Enhanced in vitro Shoot Regeneration and Biochemical Properties of Stevia rebaudiana using Chitosan

Stevia rebaudiana is a herbaceous perennial plant with great global demand due to its beneficial steviol glycosides(SGs) content. Current conventional breeding technique is unable to cater the need for more S. rebaudiana planting materials. Therefore, an improved in vitro shoot regeneration protocol was developed for S. rebaudiana by using chitosan. The highest fresh weight of plant (0.586 ± 0.176 g/explant), dry weight of plant (0.056 ± 0.02 g/explant) and plant height (4.94 ± 1.17 cm/explant) with maximum number of leaves (25.33 ± 6.95 /explant) were observed on explants grown in optimun treatment of MS basal medium supplemented with 1.0 mg/L of 6-benzyaminopurine (BAP) and 60 mg/L of low molecular weight (MW) chitosan after 4 weeks of culture. Scanning electron microscopy (SEM) analysis showed that new shoot primordia and shoot bud formation can be seen as early as day 3 and 5 of cultures on optimun treatment. Further biochemical assays showed that total phenolic acid content, total protein content and total hydrolyzed sugar content were recorded higher in explants cultured in optimun treatment as compared to control media. In contrast, total chlorophyll content and total flavonoids content were reduced in optimum treatment. Meanwhile, no significant difference in antioxidant activity was observed. All cultures from the optimal treatment were successfully regenerated, acclimatized and grew well with 100% survival rate. Taken together, an enhanced and efficient shoot regeneration protocol of S. rebaudiana was successfully developed which will be useful for rapid and large-scale micropropagation in future.

Download Full-text

In Vitro Shoot Regeneration from Callus Derived from Marubakaido Apple Rootstock under Different Aluminium Concentrations

HortScience ◽

10.21273/hortsci.33.3.460e ◽

1998 ◽

Vol 33 (3) ◽

pp. 460e-460 ◽

Cited By ~ 1

Author(s):

Marisa F. de Oliveira ◽

Gerson R. de L. Fortes ◽

João B. da Silva

Keyword(s):

Shoot Regeneration ◽

Dichlorophenoxyacetic Acid ◽

Apple Rootstock ◽

Under Five ◽

Significant Difference ◽

Previously Treated ◽

2,4 Dichlorophenoxyacetic Acid ◽

In Vitro Shoot Regeneration

The aim of this work was to evaluate the organogenesis of Marubakaido apple rootstock under different aluminium concentratons. The explants were calli derived from apple internodes treated with either 2,4-dichlorophenoxyacetic acid or pichloram at 0.5 and 1.0 μM and under five different aluminium concentrations (0, 5, 10, 15, 20 mg/L). These calli were then treated with aluminium at 0, 5, 10, 15, and 20 mg/L. It was observed shoot regeneration only for those calli previously treated with pichloram. There were no significant difference among the aluminium concentrations.

Download Full-text

PRODUKSI METABOLIT SEKUNDER STEVIOSIDA PADA KULTUR KALUS STEVIA (Stevia rebaudiana Bert. M.) DENGAN PENAMBAHAN ZPT 2,4-D DAN PEG (Polyethylene Glykol) 6000 PADA MEDIA MS (Murashige & Skoog)

el–Hayah ◽

10.18860/elha.v4i2.2627 ◽

2014 ◽

Vol 4 (2) ◽

pp. 57 ◽

Cited By ~ 2

Author(s):

Fittriya Nur Laila

Keyword(s):

Secondary Metabolites ◽

High Performance ◽

Food Additives ◽

Stevia Rebaudiana ◽

Steviol Glycosides ◽

Significance Level ◽

Peg 6000 ◽

Randomized Design ◽

Significant Difference

Stevia rebaudiana is a member of the Asteraceae family, produce steviol glycosides (steviosida, rebaudiosida A, B, C, D, E and dulcosida A) which can be used as food additives such as food flavoring or sweetener in nutritional supplements. This study aims to determine the concentration of 2,4-D combination and PEG 6000 were effective in increasing the production of secondary metabolites in callus of stevia (Stevia rebaudiana Bert. M) in vitro. PEG 6000 is a way to manipulate media in vitro to increase secondary metabolite. This research used Completely Randomized Design (CRD) with 12 factorial combination with three replications. The first factor is the provision of plant growth regulator 2,4-D (1 mg / L, 2 mg / L, and 3 mg / L) and the second factor is the provision of PEG 6000 (0 mg / L, 5 mg / L, 15 mg / L, and 25 mg / L). The emerging callus (day), percentage (%) of explants callus (g), weight of callus, callus morphology (texture and color of the callus), and secondary metabolites steviosida were measured. Data were analyzed by analysis of variance (ANOVA) two way and to find a significant difference test Duncan's Multiple Range Test (DMRT) at 5% significance level. Steviosida secondary metabolites were measured using High Performance Liquid Cromatography (HPLC). The results showed that the combination treatment of 1 mg / L 2,4-D and 25 mg / L PEG 6000 is a combination of the most efficient way to get the amount of secondary metabolites steviosida 4.792 mg / g. Observations callus morphology (texture and color of the callus) showed that the compact callus textured and colored brown due to osmotic stress of PEG 6000 has a high content of secondary metabolites steviosida

Download Full-text

Release Kinetic Studies of Stevia rebaudiana Extract Capsules from Sodium Alginate and Inulin by Ionotropic Gelation

Advances in Materials Science and Engineering ◽

10.1155/2018/6354924 ◽

2018 ◽

Vol 2018 ◽

pp. 1-8

Author(s):

Juan Pablo Quintal Martínez ◽

Jorge Carlos Ruiz Ruiz ◽

Maira Rubí Segura Campos

Keyword(s):

Sodium Alginate ◽

Release Kinetics ◽

Stevia Rebaudiana ◽

Kinetic Studies ◽

Release Kinetic ◽

Steviol Glycosides ◽

Ionotropic Gelation ◽

Diffusion Controlled ◽

Release Analysis

This study was oriented towards encapsulation of S. rebaudiana extract and the study of its release kinetics. The desired encapsulation was achieved by the ionotropic gelation method using sodium alginate and inulin of polymeric constituents. Characterization of the capsules was performed by micrometric properties, encapsulation efficiency, in vitro extract release analysis, and biological activity of released extract. The in vitro release profiles from different capsules were applied on different kinetic models. The prepared capsules were found spherical in shape with diameters ranging from 2.07 to 2.63 mm, having the encapsulation efficiencies of 43.77% and 56.53% for phenolic compounds and steviol glycosides, respectively. The best-fit model with the highest correlation coefficient was observed in the Ritger–Peppas model, indicating diffusion controlled principle. The release exponent n value obtained from the Korsmeyer–Peppas model varied between 0.2273 and 1.1719, confirming that the mechanism of S. rebaudiana extract bioactive compounds release was diffusion controlled.

Download Full-text

An Improved Protocol for Multiple Shoot Regeneration from Seedlings and Mature Explants of Citrus macroptera Mont.

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v18i1.3246 ◽

2009 ◽

Vol 18 (1) ◽

pp. 17-24

Author(s):

Md. Nesawar Miah ◽

Shahina Islam ◽

Syed Hadiuzzaman

Keyword(s):

Shoot Regeneration ◽

Multiple Shoots ◽

Multiple Shoot ◽

Nodal Explants ◽

Cow Dung ◽

Half Strength ◽

Multiple Shoot Regeneration ◽

Shoot Tip Explants ◽

In Vitro Shoot Regeneration

Efforts have been made to establish a protocol for direct multiple shoot regeneration from both in vitro grown seedlings and mature plants of Citrus macroptera. Both nodal and shoot tip explants taken from in vitro grown seedlings were cultured in MS supplemented with different concentrations of BAP and Kn either singly or in combinations. Both these explants are capable to regenerate and produce in vitro multiple shoots. Maximum number of shoots were obtained from nodal explants in MS supplemented with 1.0 mg/l BAP. BAP alone was found superior to Kn. On the other hand, only nodal explants from mature plants were used and 1.0 mg/1 BAP was also found best suitable for shoot induction and multiplication. Ex vitro rooting in pot soil (mixed with biogas slurry derived from cow-dung) was most successful compared to in vitro rooting in half strength of MS supplemented with different concentrations of NAA and IBA. Key words: In vitro, Shoot regeneration, Citrus macroptera D.O.I. 10.3329/ptcb.v18i1.3246 Plant Tissue Cult. & Biotech. 18(1): 17-24, 2008 (June)

Download Full-text

In vitro shoot regeneration of boldo from leaf explants

Ciência Rural ◽

10.1590/s0103-84782010005000173 ◽

2010 ◽

Vol 40 (10) ◽

pp. 2210-2213

Author(s):

Monalize Salete Mota ◽

Juliana de Magalhães Bandeira ◽

Eugenia Jacira Bolacel Braga ◽

Valmor João Bianchi ◽

José Antonio Peters

Keyword(s):

Shoot Regeneration ◽

Leaf Explants ◽

Shoot Development ◽

High Potential ◽

Naphthaleneacetic Acid ◽

Regeneration System ◽

Bud Induction ◽

Molecular Studies ◽

In Vitro Shoot Regeneration

A shoot regeneration system for Plectranthus neochilus was studied from leaf explants. Leaves developed under in vitro conditions were cultured on Wood Plant Medium supplemented with 0.2mg dm-3 α-naphthaleneacetic acid (NAA) and different 6-benzilaminopurine (BAP) or thidiazuron (TDZ) concentrations (0, 1.5, 3.0, 4.5 and 6.0mg dm-3). An increase in percentage of responsive explants (85.3%) and in the number of shoots developed per explant (3.2) was observed when the explants were treated with 5.3 and 4.7mg dm-3 BAP, respectively. The leaf explants cultured on media supplemented with TDZ became vitreous and did not form buds. The regeneration system used is efficient for boldo bud induction and shoot development, showing high potential for advanced cellular and molecular studies.

Download Full-text

IN VITRO SHOOT REGENERATION FROM DIFFERENT EXPLANTS OF HOSTA SIEBOLDIANA CV. GOLD STANDARD

Acta Horticulturae ◽

10.17660/actahortic.1989.251.33 ◽

1989 ◽

pp. 229-234

Author(s):

M. Lubomski

Keyword(s):

Shoot Regeneration ◽

Gold Standard ◽

In Vitro Shoot Regeneration

Download Full-text

Media composition affects seed dormancy, apical dominance and phenolic profile of Knautia sarajevensis (Dipsacaceae), Bosnian endemic

Acta Botanica Croatica ◽

10.1515/botcro-2017-0011 ◽

2018 ◽

Vol 77 (1) ◽

pp. 70-79 ◽

Cited By ~ 1

Author(s):

Erna Karalija ◽

Sanja Ćavar Zeljković ◽

Petr Tarkowski ◽

Edina Muratović ◽

Adisa Parić

Keyword(s):

Apical Dominance ◽

Germination Rate ◽

Natural Habitat ◽

Basal Medium ◽

Shoot Proliferation ◽

Mineral Nutrient ◽

Total Phenolic ◽

Endemic Plant ◽

High Concentrations

AbstractKnautia sarajevensisis an endemic plant of the Dinaric Alps and is mainly distributed on Bosnian Mountains. Due to the quite large flower heads and easy maintenance, this plant has a potential use as a substitute ornamental plant forK. arvensisin perennial beds. The current study evaluated the germination process in different treatments in an attempt to suppress dormancy and increase germination rate, and to develop a successful protocol for micropropagation. An over 60% germination rate was achieved through cultivation of seeds on MS basal medium with reduced mineral nutrient composition and the absence of sucrose. On the other hand, a below 10% germination rate was achieved with untreated seeds. Suppression of apical dominance was achieved through application of high concentrations of kinetin, apical shoot decapitation or cultivation of shoots in liquid media. Overall, liquid cultures were more successful as a micropropagation system for this plant. Shoots spontaneously developed roots on multiplication treatments and were successfully acclimatized. Moreover, phenolic compound profile was analysed in the light of the possible medicinal potential of this plant. Variable amounts of total phenolic compounds as well as individual phenolics were recorded, according to treatment and solidification of media. An increase in rosmarinic acid content was reported for kinetin treatments and acclimatized plants comparing to mother plants in natural habitat. The present study shows that choice of cytokinin concentration, explant type as well as culture type influences not only shoot proliferation and apical dominance suppression but alsoin vitroproduction of phenolics.

Download Full-text

In Vitro Effects of Different Combinations of Phytohormones on Callus Induction from Different Explants of Cabbage Brassica Oleracea Var. Capitata L. Seedlings

Iraqi Journal of Science ◽

10.24996/ijs.2021.62.10.7 ◽

2021 ◽

pp. 3476-3486

Author(s):

Alaa. M. Hasan ◽

Ekhlas. A.J. ElKaaby ◽

Rakad. M.Kh. AL-Jumaily

Keyword(s):

Brassica Oleracea ◽

Callus Induction ◽

Basal Medium ◽

Culture Conditions ◽

Control Treatment ◽

Fresh Weight ◽

Superior Result ◽

Significant Difference ◽

In Vitro Effects

The leading purpose of this work is the development of efficient culture conditions to induce calli from cabbage (Brassica oleracea var. capitata L.) under in vitro conditions. The mature seeds were surface sterilized with combinations of different concentrations of ethanol and NaOCl in different time durations and were germinated on MS basal medium. The results revealed that the best sterilization method of cabbage seeds was by using 70% ethanol for one minute, followed by 15 min in 2% (NaOCl). Seedlings were used as donor sources for hypocotyls, cotyledon leaves, true leaves, and shoot tip explants. These explants were cultured on different combinations of cytokinins (TDZ, BAP, Ad) and auxins (IAA, NAA, 2, 4-D) then implanted in Murashige and Skoog (MS) media. 4 weeks after culturing, a significant difference was found among the explants in response to plant hormones. The maximum percentage of callus induction (100%) was using the combinations of 1 BAP + 1 2, 4-D, 1 BAP + 1 NAA, and 1 BAP + 2 2,4-D mg. l-1. In addition, explants responses varied and the hypocotyls showed a superior result (85.71 %) as compared to other explants. For callus fresh weight, the combination of 0.22 TDZ + 79.9 Ad mg. l-1 had a significant effect, causing the highest fresh weight (0.2745g), while control treatment gave the lowest mean of 0.0066 g. Data showed that cotyledon explants were significantly superior in giving highest callus fresh weight with the mean of 0.1723 g. On the other hand, hypocotyl explants gave the lowest mean, reaching 0.1542 g.

Download Full-text