Concentrations Of MS Medium Research Articles

MODULATION OF CULTURE MEDIUM ON THE EX SITU CONSERVATION OF Neoregelia mucugensis Leme (BROMELIACEAE)1

ABSTRACT Bromeliads are the target of predatory extractivism and consequently many species are included in the red list of threatened species, such as those belonging to the genus Neoregelia. Although Neoregelia mucugensis has not been evaluated for the degree of threat, its exploitation is exclusively extractive and its occurrence in Chapada Diamantina-BA is subject to the action of fires that affect the region annually. In this context, applying techniques aimed at protecting this genetic resource is fundamental for both the maintenance of its natural populations and the ex situ conservation of this genetic material. Plant tissue culture techniques have been successfully applied for the conservation of several bromeliad species. One of the methods used is slow growth, which consists in reducing plant metabolism and consequently decelerating its growth, which allows the maintenance of in vitro plant collections without the need for subculture. In this context, the objective of this study was to test the reduction of salts in the culture medium and the addition of osmoregulators on the induction of slow growth of N. mucugensis. Plants were subjected to treatments composed of different concentrations of MS medium and mannitol for a period of 12 months, when then analyses were conducted to evaluate growth, chlorophyll content and regeneration capacity of shoots in vitro. It was found that the treatment containing MS ½ and 7.8 g.L-1 of mannitol is indicated for in vitro conservation of N. mucugensis with maintenance of the regenerative capacity of its tissues.

Influence of macronutrients, sucrose and LED on in vitro culture of Lomatozona artemisiifolia (Asteraceae - Eupatorieae)

Abstract Balanced levels of macronutrients and sucrose may ensure the success of micropropagation of the endangered ones. This study aimed to evaluate the effect of levels of salts in the culture medium on in vitro culture of Lomatozona artemisiifolia, as well as to determine the influence of light emitting diode (LED) on the shoot proliferation and rooting of the species. Nodal segments were used to evaluate the different macronutrient concentrations of MS medium (25, 50 and 100%), as well as sucrose concentrations (0.0 mM; 0.34 mM; 0.68 mM and 1.03 mM). Five light conditions were evaluated at shoots proliferation and rooting [100% blue (455 nm); 100% red (630 nm); 30% blue + 70% red; 30% red + 70% blue or fluorescent white]. Low levels of macronutrients in MS (50% and 25%) and sucrose (0.00 mM and 0.34 mM) resulted in plants with higher height, number of shoots and higher production of photosynthetic pigments. The 100% red light promoted rooting of 100% of the plants, and in 100% red or 70% red + 30% blue, higher plants were observed. These results demonstrate that under in vitro conditions, L. artemisiifolia has low nutritional needs, typical of plants that live in rupestrian fields.

Impact of saline solution on growth and photosystem II during in vitro cultivation of Bromelia antiacantha (Bromeliaceae)

Abstract In vitro cultivation is a technique with wide application for micropropagation. However, each species has specific mineral needs for this type of cultivation. The objective was to assess the impacts of the saline solution culture medium on the performance of the photosynthetic apparatus and growth of Bromelia antiacantha during in vitro cultivation, and thus to elucidate the mitigation of the nutritional imbalance that can interfere in the electron transport in the plants. Plants were cultivated in a salt concentration gradient of MS medium (0%, 25%, 50%, 75% or 100%). The growth traits and fluorescence a chlorophyll were analyzed. Intermediate concentrations of MS medium resulted in plants with a larger number of leaves and longer root length. The OJIP curves and results of the JIP test showed that the plants grown without MS salts presented less efficient photosystem II (PSII), as indicated by the performance index [Pi(total)]. In contrast, the intermediate concentrations (MS 25% and 50%) had a positive effect on the performance of the photosynthetic apparatus. The MS 25% medium can be used for in vitro cultivation of B. antiacantha, enabling the development of plants with suitable physiological qualities for planting in the field.

Seed analysis of Dichromanthus aurantiacus, terrestrial orchid from Toluca valley, México

Objective: The objective of this work was to analyze the viability and germination of Dichromanthus aurantiacus seeds, a terrestrial orchid from Toluca valley, México.
 Design/methodology/approach: The size and color were evaluated. Two methods determined the viability: 1) the tetrazolium test (imbibition for 24 hours in the water, 2 hours in calcium hypochlorite (CaCOCl2), and drops of Tween-80). 2) the asymbiotic seed germination by in vitro culture (imbibition for 24 hours in the water, and the concentration of MS medium plus natural extracts). 
 Results: The seeds of this specie showed approximately 0.2 mm long and 0.05 mm wide; they possess an embryo and a brownish testa. There were significant differences (P<0.05) between the treatments finding a positive effect with the tetrazolium test, achieving up to 91.4% viability. In the in vitro germination, the imbibition of the seeds favored contamination. The concentration of MS and the addition of natural extract presented significant differences (P<0.05), the 50% MS plus 10% of coconut water showed up to 92.8% of germination at 60 days.
 Study limitations/implications: The results are preliminary of a long-term experiment.
 Findings / Conclusions: The seeds of Dichromanthus aurantiacus showed brown testa and an oval embryo with dimensions of 0.2 mm long and 0.05 mm wide. The tetrazolium test’s viability showed 91.4% viability when they were soaked in sodium hypochlorite solution (CaCOCl2) for two hours, 24 hours soaking in tetrazolium solution (1%) plus two drops of Tween-80. The asymbiotic in vitro culture showed up to 92.8% germination in 60 days using MS medium at 50% enriched with 10% coconut water

PENGGUNAAN AIR KELAPA DAN ASPIRIN UNTUK PRESERVASI UBI JALAR (Ipomea batatas) SECARA IN VITRO

Sweet potato is raw locally food used as an alternative staple food that important for national food security. The objective of the reseach was to achive the suitable medium for preservation of sweet potato plants by slowly growth in in vitro medium. The factorial experiment arranged in completely randomized design with the first factor was two concentrations of MS medium: MS 1 and MS 2. The second factor was several concentrations of coconut water, i.e. 0 and 15 mg/l and the third factor was several concentrations of Aspirin (0, 5, 10, 15, 20 mg/l) . The results showed that the number of green leaves and the number of internodes were affected by concentrations of medium in six weeks after planting. The MS1 medium with 0 mg/l Aspirin and 15 mg/ coconut water could produced the higher number of green leaves (8,0) and 8,5 internodes number. The MS2 + 15 mg/l coconut water and 15 mg/l Aspirin was the best medium for sweet potato preservation. Data obtained also showed that all medium containing Aspirin produced the high number of root. Keywords : Sweet potato, coconut water, Aspirin, preservation, in vitro.

Micronutrient Delivery System for Induction of Organogenesis in Banana in vitro Cultures

Micropropagation techniques represent one of the technologies, which allows the large-scale production of banana and the culture medium composition is one of the major factors affecting in vitro propagation of plants. The objective of this study was to evaluate the efficiency of a new micronutrient delivery system based on ionic Cu and Zn for inducing in vitro organogenesis in two banana cultivars in vitro cultures, Grand Naine and Pysang Ceylon. The first experiment evaluated different concentrations of BAM-FX® (0.16, 0.32, 0.64, 1.28, and 2.56 µl ml-1) added to the MS culture medium. The concentration of 0.16 µl ml-1 BAM-FX® provided the best results for in vitro shoot and root growth and development. Therefore, a second experiment was performed to evaluate the potential of combining BAM-FX® with a reduced concentration of MS medium (¼, ½, and ¾ strength), or the use of BAM-FX® alone without MS medium. Results indicate that MS at ¾ strength combined with 0.16 µl ml-1 BAM-FX® provided proper in vitro shoot and root growth and development. The use of BAM-FX® in vitro requires additional studies to verify the feasibility of this product for efficient micropropagation of banana, as well as for other species.

Effect of medium pH and electrical conductivity on the development ofAnthurium andreanum‘Tropical’ in tissue culture

Anthurium is one of the most demanded pot plants in the European market. The effect of different pH (5.7 and 6) and electrical conductivity (EC) of MS culture medium (MS, 1/2 MS, and 1/4 MS) on growth parameters of Anthurium andreanum 'Tropical' was studied. An experiment was conducted on a factorial basis in a completely randomized design with three replications, including eight plant lets for each treatment. Leaf and root number, plant fresh weight, plant leaf area, total chlorophyll, chlorophyll a (Chl a) and chlorophyll b (Chl b) contents were significantly affected by EC of the culture medium. The largest number of leaves and roots were produced in 1/2 and 1/4 concentrations of MS medium, respectively. Full concentration MS medium resulted in the highest plant fresh weight, plant leaf area, total chlorophyll, Chl a, and Chl b content. Plant fresh weight was also significantly affected by pH of the culture medium. The interaction effect of pH and EC of medium significantly affected the carotenoid content. Plant lets grown on 1/4 MS medium had the highest and lowest carotenoid content at pH 5.7 and 6, respectively. No significant difference was observed in plant height, root or leaf number, leaf area, total chlorophyll, Chl a, or Chl b content in response to medium pH. In general, MS and 1/2 MS culture medium at pH 5.7 showed the most beneficial effects on growth parameters of anthurium 'Tropical' plant lets.

Growth and volatiles in the micropropagation of Santa Maria herb

The aims of this study were to establish micropropagation for Chenopodium ambrosioides L. and analysis of volatile constituents in vitro as a function of different salt concentrations of MS medium and sucrose. The apical buds were established on MS medium added with 3% sucrose. The vegetative growth of nodal segments was tested in different salt concentrations of MS basic medium (0.25, 0.50, 1.00, 1.50, and 2.00) and of sucrose (5, 10, 20, 30, and 40 g L-1). Nodal and apical segments were tested in vertical and horizontal orientations. Different volumes of culture medium (20; 35, 50, 65, and 80 mL) were examined in the growth of nodal segments. Plants were acclimatized in three different substrates. The apical segments collected from matrices plants were successful when established in MS medium. A multiplication rate of 8.5 was obtained with nodal and apical segments without using growth regulator. MS medium with its salt concentration reduced by half (0.50 MS) and sucrose at the concentration of 30 g L-1 provided better in vitro growth of nodal segments. Four major volatile compounds (α-terpinene, p-cymene, Z-ascaridole, and E-ascaridole) varied at different salt concentrations and only α-terpinene showed difference for sucrose concentrations. Apical segments in vertical orientation and nodal segments in horizontal with 50 mL of culture medium showed better responses for the in vitro cultivation of C. ambrosioides L.

Concentration of ms medium and cutting of seeds on in vitro establishment of baruzeiro (Dipteryx alata Vog.)

Concentration of ms medium and cutting of seeds on in vitro establishment of baruzeiro (Dipteryx alata Vog.)

Alternative supplements and ms medium concentrations in the in vitro establishment of Dipteryx alata Vog.

Dipteryx alata Vog. (barueiro) is a native fruit of the Cerrado used in food and medicine. The species has never been domesticated and is currently at risk of extinction. Plant tissue culture is an important tool for the conservation of germplasm and for quick, large-scale propagation of quality seedlings. While, this technique has never been used with barueiro, it could be a valuable tool in the domestication of this species. The objective of this work is to evaluate concentrations of MS medium salts (25, 50, 75 and 100%) supplemented with coconut water (100 mL L -1 ), dwarf banana pulp (60 g L -1 ) or the absence of any supplement in the in vitro establishment of barueiro seeds ( Dipteryx alata Vog). The experiment used a completely randomized design (CRD) with a 3x4 factorial scheme (three cultivation media types and four concentrations of MS salts) and three repetitions. The following characteristics were evaluated: germination rate , shoot length , largest root length , stem diameter , number of leaves , chlorophyll A , chlorophyll B , total chlorophyll , and the fresh and dry mass of the seedlings. The coconut water and banana pulp supplements did not improve the in vitro development of the barueiro seedlings and the full concentration of MS salts did not improve the initial development of the seedlings. However, the pure MS medium at a concentration of 25% MS salts was the best option for the in vitro establishment of Dipteyx alata .

Meristem culture for clonal micropropagation of grapevines

Experiments were carried out to determine the effects of MS medium concentrations (4/4 MS, 3/4 MS, 2/4 MS) combined with GA 3 at 0.0, 0.1 and 0.5 mg/l, and vitamin formulations (MS, Morel, thiamine-inositol) on success of meristem culture and of 15 auxin (IBA) x cytokinin (BAP) combinations on shoot and root formation for clonal rnicropropagation of the wine grape cv. Kalecik Karasi and the rootstock cv. 41 B M.G. Best results were obtained from standard MS mineral composition and vitamin formulation combined with 2.5 mg/l BAP and 0.5 mg/l GA 3 for primary meristem cultures of both genotypes; 0.0 mg/l IBA x 1.0 mg/l BAP for shoot subculture and 1.0 mg/l IBA x 0.0 mg/l BAP for root subculture of Kalecik Karasi; 0.0 mg/l IBA x 1.0 mg/l BAP for shoot subculture and 5.0 mg/l IBA x 0.0 mg/l BAP for root subculture of 41 B, considering shoot, explant, root, callus and particularly entire plant formation in the cultures of both stages.

<i>In Vitro</i> Propagation and Antibacterial Activity of <i>Ocimum basilicum</i> Linn.

The study introduced a simple, reliable procedure for the in vitro culture and standardization of indigenous plant Ocimum basilicum . A potential antibacterial activity was observed in the extracts (chloroform, methanol and petroleum ether) of Ocimum basilicum against selected bacterial strains (MTCC type strains) and their MIC (Minimal Inhibitory Concentration) values were 62.5 and 125 μg mL -1 . Existence of tannin, carbohydtares, phenolic compounds, proteins and aminoacids, anthraquinone glycosides, alkaloids were confirmed by phytochemical analysis. In vitro propagation from nodal explants and shoot tip of O. basilicum showed a higher level frequency of micro shoots on MS medium containing plant growth regulators at various combinations and concentrations. The nodal explants produced maximum shooting response (98 %); maximum shoot length (7.9 cm) and maximum multiple shoot formation (n=4.5) in BAP containing medium after 40 days. A 50% concentration of MS medium with combination of IBA and BAP showed maximum rooting response (78 %), maximum number of roots (n=12) and maximum length of root (8.67 cm) after 25 days. The regenerates were then hardened off and young plants were transferred to field. Nodal explants in MS medium with 2, 4 - D showed a fast growth and pale brown colored calli.

Micropropagation of PLUCHEA LANCEOLATA (Oliver & Hiern.) Using Nodal Explant

Abstract Pluchea lanceolata is an important medicinal plant of Asteraceae family known for its anti-arthritic and anti-inflammatory activity. A protocol was established for micropropagation of P. lanceolata using nodal explants. Nodal explants were inoculated onto Murashige and Skoog (1962) - MS medium supple–mented with 6-benzylaminopurine (BAP), kinetin (Kin), thidiazuron (TDZ) and 2iP (2-isopentenyladenine) at various concentrations (0.0, 0.5, 1.0, 1.5 and 2.0 mg·dm-3). The highest multiplication rate was obtained for nodal explants cultured on MS medium, supplemented with 0.5 mg·dm-3 thidiazuron (TDZ). In vitro raised shoots were successfully rooted on ½ mineral salt concentration of MS medium supplemented with 1.0 mg dm-3 IBA.

Influência da composição da matriz de encapsulamento de microbrotos de banana (Musa sp.) cv. Prata-Anã clone Gorutuba

A banana é a principal fruta no comércio internacional e a mais popular no mundo. A produção de sementes sintéticas vem destacando-se como uma importante técnica para a micropropagação e conservação in vitro de várias espécies. O objetivo deste estudo foi avaliar a influência da composição da matriz de encapsulamento na conversão de microbrotos de banana cv. Prata-Anã clone Gorutuba. Os microbrotos de banana cv. Prata- Anã clone Gorutuba foram encapsulados, empregando-se na matriz alginato de sódio: água destilada e MS (50% e 100%), acrescidos ou não com carvão ativado (1,5 g L-1) em duas épocas de avaliação (15 e 30 dias). Foram testados três níveis de BAP (0; 3; 6 mg L-1) adicionados ou não a 2,5 mg L-1 de ANA, em duas épocas de avaliação (15 e 30 dias). Os dois experimentos foram inteiramente casualizados, sendo um fatorial 3 x 2 x 2, com seis repetições e cinco unidades encapsuláveis por parcela. Aos quinze dias, avaliou-se a conversão, e aos 30 dias, avaliou-se conversão, altura e enraizamento das plantas. Aconstituição da cápsula quando se utilizaram diferentes concentrações de meio MS, com ou sem a presença de carvão ativado, não influenciou na taxa de conversão e na altura das plantas. As cápsulas sem adição de reguladores de crescimento resultaram em melhor conversão dos microbrotos; entretanto, a concentração de 3 mg L-1 de BAP+2,5 mg L-1 de ANA proporcionaram maior altura de plantas. A maior taxa de enraizamento ocorre nas cápsulas contendo MS 100% e carvão ativado, e nas cápsulas com 0 e 3 mg L-1 de BAP, independentemente da adição de ANA. Aos 30 dias, é obtida maior conversão das sementes sintéticas.

Optimization of the Protocol for the <i>in Vitro</i> Cultivation of <i>Piper aduncum</i> L.

Piper aduncum L. (Piperaceae) has great potential for economic exploration because of the proven use of its essential oil in the agriculture and in the human health. A technique that improves its propagation, as the tissue culture, becomes necessary. Some parameters must be determined for the successful cultivation in vitro. Thus, this study aimed to determine the salts concentration of MS medium, temperature, luminosity and light quality for in vitro culture of this species. The following treatments were conducted: 1/4MS, 1/2MS, MS and 2MS; 20℃, 25℃, 30℃ and 35℃; monochromatic blue, red and white lights and the combination of red and blue, using light emitting diodes (LEDs); luminosities of 17, 37, 48 and 73 μmol·m-2·s-1 obtained with cool white fluorescent lamp. After 35 days, the treatments were analyzed. To determine the salts concentration of MS, the variables were evaluated: number of shoots, leaves and roots, length and width of leaves, length and dry mass of root and shoots and percentage of death by necrosis. To the plantlets under different temperatures, length and dry mass of shoot and root, number of shoots, number, length and width of leaves and survival and rooting percentages were measured. The plantlets that were maintained under different luminosities were evaluated for length of shoot and root, dry mass of leaf and root. To evaluate the growth under the LEDs, the length and dry mass of shoot and root, number of shoots and roots, percentage of rooting and sprouting were assessed. The medium 1/4MS and the medium 1/2MS showed better responses for number and length of root, leaf width and shoot length. The temperature 25℃ provided the highest number of leaves, length of shoot and root, root dry mass and rooting percentage. The luminosity 73 μmol·m-2·s-1 provided higher values in length of root and dry mass of leaf and root. The red LED provided plantlets with greater growth. Thus, for the in vitro cultivation of Piper aduncum, 1/4MS, environment temperature of 25℃, light intensity of 73 μmol·m-2·s-1 and environments with red light to obtain high quality plantlets for propagation of this species are recommended.

Tipo de explante e diferentes concentrações de sais em meio de cultura MS no estabelecimento in vitro de Aniba canelilla

The peel - precious ( Aniba canelilla ) is a plant species native to the Amazon region , with economic potential in silvicultural terms , the production of perfumes and cosmetics . However , one limitation to the domestication of the species is the absence of propagation methods . The objective of this study was to evaluate the effect of explant type and different salt concentrations of MS medium in their establishment in vitro . Shoot tips ( 1.5 cm ) and nodal segments were used with two gems, isolated from bark - precious seedlings maintained in the greenhouse . The treatments consisted of two types of explants and six concentrations of salts in MS medium plus 15 or 30 g L- 1 sucrose and disinfectants . It was found that the nodal segment is less sensitive to treatments performed disinfestation and decontamination showed higher successful in survival and in vitro establishment of shell - precious.

KONSERVASI PAMELO {Citrus maxima (Burm.) Merr.} DENGAN PENURUNAN KONSENTRASI MEDIUM DAN SUKROSA

Indonesia has high variety of pummelo [ Citrus maxima (Burm.) Merr.], but only several cultivar are cultivated extensively. Others became vurnerable and the germplasm has to be conserved to prevent their extinction. One of the conservation method is in vitro conservation using a slow growth technique. Factorial experiment was used in designing the experiment, the first factor was MS medium concentration, i.e. 1/2MS and MS. The second factor was sucrose concentration, i.e. 0; 1; 2; and 3%. The results showed that low concentration of MS medium and sucrose reducing the leaf number and shoot length but increasing the root number and length. Based on inhibition of growth, the most reducing growth was planlet on MS without sucrose.

In vitro growth of Brassocattleya orchid hybrid in different concentrations of KNO3, NH4NO3 and benzylaminopurine

One of the most important applications of plant tissue culture is mass propagation of ornamental plants. This experiment evaluated the effect of different concentrations of NH4NO3 and KNO3 and BAP on the in vitro growth of orchid hybrid Brassocattleya 'Pastoral'. Seedlings of this orchid hybrid were used as explants and cultivated in medium with mineral salts and vitamins from the MS medium (Murashige & Skoog, 1962), with the macronutrients P, Ca and Mg reduced by half, and with an addition of 25 g L-1 of sucrose, 0.1 g L-1 of myo-inositol and 1.5 g L-1 of activated charcoal. Agar-agar was added (6.5 g L-1) and the pH was adjusted to 5.8. As treatments, four concentrations of the NH4NO3 and KNO3 (2x; 1x; ½ and ¼ MS medium) and three concentrations of BAP (0.0; 0.5 and 1.0 mg L-1) were assayed. The multiplication, growth in height, fresh and dry weight and sugar level in dry weight of sprouts were evaluated. There occurred a higher growth in height with 0.25x NH4NO3 and KNO3 salts concentrations of MS medium and higher rate of multiplication with combination of NH4NO3 and KNO3 reduced by half of the MS medium concentration and 1.0 mg L-1 BAP.

THE FIRST REPORT ON IN VITRO CULTURE OF MUSK ROSE

Musk rose is an important rose species for extracting rose water and rose oil, which is originated from Iran. This species has also uses in medical and food industries. Traditional methods of propagation of Musk rose (e.g. sucker, cutting, budding and grafting) are very slow and time-consuming and are usually associated with various problems such as limitation of stock plants and a prolonged production time. There is no report on in vitro propagation of Musk rose. This study was conducted on Rosa moschata to study the factors affecting micropropagation of this species for the first time. Results showed that, among the various concentrations of MS medium (MS, 1/2 MS, 1/3 MS, 1/4 MS), 1/2 MS medium was the best medium for proliferation. Comparing TDZ, BAP and KIN showed that TDZ was the best growth regulator for shoot proliferation. The highest percentage of proliferation was observed with the concentration of 1.25 mg/L TDZ. Using 1/2 MS + 1.5 mg/L TDZ + 0.1 mg/L NAA resulted in longest shoot (15 mm) in proliferation stage. The best shoot multiplication rate was observed in first subculture (4.71 shoot/explant) and thereafter this trait was decreased. The best treatment for rooting of microcuttings was obtained with 0.1 mg/L IBA + 0.1 mg/L 2,4-D. Plantlets were acclimatized in a medium consisting of 1:1 (v/v) peat moss and perlite and plantlets were successfully transferred to the greenhouse after 25 days.

Teor e composição química do óleo essencial de Melissa officinalis L. in vitro sob influência do meio de cultura

The present work was carried out with the objective of evaluating the effect of the culture medium in the content and chemical composition of the essential oil of Melissa officinalis , evaluating three concentrations of the MS culture medium. The experiment was performed in a completely randomized design, with 7 repetitions, where each repetition was represented by 20 g of fresh biomass. Oil extraction was accomplished using a Clevenger modified device for 1 hour and 30 minutes, and the chemical analysis was accomplished by gas chromatography. It was observed that the content and chemical composition of the essential oil of M. officinalis was influenced by the concentration of MS medium. The plants grown in the MS medium and MS/4 presented higher content of essential oil, and the major components of these plants were geraniol (25.23 and 16.21%, respectively) and nerol (24.5 and 20.53%, respectively). The major component present in the oil of melissa plants cultivated in MS/2 was neryl acetate (18.69%).