Murashige-Skoog Medium Research Articles

Agrobacterium tumefaciens-Mediated Stable Transformation of Daucus carota.

Daucus carota L. (carrot) is one of the ten most important vegetables cultivated and consumed worldwide and is a main source of provitamin A. Carrot storage root is rich in dietary fiber, antioxidants, and other nutrients but especially in carotenoids. It has been also used as plant model for studding embryogenesis, as well as the genetic and genomic evolution of carrots and for carotenoid synthesis regulation, among others. Research in carrot often needs genetic transformation. Here we describe a step-by-step protocol on the nuclear and stable transformation of carrot through Agrobacterium tumefaciens and somatic embryogenesis in vitro culture. Somatic embryos, induced by supplementation of Murashige-Skoog medium with the 2,4D hormone, develop into seedlings after 6months approximately when plants are ready to be transferred to a greenhouse. The protocol has over 85% of transformation efficiency.

Jasmonic acid enhances the resistance of potato plants in vitro to salt stress

The protective effect of jasmonic acid (JA) was evaluated under stress (100 mM NaCl) condition. The investigations were carried on potato plants (Solanum tuberosum L.) of the mid-season variety Lugovskoy. Plant-regenerants were grafted and cultured in test tubes on modified Murashige-Skoog agar medium in the absence (control) or in the presence of JA at concentrations of 0.001; 0.1 and 10 M under optimal growing conditions or with the addition of NaCl. After 28 days of cultivation, growth (length of stem and root, number of tiers and leaves, plant mass) and physiological (proline content and photosynthetic pigments, determination of the osmotic potential of cell exudate) of the plants were assessed. For the first time it has been shown that jasmonic acid (0.1 and 10 M) manifests a pronounced protective effect on potato plants under salt stress condition. The protective effect based on the partial removal of the salt negative effect on the main photosynthetic pigments and the maintenance of the osmotic status of cell contents during salinization.

Differential effects of synthetic media on long-term growth, starch accumulation and transcription of ADP-glucosepyrophosphorylase subunit genes in Landoltia punctata

Murashige & Skoog (MS) and Hoagland’s media were previously used for in vitro culture of Landoltia punctata. During subsequent ex vitro culture, the use of MS medium resulted in a higher growth rate, compared to Hoagland’s medium. Thus, a higher starch content of L. punctata in MS medium was previously hypothesized. Here, L. punctata strain 5632 was isolated and characterized using morphological characteristics and the atpF-atpH intergenic region. During early cultivation stage, fresh weight and relative growth rate in MS medium were lower than Hoagland’s medium. Conversely, starch content in MS medium was considerably higher than in Hoagland’s medium. Medium effects on expression of genes coding for starch-biosynthesis ADP-glucosepyrophosphorylase (AGPase) were determined. Genomic fragments of small (LeAPS) and large (LeAPL1) AGPase subunits were characterized. Differential expression between each AGPase subunit genes was observed in both media. Additionally, in MS medium, the highest correlation coefficients between starch content and gene expression was found with LeAPS (0.81) and followed by LeAPL3 (0.67), LeAPL2 (0.65) and LeAPL1 (0.28). In Hoagland’s medium, the coefficients of LeAPL3 (0.83) and LeAPL2 (0.62) were higher than LeAPS (0.18) and LeAPL1 (−0.62). This suggested different levels of contributions of these genes in starch biosynthesis in both media.

Effects of Methyl Jasmonate and Putrescine on Tryptanthrin and Indirubin Production in in vitro Cultures of Isatis demiriziana Mısırdalı

Tryptantrin and indirubin are pharmacologically active compounds used in treatment of disseases such as cancer and Alzheimer's. In this study, we investigated the influences of different concentrations of methyl jasmonate (MeJa) and putrescine (Put) on tryptanthrin and indirubin production in leaf explants and development of Isatis demiriziana Mısırdalı grown in vitro. In all media treated with methyl jasmonate, tryptanthrin production in leaves of plantlets showed an increase. The highest increase in tryptanthrin production was observed in solid Murashige-Skoog (MS) medium containing 1.0 mM MeJa (154.026 ± 0.11 µg g-1), about 2.85-fold higher than the control (untreated plantlets) (40.017 ± 0.031 µg g-1). Production of tryptanthrin decreased about 2.56-fold in the leaves of plantlets treated with Put, when compared to control. The highest indirubin production was obtained in the leaves of plantlets grown in the MS medium containing 0.1 mM MeJa (11.274 ± 0.035 µg g-1) but treatments with Put didn’t show any positive affect on the indirubin production. Analysis of tryptanthrin and indirubin were performed using high performance liquid chromatography (HPLC).

The optimal sterilizing compound and culture medium in Elaeocarpus grandiflorus L. in vitro shoot induction

Elaeocarpus grandiflorus L. is an identity plant of Salatiga City. Wherein, the plant is difficult to naturally propagation. This study would obtain propagation technique of E. grandiflorus by tissue culture. It was experimentally conducted, consists of two sub-experiments. The first aimed to find the optimal sterilization agent was conducted with a completely randomized design, namely sterilizing compound. There were two compounds studied, i.e., NaOCl and Ca(OCl)2. The second sub-experiment aimed to determine the optimal medium type and antioxidant compounds for inducing in vitro shoots. It was conducted with a completely randomized design with two factors, namely the medium type [i.e. Murashige & Skoog (MS) and Wood Plant Medium (WPM)], and antioxidant agents (i.e. activated charcoal and polyvinyl pyrrolidone/PVP). Data were analyzed by descriptive percentage, analysis of variance, and least significant difference test. The results showed that Ca(OCl)2 and NaOCl were effective to obtain the sterile and alive explants. The MS media added by PVP resulted in the most explants forming shoots, and WPM supplemented with PVP or active charcoal caused most explants regenerating callus. The results could be used as a basis for the development of in vitro shoot multiplication to reach an efficient propagation of E. grandiflorus.

Phenolic acid and flavonoid production in agar, agitated and bioreactor-grown microshoot cultures of Schisandra chinensis cv. Sadova No. 1 – a valuable medicinal plant

In vitro cultures and raw materials (fruits and leaves) of the valuable medicinal plant species – Schisandra chinensis cultivar Sadova No. 1 (SchS) – were evaluated for the production of two groups of phenolic compounds, phenolic acids and flavonoids, and their antioxidant potential. A series of experiments was conducted, aimed at optimizing culture conditions for maximum growth and phenolic production in SchS microshoots. Different concentrations of plant growth regulators (6-benzyladenine – BA and 1-naphthaleneacetic acid – NAA, from 0 to 3 mg/l) in Murashige-Skoog (MS) medium were tested in several cultivation systems (agar, agitated, bioreactor) over various growth periods (10, 20, 30, 40, 50 and 60 days). Furthermore, an elicitation experiment was conducted in which the bioreactor-grown microshoots were exposed to yeast extract. HPLC-DAD analyses confirmed the presence of eight phenolic acids – chlorogenic, cryptochlorogenic, gallic, neochlorogenic, protocatechuic, salicylic, syringic and vanillic, and two flavonoids: kaempferol and quercitrin, in the in vitro biomasses. The highest total phenolic acid (357.93 mg/100 g DW) and flavonoid (105.07 mg/100 g DW) contents were obtained in agar culture extracts cultivated for 30 days on MS medium containing 2 mg/l BA and 0.5 mg/l NAA and for 50 days on MS medium containing 0.1 mg/l BA and 2 mg/l NAA, respectively. These amounts were 1.59- and 5.95-fold lower than in parent plant leaf extracts (569.66 mg/100 g DW), and 4.30- and 1.25-fold higher than in fruit extracts (83.17 mg/100 g DW), respectively. Microshoots grown in a Plantform bioreactor also proved to be a good source of phenolic compounds, however, the elicitor treatment had no noticeable effect on their accumulation. Antioxidant capacity assessed by the Folin-Ciocalteu, FRAP, DPPH and CUPRAC assays revealed significantly higher potential in extracts from in vitro biomass and leaves of the parent plant, as compared to the parent plant fruit extracts.

A Study on In Vitro Propagation Possibilities of Some Clone Rootstocks of Prunus Species

In vitro techniques which is one of the modern breeding methods offer alternative methods to meet the growing food demands in parallel with the increasing world population by allowing rapid and intensive mass propagation of varieties and rootstocks in fruit cultivation under laboratory conditions. Clone rootstocks which are economically important and early yielding with maximum yield per unit area are preferred in fruit production. In this study, the propagation possibilities of Marianna GF 8/1, Myrobolan B, Myrobolan 29-C and St Julien A clone rootstocks used in the production of stone fruit species were investigated. In this study, shoot tips were used as explants. MS (Murashige-Skoog) media was used as the propagation media and 30 g L-1 sucrose and 7 g L-1 agar were added. The shoot tips were planted into the media and then placed in the temperature and light controlled culture chamber. Samples were placed to the subculture at 1-month intervals. Explant length, number of leaves and number of tillings were observed every 20 days. Nine different MS media were tested during the shoot growth stage. The best propagation was obtained in the 1 mg L-1 BAP + 0.5 mg L-1 IBA + 0.25 mg L-1 GA3 media. Eight different MS media were tested during the rooting stage. The best rooting was achieved in a media containing 1/2 MS + 2 mg L-1 IBA. Keywords: Marianna GF 8/1, Myrobolan B. Julien A, Myrobolan 29-C, micropropagation, plant growth regulators. DOI: 10.7176/JSTR/5-9-10

Introduction to in vitro culture and callus initiation in Salvia hispanica L. (chia)

Aim. Preparation of aseptic seedlings Salvia hispanica L., callus initiation in vitro and establishment of primary explants suitable for the callus production. Methods. Seeds are sprouted on our own modification of conventional methods. The non-hormonal Murashige-Skoog agarized nutrient medium was used as basic medium for the experiments. Parts of one-month seedlings (roots, hypocotyl, cotyledon leaves) were used as explants for the use of the colza. We added growth regulators (BAP, 2,4-D) in different concentration combinations into the nutrient medium for callus initiation. Statistical processing was performed in Microsoft Office Excel. Results. Aseptic S. hispanica seedlings have been obtained. The callus growth was initiated on all types of explants, the dependence of the callus intensity on the type of explants and the growth regulators content in the nutrient medium was established. Morphogenic callus and root-regenerants have been obtained. Conclusions. Hypocotyl was the most suitable primary explant for callus growth. Seedlings, leaves and roots showed low morphogenetic capacity. The nutrient medium with an elevated 2,4-D content was the most effective for initiation of callus genesis and proliferation of non-morphogenous callus. A high concentration of 2,4-D in the medium improves S. hispanica callus growth but suppresses its morphogenic ability.Keywords: Salvia hispanica (Chia), in vitro culture, callus.

Multiplicação in vitro de Xylopia aromatica em diferentes meios de cultura e concentrações de BAP

Este estudo teve por objetivo desenvolver uma metodologia de multiplicação in vitro para Xylopia aromatica, visando a produção de mudas, avaliando-se meios de cultura e concentrações de reguladores de crescimento mais adequados. Foram utilizados, como explantes, segmentos nodais, provindos de brotações de mudas produzidas via seminal e mantidas em viveiro. Foram testados os meios de cultura Murashige Skoog (MS) e Wood Plant Medium (WPM) suplementados com 800 mg L-1 de polivinilpirrolidona (PVP), 30 g L-1 de sacarose, 0,054 µM de ácido naftalenoacético (ANA) e 6 g L-1 de ágar e concentrações de 2,22 e 3,55 µM de benzilaminopurina (BAP) para a multiplicação in vitro. Aos trinta dias, avaliou-se o número de brotações laterais emitidas e o percentual de explantes formando calos na base. As maiores taxas de multiplicação foram obtidas com o meio MS acrescido de 3,55 µM de BAP. Em relação à formação de calos na base dos explantes, o meio MS promoveu maior formação quando comparado ao WPM, no entanto não comprometeu a formação de brotos. Foi possível estabelecer as fases de introdução e multiplicação, definindo assim o meio de cultura e concentrações de reguladores de crescimento mais adequados ao cultivo in vitro de Xylopia aromatica.

Efficient low cost seesap (CSUP) technique for micropropagation of newly introduced sweetener plant, Stevia rebaudiana Bertoni

Stevia rebaudiana is a non-caloric sweetening herb, which has a high potential for commercialization. During this study low cost media sterilization, seesap (CSUP) technique which applies sodium hypochlorite was used as an alternative to autoclaving. In vitro cultures were established using nodal segments collected from stevia mother plants. These in vitro shoots were cultured on Murashige & Skoog (MS) medium supplemented with 16 combinations of 6- benzyl amino purine (BAP) and kinetin (KN) with 3% sugar at 5.8 pH, to investigate the in vitro shoot multiplication. The developed shoots were transferred to half MS medium supplemented with 0, 1 and 2 mg/L Indole butyric acid (IBA) with 3, 4 and 5% sugar in nine combinations at 5.8 pH, to investigate the rooting. The well rooted stevia plantlets were transferred to Jiffy™ bags for hardening. The results showed that the combination of 1.0 mg/L BAP and 1.5 mg/L KN resulted in the highest number of shoots (11.8 shoots/shoot). The combination of 2.0 mg/L IBA with 5% sugar in half MS medium resulted in the highest root induction (7.2 roots / plant) and root elongation (1.02 cm). The plants grown in culture medium containing 5% sucrose gave 100% survival rate at acclimatization. Key words: Stevia, micropropagation, shoot induction, root induction, acclimatization, low cost seesap (CSUP).

Production of Stilbenes in Callus Cultures of the Maltese Indigenous Grapevine Variety, Ġellewża.

The production of secondary metabolites in tissue culture has been considered as an alternative to the cultivation and harvesting of crops intended for this purpose. The present study was aimed at the growth of callus and production of polyphenolic compound of callus derived from a Maltese indigenous grapevine variety, Ġellewża. Callus was inoculated onto plant growth regulators-enriched Murashige Skoog media (MSm) to determine whether polyphenols are produced in vitro as well as to determine the best combination of plant growth regulators needed for the production of these metabolites. From results obtained, it was observed that the best callus production was obtained by auxin-enriched MSm. In fact, indole acetic acid and indole acetic acid /6-benzyl aminopurine enhanced biomass accumulation (3.04 g and 3.39 g) as opposed to the others (<1.97 g). On the other hand, parameters showing the presence of flavonoids (tonality, 3.80), particularly anthocyanins (24.09 mg/kg) and total polyphenols (1.42 mg/g), were optimum in the presence of cytokinins, particularly 6-benzyl aminopurine. Analysis for single polyphenols revealed a high amount a particular stilbene: polydatin (glucoside of resveratrol). Resveratrol and other typical polyphenols, found in mature berries, were also found in significant quantities, while the other polyphenolic compounds were found in minimal quantities. This is the first study to describe the production and composition of polyphenols in Ġellewża callus cultures. From the results obtained, it can be seen that this grape tissue is an excellent alternative for the production of polyphenols from the stilbene group, which can be upscaled and exploited commercially.

Influence of Ribavirin on Prunus Domestica L. Regeneration, Genome Stability and Virus Eradication In Vitro

Abstract Prunus necrotic ring spot ilarvirus (PNRSV) and Apple chlorotic leaf spot trichovirus (ACLSV) are common in plum orchards. The aim of the study was to obtain virus-free planting material of Prunus domestica L. by chemotherapy in vitro. Ribavirin at concentrations of 10 to 50 mg·l−1 was added to Murashige–Skoog (MS) nutrition medium for virus eradication from microshoots. After a two-week period of chemotherapy, meristems were subcultured monthly on MS medium and proliferation index of shoots was estimated. Microshoots were retested by reverse transcription polymerase chain reaction for presence of virus. At lowest concentrations of 10 mg·l−1 ribavirin was entirely ineffective for ACLSV and 10 to 30 mg·l−1 was ineffective for PNRSV elimination. Ribavirin concentrations of 40 and 50 mg·l−1 destroyed both pathogens. However, at higher concentrations of 40 and 50 mg·l−1 ribavirin exhibited some signs of phytotoxicity on microshoots in the first sub-cultivation period. In order to test the genetic stability of the microplants after chemotherapy the amplified fragment length polymorphism (AFLP) method was applied. Plant genome stability in ‘Magna Glauca’ at concentrations of 40 mg·l−1 was damaged, as the presence of polymorphic AFLP markers were observed.

Growth regulators induced shoot regeneration and volatile compound production in Lippia rotundifolia Cham., a threatened medicinal plant

Lippia rotundifolia Cham. is an endemic and endangered plant found in the savanna that is aromatic and rich in monoterpenes. The aims of the study were to evaluate the influence of plant growth regulators on the growth and chemical analysis of L. rotundifolia by headspace/GC–MS. Nodal segments (1.0 cm) were cultured on basal Murashige & Skoog (MS) medium supplemented with the following amounts of growth regulators: 0.0, 2.22, 6.66, 11.10, and 15.54 μM 6-benzylaminopurine (BAP) combined with 0.0, 2.68, and 5.36 μM naphthaleneacetic acid (NAA) or 0.0, 2.27, 4.54, 6.81, and 9.08 μM thidiazuron (TDZ) combined with 0.0, 2.68, and 5.36 μM NAA. The most effective concentrations were 11.10 μM BAP + 2.68 μM NAA, which induced 11 shoots explant−1, and 15.54 μM BAP + 2.68 μM NAA, with 11.6 shoots explant−1. The TDZ at a concentration of 6.81 μM with 2.68 μM NAA resulted in 20 shoots explant−1, 9.08 μM TDZ without NAA in 22.8 shoots explant−1, and 9.08 μM TDZ + 2.68 μM NAA in 18.1 shoots explant−1. Rooting took place on MS medium free of plant growth regulators, and acclimatization was successful (95%). The influence of plant growth regulators on the fraction of volatile compounds appeared to be quite variable. The cytokinin concentration was found to significantly influence the production of volatile compounds in L. rotundifolia shoot cultures. These findings indicate that shoots regenerated from nodal segments on MS medium containing higher concentrations of BAP and NAA have increased myrcenone and limonene. Ocimenone contents increased only at low concentrations of BAP and NAA.

Substituting plant vegetative parts with callus cell extracts: Case study with Woodfordia fruticosa Kurz. – A potent ingredient in skin care formulations

Herbal pharmaceutical companies are dependent on plant parts which are used as raw materials in their products. The naturally available herbal raw material does not meet the demand of industries. This can lead to exploitation of nature and affect the biodiversity in various ways. In vitro plant micropropagation and conservation can be a successful alternative to minimize this environmental exploitation. Woodfordia fruticosa Kurz. (Lythraceae) is a plant endemic to India, known for its anti-aging benefits. Apical leaf explants were used to generate calli which was achieved in Murashige Skoog medium with 0.25 mg-l 2,4,5-T and 0.10 mg-l 6-BAP. Callus was sub cultured continuously for six months and retrieved every month for in vitro studies (WSC 1-WSC 6). Among them, WSC-3 (callus extract of third month) increased the collagen production by 60.67 ± 4.2% in human dermal fibroblasts (HDF). Collagen-I and elastin gene expression was increased in WSC-3 treated HDF. WSC-3 inhibited hydrogen peroxide (H2O2) induced cellular senescence in HDF. LC–MS analysis of WSC-3 upheld the presence of polyphenols like gallic acid, 1,2,3,4,6-pentagalloyl glucose, glucogallin, and other phytoconstituents. The results of the in vitro experiments were on par with the extracts obtained from flowers of W. fruticosa. IC50 values in HDF showed that callus cell extract is non-toxic (IC50: >1000 μg/ml) whereas, the flower extracts were moderately toxic (IC50 278.25 ± 1.85 μg/ml). Woodfordia fruticosa callus extract is a potential ingredient for skin care formulations, can be a strong substitute to conventional flower extract and warrants further clinical studies.

Pepper plants response to metal nanoparticles and chitosan in nutrient media

Neutral nanoparticles (NPs) of iron, zinc, copper or their compositions were introduced into nutrient Murashige-Skoog (MS) medium instead of traditional metal salts with/without chitosan supplements to obtain healthy and crop productive plants. Metal NPs concentrations were: Fe NPs (0.06, 0.3, 3.0); Zn NPs (0.016, 0.08, 0.4); Cu NPs (0.004, 0.0008, 0.00016) (mg L-1). Pepper (Capsicum annuum) cultivar LJ-King was grown on modified media in aseptic conditions. After 45 days, root and stem length, root activity, green mass gain and chlorophyll content in leaves were measured. Pepper plants grown on media with metal NPs showed increased root length and activity (for Fe, Zn, Cu NPs and their composition) and chlorophyll content in leaves (for NPs Fe and Cu). However, addition of Chitosan modified the effects. Monitoring of pepper grown on MS with NPs composition (Fe: Zn: Cu as 3.0:1.0:0.004 mg L-1) with electron microscope demonstrated no NPs accumulation in roots, sprouts and leaves.

An Efficient Somatic Embryo Liquid Culture System for Potential Use in Large-Scale and Synchronic Production of Anthurium andraeanum Seedlings.

Anthurium andraeanum Lind. is the second most important tropical flower in the world flower market. Somatic embryogenesis and plant regeneration in Anthurium has been reported previously; however, a stable and effective method for its commercial use has not been available. In this study, an efficient somatic embryogenesis and liquid culture system for large-scale production of A. andraeanum seedlings was achieved. Building on previous research, this study investigated the main factors for proembryogenic mass (PEM) proliferation, somatic embryo (SE) development, and SE germination in Anthurium. The results showed that relatively low concentrations of plant growth regulators, mineral nutrition, and sucrose promoted PEM proliferation, SE formation, and germination in a liquid culture system. This system can be described as follows: PEMs were induced from leaf blade explants on Murashige & Skoog (MS) medium with half-strength MS macronutrients (1/2 MS) containing 2.0 mg L-1 2,4-dichlorophenoxyacetic acid (2,4-D), 0.5 mg L-1 kinetin (KT), and 3% sucrose and were proliferated in ½ MS liquid medium containing 1.0 mg L-1 2,4-D, 0.5 mg L-1 KT, and 3% sucrose. The highest proliferation coefficients were 5.11–5.16. PEMs were then transferred to MS medium with 1/8 MS macronutrients (1/8 MS) liquid medium containing 1% sucrose to develop into globular embryos and mature embryos. Finally, the mature embryos were placed on four layers of absorbent filter paper saturated with 1/8 MS liquid medium containing 1% sucrose for germination, and an average of 60 seedlings per gram SEs was obtained. This liquid culture system can be used in large-scale and synchronic production of Anthurium seedlings.

Micropropagation and Genetic Fidelity of in vitro Grown Plantlets of Begonia malabarica Lam.

Begonias are ornamental indoor plants and also cultivated for landscaping. Begonia malabarica Lam., found naturally in the hilly regions of South India and Sri Lanka called by the tribals as Narayana Sanjeevi, i.e. possess miraculous therapeutic properties. The present study targets in vitro culture, field establishment and evaluation of its genetic uniformity using anatomical, molecular markers followed by callus induction and establishment of cell suspension. Explants such as shoot tip and leaves from in vitro germinated seeds were cultured on Murashige-Skoog medium. Maximum callus induction was obtained in Murashige-Skoog medium with 0.3 mg L–1 2, 4-D + 0.5 mg L–1 BAP, i.e. 90.8 ± 2.8% with 2.5 ± 0.1 g callus growth. In vitro suspension of cells and their growth rate were also analysed. RAPD and ISSR experiments were carried to confirm the genetic fidelity among in vitro and in vivo plants. Twelve RAPD and seven ISSR primers raised 119 amplicons. Genomic DNA amplification showed similarity between the in vivo and the in vitro generated plantlets. The overall results revealed the uniformity of the in vitro raised B. malabarica plantlets in terms of histoanatomical genetical features and substantiated the assumption that in vitro micropropagation is the secure mode for mass propagation of true to type plants without any somaclonal variations.

Multiple Shoot Regeneration in Seed-derived Immature Leaflet Explants of Red Dragon Fruit (Hylocereus costaricensis)

Dragon fruit which belongs to Cacteceae family was once widely grown as ornamental plant and is now considered as fruit crop. Though there were many reports on propagating dragon fruit inadequate information is available on obtaining best quality pant material. Hence the present work is carried out to find the effect of phyto harmones, 6-Benzylaminopurine (BAP) and Naphthalene acetic acid (NAA) on callus formation, shoot and root regenerations from dragon fruit in vitro using stem cuttings as explants. Stem explants in Murashige Skoog (MS) medium containing 3mg/l BAP and 0.5mg/l NAA exhibited highest in vitro response for callus formation. Further, maximum number of shoots (13.2) was regenerated from explants when MS basal medium was supplemented with 3mg/l BAP and 0.5mg/l NAA. Within four weeks somatic shoots were raised from explants. It took 70 days for shoots to grow 2.0cm long. Subsequently regenerated shoots were transferred to rooting medium supplemented with 3mg/l BAP and 0.5mg/l NAA which proved to be effective concentrations for the growth and development of roots (4.8) in vitro.

Comparison of Volatile Profile and Antioxidant Activity of Piper divaricatum G. Meyer (Piperaceae) Using Cuttings and Cell Tissue

In this study, P. divaricatum (Piperaceae) plants were subcultured by cuttings and cell tissue in order to carry out a comparison of the volatile profile, phenolic compounds, and antioxidant activity. Propagation by cuttings used vermiculite substrate and Murashige-Skoog (MS) medium in the absence of growth regulators and rooting started at 15 days of growth providing a higher number of plants after 90 days. In vitro propagation was performed using shoot apices as explants in MS media supplemented with 0.5 mg mL-1 BA (6-benzyladenine). Volatile profiles analyzed by gas chromatography-mass spectrometry (GC-MS) showed as main compounds methyl eugenol, E-β-ocimene, and β-elemene in the in vivo and in vitro cultures. Phenolic contents determined by the Folin-Ciocalteu method had no significant difference at the end of 90 days of growth displaying a good linear correlation with antioxidant activity and phenylpropanoids amounts (r > 0.7). However, the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging was higher for in vitro plants grown in comparison to in vivo plants cultured.

In Vitro Propagation of Wetland Medicinal Plant Lythrum salicaria L.

The aim of this experiment is to develop a protocol to establish the in vitro propagation potential of Lythrum salicaria L (purple loosestrife). Seeds of L. salicaria were germinated and grown in Murashige Skoog (MS) medium for 30 days. At the end of 30 days, shoot tips of L. salicaria were cultured on MS medium supplemented with different combinations and concentrations of 6-benzylaminopurine (BAP; 1, 2 and 3 mg/L) and 1-naphthaleneacetic acid (NAA; 0, 0.1 and 0.5 mg/L). Regenerated shoots were transferred to MS supplemented with varying concentrations of different auxins: NAA (0.5 and 1 mg/L) and indole-3-butyric acid (IBA; 0.5 and 1.0 mg/L) for rooting. Among the cytokinins investigated, BAP indicated the best response to initiate shoot elongation and multiple shoot formation alone. The highest number of shoot per explant (37±3.38) was obtained in MS medium with 1 mg/L BAP. Although, the highest shoot length (3.1 cm ± 0.48) was found on medium with 1 mg/L BAP+0.5 mg/L NAA, there were no statistically differences among all types of medium. Percentage of rooting, shoot length and root length were altered significantly with different concentrations of NAA and IBA. Thus, shoot length (9.5 cm±0.46), root length (4.1 cm±0.13) and the percentage of rooting (100%) increased significantly with a concentration of 1 mg/L NAA. It was concluded that MS medium with 1 mg/L NAA has been superior for root development compared to other hormone concentrations.