In vitro Growth of Genovese Basil in Response to Different Concentrations of Salts and Interaction of Sucrose and Activated Carbon

2018 ◽  
Vol 10 (9) ◽  
pp. 142 ◽  
Author(s):  
J. R. Trettel ◽  
A. B. Nascimento ◽  
L. N. Barbosa ◽  
H. M. Magalhães

Genovese basil has great economic potential; however, there is no established micropropagation protocol for this species. Therefore, this study aimed at assessing the in vitro growth of Genovese basil in response to different concentrations of salts in the Murashige and Skoog medium (MS) and interaction of sucrose and activated carbon. Two assays were conducted independently in an in vitro environment using the MS medium, regulators, agar, and five salt concentrations (0, 25, 50, 70 and 100%). In the second assay, two concentrations of sucrose (30 and 60 g L-1) and three concentrations of activated carbon (0, 3.0, and 4.5 g L-1) were tested.In addition, copper and zinc were quantified in the roots. The results showed that shoots were favored when the medium was at its full strength (100% salts), with seedlings forming more leaves.This result may be associated with a higher demand for nitrogen and because of the ionic balance between NH4+ and NO3-. High concentrations of salts affected the roots, but a reduction to 70% salt favored root development. Doubling the usual dose of sucrose (60 g L-1) damaged the growth of the seedlings. Damage caused by osmotic and oxidative potentials, and by toxic compounds may be related to the observed results. The amount of copper and zinc in the root increased with increased concentrations of activated carbon in the medium. The presence of activated carbon reduced callus formation but did not mitigate the effects of increased sucrose concentration.

2012 ◽  
Vol 30 (1) ◽  
pp. 119-124 ◽  
Author(s):  
Armando R Tavares ◽  
Jorge Luiz M Young ◽  
Sandra S Ori ◽  
Shoey Kanashiro ◽  
Giuseppina PP Lima ◽  
...  

In vitro cultivation is the main propagation method for the family Orchidaceae, whereas nitrogen is the most important nutrient in the culture media. This work was carried out to study the influence of different nitrogen concentrations on the in vitro growth of the orchid Phalaenopsis amabilis. Nitrogen concentrations varied by altering the ionic balance of the Murashige & Skoog (MS) culture medium. Plants, 360 days old, were cultivated in liquid MS, modified with 7.5, 15, 30, 45, and 60 mM N. After 180 days, we assessed plant and root length, number of leaves and roots, and fresh and dry weight of leaves, roots and plants. Treatments were assigned to completely randomized plots, with four replications. Plots consisted of five three-plant flasks. The lowest nitrogen level (7.5 mM) in the medium induced root development in length, number, and fresh and dry weight. The concentration 30 mM N stimulated both emission and dry weight accumulation of leaves. The original nitrogen concentration in the MS medium (60 mM) was excessive for the in vitro growth of P. amabilis.


2007 ◽  
Vol 2 (2) ◽  
pp. 297-306 ◽  
Author(s):  
Manuel Cantos ◽  
Juana Linán ◽  
José García ◽  
María García-Linán ◽  
Miguel Domínguez ◽  
...  

AbstractRhododendron ponticum subsp. baeticum is endemic in the southern region of the Iberian Peninsula. The relict populations of this species are vulnerable, due mainly to difficult conditions for the establishment of seedlings, resulting in a virtual lack of sexual recruitment. In order to preserve the surviving populations, in vitro culture methods have been applied for both the sexual and the agamic propagation of the species. The in vitro germination of seeds was high when conducted with Anderson’s medium without plant growth regulators. The self-rooted seedlings obtained were easily transplanted to outside conditions. The presence of growth regulators in the medium interfered with the development of the seedlings, causing heavy callus formation. The in vitro growth of explants took place readily in Anderson’s medium plus 0.072 mg L−1 of BA and 0.036 mg L−1 of NAA although the explants did not form roots. Rooting was achieved by the basal dipping of the explants in hydroalcoholic solutions of 500 mg L−1 IAA during the outside transplanting process. Therefore, the combination of in vitro grown explants together with ex vitro rooting, results in a good method for the agamic propagation of Rhododendron ponticum subsp. baeticum.


2017 ◽  
Vol 14 (2) ◽  
pp. 607-614
Author(s):  
Hossein Nazarian ◽  
Maryam Beigi Harchegani ◽  
Mahmoud Otroshy ◽  
Ali Motamedi

ABSTRACT: This study was designed in order to optimize the indirect organogenesis (during callus induction and regeneration) of Alstroemeria cv. ‘Balance’ through tissue culture technique in two phases; the first stage: callus induction by rhizome segments, leaf and nodal stem which in the start, callus formation media were examined using two types of auxins; 2,4-D and NAA and a cytokinin; BAP in four different experimentations. In the second stage, calli derived from rhizome segments and nodal stem explants were transferred to regeneration media. The results revealed that 2,4-D in combination with BAP in the rhizome segments and nodal stem explants were efficient as compared to NAA. The highest yield of callus formation was also obtained in the rhizome segments explants. According to the results, it can be suggested that NAA as auxin, does not have direct positive effect on cell division in Alstroemeria. The 2,4-D is toxic at high concentrations and may bring about cell death. Eventually, the composition of 0.5 mg/l NAA with 3 mg/l BAP and callus derived from nodal stem explants may be introduced as the best combination for regeneration. These results indicate the necessity of the BAP cytokinin presence for regeneration. In addition, the maximum length of the shoot was obtained from combination of BAP with nodal stem explants, without the presence of NAA.


2017 ◽  
Vol 59 (2) ◽  
pp. 31-39 ◽  
Author(s):  
Csongor Freytag ◽  
Sándor Attila Pabar ◽  
Zita Demeter ◽  
Ádám Simon ◽  
Anna Resetár ◽  
...  

AbstractWe aimed to produce tissue cultures and plant regeneration from endangered Crocus species: C. scepusiensis, C. tommasinianus, C. vittatus (“Verni” series of the genus) and C. banaticus. For initiation of cultures we used a plant growth regulator (PGR) combination used for in vitro culture of saffron and its relatives: 10 mg L-1 α-naphthaleneacetic acid (NAA) and 1 mg L-1 6-benzyladenine (BA). Shoot tips of young seedlings (C. scepusiensis) and corms (for the rest of species) were used as explants. C. scepusiensis explants developed into organogenic calli. On media with decreased NAA and with or without increased BA concentration, calli produced stigma-like structures and/or shoots and whole plants. In the other species, callus initiation medium induced callus formation with abundant somatic embryos. In C. tommasinianus, embryos developed shoots when auxin content of medium was decreased. In C. banaticus, a decrease of auxin with or without an increase in cytokinin content led to shoot or whole plant regeneration, as in C. scepusiensis. In the case of C. vittatus and C. banaticus, initiation and/or maintenance of cultures on indole-3-butyric acid (IBA) and increased sucrose concentration stimulated whole plant regeneration and in vitro cormlet development. C. scepusiensis and the rest of cultures (organogenic vs. embryogenic) differed at the biochemical level: C. scepusiensis cultures had higher (yet still low) enzymatic antioxidant (catalase, peroxidase) activities. With respect to catalase isoenzyme patterns, C. banaticus was different from the rest of cultures, demonstrating its distinct taxonomical position. Besides germplasm preservation use of the present cultures, they have a potential biotechnological value.


Plants ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 1768
Author(s):  
Agnieszka Wojtania ◽  
Monika Mieszczakowska-Frąc

Culinary rhubarb is a popular vegetable crop, valued for its long, thickened stalks, very rich in different natural bioactive ingredients. Tissue cultures are a useful tool for vegetative propagation of virus-free rhubarb plants and rapid multiplication of valuable selected genotypes. The aim of this study was to develop an effective method for in vitro propagation of selected genotypes of Polish rhubarb ‘Malinowy’ characterized by high yield and straight, thick and intensive red stalks. Identification and quantification of anthocyanins and soluble sugars by the HPLC method in shoot cultures and ex vitro established plantlets were also performed. Shoot cultures were established from axillary buds isolated from dormant, eight-year-old rhizomes. Effective shoot multiplication of rhubarb ‘Malinowy’ was obtained in the presence of 6.6 µM benzylaminopurine or 12.4 µM meta-topolin. Both cytokinins stimulated shoot formation in a manner that depended on sucrose concentration. Increasing the sucrose concentration from 59 to 175 mM decreased the production of shoots and outgrowth of leaves by 3-fold but enhanced shoot length, single shoot mass and callus formation at the base of shoots. This coincided with increased accumulation of soluble sugars (fructose, glucose) and anthocyanins-cyanidin-3-O-rutinoside (max. 208.2 mg·100 g−1 DM) and cyanidin-3-O-glucoside (max. 47.7 mg·100 g−1 DM). The highest rooting frequency (94.9%) and further successful ex vitro establishment (100%) were observed for shoots that were earlier rooted in vitro in the presence of 4.9 µM indole-3-butyric acid. Our results indicated that anthocyanin contents in leaf petioles were influenced by developmental stage. Under in vitro conditions, it is possible to elicit those pigments by sucrose at high concentration and meta-topolin.


2004 ◽  
Vol 22 (4) ◽  
pp. 780-783 ◽  
Author(s):  
Ricardo T. de Faria ◽  
Fabiana N. Rodrigues ◽  
Luciana do V.R. Oliveira ◽  
Cláudio Müller

Sucrose is a very important component in in vitro culture media, serving as a source of carbon and energy. In this paper, the rooting and in vitro growth of Dendrobium nobile Lindl (Orchidaceae) were studied using different sucrose concentrations (0 g L-1; 5 g L-1; 10 g L-1; 20 g L-1; 30 g L-1 and 60 g L-1), in a modified MS medium containing half the regular concentration of macronutrients at pH 5.8. Greater increases in plant height (4.21±0.6 cm) and high seedling multiplication (1:4) were observed in the 60 g L-1 sucrose treatment, even without the addition of plant hormones. Sucrose concentration in the culture medium did not influence in vitro plant rooting.


2013 ◽  
Vol 49 (1-2) ◽  
pp. 131-140 ◽  
Author(s):  
Eleonora Gabryszewska

The influence of temperature, daylength and sucrose concentration on the growth and development of <em>Alstroemeria</em> 'Zebra' <em>in vitro</em> was investigated. Only slightly more lateral rhizomes were formed at 25°C in comparison with 17°C on the medium with BA. Presence of BA in the medium strongly increased number of upright growing shoots and more shoots were formed in temperature 25°C than in temperature 17°C. Rhizome multiplication and formation of upright growing shoots were not significantly influenced by daylength. Sucrose was required for the formation of lateral rhizomes, upright growing shoots and roots. The highest number Of lateral rhizomes was observed on the medium containing 60 and 80 gl<sup>-1</sup> sucrose and BA. Presence of BA in the medium with different sucrose concentrations markedly influenced the formation of upright growing shoots; the highest number of shoots was found on the explants cultured on the media with 20 and 30 gl<sup>-1</sup> sucrose. Low and high concentrations of sucrose inhibited upright growing shoots formation and their elongation. Rhizogenesis was not observed on rhizomes growing on the medium without sucrose and with 10 gl<sup>-1</sup> sucrose, either in the presence or absence of NAA„ The number of roots per plantlet on the medium with NAA increased with increasing sucrose levels. The highest sucrose concentrations slightly inhibited roots formation.


2020 ◽  
pp. 395-403
Author(s):  
Zhanna Aleksandrovna Koh ◽  
Yuliya Aleksandrovna Litovka ◽  
Polina Vasil'yevna Makolova ◽  
Kseniya Aleksandrovna Shabanova ◽  
Igor' Nikolayevich Pavlov

The methods of sterilization of annual shoots Pinus sibirica Du Tour and the conditions for their introduction into in vitro culture were studied. Induction of callusogenesis of aseptically viable explants of P. sibirica proceeds more intensively on the modified Murasige-Skoog medium: hormonal supply of 0.4% kinetin and 0.25% 6-benzylaminopurine; reduced sucrose concentration of 1.5%. The frequency of callus formation was 83%. Close quantitative indicators of extractive substances were established (36 and 33% of absolutely dry weight for callus and explant, respectively); easily hydrolyzable polysaccharides (18 and 16%) and proteins (11 and 10%).Callus P. sibirica has a higher content of ascorbic acid, flavanoids, tocopherols and ash elements compared to explants and a low amount of hard-hydrolyzable polysaccharides, lipids, tannins, pigments, and essential oils.The electrophoretic spectrum of water-soluble callus proteins is represented by eleven fractions: 63% of the total water-soluble proteins are fractions with a molecular weight of 33 kD and above. Fractions with molecular weights of 50 and 62 kD (20 and 17%, respectively) are represented as much as possible.In the explants of P. sibirica, low molecular weight fractions of proteins with molecular masses of 5 kD and lower (59%) predominate. The amino acid composition of calli and explants of P. sibirica is identical and is represented by fifteen individual amino acids. Callus tissue has a higher content of glutamic acid and two hydrophobic amino acids (proline and isoleucine) compared to the vegetative part of the plant and low tyrosine content.


Author(s):  
María A. Aguilar Morales ◽  
Armandina De la Cruz Olvera ◽  
E. Archundia-Garduño ◽  
Rosy G. Cruz Monterrosa ◽  
Mayra Díaz-Ramírez ◽  
...  

Objective: The objective of this study was to establish the method of propagation of Oryganum vulgare and Lippia graveolens employing a plant tissue culture technique that decreased the phenolization percentages and increased the multiplication coefficients. Design/ methodology/ approach: The in vitro germination percentage was evaluated in both MS and MS medium + activated carbon. Microcuttings (small shoots) of both species were established in base medium added with different antioxidant agents to decrease the phenolization of explants; the treatments were arranged in a completely randomized block  design. For the propagation phase, a completely randomized factorial design was used, where the auxin/cytokinin phytoregulators, type of explants (axillary buds and leaves), and the species (Lippia graveolens and Oryganum vulgare)  were considered as factors. Results: maximum germination (63.3% ±12.5) was obtained on day 15 ​​in both culture media for L. graveolens and O. vulgare. The use of antioxidant agents mainly activated carbon, increased the in vitro establishment and activation of vegetative buds in both species by up to 90%. There were significant differences in the variables evaluated regarding the treatments, the explant, and the species in the multiplication phase. The combination 1.0/ 0.5 mg L-1 BA/AIB induces callus formation for both species. When used as leaf explants, callus formation was potentiated. Study Limitations / Implications: The results presented are advances from a long-term experiment. Findings/conclusions: The germination of L. graveolens seeds can be achieved in MS medium after 15 days. Microcuttings of both L. graveolens and O. vulgare were successfully established in MS basal medium enriched with 1 g L-1 charcoal that showed low oxidation percentages and induced up to 90% the production of shoots in the explants. The mixture of 1.0/0.5 mg L-1 BA/AIB induces callus formation for both species; when this medium is in contact with leaves as an explant, its formation is potentiated, achieving diameters up to 15 mm. In order to achieve the induction of shoots and roots, buds should be established in MS medium enriched with 0.5 mg L-1 IBA for both species; this mixture encreased the multiplication coefficients


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