) with 5.0 µM BA and 2.5 µM NAA exhibited the highest percentage (97.1%) of regeneration and the maximum number (19.3) of shoots per explant. Individual elongated shoots were rooted on half strength MS basal medium containing 0.25 µM indole 3-butyric acid (IBA) after 4 weeks of culture, producing 5.3 roots/shootlets with a root induction frequency of 93.7%. First time genetic stability of in vitro raised P. amboinicus plants was determined using SPAR markers, such as DAMD and ISSR, as well as flow cytometric tests, assuring the availability of authenticated raw materials for commercial production of the plant and its bioactive components.

Effects of elicitors on secondary metabolite (SM) production and antioxidant activity in sweet basil (Ocimum basilicum L.) cell suspension cultures

Abstract: Elicitor treatments play an important role in inducing some protective signal transmitter enzymes in cells and regulating phenylalanine ammonia-lyase (PAL) activity. The aim of this study was to determine the effects of elicitors [silver nitrate (AgNO3), salicylic acid (SA) and yeast extract (YE)], which were added individually or in combination to Ocimum basilicum L. cell suspension cultures, on the production and antioxidant activity of secondary metabolites (SMs). Calluses were obtained from the leaves of O. basilicum kept on Murashige and Skoog (MS) medium containing 0.5 mg/l kinetin (KIN) + 2.5 mg/l naphthalene-acetic acid (NAA) and cell suspension cultures were initiated. Then elicitors were applied to the cell suspension cultures individually or in combination, and cells were harvested at the end of the second, fourth, and eighth days. Compared with the control culture, the maximum rosmarinic and chicoric acid production was obtained at the end of the 4th day from SA (24 µM) + YE (80 mg/l) treatment as 20.19 mg/g DW (118%) and 7.55 mg/g DW (123%), respectively. The maximum biosynthesis of isoquercetin and rutin compared with the control culture was 3.88 mg/g DW [YE (80 mg/l)] with a 1.6-fold increase and 6.35 mg/g DW [YE (80 mg/l) + AgNO3 (6 µM)] with a 1.76-fold increase, respectively. Estragole and linalool's highest values compared with the control culture were 4.50 μg/g DW [AgNO3 (6µM) + SA (24 µM)] and 3.02 μg/g DW [SA (24 µM)], respectively. Results clearly show that the elicitor treatment could enhance the biosynthesis of phenolic compounds and terpenoid content in cell suspension cultures of O. basilicum and may be used for commercial supply in the future for therapeutic applications.

Shoot Induction, Multiplication, Rooting and Acclimatization of Black Turmeric (Curcuma caesia Roxb.): An Important and Endangered Curcuma Species

Abstract: Curcuma caesia Roxb., commonly known as Kali Haldi or black turmeric, is one of the important species in the genus Curcuma. This species has been classified as one of the endangered Curcuma species due to the drastic decrement of this plant in its natural habitat. C. caesia has been overharvested for various purposes, including bioactive compound extraction to fulfill the pharmaceutical industry demand. Hence, this study was conducted to establish a protocol for the propagation of C. caesia via plant tissue culture techniques. In the shoot induction stage, three basal medium formulations, including Murashige and Skoog (MS medium), the combination of Murashige and Skoog macronutrients and B5 micronutrients (MSB5 medium) and woody plant medium (WPM medium) supplemented with 15 μM of 6-benzylaminopurine (BAP), were used. The results found that the MSB5 medium was the most suitable basal medium formulation for shoot induction of C. caesia. In the subsequent experiment, different types of cytokinin, including BAP, kinetin and 2-iP at concentrations of 5, 10, 15 and 20 μM, were fortified in the MSB5 medium for shoot multiplication. The shoot multiplication was further enhanced by supplementing the MSB5 medium with indole-3-butyric acid (IBA) or 1-napthaleneacetic acid (NAA) at the concentrations of 2, 4, 6 and 8 μM. The results showed that a combination of 15 μM of BAP and 6 μM of IBA significantly increased the shoot multiplication with 100% shoot induction, 3.53 shoots/explant, 10.81 cm of shoot length, 9.57 leaves, 0.486 g of leaves fresh weight and 0.039 g of leaves dry weight. After the multiplication, the rooting stage was carried out by altering the basal medium strength into half and full strength and supplementing with 2.5, 5, 7.5 and 10 μM of indole-3-acetic acid (IAA). The full strength of MSB5 medium supplemented with 5 μM of IAA exhibited the highest number of roots and length of roots, with 6.13 roots and 5.37 cm, respectively. After the rooting stage, the plantlets were successfully acclimatized in the potting medium with the combination of cocopeat and peatmoss, and the ratio of 1:1 was found to produce the highest survival rate with 77.78%. In conclusion, the protocol established in this study could be useful for large-scale raw material production, either for conservation or bioactive compound extraction.

Screening on the Presence of Plant Growth Regulators in High Biomass Forming Seaweeds from the Ionian Sea (Mediterranean Sea)

Abstract: The use of seaweed as plant biostimulants is a solution for sustainable agriculture. The present study aims to quantify and compare the presence of plant growth regulators (PGRs) in four genetically labeled macroalgae growing in the Ionian Sea. Species were selected because they produce abundant biomass, disturbing ecological equilibrium and anthropic activities. We measured the content of gibberellic acid (GA3), kinetin (KN), indoleacetic acid (IAA), abscisic acid (ABA) and indole butyric acid (IBA). The method applied was modified from the literature to obtain simultaneously different PGRs from seaweed biomass in a shorter period of time. Among results, it is notable that Hypnea corona Huisman et Petrocelli (Rhodophyta) showed higher GA3 concentration, while in Spyridia filamentosa (Wulfen) Harvey (Rhodophyta), higher KN, IBA, IAA and ABA contents were recorded. The latter species displayed an interesting profile of PGRs, with an IAA value comparable with that reported in Ascophyllum nodosum (Linnaeus) Le Jolis (Ochrophyta), which is currently used as a source of plant biostimulants in agriculture. Macroalgae thrive abundantly in nutrient-rich environments, such as anthropized coastal areas affecting human economic activities. Consequently, environmental agencies are forced to dredge algal thalli and discard them as waste. Any use of unwanted biomass as an economic product is highly desirable in the perspective of ecosustainable development.