In vitro regeneration of Melastoma malabatricum Linn. through organogenesis and assessment of clonal and biochemical fidelity using RAPD and HPLC

Ghimire, Bimal Kumar; Seong, Eun Soo; Nguyen, Truong Xuan; Yu, Chang Yeon; Kim, Seung Hyun; Chung, Ill‐Min

doi:10.1007/s11240-015-0911-3

Cited by 16 publications

(6 citation statements)

References 34 publications

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“…The effectiveness of TDZ on shoot regeneration may be explained by the capability of this PGR to stimulate the biosynthesis of endogenous adenine-type cytokinin (Huetteman and Preece, 1993) and/or ability to work as an efficient bioregulator of in vitro morphogenesis (Murthy et al, 1998). Similar to the findings of this study, the formation of many shoots with shorter lengths on medium supplemented with TDZ and auxin has been documented for many plant species including Rhododendron (Ghimire et al, 2016), Exacum travancoricum (Kannan et al, 2007), and Levisticum officinale (Wang et al, 2014). However, the results obtained in this study are not consistent with those obtained by (Ledbetter and Preece, 2004) for in vitro regeneration of Hydrangea quercifolia from leaf explants, which showed that a higher level of TDZ in combination with IBA produced many short shoots, whereas a lower level of TDZ induced fewer and longer shoots.…”

Section: Discussionsupporting

confidence: 80%

Ascorbic Acid Controls Lethal Browning and Pluronic F-68 Promotes High-frequency Multiple Shoot Regeneration from Cotyldonary Node Explant of Okra (Abelmoschus esculentus L.)

Irshad¹,

Rizwan²,

Debnath³

et al. 2018

horts

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The regeneration frequency of okra (Abelmoschus esculentus) is greatly influenced by its genetic makeup and recalcitrant nature. Phenolic secretion, in particular, is a major problem in okra tissue culture. This study describes a reproducible, rapid, and more efficient in vitro regeneration method using cotyledonary node explants of okra. Explants were incubated on Murashige and Skoog (MS) medium containing different concentrations and combinations of various plant growth regulators (PGRs) [benzyladenine (BA), thidiazuron (TDZ), and α-naphthylacetic acid (NAA)], and regeneration enhancers [silver nitrate (AgNO3) and Pluronic F-68]. Cut ends of cotyledonary node segments rapidly turned brown and cultures failed to establish. Antibrowning additives, such as activated charcoal (AC), ascorbic acid (AA), and AgNO3 at various concentrations in PGR-free MS basal medium were tested for their ability to control phenolic secretion from explants. Among these additives, 15 mg·L−1 AA was found to be optimal for controlling phenolic secretion, resulting in healthy explants and culture establishment. The highest number of shoots (a mean of 9.3 ± 0.9 shoots per cotyledonary node explant) was obtained on MS media containing 0.5 mg·L−1 NAA + 1 mg·L−1 TDZ + 0.1% Pluronic F-68. Individual shoots were elongated on MS medium + 1 mg·L−1 BA + 0.1 mg·L−1 gibberellic acid (GA3) (shoot length 5.3 ± 0.2 cm) and rooted on ½ MS medium + 1 mg·L−1 indole-3-butyric acid (IBA) and 200 mg·L−1 AC (5.3 ± 0.2 roots per shoot). Rooted plantlets were acclimatized in plastic pots inside a plant growth chamber at 25 ± 2 °C and 70% relative humidity, with an 80% survival rate. This optimized protocol can be used for producing transgenic plants of commercial okra cultivars through genetic transformation.

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Section: Discussionsupporting

confidence: 80%

Ascorbic Acid Controls Lethal Browning and Pluronic F-68 Promotes High-frequency Multiple Shoot Regeneration from Cotyldonary Node Explant of Okra (Abelmoschus esculentus L.)

Irshad¹,

Rizwan²,

Debnath³

et al. 2018

horts

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“…Similar response of BAP in combination with 2-iP for shoot multiplication has also been reported in Croton scabiosus (Salamma and Rao 2014 ). On contrary to the above result, optimum shoot multiplication was obtained on MS medium fortified with a combination of BAP and Kn in Momordica balsamina and T. cordifolia (Thakur et al 2011 ; Sivakumar et al 2014 ), TDZ and NAA in Melastoma malabatricum (Ghimire et al 2016 ). In vitro raised elongated shoots (9.3 ± 0.48 cm) were transferred onto root induction medium composed of full, ½ and ¼ strength of MS salts.…”

Section: Resultsmentioning

confidence: 87%

Enhanced production of berberine in In vitro regenerated cell of Tinospora cordifolia and its analysis through LCMS QToF

Mittal

Sharma

2017

3 Biotech

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Tinospora cordifolia is a prioritized medicinal plant and having an immense medicinal importance especially in Indian medicinal system. But this plant needs a regeneration protocol for its rapid propagation. An efficient regeneration protocol was developed for T. cordifolia using nodal explants. High frequency of multiple shoot formation was induced when the nodal segments were cultured on MS medium supplemented with BAP (1.0 mg L−1) and 2-iP (0.5 mg L−1). The highest mean number of shoots per nodal explant (7.9 ± 0.45) with highest shoot length (9.3 ± 0.48 cm) and 86% response were achieved on this media and hormonal concentration. The optimum rooting was obtained on ½ strength of MS medium augmented with IBA (0.5 mg L−1) with 8.3 ± 0.46 cm root length and 89% response. Micropropagated plantlets were found to be identical with the mother plant when clonal fidelity of these plantlets were analyzed with inter simple sequence repeat (ISSR) marker. The berberine content was analyzed through LCMS QToF and the highest amount was found in in vitro callus (19.8 µg/gm) followed by stem (9.3 µg/gm) and leaves of field-grown plants (8.4 µg/gm). Further, presence of berberine was confirmed by ESI–MS spectra with protonated molecular ions ([M + H]+) at m/z 336. Furthermore, MS–MS fragmentation pattern confirmed for the presence of berberine in both the samples. Both the spectra (standard and samples) showed common peaks for berberine in the form of protonated molecular ions ([M + H]+) at m/z 320, m/z 304, m/z 292, m/z 278 in MS/MS mode. The study revealed that developed protocol is potent for rapid mass propagation of this plant species with high accumulation of important secondary metabolite berberine.

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“…It significantly differed from the control and other vegetative strains used in all the tested nutrient media. As for the effect of the explant type, the results show that the stems were significantly superior to the leaves and roots in all the tested nutrient media, as they needed the lowest number of days for the differentiation of somatic embryos into cultivable buds to start; this is attributed to the rate of nutrient accumulation in the stems and the consistency of their plant hormone content with the amount of the medium, in addition to the potential energy in the cells forming the buds and their ability to divide, which stimulated their growth and development (Ghimire et al, 2016). On the other hand, the superiority of the MS medium may be attributed to its richness in certain nutrients, especially macro-elements, such as magnesium and calcium; the importance of these elements in vital reactions in the plant; and the balance of the concentrations of the medium's elements with the explant content of the nutrients not having a toxic effect; this, in turn, leads to the ideal concentrations for the growth, development, and exposure of the cultivated plant part (Mineo, 1990).…”

Section: Days Required For Somatic Embryos To Differentiate Into Vege...mentioning

confidence: 97%

Effects of basic media and plant growth regulators on Direct-somatic embryogenesis induction in vegetative strains obtained from the tissue culture of digitalis purpurea

2023

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This study aimed to investigate the effects of certain nutrient medium types enhanced with plant-growth regulators on the direct inductionof somatic embryogenesis in vegetative strains produced and propagated from a tissue culture of Digitalis purpurea. The results showthat MS medium was superior to other tested nutrient media-B5, LS, and 5C01—in terms of the days required to initiate direct somaticembryogenesis induction and induction rate. In contrast, the LS medium resulted in the highest number of embryos. Moreover, 2 mg/LBA (benzyl adenine) was superior to similar concentrations of BAP (benzyl aminopurine) and Kin (kinetin) in the MS medium, whereasBAP performed better in LS and B5. On the other hand, auxin 2.4D (dichlorophenoxy acetic acid) inhibited somatic embryogenesis, as didNAA (naphthalene acetic acid) and IAA (indole acetic acid) when added individually. Regarding the type of explant, the stems showedthe induction rates and numbers of direct-somatic embryos produced in all the basic media.Keywords: Digitalis purpurea; Vegetative strains; Direct-somatic embryogenesis; Explant; Basic media

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In vitro regeneration of Melastoma malabatricum Linn. through organogenesis and assessment of clonal and biochemical fidelity using RAPD and HPLC

Cited by 16 publications

References 34 publications

Ascorbic Acid Controls Lethal Browning and Pluronic F-68 Promotes High-frequency Multiple Shoot Regeneration from Cotyldonary Node Explant of Okra (Abelmoschus esculentus L.)

Ascorbic Acid Controls Lethal Browning and Pluronic F-68 Promotes High-frequency Multiple Shoot Regeneration from Cotyldonary Node Explant of Okra (Abelmoschus esculentus L.)

Enhanced production of berberine in In vitro regenerated cell of Tinospora cordifolia and its analysis through LCMS QToF

Effects of basic media and plant growth regulators on Direct-somatic embryogenesis induction in vegetative strains obtained from the tissue culture of digitalis purpurea

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