3D Tissue cultures

Narcissus tazetta bulbs were fertilized at 0, 4, 8 and 12 kg/da with DAP (diamonium phosphate) and the effects on bulb growth and flower quality criteria were studied. The highest dose applied in fertilization, 12 kg/da, yielded 3.93 bulb efficiency, 36.82 mm of bulb diameter, 212.45 mm of flower height, 18.38 mm of bunch height, 5.56 mm of stem thickness and 196.85 mm of leaf length,while the highest value of average bunch number, 2.69, was obtained at 0 kg/da of fertilization dose and the highest main bulb weight of 27.68 g was yielded at 4 kg/da.

The plants used in the phyto-pharmaceutical preparations are obtained mainly from the natural growing areas. With the increasing demand for the crude drugs, the plants are being overexploited, threatening the survival of many rare species. In addition, agriculture land decreasing day by day due to the real estate, industrialization, and roads for the betterment of human beings. To maintain the required demand of the important secondary metabolites and their sources; several research institutions and pharmaceutical industries using advanced biotechnological tools, this includes culturing of plant cells, genetic manipulation aiming to restore the germplasm, insertion of interest of genes for the production of important active principle. The present review article covering the in vitro micropropagation and production of selected secondary metabolites through biotechnological intervention viz. Alliin, Artimisnin, Podophyllotoxin, and Taxol.

Elimination of sugarcane grassy shoot disease (SGSD) through apical meristem culture technique for producing clean planting material of sugarcane has been attempted in the present study. The results showed that meristems length of 2 and 3 mm were free from the SGSD pathogen at higher frequency than larger meristem length of 4 mm. However, the frequency of survival of explants during initiation of shoot cultures was higher in larger meristems (60%) in comparison to smaller ones (40%). The micropropagated plantlets raised from meristem culture were confirmed for disease-free by nested polymerase chain reaction (PCR) analysis at monthly interval up to 6 months. This is the first report on the elimination of SGSD phytoplasma through meristem culture in India.

Background: The escalating global threat of antimicrobial resistance (AMR) necessitates the development of novel antimicrobial agents, innovative strategies, and representative infection models to combat AMR bacterial infections. Host defence peptides (HDPs) and their derivatives have been proposed as complements to conventional antibiotics due to their antibacterial activity and modulation of the immune response.
Objectives: This study investigated the novel use of the HDP-derived synthetic antibacterial and anti-biofilm peptide (SAAP)-148 as a pretreatment in epithelial tissue models to prevent colonization by AMR bacteria. The combined activities of SAAP-148 pretreatment with post-infection halicin to treat infections were also explored.
Methods: Employing cultured human skin equivalents (HSEs) and primary bronchial epithelial cells (PBECs) as models of tissue infection, we examined the prophylactic and therapeutic effects of SAAP-148, both singularly and in combination with the repurposed antibiotic halicin, against AMR bacteria. We additionally interrogated the response of HSE and PBEC cultures to SAAP-148 treatment via confocal microscopy and quantitative PCR of native HDPs and inflammatory cytokine genes.
Results: Our findings demonstrated that pretreatment with SAAP-148 significantly reduces colonization of HSEs and PBECs by AMR Staphylococcus aureus and Pseudomonas aeruginosa. Confocal microscopy revealed differential uptake and localization of SAAP-148 in these tissues, correlating with its distinct activity in these tissues. SAAP-148 exposure temporarily increased expression of the HDPs cathelicidin (CAMP) and β-defensin 1 (DEFB1), and the cytokine IL-8 (CXCL8), which did not correlate with the transient antibacterial activity observed. Sequential treatment with SAAP-148 prior to infection with AMR S. aureus and post-infection halicin treatment demonstrated synergistic activity in HSEs, whereas this combined activity was indifferent in PBEC cultures.
Conclusions: These results support SAAP-148 as a candidate for pre-infection prophylaxis and synergistic antibiotic therapy with halicin in skin, broadening the potential of both agents to address AMR bacterial infection.

Background/Aim: Malignant pleural mesothelioma (MPM) is a therapy-resistant neoplasm of the pleura. Standard chemotherapy consists of a combination of cisplatin (CPDD) and pemetrexed (PEM). The aim of this study was to assess whether inhibition of aerobic glycolysis by 2-deoxyglucose (2DG) would enhance the effects of standard chemotherapy. Materials and Methods: MeT-5A, M14K, MSTO and ZL34 cell lines were used. Cell viability with 2DG and cell proliferation and spheroid formation with CPDD+PEM alone and with 2-DG were tested. Results: Viability with 2-DG was dose-dependent. Cell proliferation with CPDD+PEM on 2D surface was reduced in all cell types, 2-DG inclusion demonstrated a synergistic effect in MSTO and ZL34 cells. Spheroid growth in 3D with CPDD+PEM or CPDD+PEM+2-DG lowered spheroid growth in all cell types. Conclusion: 2-DG synergizes with CPDD+PEM in lowering MPM cell proliferation in 2D to <20%. In 3D MPM spheroid growth 2-DG synergism with CPDD+PEM treatment is not maintained.

Coleus spp. have long been prized for their colorful foliage, which may combine shades of green, yellow, pink, red and maroon. They are plants for all locations. Besides their ornamental nature, Coleus spp. are also valued in folk medicine. C. blumei Benth., a natural hybrid of several Coleus spp. is an ornamental plant growing all over the world in an enormous number of different cultivars that vary in color and shape of the leaves, and is also known for its medicinal properties, in particular rosmarinic acid, one of its most important secondary compounds. Rosmarinic acid has been shown to exhibit antioxidative, anti-inflammatory and antiallergic activities in mammalian systems. C. forskohlii Briq. is a medicinally important plant valued for the production of forskolin, a diterpenoid, present in root tubers of the plant and known to lower blood pressure and intraocular pressure. C. parviflorus Benth. (country potato) has the potential of developing into a crop similar to potato. An...

Sugarcane (Saccharum officinarum L.) is monocotyledonous crop plant that belongs to the Poaceae family that mostly propagates through conventional methods. However, conventional propagation lacks rapid multiplication procedures to commercialize newly released varieties within a short period of time. Hence, the objective of this work was to optimize in vitro micro propagation protocol for two sugarcane varieties (C 86-165 and C 86-12) using shoot apical meristem explants. The two varieties were cultured on MS medium supplemented with different concentrations of growth regulators on shoot initiation, multiplication and root induction stages. Analysis of variance (ANOVA) revealed that the two varieties showed statistically significant difference in their response to the various hormonal treatments with regard to the parameters measured. For initiation stage vars. C 86-165 and C 86-12 performed best on 1.0 mg/l and 0.5 mg/l of BAP, respectively. On the other hand, multiplication stage was best in MS media enriched with 2.0 mg/l BAP + 0.5 mg/l NAA and 1.5 mg/l BAP + 0.5 NAA as manifested in terms of number of shoots and shoot length per explants after four weeks of culture for vars. C 86-165 and C 86-12, respectively. With regard to root induction, best rooting response in terms of mean root number, mean root length, mean fresh weight and mean dry weight was achieved in 1/2 MS media enriched with 2.0.mg/l NAA and 1.0 mg/l NAA after four weeks of culture for vars. C 86-165 and C 86-12, respectively.

Having a great genetic diversity, shape and color, Narcissus is a bulbous ornamental geophyte scattered from Europe to East Asia. Beside their ornamental value, Narcissus species produce alkaloids which can be used for the treatment of neurological diseases such as Alzheimer's by biotechnological approaches. The main issue in commercializing this plant is its very slow natural vegetative propagation rate. Manipulated vegetative propagation approaches such as chipping and twin scaling are time consuming and usually altered with degeneration and virus and disease infection. Therefore producers are led to micropropagation techniques which are more productive and efficient. In this chapter many biotechnological applications for Narcissus such as in vitro culture, somatic embryogenesis, gene transfer and molecular markers besides mutation induction, virus-free stock production and cytogenetics of Narcissus are discussed.

Biology, damage pattern, incidence levels and status of the leaf miner Aphanisticus aeneus Kerremans (Coleoptera: Buprestidae) were studied in accessions of Saccharum spp. Egg, grub and pupal periods of the miner lasted 6.0 § 0.8, 18.5 § 1.9 and 7.0 § 1.3 days, respectively; adults lived for 8À10 days. The grub tunneled the leaf on the lower surface just above the epidermis forming a linear channel; infested tissues showed brownish discoloration. The extent of leaf miner attack among the 331 accessions of Saccharum spp. ranged 4.00%À76.47% on cane basis and 7.14%À100.00% on leaf basis; the mined leaf area ranged 3.22À11.00 cm 2. When the accessions were grouped on the basis of an infestation index developed with the above three parameters, 267 occupied very low category, 51 were in low category, 11 were in moderate category and 2 were placed in high incidence category. All accessions showed miner attack in at least one study year. Within each of the four Saccharum spp., accessions differed significantly among the three study years for all infestation parameters. Among the four species, while S. robustum accessions recorded significantly highest infestation index, the other three species remained on par.

The purpose of this study was to determine the antibacterial activity of C. officinalis callus derived from cotyledon explants. Cotyledons excised from in vitro germinated seedlings were used as explants. Explants were transferred on MS medium supplemented with benzil amino purine (BAP; 2 mg l-1), α-naphthalene-acetic acid (NAA; 2 mg l-1) for callus studies. The cultures were maintained on the same media compositions and were subcultured at an interval of 4 weeks. Callus cultures were harvested at the end of the 16th week. Calli were dried at 40̊ C in the dark for antimicrobial studies. Calendula officinalis callus extracts were tested for their antibacterial activities by using agar well diffusion method. Ethanol and chloroform extracts from these plants were assayed against nine bacteria species (

The purpose of this study was to determine the antibacterial activity of C. officinalis callus derived from cotyledon explants. Cotyledons excised from in vitro germinated seedlings were used as explants. Explants were transferred on MS medium supplemented with benzil amino purine (BAP; 2 mg l-1), α-naphthalene-acetic acid (NAA; 2 mg l-1) for callus studies. The cultures were maintained on the same media compositions and were subcultured at an interval of 4 weeks. Callus cultures were harvested at the end of the 16th week. Calli were dried at 40̊ C in the dark for antimicrobial studies. Calendula officinalis callus extracts were tested for their antibacterial activities by using agar well diffusion method. Ethanol and chloroform extracts from these plants were assayed against nine bacteria species (

In a bioassay-directed screening programme of plants for the identification of active constituents the following steroids were isolated. Chonemorphine was identified as the antiamoebic and antitrichomonad principle of Chonemornha fraarans. St&master01 from Coleus forskohlii and ecdysterone from Dioloclisia glaucescens were inactive constituents isolated during the process of purifying the active principles of the plants. D. slaucescens roots are a high-yielding source (0.5% of the dry root weight) of ecdysterone.

Propagation of some ornamental plants has increased tremendously due to the demand for them as cut flowers, in addition to their usage for interior and exterior landscaping purposes. Geophytes (bulbous-tuberous) are the most preferred group among the ornamental plants due to their aesthetic features, suitability to be cut flowers and their fragrance. These plants are highly propagated and consumed. The geophyte species which are highly profitable globally traded and constituting 90% of the flower bulb market are Tulipa (tulip), Lilium (lily), Narcissus (daffodil), Gladiolus (gladioli), Iris (iris) and Hyacinthus (hyacinth). In vitro propagation techniques, which provide disease-free mass production options, have started to be used increasingly to fulfil the demand for these species in the market. In this study, the results of in vitro propagation studies for some economically valuable tulip, lily, daffodil, gladiolus, iris and hyacinth species are provided.

Twin scale explants of Pancratium maritimum were cultured on MS supplemented with BA and NAA, each at concentrations of 0, 5, 10 or 20 µM, alone or in combination. Each explant produced 3-4 adventitious bulblets when cultured on MS with or without BA, whereas, in all combinations of BA and NAA (or NAA alone) the number of bulblets per explant was reduced. Bulblets derived from explants without growth regulators were either transferred to the soil or subcultured in vitro on MS supplemented with various combinations of BA and NAA, each at concentrations of 0, 0.5 or 5 µM. Plantlets with leaves and roots were produced on MS with the addition of 5 µM NAA alone or with 0.5 µM BA, which then were transferred to the soil. Only 15% of the plantlets survived after transplantation, while 100% of the bulblets of more than 200 mg and 69% of the bulblets of 100-200 mg survived for at least 12 months.

Secretion of phenolic compounds is a major limitation for sugarcane in vitro shoot culture, causing a loss of regenerative capacity and subsequent cell death. In this study, micropropagation and phenolic secretion of four Saccharum genotypes were evaluated in presence of different antioxidants. Aseptic cultures of S. officinarum (PI 184794 and PI 88652), S. sinense (PI 29109) and S. robustum (UNK R65P35) were propagated on medium containing antioxidants, citric acid (100 mg/L), L-cysteine (100 mg/L), polyvynylpirrolidone (300 mg/L) and L-glutathione (50 mg/L) in two consecutive subculture cycles. Interaction between genotypes and antioxidants was significant in both cycles. All genotypes showed good shoot formation, shoot vigor and color, except in PI 88652 which had less shoot development in both the presence and absence of the antioxidants tested. PI 184794 displayed the highest shoot proliferation in the presence of citric acid, and UNK R65P35 produced more shoots per explant in ...

The influence of varied concentrations of sucrose and ammonical (NH 4 ?) nitrogen on in vitro induction and expression of anthocyanin pigments from Rosa hybrida cv. 'Pusa Ajay' was investigated. Of two explants (petal and leaf discs) selected and cultured under two different conditions (light and dark), leaf discs were found to be most suitable for callus initiation. Profuse and early callus induction was observed when leaf discs of rose were cultured under total dark conditions on solid Murashige and Skoog (MS) medium supplemented with 4.0 mg l-1 2,4-dichlorophenoxyacetic acid (2,4-D). Early pigment initiation, enhancement and maximum anthocyanin production from calluses were recorded when leaf discs were cultured on Euphorbia millii (EM) medium supplemented with 7% sucrose compared with calluses cultured at 4% sucrose concentration under 16/8 h (light/dark) photoperiod regime. Reducing the concentration of NH 4 ? nitrogen in the solid MS medium led to slight improvement in anthocyanin production in rose leaf calluses. Keywords Ammonical nitrogen Á EM medium Á Sucrose concentration Á Rosa hybrida Á Anthocyanin pigments Abbreviations 2,4-D 2,4-Dichlorophenoxyacetic acid ANOVA Analysis of variance EM Euphorbia millii medium FW Fresh weight MS Murashige and Skoog medium NH 4 ?

For a long period of time, nature has been a great source of medicinal agents. In present study, antibacterial and antioxidant activity of different extracts of Spilanthes acmella L. were evaluated in-vitro by using Agar-well diffusion method and DPPH free radical scavenging assay respectively. Acetone, methanol and water extracts of different plant parts (leaf, flower and stem) were screened for their antibacterial activity against four pathogenic bacteria (Bacillus cereus, Escherichia coli, Salmonella typhi and Staphylococcus aureus) using different concentrations i.e. 25, 50, 75 and 100%. Results showed low to significant antibacterial activity against tested bacterial species. S. typhi was the most susceptible bacteria. In addition, the antioxidant potential of the different extracts of this plant was evaluated by DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging assay. Methanol extracts of all plant parts exhibited higher antioxidant capacity with lesser IC 50 values than acetone and water extracts. The results provided evidence confirming the fact that the studied plant could act as a potent source of antimicrobials and antioxidants.

In vitro bulbification is a method by which bulb formation is stimulated and speeded up in the species that are propagated by bulbs and which need, in the conditions of a classicel culture, a longer period for the formation of flower-bearing bulbs. In this article, the results of research is presented, regarding the conservation, multiplication and maintenance of this vulnerable species included in the Red Book of the Republic of Moldova. This study was carried out in order to re-introduce this species into its native habitats. Optimal media were elaborated for the initiation, multiplication, rhyzogenesis and the maintenance of this species in in vitro conditions.

ABSTRACTHydrogenated diamond-like-carbon (a-C:H) and hydrogen-free amorphous carbon (a-C) coatings are known to be biocompatible and have good chemical inertness. For this reason, both of these materials are strong candidates to be used as a matrix that embeds metallic elements with antimicrobial effect. In this comparative study, we have incorporated silver into standard diamond-like carbon (DLC) coatings by plasma ion implantation and deposition (PIII&D) using methane (CH4) plasma and simultaneously depositing Ag from a pulsed cathodic arc source. In addition, we have grown amorphous carbon – silver composite coatings using a dual-cathode pulsed filtered cathodic-arc (FCA) source. The silver atomic content of the deposited samples was analyzed using glow discharge optical spectroscopy (GDOES). In both cases, the arc pulse frequency of the silver cathode was adjusted in order to obtain samples with approximately 5 at.% of Ag. Surface hardness of the deposited films was analyzed usi...

Produksi metabolit sekunder pada tanaman biasanya menghasilkan kadar yang rendah. Metode bioteknologi telah terbuktidapat meningkatkan produksi beberapa metabolit sekunder pada tanaman. Untuk meningkatkan perolehan metabolit sekunder telah digunakan teknik kultur jaringan dan transformasi genetik dengan induksi Agrobacterium rhizogenes. Penelitian ini bertujuan untuk meningkatkan kandungan metabolit sekunder dari kultur kalus dan akar rambut dari tanaman Artemisia annua hasil transformasi genetik menggunakan A. rhizogenes. Kultur kalus dan akar rambut hasil transformasi genetika mengandung senyawa artemisinin lebih tinggi dibanding dengan kultur kalus dan akar tanpa transformasi.Kata Kunci : Artemisia annua, kultur kalus, akar rambut Agrobacterium rhizogenes, artemisinin. The production of secondary metabolites of plant is usually low. Biotechnological methods have been proved to enhance the production of some of plant&#39;s secondary metabolites. To enhance the production of second...

Stevia rebaudiana is a perennial herb belonging to Asteraceae family and an excellent substitute of artificial sugar. Increasing demand of quality panting material has emphasized the need of rapid and mass multiplication through plant tissue culture technology in limited time duration. Sterilized in vivo leaves were used as leaf explants on different concentration of BAP, Kin and TDZ. In all media, shoot induction was not observed at any concentration from in vivo leaf explants. Cold treatment (4 oC) of different time duration (0-72 hr) was applied on in vitro leaf explants and cultured on various concentration of TDZ (1.0-5.0 mg/l). Highest shoot induction response (42.40%) was occurred at 4.0 mg/l TDZ from 72 hrs cold treated in vitro leaf. Highest number of shoots per explant (25.80) was observed at 3.0 mg/l TDZ from 48 hrs cold treated in vitro leaf explants. Highest shoot length (1.36 cm) was observed at 3.0 mg/l TDZ from 72 hrs cold treated in vitro leaf. Cold treatment was us...

The present investigation was carried out at Anand Agricultural University, Gujarat during the year 2016 to develop a micropropagation protocol for mass multiplication of medicinally rich stevia plants. Axenic culture was established by sequential application of 200 mg/l Kanamycin, 200 mg/l Carbendazim-50% and 1000 mg/l HgCl2. Out of two different basal media, viz. Gamborg (B5) and Murashige and Skoog (MS) used, full strength MS gave highest (98%) culture establishment. Out of three different basal media, viz. Gamborg’s (B5), Linsmair and Skoog (LS), Murashige and Skoog (MS) and four different cytokinins, viz. 6-benzylaminopurine (BAP), Kinetin (Kn), Thiadiazuron (TDZ), Zeatin (Zn) tested for multiple shoot induction, full strength MS medium and 2 mg/l BAP, respectively, promoted maximum shoot formation. In the experiment to study the effect of varying strength of MS media and various plant growth regulators, viz. Indole-3-acetic acid (IAA), Indole-3-butyric acid (IBA), Naphthalene ...

Agarwood (Aquilaria malaccensis) is a scare plant with high economic value and many benefits. Tissue culture is an effective way of producing plants in large quantities, in a short time with uniform results. This study aims to get the proper concentration of IAA and yeast extract for agarwood growth in vitro. The research was conducted in

Plants are one of the resources, part of life, part of the ecosystem which is very important in efforts to improve and maintain public health. Even today, according to estimates (WHO), 80% of the world's population still depend on traditional medicine, including the use of drugs derived from plants. Until now, a quarter of modern medicines circulating in the world are derived from active ingredients isolated and developed from plants. Currently it is known that around 30% of drug sales worldwide are medicines derived from natural ingredients. In the United States, for example, about 25% of prescription drugs are mainly composed of natural products derived from plants and their derivatives.

Brief Reports 855 Diploicin, a chlorinated depsidone related to orcinol, previously not reported to be present in Pannariaceae, was found in P~w~m a dimorpbum and leprolomin, a diary1 ether previously described (4), was isolated from P. dimmphum and Psmmapallidum. All these compounds were characterized from 'H-nmr, ms, and ir spectral data and by comparison with authentic samples. EXPERIMENTAL GENERAL EXPERIMENTAL PRocEDURES.-Melting points were determined on a Kotler hot plate. Spectra were recorded with the following instruments: ir, Perkin-Elmer model 683; 'H nmr, Varian T-60; ms, Varian MAT CH-7.

Germplasm conservation of elite Stevia rebaudiana Bert. was attempted by using synthetic seed technology and medium with added osmotic agents. Explants of nodal segments containing single axillary bud were excised from in vitro proliferated shoot cultures and encapsulated in high-density sodium alginate (5%) hardened by 50 mM CaCl 2. The encapsulated and non-encapsulated nodal segments were stored at 5, 15 and 25°C for 4, 8 and 12 months and monitored for the re-growth and survival frequency under the tissue culture conditions on Murashige and Skoog (MS) medium supplemented with thidiazuron (TDZ 0.2 mg L-1). Encapsulated nodal segments could be stored at 25°C up to eight months with maximum regrowth ability and survival frequency of 77%. The highest re-growth in non-encapsulated cultures was observed in the explants kept at 25°C without osmotic agents, however, with a lower survival frequency. The effect of osmotic agents dextrose, mannitol and sorbitol on non-encapsulated shoot cultures was also evaluated. A considerable decrease in re-growth and survival was observed in the cultures treated with osmotic agents. Among the cultures treated with osmotic agents, the highest rate of regrowth and survival was observed at 2% sorbitol followed by 2% sorbitol + 2% dextrose. Well-developed plantlets regenerated from encapsulated nodal segments were successfully acclimatized inside the growing room with 87% survival. Highperformance liquid chromatography was used to assess the stability in chemical profile and quantification of rebaudioside A and stevioside content of mother plant (MP) and the plants grown through synthetic seeds after 4, 8 and 12 months of storage under slow growth conditions. Our results show no significant differences (p < 0.05) in rebaudioside A and stevioside content between the mother and re-grown plants following in vitro storage.

Respiratory viral infections, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), are among the most common illnesses and a leading cause of morbidity and mortality worldwide. Due to the severe effects on health, the need of new tools to study the pathogenesis of respiratory viruses as well as to test for new antiviral drugs and vaccines is urgent. In vitro culture model systems, such as three-dimensional (3D) cultures, are emerging as a desirable approach to understand the virus host interactions and to identify novel therapeutic agents. In the first part of the article, we address the various scaffold-free and scaffold-based 3D culture models such as hydrogels, bioreactors, spheroids and 3D bioprinting as well as present their properties and advantages over conventional 2D methods. Then, we review the 3D models that have been used to study the most common respiratory viruses including influenza, parainfluenza, respiratory syncytial virus (RSV) and coronaviruses...

Natural product that has developed as anticancer drugs are consists from secondary metabolites that has isolated from plant, ocean plant and microorganisms. To maintain its productivity, must be concerned to the cultivation of the medicinal plants. Medicinal plants need time to grow and producing secondary metabolites that would be isolated, so that the production of secondary metabolites compound from plant is very limited. The plants that infected by endophytic fungi be able to grow faster than the normal plants. Endophytic fungi also have ability producing secondary metabolites compound that produced by the host plant. Taxol, podophyllotoxin and camptothecin are compounds that proved has anticancer activity. Endophytic fungi have also proved increasing the production of the compound. Key Word: Metabolites Compound, anti cancer, endophytic fungi

An efficient high frequency plant regeneration protocol through direct organogenesis was developed for Sevia rebaudiana Bert. Nodal segments containing axillary buds were used as an explant and inoculated on Murashige and Skoog's (MS) medium containing 3% (w/v) sucrose, 0.8% (w/v) agar supplemented with various concentrations of benzyladenine (BA), kinetin (Kn) and thidiazuron (TDZ) ranging from 1.00 to 9.00 µM. Maximum multiple shoots (96%) were obtained in MS medium supplemented with 1.0 µM TDZ with an average of 60 shoots per culture, having an average shoot length of 6.0 cm. The best in vitro root induction (89%) was achieved on half strength MS medium without any growth regulator with an average of 24 roots per culture and root length of 7 cm. The rooted plantlets were successfully established in soil and grown to maturity at the survival rate of 95% in the indoor grow room. High-performance liquid chromatography was used to assess the stability in chemical profile and quantification of stevioside and rebaudioside A content of in vitro propagated S. rebaudiana plants and compared with their mother plant at the peak vegetative stage. Our results show no significant differences (p < 0.05) between the mother and in vitro propagated plants. Furthermore, fully developed in vitro propagated S. rebaudiana plants were also compared with mother plant for their gas and water vapour exchange characteristics and leaf anatomy. The results show that in vitro propagated and hardened plants of S. rebaudiana are morphologically as well as functionally comparable to each other and to their mother plant.

The effect of different culture media with different growth regulator and nitrate/ammonium ratios on split garlic shoots was studied to obtain an optimized micropropagation protocol. Three basal media: MS, BDS and BLM (i.e. BDS modi®ed to obtain a nitrate:ammonium ratio of 35 mM:8 mM) combined with two concentrations of NAA (0.5 or 5 mM) and BAP (1 or 10 mM) were evaluated in three Colorado garlic clones: Espan Äol Seleccio Ân Ascasubi (ESA), Espan Äol Seleccio Ân Me Âdanos (ESM) and I 50. In BLM medium with 5 mM NAA and 10 mM BAP the highest multiplication rates were 140, 542 and 743 shoots per split shoot after three subcultures in 140 days for clones ESA, ESM and I 50, respectively. Comparing the mean rates obtained in MS medium with those obtained in BDS medium in the same time, the latter produced 1.5-, 4-and 2.2-fold increases in the mean multiplication rates for ESA, ESM and I 50 clones, respectively; while BLM produced 5-, 3-and 10-fold increases in the mean multiplication rates. Since BDS and BLM media only differ in their NO 3 À :NH 4 ratio we can conclude that the differences in the multiplication rates could probably be due to an increase in the NO 3 À level of the BLM medium and so the multiplication ratios for these garlic clones can be greatly improved by the use of higher levels of nitrogen supplied as nitrate (35 mM NO 3 À :8 mM NH 4 ratio) with higher concentrations of growth regulators such as 5 mM NAA and 10 mM BAP.

The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content. This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden.

An efficient high frequency plant regeneration protocol through direct organogenesis was developed for Sevia rebaudiana Bert. Nodal segments containing axillary buds were used as an explant and inoculated on Murashige and Skoog's (MS) medium containing 3% (w/v) sucrose, 0.8% (w/v) agar supplemented with various concentrations of benzyladenine (BA), kinetin (Kn) and thidiazuron (TDZ) ranging from 1.00 to 9.00 µM. Maximum multiple shoots (96%) were obtained in MS medium supplemented with 1.0 µM TDZ with an average of 60 shoots per culture, having an average shoot length of 6.0 cm. The best in vitro root induction (89%) was achieved on half strength MS medium without any growth regulator with an average of 24 roots per culture and root length of 7 cm. The rooted plantlets were successfully established in soil and grown to maturity at the survival rate of 95% in the indoor grow room. High-performance liquid chromatography was used to assess the stability in chemical profile and quantification of stevioside and rebaudioside A content of in vitro propagated S. rebaudiana plants and compared with their mother plant at the peak vegetative stage. Our results show no significant differences (p < 0.05) between the mother and in vitro propagated plants. Furthermore, fully developed in vitro propagated S. rebaudiana plants were also compared with mother plant for their gas and water vapour exchange characteristics and leaf anatomy. The results show that in vitro propagated and hardened plants of S. rebaudiana are morphologically as well as functionally comparable to each other and to their mother plant.

Stevia rebaudiana Bertoni is a medicinal plants and commercially use as non-caloric sweetener for diabetic patient. In the present study, a protocol was developed for in vitro micropropagation using 6-benzylamino purine (BAP) and Kinetin (Kn) for the formation of multiple shoot proliferation and Indole-3-acetic acid (IAA), Indole-3-butyric acid (IBA) and 1-Naphthaleneacetic acid (NAA) for the induction of roots. Maximum shoot formation (7.82 ± 0.7 shoots per explants) was observed on a Murashige and Skoog (MS) medium supplemented with 0.5 mg L-1 BAP and 0.25 mg L-1 Kn. The maximum number of roots (30.12 ± 2.1 roots per explants) was obtained on a MS medium containing 1.0 mg L-1 IBA. The well rooted plantlets were successfully weaned and acclimatized in plant soil with survival rate of 83.3 %.

A procedure for in vitro production of bulbs of Narcissus bulbocodium was developed. For the induction of shoots, twin-scales were used as primary explants and four media were compared. Using the basal MS medium supplemented with BAP (4 mg/l)NAA (0.12 mg/l) or BAP (2 mg/l) IBA (1 mg/l) resulted in shoot initiation and leaf development. The first medium also caused the production of amorphous achlorophyllous tissue at the base of the shoots to a greater extent than the second medium. Both media were also appropriate for abundant shoot proliferation and when shoot clumps were cultured on the medium with BAPIBA for a long period (70 days) tiny bulbs developed at the base of senescent leaves. The growth of these bulbs was achieved after two additional periods of culture on similar basal medium with or without NAA (0.12 mg/l), both with an increased sucrose content (9%). The presence of NAA did not significantly increase the final size of the bulbs but caused the development of a better root system. Bulbs grown on the media with 9% sucrose with 8.0 to 14.5 mm in diameter, after fungicide treatment, were transplanted into a soil mixture and grown for 1 month under a 128C/98C regime to release the dormancy. Then, they were transferred to a glasshouse where they produced normal plants. The evaluation in three experiments, involving 3600±3720 bulbs, gave a figure of 92%±95% transplantation success and 90% of the bulbs flowered during this first growing season. The procedure described here allows an in vitro bulb production adapted to a successful transplantation and with the capacity to flower in the first growing season.

The present investigation was carried out with the objective to the assess the nature and magnitude of variability and extent of genetic divergence among different genotypes of narcissus on the basis of vegetative, floral and bulbous traits. Experimental results revealed significant differences among nine genotypes for different traits under study. Earliest scape emergence (64.43 days) and flowering (80.57 days) and highest bulb size (4.22 cm), weight of bulb (67.86 g) and bulblets (14.27 g) was recorded in the genotype Cumlaude, while largest flower size (8.37 cm) was observed in the cultivar Carlton. Further, significant positive correlations of scape length (0.825), weight of bulb (0.914) and root length (0.876) was recorded with bulb size. The principal component analysis (PCA) revealed that 90.97 % of total variations were explained by first four components. Further, loading of different traits based on first two principal components indicated that days to goose neck stage, bul...

The objective of this study is to improve total alkaloid contents of Narcissus tazetta var. italicus tissue cultures by using different types of growth regulators and fungal elicitors. Seven and six species belong to four genera were isolated from both rhizosphere and bulbs of N. tazetta var. italicus, respectively. Aspergillus niger, A. ochraceus, A. oryzae, Fusarium oxysporum, F. sporotrichioides, Penicillium. chrysogenum and Trichoderma viride were isolated from the rhizosphere. On the other hand, A. flavus, A. niger, F. oxysporum, F. semitectum, P. chrysogenum and T. viride were isolated from the bulbs. Explants derived from bulbs and leaves of N. tazetta var. italicus were cultured on Murashige and Skoog medium (MS) supplemented with different combinations and concentrations of plant growth regulators. Calli derived from both bulb and leaf explants, and bulbs derived from bulb explants, had maximum growth and highest amount of total intracellular alkaloids, when cultured on MS4, MS6 and MS2 media, respectively (without the addition of fungal elicitor). MS4 contained benzyl adenine (BA 1.5 mg/l) and naphthalene acetic acid (NAA 3 mg/l), MS6, contained BA (2 mg/l) and indole acetic acid (IAA 0.5 mg/l), whereas, MS2 contained BA (2 mg/l) and indole butyric acid (IBA 1mg/l). The isolated fungi were screened for their potentiality to elicit alkaloids production. F. sporotrichioides culture filtrate was the most effective elicitor in enhancing growth and total intracellular alkaloids production when incubated on MS4 medium for 10 days with 4 week old callus cultures of N. tazetta var. italicus.

To optimize conditions for micropropagating Galanthus transcaucasicus Fomin, the effect of explant type, different concentrations of sucrose, type and different concentrations of auxin were examined on bulblet production. For the induction of bulblets, three types of bulb scales were usesd as explants. A two-step sterilization procedure was applied. Assessments were made after incubation for 20 weeks at 23±1oC with 16h photoperiods. Highest number of bulblets were proliferated culturing bulb scales on  modified MS medium supplemented with 2.0 mg.l-1 BA and 2.0 mg.l-1 IBA. About  27% of bulblets were bigger than 5 mm in diameter, suitable for acclimatization. With the increase of sucrose concentration, number of bulblets decreased, but their size increased. This protocol provided a basis for future study on large-scale multiplication system for commercial nurseries of Galanthus transcaucasicus.

Respiratory viral infections, including severe acute respiratory syndrome coronavirus 2
(SARS-CoV-2), are among the most common illnesses and a leading cause of morbidity and mortality
worldwide. Due to the severe effects on health, the need of new tools to study the pathogenesis of
respiratory viruses as well as to test for new antiviral drugs and vaccines is urgent. In vitro culture
model systems, such as three-dimensional (3D) cultures, are emerging as a desirable approach to
understand the virus host interactions and to identify novel therapeutic agents. In the first part of the
article, we address the various scaffold-free and scaffold-based 3D culture models such as hydrogels,
bioreactors, spheroids and 3D bioprinting as well as present their properties and advantages over
conventional 2D methods. Then, we review the 3D models that have been used to study the most
common respiratory viruses including influenza, parainfluenza, respiratory syncytial virus (RSV)
and coronaviruses. Herein, we also explain how 3D models have been applied to understand the
novel SARS-CoV-2 infectivity and to develop potential therapies.

Hydrogenated diamond-like-carbon (a-C:H) and hydrogen-free amorphous carbon (a-C) coatings are known to be biocompatible and have good chemical inertness. For this reason, both of these materials are strong candidates to be used as a matrix that embeds metallic elements with antimicrobial effect. In this comparative study, we have incorporated silver into diamond-like carbon (DLC) coatings by plasma based ion implantation and deposition (PBII&D) using methane (CH 4) plasma and simultaneously depositing Ag from a pulsed cathodic arc source. In addition, we have grown amorphous carbon-silver composite coatings using a dualcathode pulsed filtered cathodic-arc (FCA) source. The silver atomic content of the deposited samples was analyzed using glow discharge optical spectroscopy (GDOES). In both cases, the arc pulse frequency of the silver cathode was adjusted in order to obtain samples with approximately 5 at.% of Ag. Surface hardness of the deposited films was analyzed using the nanoindentation technique. Cell viability for both a-C:H/Ag and a-C:/Ag samples deposited on 24-well tissue culture plates has been evaluated.

Stevia rebaudiana (Bertoni), known to be an anti-diabetic medicinal herb, belongs to Asteraceae family which has bioactive compounds stevioside and rebaudioside in its leaves that taste about 300-350 times sweeter than sucrose. Nodal segments were cultured in half-strength Murashige and Skoog (MS) medium supplemented with kinetin (Kn) and benzylaminopurine (BAP), alone and in combination (0.2, 0.5, 1.0, 2.0 and 3.0 mg L-1), for shoot proliferation while various concentrations of auxins i.e. indole acetic acid (IAA) and indole butyric acid (IBA) (0.2, 0.5, 1.0, 2.0 and 3.0 mg L-1), alone or in combination with different cytokinins, for rooting. Almost in all the cultures shoot primordia initiation was observed 2-5 days after the inoculation. Overall, from the economical point of view, half-strength MS media with 0.2 mg L-1 Kn was the best producing maximum number of shoot (3.00 ± 0.00), shoot length (7.13 ± 0.43) and number of leaves (27.75±0.85), after 60 days of inoculation. However, after 30 days of transfer in rooting media supplemented with combined IBA (2.0 mg L-1) and Kn (0.5 mg L-1) showed best response in number of roots (10.00 ± 0.41) and length of root (cm) (6.73 ± 0.09), respectively. Regenerated plantlets were hardened in potting mixture containing perlite: sand: soil (1:1:1) for 2 months inside culture room with 70% relative humidity and later transferred in green house for 8 weeks before final transfer into the field. This micro-propagation method using half-strength MS media could be economical and effectively used for mass production of Stevia rebaudiana under in vitro condition.

Stevia rebaudianaisnative plant of tropical climate of Brazil and Paraguay. Its leaves produce a sweetener -stevioside which is 150-300 times sweeter than cane sugar. An attempt is made to grow this plant in hot semi arid climate of Lahore region of Pakistan through tissueculture technique .A completely randomized design was used for the experiment with three replicates. The data was analyzed by applying on

A procedure for in vitro production of bulbs of Narcissus bulbocodium was developed. For the induction of shoots, twin-scales were used as primary explants and four media were compared. Using the basal MS medium supplemented with BAP (4 mg/l)+NAA (0.12 mg/l) or BAP (2 mg/l)+IBA (1 mg/l) resulted in shoot initiation and leaf development. The first medium also caused the production of amorphous achlorophyllous tissue at the base of the shoots to a greater extent than the second medium. Both media were also appropriate for abundant shoot proliferation and when shoot clumps were cultured on the medium with BAP+IBA for a long period (70 days) tiny bulbs developed at the base of senescent leaves. The growth of these bulbs was achieved after two additional periods of culture on similar basal medium with or without NAA (0.12 mg/l), both with an increased sucrose content (9%). The presence of NAA did not significantly increase the final size of the bulbs but caused the development of a bette...

Narcissus tazetta L., a bulbous plant belongs to the Amaryllidaceae family, contains alkaloid galantamine (GAL) with acetylcholinesterase inhibitory activity which has been recently considered to treat Alzheimer's disease (AD). In the current work, the effect of photoperiod (16/8 h light/dark and 24 h dark) and various concentrations of NAA, BAP, and GA 3 (0, 0.5, 1 and 2 mg l-1) on the in vitro mass bulblet regeneration of N. tazetta was studied. The GAL production ability of the regenerated bulblets was assessed by HPLC-UV-MS. Light treatments significantly affected the number of bulblet and leaf, the ratio of bulblet/leaf, and leaf length. The maximum number of bulblet (31.0 ± 1.58) and leaf (13.3 ± 1.33) was recorded from the cultures fortified with NAA and BAP (2 mg l-1) kept in 16/8 h light/dark, while the maximum leaf length (2.1 ± 0.92 cm) was measured on the MS medium containing 0.5 mg l-1 NAA and 2 mg l-1 BAP incubated in the same photoperiod. The average ratio of bulblet proliferation per explant was significantly different between studied photoperiod (1.1 ± 0.86) and 24 h dark (0.62 ± 0.31). The regenerated bulblets contained 40 and 20 µg g-1 DW GAL underexposed photoperiod and 24 h dark, respectively. This information could be useful in the commercial production of GAL as a valuable anti-AD compound through in vitro mass bulblet proliferation of N. tazetta. Key message The regenerated mass bulblets of Narcissus tazetta (Amaryllidaceae) on MS medium containing 2 mg l-1 NAA and BAP kept in 16/8 h light/dark are recommended to produce galanatamine and lycorine.

Narcissus tazetta bulbs were fertilized at 0, 4, 8 and 12 kg/da with DAP (diamonium phosphate) and the effects on bulb growth and flower quality criteria were studied. The highest dose applied in fertilization, 12 kg/da, yielded 3.93 bulb efficiency, 36.82 mm of bulb diameter, 212.45 mm of flower height, 18.38 mm of bunch height, 5.56 mm of stem thickness and 196.85 mm of leaf length,while the highest value of average bunch number, 2.69, was obtained at 0 kg/da of fertilization dose and the highest main bulb weight of 27.68 g was yielded at 4 kg/da.

From immature embryos of Azadirachta indica A. Juss, callus was developed with 100% response on MS medium supplemented with (a) 2.0 mg J-1 NAA + 0.5 mg 1·1 BA, (b) 2 mg J-1 BA and (c) 1.0 mg J-lAA + 0.5 mg J-1 BA. For optimum multiple shoot formation, medium containing (a) 2.0 mg )·1 BA and (b) 0.5 mg J·l BA + 0.1 kgl· NAA was used which rooted in 1/2 strength MS medium supplemented with 0.5 mgJ-1 IBA. Azadirachtin and nimbin content were analysed in callus, in vitro cultured roots and shoots. Roots had 0.004 mg/g azadirachtin and shoots had 0.008 mg/g azadirachtin and 0.003 mg/g nimbin. Callus had negligible amount of azadirachtin.

Seedling tissues of Artemisia annua L., a medicinally important plant in Iran, were infected with Agrobacterium rhizogenes strains AR15834 for induction of hairy roots. The produced neoplastic roots was characterized by high growth rate, genetic stability and producing higher levels of secondary metabolites. Hairy roots induction in this plant was investigated using leaves of 2 weeks old and 2 months old of A. annua as explants and A. rhizogenesstrain led to the induction of hairy roots in the explants. Transformation frequency of the 2 weeks old and the 2 months old leaf explants were 75.60% and 6.66% respectively. The morphological identification of the transformed hairy roots was confirmed by PCR using rolB gene specific primers. The obtained hairy root lines of A. annua showed 3 different morphological phenotypes: typical transformed roots, callus and callus-like roots. The typical transformed hairy roots were established and grown in liquid medium for elicitation treatments with different concentration of chitosan (50,100 and 150 mg/L) and jasmonic acid (50, 100 and 200 µm/L). The results revealed that, these elicitors did not show any significant effect on the biomass of obtained hairy roots but there was significant difference between hairy roots and non-hairy roots biomass.

To optimize conditions for micropropagating Galanthus transcaucasicus Fomin, the effect of explant type, different concentrations of sucrose, type and different concentrations of auxin were examined on bulblet production. For the induction of bulblets, three types of bulb scales were usesd as explants. A two-step sterilization procedure was applied. Assessments were made after incubation for 20 weeks at 23±1oC with 16h photoperiods. Highest number of bulblets were proliferated culturing bulb scales on  modified MS medium supplemented with 2.0 mg.l-1 BA and 2.0 mg.l-1 IBA. About  27% of bulblets were bigger than 5 mm in diameter, suitable for acclimatization. With the increase of sucrose concentration, number of bulblets decreased, but their size increased. This protocol provided a basis for future study on large-scale multiplication system for commercial nurseries of Galanthus transcaucasicus.