Micropropagation of Withania somnifera from germinating seeds and shoot tips
TL;DR: Shoot multiplication was achieved in vitro from shoot tips of aseptically germinated seedlings of Withania somnifera L. with direct multiple shoot initiation from germinating seeds in the presence of BA alone.
Abstract: Shoot multiplication was achieved in vitro from shoot tips of aseptically germinated seedlings of Withania somnifera L. using low concentrations of 6-benzyladenine (BA), viz. 2.2, 4.4 and 8.9 μM. Maximum number of shoots were obtained when 2.3 μM 2,4-dichlorophenoxyacetic acid (2,4-D) or 2.5 μM indolebutyric acid (IBA) was added to medium containing 4.4 μM BA during initiation of shoot multiplication, but not when added later. Direct multiple shoot initiation was also obtained from germinating seeds in the presence of BA alone. Rooting was successful in excised shoots grown on growth regulator-free MS medium. Rooted shoots were successfully established in soil in a greenhouse.
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TL;DR: An efficient in vitro protocol for high frequency regeneration has been developed using mature seeds as explant in W. somnifera using mature seed as an explant and it was found that MS medium supplemented with GA3 and IBA alone was suited for shoot elongation and rhizogenesis respectively.
Abstract: Withania somnifera (L.) Dunal a member of the Solanaceae family, is a traditional medicinal plant commonly known in India as Ashwagandha. It is used for different diseases such as hiccup, cough, rheumatism, tuberculosis, and exhibits excellent antitumor and anti-bacterial activities as well. Direct organogenesis of plants using mature seeds provides faster response and is also a time saving approach, thus the present study was conducted to investigate the optimal concentrations and combinations of plant growth regulators with MS medium for the establishment of an efficient regeneration system in W. somnifera using mature seed as an explant. Therefore, an efficient in vitro protocol for high frequency regeneration has been developed using mature seeds as explant. In the present study, the multiple shoots along with embryogenic callus induction was best seen in MS medium supplemented with BAP (1.5 mg/L) and IAA (0.5 mg/L). Furthermore, MS medium fortified with GA3 (0.3 mg/L) and IBA (3.0 mg/L) alone was suited for shoot elongation and rhizogenesis respectively. The rooted plantlets were hardened and successfully established in the soil. The establishment of a highly reproducible regeneration system would greatly influence the efforts of improvement of the hereby studied medicinal plant species through useful gene transfer technology.
1 citations
Cites background or result from "Micropropagation of Withania somnif..."
...Very few reports are available in regard with in vitro plant regeneration system of Withania somnifera using mature seeds and have been published (Sen and Sharma, 1991; Supe et al., 2006; Sharma et al., 2015)....
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...Hence, this was opted as the most suitable medium for mass propagation of Withania somnifera through seeds when compared to the previous reports of Sen and Sharma (1991), Supe et al. (2006) and Sharma et al. (2015)....
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01 Jan 2014
TL;DR: An efficient plant propagation system through rhizomal explants was established in Curcuma caesia Roxb.
Abstract: An efficient plant propagation system through rhizomal explants was established in Curcuma caesia Roxb., a medicinally important herbaceous annual herb belonging to the family Zingiberaceae. Here we report a rapid and reliable method for high fidelity micro-propagation. Rhizomal explants from two months old seedlings were cultured on Murashige and Skoogs (MS) medium supplemented with different concentrations of N6benzyladenine (BA) (0.5 5.0 mg/l), Napthalenic acetic acid (NAA) (0.5 5.0 mg/l) and Indole 3 butyric acid (IBA) (0.5 5.0 mg/l). During the first culture on 1.5 mg/l of 6-benzylamino purine (BAP) and 1 mg/l of Napthalenic acetic acid (NAA) maximum 15.40±0.40 shoots with an average shoot-length of 8.46±0.06 were produced. The elongated shoots produced a maximum of 12.00±0.00 roots on half-strength MS liquid medium supplemented with 1 mg/l of Indole 3 butyric acid (IBA). The plantlets were acclimatized by transferring them first to peat moss: compost (1:1) mixture followed by sand: soil (2:1) mixture, recording 95% survival. Genetic fidelity was assessed by DNA fingerprinting using random amplified polymorphic DNA (RAPD) of in vitro and in vivo plants. Five arbitrary decamers displayed same banding profile showed no genomic alterations, indicating homogeneity among the tissue culture regenerates and genetic uniformity with that of donor plants. The present study provides high genetic fidelity micropropagated system for efficient and rapid micropropagation protocol of this important medicinal plant and great use in conserving without risk of genetic instability. 2013 Trade Science Inc. INDIA
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Journal Article•
TL;DR: The regenerated plantlets were sequentially transferred to liquid basal medium having a gradual decrease in sucrose concentration and successfully hardened and transferred to sand-soil and farmyard manure mixture, where 90% of C. borivilianum regenerants survived after successful hardening.
Abstract: The potential for photoautotrophic growth was studied in in vitro cultures of Chlorophytum borivilianum Sant. et Fernand. Best in vitro shoot differentiation was observed on MS medium supplemented with BAP 5 mg L-1, whereas MS + IBA 2 mg L-1 produced maximum root number and root length. The regenerated plantlets were then sequentially transferred to liquid basal medium having a gradual decrease in sucrose concentration [3%, 2%, 1.5%, 1%, 0.5% and 0%] after 48 hours stay in each. The plantlets thus formed were successfully hardened and transferred to sand-soil and farmyard manure mixture [1:1:1]. Approx. 90% of C. borivilianum regenerants survived after successful hardening.
1 citations
01 Jan 2007
TL;DR: This paper describes efficient propagation of Artemisia vulgaris using shoot tip explants isolated from 35 days old in vitro grown seedlings and optimal proliferation obtained on Murashige and Skoog's salts and B5 vitamins supplemented with 3% sucrose, 4.44 µM BA, and 0.7% agar.
Abstract: This paper describes efficient propagation of Artemisia vulgaris using shoot tip explants isolated from 35 days old in vitro grown seedlings. Optimum proliferation was obtained on Murashige and Skoog's salts and B5 vitamins supplemented with 3% sucrose, 4.44 µM BA, and 0.7% agar. Shoot proliferation was maximal (99.8%) with 14-23 shoots per explant after 6 weeks of culture. Shoots with a minimum length of 1.5 cm were transferred to shoot elongation medium supplemented with 0.44 µM BA and 1.44 µM GA3. The successfully elongated shoots with a height of 7.2-12.1 cm were transferred to rooting medium augmented with 8.56 µM IAA. Rooted plantlets were transferred to plastic cups containing autoclaved garden soil, farmyard soil and sand (2:1:1) for hardening. Plantlets were initially maintained under culture room conditions (5 weeks), followed by normal laboratory conditions (4 weeks) and finally transferred to a Botanical Evaluation Garden and maintained there.
1 citations
01 Jan 2012
1 citations
Cites background from "Micropropagation of Withania somnif..."
...The shoot tip, nodal culture and callus culture could be valuable technique for the production of steroidal alkaloids in large scale (Girija et al., 2006, Sen and Sharma, 1999)....
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References
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TL;DR: In vivo redox biosensing resolves the spatiotemporal dynamics of compartmental responses to local ROS generation and provide a basis for understanding how compartment-specific redox dynamics may operate in retrograde signaling and stress 67 acclimation in plants.
Abstract: In experiments with tobacco tissue cultured on White's modified medium (basal meditmi hi Tnhles 1 and 2) supplemenk'd with kiticthi and hidoleacctic acid, a slrikin^' fourlo (ive-told intTease iu yield was ohtaitu-d within a three to Tour week j^rowth period on addition of an aqtteotis exlrarl of tobacco leaves (Fi^'ures 1 and 2). Subse(iueutly it was found Ihiit this jnoniotiou oi' f^rowih was due mainly though nol entirely to inorj^auic rather than organic con.stitttenls in the extract. In the isolation of Rrowth factors from plant tissues and other sources inorj '̂anic salts are fre(|uently carried along with fhe organic fraclioits. When tissue cultures are used for bioassays, therefore, il is necessary lo lake into account increases in growth which may result from nutrient elements or other known constituents of the medium which may he present in the te.st materials. To minimize interference trom rontaminaitis of this type, an altempt has heen made to de\\eh)p a nieditmi with such adequate supplies of all re(iuired tnineral nutrients and cotntnott orgattic cottslitueitls that no apprecial»le change in growth rate or yield will result from the inlroduclion of additional amounts in the range ordinarily expected to be present in tnaterials to be assayed. As a point of referetice for this work some of the culture media in mc)st common current use will he cotisidered briefly. For ease of comparis4)n Iheir mineral compositions are listed in Tables 1 and 2. White's nutrient .solution, designed originally for excised root cultures, was based on Uspeuski and Uspetiskaia's medium for algae and Trelease and Trelease's micronutrieni solution. This medium also was employed successfully in the original cttltivation of callus from the tobacco Iiybrid Nicotiana gtauca x A', tanijadorffii, atitl as further modified by White in 194̂ ^ and by others it has been used for the
63,098 citations
01 Jan 1962
16,251 citations
01 Jan 1985
TL;DR: From Metabolism and Osmotic Work to Totipotency and Morphogenesis: A Study of Limitations Versus Multiple Interactions.
Abstract: I From Metabolism and Osmotic Work to Totipotency and Morphogenesis: A Study of Limitations Versus Multiple Interactions.- II Primary Metabolism.- Photosynthetic Carbon Metabolism in Photoautotrophic Cell Suspension Cultures of Chenopodium rubrum L..- On the Photosynthetic System and Assimilate Metabolism of Daucus and Arachis Cell Cultures.- Regulation of Carbon and Nitrogen Assimilation Pathways in Tobacco Cell Suspension Cultures in Relation with Ultrastructural and Biochemical Development of the Photosynthetic Apparatus.- Application of Gas Analysis to Continuous Culture.- Carbohydrate Source, Biomass Productivity and Natural Product Yield in Cell Suspension Cultures.- Nitrogen Metabolism of Leaf and Microspore Callus of Betula pendula.- III Secondary Metabolism.- 4-Coumarate: CoA Ligase in Wild Carrot Cell Culture Clones Which Accumulate Different Amounts of Anthocyanin.- Induction of Anthocyanin Synthesis in Relation to Embryogenesis in a Carrot Suspension Culture - A Model System for the Study of Expression and Repression of Secondary Metabolism.- Metabolism of Quinolizidine Alkaloids in Plants and Cell Suspension Cultures: Induction and Degradation.- Production of Alkaloids by Ergot (Claviceps fusiformis Lov.) on Pennisetum typhoides (Burm.) Stapf and Hubb. in Vitro.- Compartmentation of Alkaloids in a Cell Suspension of Catharanthus roseus: A Reappraisal of the Role of pH Gradients.- Studies on Variability of Plant Tissue Cultures for Alkaloid Production in Catharanthus roseus and Papaver somniferum Callus Cultures.- Biosynthesis and Accumulation of Indole Alkaloids in Cell Suspension Cultures of Catharanthus roseus Cultivars.- Formation of Cardenolides in Cell and Organ Cultures of Digitalis lanata.- Metabolism of Caffeoyl Derivatives in Plant Cell Cultures.- Metabolic Relationships of Putrescine, GABA and Alkaloids in Cell and Root Cultures of Solanaceae.- Metabolism and Degradation of Nicotinic Acid in Plant Cell Cultures.- Plant Cell and Tissue Culture of Cinchona Species.- The Production of Pyrethrins by Chrysanthemum cinerariaefolium (Trev) Bocc..- Biosynthesis of Chorismate-Derived Quinones in Plant Cell Cultures.- The Role of Leucine in Terpenoid Metabolism: Incorporation of Leucine into Sesquiterpenoids and Phytosterols by Andrographis Tissue Cultures.- Accumulation of Antineoplastic Agents by Plant Tissue Cultures.- Induction of Enzymes of Phytoalexin Synthesis in Soybean Cells by Fungal Elicitor.- Protoplast Fusion of Secondary Metabolite-Producing Cells.- Conventional and New Approaches to Increase the Alkaloid Production of Plant Cell Cultures.- Multiple Shoot Cultures: A Viable Alternative in Vitro System for the Production of Known and New Biologically Active Plant Constituents.- IV Fermentation and Cryopreservation.- Large-Scale Production of Rosmarinic Acid from Plant Cell Cultures of Coleus blumei Benth..- Immobilised Plant Cell Culture Systems.- Biotransformation of Cardiac Glycosides by Digitalis Cell Cultures in Airlift Reactors.- Cryopreservation of Plant Cell Cultures.- V Herbicides.- Altered Amino Acid Biosynthesis in Amino Acid Analog and Herbicide-Resistant Cells.- Acetohydroxyacid Synthase Inhibitors as Herbicides.- A Glyphosate-Tolerant Plant Tissue Culture.- VI Plant Cell Culture - Future Perspectives.
321 citations
TL;DR: In this paper, the inheritance characteristics of various substituents on the withanolide skeleton were analyzed based on the occurrence in per cent of each substituent in relation to the total withanolides content in the hybrid plants and their respective parents.
43 citations
TL;DR: Of the cytokinins tested, benzylaminopurine (BAP) was more effective than either 2-isopentenyladenine (2iP) or kinetin (Kin) and produced maximum proliferation when used at 5×10-6M.
Abstract: Successful vegetative propagation of seedling jackfruit (Artocarpus heterophyllus Lam.) has been achieved by in vitro methods. Proliferation from nodal explants was greater than from shoot tips. Of the cytokinins tested, benzylaminopurine (BAP) was more effective than either 2-isopentenyladenine (2iP) or kinetin (Kin) and produced maximum proliferation when used at 5×10-6M. Shoot proliferation was optimal at 30°C with a 12 h photoperiod. Optimal rooting of shoots in vitro was obtained with indolebutyric acid (IBA) at 10-6M. The number and length of roots was significantly increased in 12 h light as compared with the dark.
40 citations