scholarly journals High Frequency Plant Regeneration System from Transverse Thin Cell Layer Section of In vitro Derived ‘Nadia’ Ginger Microrhizome

2014 ◽  
Vol 6 (1) ◽  
pp. 85-91
Author(s):  
Dikash Singh THINGBAIJAM ◽  
Devi Sunitibala HUIDROM

An efficient and reproducible procedure is outlined for rapid in vitro multiplication of Zingiber officinale var. ‘Nadia’ through high frequency shoot proliferation from transverse thin cell layer (tTCL) sections of in vitro derived microrhizome. In vitro derived microrhizome of size 500 μm in thickness was used as initial explants for induction of somatic embryos. Among the different phytohormones tested, tTCL explants shows maximum calli proliferation in medium containing 2 mg/L 2,4-Dichlorophenoxyacetic acid (88.30±0.11%). Reduced concentration of 2,4 Dichlorophenoxyacetic acid was supplemented with different cytokinins for regeneration of callus. Among the different medium tested, optimum redifferentiation of somatic embryos were observed in medium containing 0.2 mg/L 2,4 Dichlorophenoxyacetic acid and 6.0 mg/L BAP (141.08±0.25). Clump of regenerated plantlets were further subculture and transfer into microrhizome inducing medium containing high sucrose concentration (8%). Plantlets with well developed microrhizome were successfully acclimatized and eventually transferred to the field. The application of studying embryo section for regeneration of plants might be useful alternative to ginger improvement programme. Histological analysis showed formation of somatic embryos and regenerated adventitious shoot.

2016 ◽  
Vol 14 (1) ◽  
pp. 63-73
Author(s):  
Vu Thi Hien ◽  
Nguyen Phuc Huy ◽  
Bui Van The Vinh ◽  
Hoang Xuan Chien ◽  
Hoang Thanh Tung ◽  
...  

No report on plant regeneration via somatic embryogenesis of P. vietnamensis has been previously published. In the present study, somatic embryogenesis via callus formation from cultures of leaf transverse thin cell layers (tTCLs) of Vietnamese ginseng (Panax vietnamensis Ha et Grushv.) was investigated. α-naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BA) and thidiazuron (TDZ) were added separately and in combination into the culture media. Explant necrosis or low callogenesis rates were observed when 1-mm wide leaf tTCLs were cultured on media with TDZ, BA, 2,4-D or NAA. On the other hand, calli were successfully induced from the tTCL explants cultured on medium supplemented with either 2,4-D and BA or 2,4-D and TDZ. Callogenesis was observed under both light and dark conditions. The highest callogenesis rate (100%) was obtained on Murashige and Skoog (MS) basal medium supplemented with 1.0 mg l-1 2,4-D in combination with 0.1 mg l-1 TDZ in darkness after eight weeks of culture. White calli were cut into small pieces (1.0 x 1.0 cm dimension) and placed on MS media containing 1.0 mg l-1 2,4-D, 0.5 mg l-1 NAA and TDZ at various concentrations (0.01; 0.1; 0.2; and 0.5 mg l-1), and the best callus proliferation was recorded on medium containing 1.0 mg l-1 2,4-D and 0.2 mg l-1 TDZ. Somatic embryogenesis, with a success rate of 53.3% and 35 embryos per explant, was achieved when calli were subcultured onto MS medium supplemented with 1.0 mg l-1 2,4-D, 0.5 mg l-1 NAA and 0.2 mg l-1 TDZ.


HortScience ◽  
1990 ◽  
Vol 25 (7) ◽  
pp. 792-793 ◽  
Author(s):  
Paula P. Chee

A simple procedure for regeneration of cucumber plants (Cucumis sativus L. cv. Poinsett 76) from cotyledon and hypocotyl explants has been developed. Somatic embryogenesis was induced on Murashige and Skoog (MS) salts and vitamins medium supplemented with 2,4-D at 2.0 mg·liter-1 and kinetin at 0.5 mg·liter-1. Development of embryos was accomplished on MS medium with NAA at 1.0 mg·liter-1 and kinetin at 0.5 mg·liter-1. Eighty-five percent of the mature somatic embryos formed showed a typical bipolar structure. All developed into morphologically normal plantlets when transferred to MS medium containing no growth regulators. Chemical name used: 2,4-dichlorophenoxyacetic acid (2,4-D).


1993 ◽  
Vol 23 (5) ◽  
pp. 835-838 ◽  
Author(s):  
Paula M. Pijut

Immature cotyledonary explants excised from developing fruits of Juglanscinerea L. were cultured in vitro to induce regeneration of somatic embryos. Somatic embryos were initiated directly on cotyledons collected 9 weeks postanthesis and cultured on a Driver and Kuniyuki medium supplemented with 250 mg/L L-glutamine, 0.01 mg/L indole-3-butyric acid, 1 mg/L 6-benzylaminopurine, and 2 mg/L kinetin for 3 weeks, prior to transfer to hormone-free Driverand Kuniyuki medium. Embryogenic callus was initiated on explants collected 8–11 weeks postanthesis and cultured on two different media formulations containing 0.25 mg/L 6-benzylaminopurine and 2 mg/L 2,4-dichlorophenoxyacetic acid for 3 weeks, prior to transfer to hormone-free media. Globular to mature somatic embryos were differentiated, and conversion of somatic embryos into whole plants was incomplete. Competence of embryogenic callus was maintained for 1 year with regular subculturing on hormone-free media.


2015 ◽  
Vol 21 (1) ◽  
Author(s):  
Esther Julia Naranjo ◽  
Osman Fernandez Betin ◽  
Aura Inés Urrea Trujillo ◽  
Ricardo Callejas Posada ◽  
Lucía Atehortúa Garcés

<p class="p1"><strong>ABSTRACT</strong></p><p class="p2"><em>Stevia rebaudiana</em> (Asteraceae) is a plant of economic importance because of its medicinal properties and the presence of sweetener compounds on its leaves. These compounds can be a substitute for sucrose in a wide variety of products used by persons with diabetes and obesity problems. To standardize an efficient and effective propagation method for the different Stevia genotypes grown in Colombia, this study evaluated the effect of different combinations of the plant growth regulators 2,4-dichlorophenoxyacetic acid (2,4-D), indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), 6-(gamma, gamma-dimethylallylamino) purine (2iP) and Zeatin on the induction and development of somatic embryos. Adenine and coconut water were also evaluated as supplements in the basal culture medium Murashige and Skoog Basal Salt Mixture (MS) with glutamine. The combination of 2,4-D (18.09 μM) and 2iP (7.38 μM) produced the highest number of somatic embryos per explant, which had well-defined characteristics. The genotype showed a significant effect on the embryogenic response. In the “SRQ-93” genotype, the formation and development of somatic embryos was achieved, whereas the genotypes “Bertoni” and “Morita II” only yielded embryogenic and non-embryogenic calli, respectively. The conversion to seedlings was achieved on the regeneration medium containing gibberellic acid (GA<span class="s1">3</span>) (0.29 μM) and activated charcoal. </p><p class="p1"><strong>RESUMEN</strong></p><p class="p2"><em>Stevia rebaudiana</em> (Asteraceae), es una planta de gran importancia económica debido a sus propiedades medicinales y a la presencia de compuestos endulzantes en sus hojas, los cuales pueden sustituir la sacarosa en gran variedad de productos utilizados por personas con problemas de diabetes y obesidad. Con el propósito de estandarizar un método de propagación eficiente y efectivo para diferentes genotipos de Stevia cultivados en Colombia, en la presente investigación se evaluó el efecto sobre la inducción y desarrollo de embriones somáticos de diferentes combinaciones de los reguladores de crecimiento vegetal 2,4-D, IAA, IBA, 2iP y Zeatina, además de los suplementos adenina y agua de coco en el medio de cultivo basal Murashige y Skoog (1962), adicionado con glutamina. Con la combinación 2,4-D (18.09 μM) y 2iP (7.38 μM) se obtuvo el mayor número embriones somáticos por explante con características bien definidas. El genotipo tuvo un efecto significativo en la repuesta embriogénica, en el genotipo “SRQ-93” se logró la formación y el desarrollo de embriones somáticos, mientras que en los genotipos “Bertoni” y “Morita II”, solo se obtuvo callo embriogénico y no embriogénico respectivamente. La conversión a plántulas se alcanzó en el medio de regeneración conteniendo GA3 (0.29 μM) y carbón activado.</p><p class="p2"> </p>


2021 ◽  
Vol 12 (4) ◽  
pp. 4731-4746

The Crocus sativus, an endangered medicinal and aromatic plant in Morocco, has a low propagation rate in natural conditions and, therefore, an efficient method for in vitro propagation is required. This study investigated the effects of various hormones on the induction of callogenesis and callogenesis in C. sativus corms using the Box-Behnken experimental design. The best shoot formation was obtained with Murashige and Skoog fortified with 3 mg/L 6-Benzylaminopurine. On the other hand, callus formation was obtained with 3 mg/L 1-Naphthaleneacetic Acid or 3 mg/L 2,4-Dichlorophenoxyacetic Acid. However, a combination of 3 mg/L 6-Benzylaminopurine, 1.056 mg/L Indole Butyric Acid, and 3 mg/L 2,4-Dichlorophenoxyacetic Acid allows 50% caulogenesis and 60% callogenesis. The in vitro regeneration system could be utilized for both conservation and largescale multiplication of Crocus sativus corms.


2012 ◽  
Vol 2 (4) ◽  
pp. 184-189
Author(s):  
Kavita Kshirsagar ◽  
V. J. Braganza

A basic factor underlying the success of large‐scale micropropagation and genetic transformation of any plant species is regeneration. In order to regenerate propagules of Rosa damascena Mill. on a large scale, an efficient and improved in vitro propagation system has been established using transverse thin cell layer culture (tTCL). By optimizing the position of the tissue and applying an improved selection procedure, in vitro shoots were elongated in 8 weeks of culture. Modified Murashige and Skoog (1962)(MS) medium fortified with 4.0 mg l‐1 6‐benzylaminopurine (BAP) and 0.4mg l‐1 anaphthalene acetic acid (NAA) gave optimal shoot regeneration. The explant was inoculated on this medium in the upright position and exhibited a high frequency of shoot regeneration (~96.66%), and it also gave the highest number of shoots (22.33/explant). The horizontally placed explant on an average 7.66 shoots/explant. Our experiments indicate that explant orientation strongly influences the organogenesis response. The frequency of shoot initiation and the number of multiple shoots produced from each explant were significantly dependent on the plant source, concentration of plant growth regulators and the orientation of the explants and contributed significantly to in vitro regeneration. Rooting of well developed shoots was achieved on hormone free ¼ strength MS medium with 4% sucrose.


Sign in / Sign up

Export Citation Format

Share Document