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 protocol for indirect shoot organogenesis and plantlets regeneration of Withania somnifera (L.) Dunal is reported and leaves contained higher contents of withanolides and phenolics than roots or stems, whereas roots contained the highest contents of flavonoids and polysacharides.
Abstract: The present study reports an efficient protocol for indirect shoot organogenesis and plantlets regeneration of Withania somnifera (L.) Dunal. Leaf explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of 6-benzylaminopurine (BAP) and indole-3-acetic acid (IAA). The highest callus induction rate (89.5 %) and shoot regeneration rate (92 %) were obtained when 2 mg dm−3 BAP was combined with 0.5 mg dm−3 IAA. Three major withanolides (withaferine A, 12-deoxywithastramonolide and withanolide A) were investigated in different plant organs from in vitro and greenhouse grown plants. Leaves contained higher contents of withanolides and phenolics than roots or stems, whereas roots contained the highest contents of flavonoids and polysacharides. In vitro grown plants contained greater contents of phenolics, flavonoids and polysaccharides while lower contents of withanolides than greenhouse grown plants.
37 citations
Cites background from "Micropropagation of Withania somnif..."
...1991), micropropagation from shoot tips and nodal explants (Sen and Sharma 1991), androgenesis (Vishnoi et al....
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...…propagation of Withania include multiple shoot formation from axillary buds (Roja et al. 1991), micropropagation from shoot tips and nodal explants (Sen and Sharma 1991), androgenesis (Vishnoi et al. 1979), direct regeneration from leaf explants (Kulkarni et al.1996) and from node, internode,…...
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TL;DR: N6-benzyladenine used at 15 μM was the most effective in inducing bud break and growth, and in initiating multiple shoot proliferation at the rate of 43 microshoots per nodal explant with axillary buds, after 30 d of eulture.
Abstract: An efficient, rapid and large-scale propagation of the woody, aromatic and medicinal shrub, Holarrhena antidysenterica, through in vitro culture of nodal segments with axillary buds, is described. N6-benzyladenine used at 15 μM was the most effective in inducing bud break and growth, and also in initiating multiple shoot proliferation at the rate of 43 microshoots per nodal explant with axillary buds, after 30 d of eulture. By repeated subculturing of nodal explants with axillary buds, a high-frequency multiplication rate was established. Efficient rooting was achieved with 35 μM indole-3-butyric acid which was the most effective in inducing roots, as 80% of the microshoots produced roots. Plantlets went through a bardening phase in a controlled plant growth chamber, prior to ex vitro transfer Micropropagated plants established in garden soil were uniform and identical to donor plants with respect to growth characteristics and vegetative morphology.
36 citations
TL;DR: Histological studies revealed the origin of shoot tip in the callus during regeneration of Withania somnifera (L.) Dunal, an important medicinal plant being the source of extremely important compounds like withanolides and withaferin.
Abstract: Withania somnifera (L.) Dunal, is an important medicinal plant being the source of extremely important compounds like withanolides and withaferin. Influence of different plant growth regulators (PGRs) were evaluated for induction of callus, callus mediated regeneration and production of secondary metabolites in them. Explants for callusing were collected from plants grown in vitro and maximum callusing (98 %) was obtained on MS medium supplemented with a combination of 2,4-dichlorophenoxy acetic acid (2,4-D) (0.5 mg l-1) and kinetin (KN) (0.2 mg l−1). Among different types of calli, best shoot regeneration was observed on green, compact calli produced on MS medium with a combination of 6-benzylamino purine (BAP) and indole butyric acid (IBA). MS medium supplemented with BAP (2 mg l−1) showed highest frequency (98 %) of shoot bud regeneration. The micro-shoots were efficiently rooted on MS media supplemented with 0.5 mg l−1 IBA. Rooted plants were transferred to soil-vermi-compost (1:3; w/w) medium in greenhouse for acclimatization. Presence of withanolide A and withaferin A in calli was validated through high performance thin layer chromatography (HPTLC). It was interesting to observe that the PGRs showed significant influence on the secondary metabolites production in callus and 2,4-D having the least effect. Histological studies revealed the origin of shoot tip in the callus during regeneration.
35 citations
01 Jan 2005
TL;DR: Micropropagated plants established in a mixture garden soil, farmyard and sand were uniform and identical to the donor plant with respect to growth characteristics as well as floral features and grew normally without showing any morphological variation.
Abstract: An efficient, rapid and large-scale in vitro clonal propagation of the valuable medicinal herb Eclipta alba (Asteraceae) by enhanced auxillary shoot proliferation in cotyledonary nodal segments was designed. The medium type, various carbon sources, plant growth regulators markedly influenced in vitro propagation of E. alba. The in vitro plantlet production system was investigated on Murashige and Skoog (MS) medium with the synergistic combination of BA (4.4µM), Kin (9.2µM), 2iP (2.4µM) and 3% sucrose which induced maximum number of shoots as well as beneficial shoot length. Subculturing of cotyledonary nodal segments on similar medium enabled continuous production of healthy shoots with similar frequency. Rooting was highest (94.3%) on full strength MS medium containing 9.8µM IBA. Micropropagated plants established in a mixture garden soil, farmyard (manure) and sand (2:1:1) were uniform and identical to the donor plant with respect to growth characteristics as well as floral features. These plants grew normally without showing any morphological variation.
31 citations
Cites background from "Micropropagation of Withania somnif..."
...Micropropagation via shoot culture, often utilized to maintain clonal fidelity, would be a specially appropriate in this respect [10]....
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TL;DR: The present study has established the rapid micropropagation protocol for the first time and will be of great use in conservation of C. panchganiensis with low risk of genetic instability and the rapid amplified polymorphic DNA (RAPD) and inter-simple sequence repeats (ISSR) markers.
31 citations
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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