Micropropagation of Withania somnifera from germinating seeds and shoot tips

Micropropagation of Vitex negundo L., a woody aromatic medicinal shrub, through high-frequency axillary shoot proliferation.

Abstract: A protocol is described for rapid and large-scale propagation of the woody aromatic and medicinal shrub Vitex negundo by in vitro culture of nodal segments from mature plants. Of the three different cytokinins – N6-benzyladenine (BA), kinetin, and thidiazuron – evaluated as supplements to Murashige and Skoog (MS) medium, BA at an optimal concentration of 2.0 mg/l was most effective in inducing bud break. Although callus-free multiple-shoot formation was a function of cytokinin activity alone, faster bud break coupled with an enhanced frequency of shoot development (92%) and internode elongation were dependent on the synergistic influence of gibberellic acid (GA3) when used at an optimal concentration (0.4 mg/l) along with BA (2.0 mg/l). The frequency of shoot proliferation was markedly influenced by the explanting season. By repeated subculturing of nodal segments harvested from the in vitro-formed axenic shoots on MS containing 1.0 mg/l BA and 0.4 mg/l GA3, prolific shoot cultures free from proximal callusing and showing a high-frequency multiplication rate were established. The percentage shoot multiplication (98–100%) as well as the number of shoots per node (six to eight) were highest during the first three culture passages, after which there was a gradual decline in shoot development. Rooting was best induced (94%) in shoots excised from proliferated shoot cultures on half-strength MS medium augmented with an optimal combination of indole-3-acetic acid and indole-3-butyric acid each at 1.0 mg/l. Vermi-compost was the most suitable planting substrate for hardening inside a plant growth chamber and its use ensured high-frequency survival (93%) of regenerated plants prior to outdoor transfer. Micropropagated plants established in garden soil were uniform and identical to the donor plant with respect to growth characteristics as well as vegetative and floral morphology.

Photon flux density and light quality induce changes in growth, stomatal development, photosynthesis and transpiration of Withania Somnifera (L.) Dunal. plantlets

Abstract: The aim of the study was to establish whether the quantity and the quality of light affect growth and development of Withania somnifera plantlets. We have studied growth and histo-physiological parameters [stomatal characteristics, chloroplastic pigments concentrations, photosynthesis, and transpiration (E)] of W. somnifera plantlets regenerated under various light intensities, or monochromatic light or under a mixture of two colors of light in tissue culture conditions. Plantlets grown under a photon flux density (PFD) of 30 μmol m-2 s-1 showed greater growth and development than those raised under other PFDs. Chlorophylls and carotenoids, numbers of stomata, rate of photosynthesis (PN) and transpiration (E), stomatal conductance (gs), and water use efficiency (WUE) increased with increasing the PFD up to 60 μmol m-2 s-1. Light quality also affected plantlets growth and physiology. Highest growth was observed under fluorescent and in a mixture of blue and red light. Very few stomata were developed in any of the monochromatic light but under fluorescent or under a mixture of two colors stomatal numbers increased. Similarly, gs, E, PN, and WUE were also higher under fluorescent light and under a mixture of red and blue light. Regressional analysis showed a linear relationship between PN (r2 = 70) and gs and between E (r2 = 0.95) and gs. In conclusion, both the quality and the quantity of light affect growth of plantlets, development of stomata and physiological responses differently depending on the intensity and the wavelength of light.

In vitro propagation of the medicinal herbs Ocimum americanum L. syn. O. canum Sims. (hoary basil) and Ocimum sanctum L. (holy basil).

Abstract: A procedure is outlined for in vitro propagation of two medicinal herbs, Ocimum americanum L. syn. O. canum Sims (hoary basil) and Ocimum sanctum L. (holy basil), using axillary shoot buds. Multiple shoot formation was induced from shoot bud explants of both species on Murashige and Skoog medium (MS) supplemented with benzyladenine (BA). The optimum BA concentrations for shoot proliferation were 0.25 mg/l for O. americanum and 1.0 mg/l for O. sanctum. Incorporation of 0.5 mg/l gibberellic acid (GA3) along with BA in the culture medium resulted in a marked increase in the frequency of axillary branching as well as multiple shoot formation. Shoot buds collected between September through December were most responsive in culture. Shoots of O. americanum were rooted on half-strength MS supplemented with 1.0 mg/l indole-3-butyric acid (IBA), whereas O. sanctum rooted best on medium with 1.0 mg/l naphthaleneacetic acid (NAA). The plantlets were hardened off and successfully established in natural soil, where they grew and matured normally.

Current trends in

Abstract: Silicosis — Continued Silicosis is a chronic lung disease associated with the inhalation and pulmonary deposition of dust that contains crystalline silica. Through the Sentinel Event Notification System for Occupational Risks (SENSOR)* program, CDC's National Institute for Occupational Safety and Health (NIOSH) is assessing practical models for implementing state-based surveillance of silicosis and linking follow-up intervention activities to surveillance reports. From 1989 through 1992, the Ohio Department of Health (ODH) SENSOR program identified silicosis cases through reports of Bureau of Workers' Compensation (BWC) claims, physician reports, and death certificates. The addition in 1993 of hospital discharge reports as an ascertainment source resulted in a substantial increase in the number of silicosis case reports identified annually (Table 1). This report describes the investigation of a case of occupational silicosis in Ohio and summarizes the impact of hospital-based reporting on surveillance for silicosis in Case Report In September 1991, a case report † was sent to ODH by an infectious disease specialist who was treating a 55-year-old sandblaster with advanced silicosis and an associated Mycobacterium kansasii infection § (2). In January 1992, NIOSH and ODH conducted a joint investigation at the worker's place of employment—a metal preparation shop—to evaluate current levels of exposure to respirable crystalline silica and to screen co-workers for silicosis. The investigation detected excessive exposures to respirable crystalline silica (2–5). Chest radiology revealed radiographic abnormalities consistent with pneumoconiosis in four of 16 current and former workers *SENSOR is a program of cooperative agreements with state health departments to develop surveillance and intervention strategies for selected occupational conditions. The National Institute for Occupational Safety and Health currently supports SENSOR silicosis programs in seven states and Wisconsin). † Case reports should be submitted for persons with a physician's provisional diagnosis of silicosis, a chest radiograph interpreted as consistent with silicosis, or pathologic findings consistent with silicosis (1).

In vitro callus induction and regeneration studies in Withania somnifera

Abstract: Callus cultures were initiated from axillary leaves, axillary shoots, hypocotyls, and root segments on Murashige and Skoog (MS) (1962) medium supplemented with 2,4-D (2 mg l−1) and KN (0.2 mg l−1). Shoots differentiated best from axillary shoot base callus on MS medium containing BA (2 mg l−1). Regenerated shoots rooted best on MS medium containing IBA (2 mg l−1) alone, and IBA (2 mg l−1) with IAA (2 mg l−1). Plantlets were transferred to pots containing sand and soil mixture, acclimatized in a culture room and afterwards transferred to the glasshouse.

A revised medium for rapid growth and bio assays with tobacco tissue cultures

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

Primary and secondary metabolism of plant cell cultures

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.

Chemistry and genetics of withanolides in Withania somnifera hybrids

Abstract: Hybrid plants of Withania somnifera from cross-pollinations of either chemotypes II or III (Israel) and Indian I (Delhi) have been examined. From both hybrids, 14β-hydroxywithanone (5α,14β,17α-trihydroxy-6α,7α-epoxy-1-oxo-22 R-witha-2,24-dienolide) has been identified. This compound is the first example of a 14β-substitution among withanolides. From the second hybrid three additional new compounds were characterized: a 2,4,6-trien-1-one (14α,2OαF-dihydroxy-i-oxo-22 R-witha-2,4,6,24-tetraenolide), a 14α-hydroxywithanone (5α,14α,17α-trihydroxy-6α,7α-epoxy-1-oxo-22 R-witha-2,24-dienolide) and a 6β,7β-epoxywithanone (5α,14α,17α-trihydroxy-6β,7β-epoxy-1-oxo-22 R-witha-2,24-dienolide). An analysis of the inheritance characteristics of various substituents on the withanolide skeleton was based on the occurrence in per cent of each substituent in relation to the total withanolide content in the hybrid plants and their respective parents.

Factors affecting in vitro proliferation and rooting of shoots of jackfruit ( Artocarpus heterophyllus Lam.)

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.