scholarly journals A fast and effective protocol for obtaining genetically diverse stevia (Stevia rebaudiana Bertoni) regenerants through indirect organogenesis

2021 ◽  
Vol 76 (4) ◽  
pp. 47-62
Author(s):  
Magdalena Dyduch-Siemińska
Keyword(s):  
Callus Tissue ◽  
Phenotypic Variability ◽  
Leaf Explants ◽  
Stevia Rebaudiana ◽  
Nodal Segments ◽  
Naphthaleneacetic Acid ◽  
Indirect Organogenesis ◽  
Stevia Rebaudiana Bertoni ◽  
One Step ◽  
The Mean

Plant regeneration through indirect organogenesis allows obtaining genetic variability that can be used in the creation of new cultivars. The study presents a fast and effective protocol of one-step preparation of stevia (Stevia rebaudiana Bertoni) regenerants. To obtain callus tissue and shoot regeneration, leaves and nodal segments were used as primary explants, which were placed on MS (Murashige and Skoog) medium supplemented with plant growth regulators (PGRs): NAA (1-naphthaleneacetic acid – 2.0 mg·dm–3, BA (6-benzylaminopurine – 4.0 mg·dm–3), 2,4‑D (2,4-dichlorophenoxyacetic – 2.0 mg·dm–3). Callus tissue was formed on both types of explants, however, only those derived from nodal segments were proliferating. An average of 3.92 shoots per explant were obtained from leaf explants on the applied medium after 6 weeks of culture. The analysis of the morphogenetic capacity of the obtained regenerants was carried out on MS medium supplemented with PGRs – kinetin (0.25 mg·dm–3), BA (0.5 mg·dm–3). The evaluation of the mean number of shoots, mean shoot length (cm), and the mean number of nodes per shoot indicated phenotypic variability of regenerants. The use of RAPD (randomly amplified polymorphic DNA) markers confirmed the differences also at the DNA level. The proposed one-step indirect organogenesis regeneration protocol induced somaclonal variation of Stevia rebaudiana Bertoni and the obtained regenerants, after selection, could be used in the breeding of this species.

scholarly journals Comparative analyses of stevioside between fresh leaves and in-vitro derived callus tissue from Stevia rebaudiana Bert. using HPLC

2015 ◽  
Vol 49 (4) ◽  
pp. 199-204 ◽  
Author(s):  
S Mahmud ◽  
S Akter ◽  
IA Jahan ◽  
S Khan ◽  
A Khaleque ◽  
...  
Keyword(s):  
Retention Time ◽  
Callus Tissue ◽  
Callus Formation ◽  
Leaf Explants ◽  
Stevia Rebaudiana ◽  
Poor Performance ◽  
The Other ◽  
Ms Medium ◽  
Green Callus

A protocol was developed to produce large amount of callus in short a period of time from leaf explants of Stevia rebaudiana Bert. The highest amount of white callus was obtained on MS medium supplemented with 2.5 mg/l 2, 4-D and 0.5 mg/l BAP after 3 weeks of inoculating leaf segments. On the other hand, 0.5 mg/l BAP and 1.0 mg/l Kn exhibits poor performance towards callus formation while after using 1.0 mg/l Kn alone did not develop any callus. In this experiment, highest amount of green callus was obtained when MS medium supplemented with 2.5 mg/l NAA and 10% coconut water was used. An improved analytical method HPLC was applied to analyze stevioside extracted from the leaf and callus of Stevia rebaudiana. The stevioside in each sample were analyzed by comparing their retention times with those of the standards. The retention time (RT) of stevioside for leaves were found 14.96 and for callus 13.81 mins. The percentage of stevioside content from leaves and callus was 12.19% and 12.62% respectively DOI: http://dx.doi.org/10.3329/bjsir.v49i4.22621 Bangladesh J. Sci. Ind. Res. 49(4), 199-204, 2014

scholarly journals Regeneration of Plants from Apical Meristem Tips and Nodal Segments of Arachis pintoi

2003 ◽  
Vol 30 (2) ◽  
pp. 75-79 ◽  
Author(s):  
H. Y. Rey ◽  
L. A. Mroginski
Keyword(s):  
Apical Meristem ◽  
In Vitro Regeneration ◽  
Nodal Segments ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Regeneration Potential ◽  
Arachis Pintoi ◽  
Solid Media ◽  
One Step

Abstract The in vitro regeneration potential of shoot apical tips (2 to 3 mm in length), meristems (0.3 to 0.5 mm in length), and nodal segments (4 to 7 mm long with an axillary bud) of diploid (2n = 2x = 20) and triploid (2n = 3x = 30) cytotypes of Arachis pintoi was evaluated. Explants were cultured on MS medium supplemented with different concentrations and combinations of naphthaleneacetic acid (NAA) and benzyladenine (BA). In one experiment the effect of gibberellic acid was tested. The cultures were done in liquid and solid media. Plant regeneration can be readily achieved from all explants in one step of 30 d culture on MS + 0.01 mg/L each of NAA and BA or two steps consisting of 1) shoots regeneration through culture of explants on MS + 0.01 mg/L each of NAA and BA, and 2) induction of rooting in regenerated shoots by reculture on MS + 0.01 mg/L NAA. The plantlets were successfully transferred to pots in a greenhouse.

scholarly journals In Vitro Micropropagation of Stevia rebaudiana Bertoni

2019 ◽  
Vol 29 (2) ◽  
pp. 277-284
Author(s):  
Sabina Yesmin
Keyword(s):  
Morphological Variation ◽  
Stevia Rebaudiana ◽  
Multiple Shoots ◽  
Nodal Segments ◽  
Root Induction ◽  
Full Strength ◽  
Stevia Rebaudiana Bertoni ◽  
Regenerated Plants ◽  
In Vitro Micropropagation

Multiple shoots were obtained from both shoot tips and nodal segments cultured on MS fortified with different concentrations of BAP and Kn singly or in combination with low concentration of NAA. Maximum number of shoots (9.28 ± 0.61) were found on MS supplemented with 1.5 mg/l BAP and 05 mg/l NAA. In vitro regenerated shoots were separated and transferred onto half and full-strength MS supplemented with different concentration of IBA, IAA and NAA for root induction. Full strength MS containing 0.2 mg/l IBA was found to be best for root induction where 93.33% shoots were rooted. In vitro regenerated plants grew normally without showing any morphological variation and flowered after 40 days of transplantation.

scholarly journals In vitro micropropagation of Rauvolfia serpentina (L.) Benth through induction of direct and indirect organogenesis

2013 ◽  
pp. 01-09
Author(s):  
SK Bhadra ◽  
TK Bhowmik ◽  
P Singh
Keyword(s):  
Callus Tissue ◽  
Multiple Shoot ◽  
Nodal Segments ◽  
Nodal Segment ◽  
Ms Medium ◽  
Indirect Organogenesis ◽  
Rauvolfia Serpentina ◽  
Shoot Buds ◽  
Wide Range ◽  
Multiple Shoot Buds

Leaf and nodal segments of two months old field grown seedlings of Rauvolfia serpentina (L.) Benth were aseptically cultured on agar solidified MS medium supplemented with various combinations and concentrations of auxins (NAA, IAA, 2,4-D and picloram) and cytokinins (BAP and Kn). The nodal segments produced highest number of multiple shoot buds (5.85/explant) on MS medium supplemented with 2.0 mgl-1 BAP + 0.2 mgl-1 NAA or 2.0 mgl-1 BAP + 0.1 mgl-1 IAA. Whereas nodal segment produced callus tissue of different nature on MS medium supplemented with 1.5 mgl-1 BAP + 0.5 mgl-1 IAA+ 1.5 mgl-1 2,4-D; 3.0 mgl-1 BAP + 1.0 mgl-1 NAA + 1.5 mgl-1 Kn and 0.1 mgl-1 Pic + 1.0 mgl-1 Kn. The callus tissue of light green and nodular nature on further subculture in a wide range of plant growth regulators (PGRs) supplemented media, differentiated into multiple shoot buds that underwent rapid elongation on 2.0 mg/l BAP and 0.2 mg/l NAA supplemented media. The elongated shoot buds on further subculture in rooting media produced strong and stout roots. Half strength MS with 1.5% (w/v) sucrose was most effective for enhancing rooting. Finally those plantlets were acclimatized in field. Thus a protocol was established for rapid micropropagation of this medicinal plant through induction of direct and indirect organogenesis from nodal explant. DOI: http://dx.doi.org/10.3329/cujbs.v3i1.13401 The Chittagong Univ. J. B. Sci.,Vol. 3(1&2):01-09, 2008

scholarly journals The Success Rate of Two Explant Types of Stevia (Stevia rebaudiana Bertoni) in Various Sterilant Formulas

2020 ◽  
Vol 3 (2) ◽  
pp. 92
Author(s):  
Riedha Kariena ◽  
Nofia Hardarani ◽  
Hilda Susanti
Keyword(s):  
Success Rate ◽  
Leaf Explants ◽  
Stevia Rebaudiana ◽  
Stevia Rebaudiana Bertoni ◽  
Randomized Design ◽  
Two Factors

This study aims to determine the effect of interaction between several sterilants and types of explants and determine the best interaction with stevia culture's success rate. This study was designed using a Factorial Complete Randomized Design with two factors and three replications. The first factor is sterilant formulas i.e: fungicide 3% + alcohol 70% + Bayclin 5%; fungicide 3% + bactericide 6% + 70% alcohol + Bayclin 5%; sublimate 0.1% + 70% alcohol + Bayclin 5%; and fungicide 3% + bactericide 6% + sublimate 0.1% + alcohol 70% + Bayclin 5%. The second factor is explant types, i.e., stevia nodes and leaves. The variables observed are the percentage of contamination (%), percentage of alive explants (%), and browning percentage.  The interaction between sterilants and explant types only had a significant effect on the percentage of alive explants. The best interactions of sterilant formulas and explant types on the percentage of alive explants are 0.1% sublimate + 70% alcohol + 5% Bayclin and leaf explants.

scholarly journals Plant Regeneration from Leaf Explants of the Medicinal Herb Wedelia chinensis

Horticulturae ◽  
2021 ◽  
Vol 7 (10) ◽  
pp. 407
Author(s):  
Yung-Ting Tsai ◽  
Kin-Ying To
Keyword(s):  
Plant Regeneration ◽  
In Vitro Propagation ◽  
Liver Toxicity ◽  
Leaf Explants ◽  
Basal Medium ◽  
Nodal Segments ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Average Percentage

Wedelia chinensis, belonging to the Asteraceae family, has been used in folk medicine in East and South Asia for the treatment of common inflammatory diseases and protection against liver toxicity. Previously, in vitro propagation through different tissue explants has been reported, including through nodal segments, axillary buds, and shoot tips, whereas leaf segments failed to proliferate. Here, we report on the in vitro propagation of W. chinensis by culturing young leaf explants in MS medium supplemented with 0.5 mg/L α-naphthaleneacetic acid (NAA), 0.75 mg/L thidiazuron (TDZ), 1 mg/L gibberellic acid (GA3), 3.75 mg/L adenine, 3% sucrose, and 0.8% agar at pH 5.8. Calli were observed in all explants derived from the youngest top two leaves, and the average percentage of shoot regeneration was 23% from three independent experiments. Then, several shoots were excised, transferred onto MS basal medium supplemented with 3% sucrose and 0.8% agar at pH 5.8, and cultured in a growth chamber for 1 to 2 months. Roots were easily induced. Finally, plantlets carrying shoots and roots were transferred into soil, and all of them grew healthily in a greenhouse. No morphological variation was observed between the regenerated plantlets and the donor wild-type plants. In addition, we also established root cultures of W. chinensis in culture medium (MS medium, 3 mg/L NAA, 3% sucrose, pH 5.8) with or without 0.8% agar. To the best of our knowledge, this is the first paper reporting plant regeneration from leaf explants in the herbal plant W. chinensis.

scholarly journals Vegetative Propagation of Hoya imperialis and Hoya coronaria by Stem Cutting and Micropropagation

2021 ◽  
Vol 32 (3) ◽  
pp. 1-23
Author(s):  
Siti Madihah Mohd Don ◽  
Nur Maziyyah Abdul Hamid ◽  
Hussein Taha ◽  
Rahayu Sukmaria Sukri ◽  
Faizah Metali
Keyword(s):  
Leaf Explants ◽  
Hormone Treatment ◽  
Stem Cutting ◽  
Ex Situ ◽  
Peat Moss ◽  
Naphthaleneacetic Acid ◽  
Stem Cuttings ◽  
Brunei Darussalam ◽  
The Mean ◽  
Propagation Methods

Hoya imperialis (H. imperialis) and H. coronaria (Apocynaceae) are known to have ornamental value due to their beautiful flowers; however, the feasibility of propagating these plants have not been reported despite the wild populations in Brunei Darussalam being highly threatened due to habitat loss and overcollection. Thus, the present study aimed to conduct a preliminary study of the feasibility of two alternative propagation methods, stem cutting and micropropagation, as a potential approach for their ex situ conservation. Hoya stem cuttings were treated with either indole-3-butyric acid (IBA) or 1-naphthaleneacetic acid (NAA) (0–2000 mg/L), and then propagated onto a mixture of peat moss and perlite. For micropropagation, Hoya leaf explants were cultured onto Murashige and Skoog (MS) agar media that were supplemented with IBA and/or kinetin (KN) (0–10.0 mg/L). This present study shows that both Hoya species were successfully propagated by stem cutting even without hormone treatment. However, interestingly, in H. imperialis, when compared with control, the mean number of new leaves (6.3 ± 1.0) and the mean relative growth rate (RGR) based on stem diameter (0.004 ± 0.0007 cm cm−1 day−1) significantly increased when treated with 500 mg/L NAA and 2000 mg/L IBA, respectively. Meanwhile, in H. coronaria, significantly higher mean number of roots was achieved by treating with 1000 mg/L NAA (16.6 ± 1.4) or 2000 mg/L IBA (17.5 ± 2.7) compared with control. For micropropagation, callus induction was not promising and could only be observed at specific concentrations of both IBA and KN, with H. imperialis appearing to be more responsive towards these hormones in comparison to H. coronaria. The present study showed that stem cutting appeared more feasible in propagating both Hoya species.

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