RNA polymerase activity in isolated nuclei of Nicotiana sanderae callus: Characteristics and modulation during differentiation

Isolated nuclei from differentiating cultures of Nicotiana sanderae showed increased levels of RNA polymerase activity as compared to the nuclei from callus cultures. The RNA synthetic activity was dependent on nucleotide triphosphates and Mg2+ and was destroyed by RNase. Maximum activity was obtained in the presence of 50 mM (NH4)2 SO4 and α-amanitin inhibited 40% and 55% of the activity in the nuclei from callus and differentiating tissue respectively. The nuclei from differentiating tissue elicited a 3-fold increase in RNA polymerase I and a 4-fold augmentation in RNA polymerase II activities.

Differentiation of embryoids and plantlets from shoot callus of sandalwood

Callus cultures of sandalwood (Santalum album L.) were established from shoot segments and shoot tips of trees over 20 years old. Shoots were induced directly from shoot tip callus, while in shoot segments embryoids developed from the callus within 4 weeks after subculturing on to a medium supplemented with gibberellic acid (GA). Embryoids of 4–5 mm were transferred to basal medium or basal medium supplemented with low concentrations of auxin showed plantlet development.

Peanut agglutinin from callus and cell suspension cultures of Arachis hypogaea L

Synthesis of peanut agglutinin was induced in callus and cell suspension cultures of cotyledons of peanut (Arachis hypogaea L.). The lectin was synthesised in cultures through several passages. Biosynthesis of peanut agglutinin was regulated by the type and concentration of exogenous growth regulators and was positively correlated to the growth of the cultures,indicating that the agglutinin may have a role to play during cell growth. Movement of agglutinin from the cells into the medium not only facilitated easy isolation of the lectin but also provided a clue that it may probably serve as a defence molecule. The synthesized lectin purified from culture, was found to be biologically active, and was found to be comparable with the lectin from seeds, in terms of its electrophoretic mobility.

Nanoparticles of SrTiO3 prepared by gel to crystallite conversion and their photocatalytic activity in the mineralization of phenol

Nanometre-sized powders of SrTiO3 were prepared at 70-100 degrees C by the wet-chemical method of gel to crystallite (G-C) conversion. The crystallite sizes obtained were in the range 5-13 nm, as estimated by transmission electron microscopy (TEM) studies. The photocatalytic activities of these powders in the mineralization of phenol were evaluated in comparison with Degussa P25 (TiO2). The maximum photocatalytic activity was observed for powders annealed in the range 1100-1300 degrees C. The optical spectra of the particle suspensions in water showed broadened absorption around the band gap region, together with the appearance of an absorption maximum in the UV region. The effect of inorganic oxidizing species as electron scavengers on the rate of the photocatalytic degradation of phenol was studied. The influence of bulk and surface defects, which participate in the charge transfer process during photocatalysis, was investigated systematically.

Some microbiological aspects of bauxite mineralization and beneficiation

A microbial survey of Jamnagar bauxite mines in Gujarat, India, revealed the indigenous presence of a variety of autotrophic and heterotrophic bacteria and fungi associated with the ore body and water ponds in the vicinity. Among these, bacteria belonging to the genera Thiobacillus, Bacillus and Pseudomonas are implicated in the weathering of aluminosilicates; the precipitation of iron oxyhydroxides; the dissolution and conversion of alkaline metal species; and the formation of alumina, silica and calcite minerals. Fungi belonging to the genus Cladosporium can reduce ferric iron and dissolve alumina silicates. Biogenesis thus plays a significant role in bauxite mineralization. Various types of bacteria and fungi, such as Bacillus polymyxa, Bacillus coagulans and Aspergillus niger, were found to be efficient in significant calcium solubilization and partial iron removal from bauxite ore. Probable mechanisms in the biobeneficiation process are analyzed. Biobeneficiation is shown to be an effective technique for the removal of iron and calcium from bauxite ores for use in refractories and ceramics.

In vitro osteogenic cell proliferation, mineralization, and in vivo osseointegration of injection molded high-density polyethylene-based hybrid composites in rabbit animal model

The present work reports the biocompatibility property of injection molded HDPE-HA-Al2O3 hybrid composites. In vitro cytocompatibility results reveal that osteogenic cell viability and bone mineralization are favorably supported in a statistically significant manner on HDPE-20% HA-20% Al2O3 composite, in comparison to HDPE-40 wt.% HA or HDPE-40 wt.% Al2O3. The difference in cytocompatibility property is explained in terms of difference in substrate wettability/surface energy and importantly, both the cell proliferation at 7 days or bone mineralization at 21 days on HDPE-20% HA-20% Al2O3 composite are either comparable or better than sintered HA. The progressive healing of cylindrical femoral bone defects in rabbit animal model was assessed by implantation experiments over 1, 4 and 12 weeks. Based on the histological analysis as well as histomorphometrical evaluation, a better efficacy of HDPE-20% HA-20% Al2O3 over high-density polyethylene (HDPE) for bone regeneration and neobone formation at host bone-implant interface was established. Taken together, the present study unequivocally establishes that despite the presence of 20% Al2O3, HDPE-based hybrid composites are as biocompatible as HA in vitro or better than HDPE in vivo.

In Vitro Production of Echioidinin, 7-O-Methywogonin from Callus Cultures of Andrographis lineata and Their Cytotoxicity on Cancer Cells

Andrographis lineata is an herbal medicinal plant used in traditional medicine as a substitute for Andrographis paniculata. Here, using mature leaf explants of A. lineata we demonstrate for the first time the callus induction established on MS medium containing 1.0 mg l(-1) IAA. Dried callus was subjected to solvent extraction with acetone. Further the acetone residue was separated by silica gel column chromatography, crystallized and characterized on the basis of nuclear magnetic resonance (proton and c13) and liquid chromatographic mass spectroscopy. This analysis revealed the occurrence of two known flavones namely, 7-O-methylwogonin (MW) and Echioidinin (ED). Furthermore, these compounds were tested for their cytotoxicity against leukemic cell line, CEM. We identify that ED and MW induced cytotoxicity in a time-and concentration-dependent manner. Further increase in the LDH release upon treatment with ED and MW further confirmed our cytotoxicity results against leukemic cell line. Strikingly, MW was more potent than ED when compared by trypan blue and MTT assays. Our results recapitulate the utility of callus cultures for the production of plant specific bioactive secondary metabolites instead of using wild plants. Together, our in vitro studies provide new insights of A. lineata callus cultures serving as a source for cancer chemotherapeutic agents.

Triploid plants from endosperm cultures of sandalwood by experimental embryogenesis

Embryogenesis has been induced from endosperm callus cultures of sandalwood (Santalum album L.). Viable plantlets developed from the embryoids on subculture to White's basal medium supplemented with 0.5 mg/l of indole acetic acid. Chromosomal analysis of the root tips showed the triploid number 3n = 30.

In vitro culture of the allergenic weed Parthenium hysterophorus L

Excised stem, leaf segments and whole flower of the allergenic weed P. hysterophorus were cultured on Murashighe and Skoog's basal medium supplemented with hormones. Shoot buds readily formed in the stem callus cultured on MS Medium supplemented with IAA and BAP or Kinetin. The leaf callus formed roots alone in a wide variety of media. Suspension cultures were initiated from the leaf and stem callus. The leaf callus elicited a positive patch test response for delayed hypersensitivity in 4 patients suffering from Parthenium dermatitis, thus indicating its ability to synthesise the allergenic principle(s).

Plant regeneration from protoplasts of Capsicum annuum L cv. California Wonder

Plant regeneration from mesophyll protoplasts of pepper, Capsicum annuum L. cv. California Wonder has been demonstrated via shoot organogenesis, Protoplasts isolated from fully expanded leaves of 3-week-old axenic shoots when cultured in TM medium supplemented with 1 mgl(-1) NAA, 1 mgl(-1) 2, 4-D, 0.5 mgl(-1) BAP (CM 1) resulted in divisions with a frequency ranging from 20-25%. Antioxidant ascorbic acid and polyvinylpyrrolidone (PVP) in the medium and incubation in the dark helped overcome browning of protoplasts. Microcalli and macrocalli were formed in TM medium containing 2 mgl(-1) NAA and 0.5 mgl(-1) BAP (CM II) and MS gelled medium containing 2 mgl(-1) NAA and 0.5 mgl(-1) BAP (CM III), respectively, Regeneration of plantlets was possible via caulogenesis, Microshoots, 2-5 per callus appeared on MS gelled medium enriched with 0.5 mgl(-1) IAA, 2 mgl(-1) GA and 10 mgl(-1) BAP (CM IVc). Rooting of microshoots was obtained on half strength gelled medium containing 1 mgl(-1) NAA and 0.5 mgl(-1) BAP, Protoplasts isolated from cotyledons failed to divide and degenerated eventually.

Characterization of Somatic Embryogenesis in Sandalwood (Santalum album L.)

In the synchronous embryogenesis system of sandalwood developed in our laboratory, we observed that the early events of differentiation from freshly induced callus (stage 0) are accomplished in three distinct stages viz., preglobular masses (stage 1), globular embryos (stage 2), and bipolar embryos (stage 3). Transition from stage 0 to 1 was accomplished using 2,4-D and involves a stage specific appearance of two polypeptides of 15 and 30 kDa molecular weight. A 24 kDa polypeptide that was detected as a marked band in extracts of primary callus was not detected in stages 1, 2, and 3. Further, the tissue level of a 50 kDa glycoprotein decreased during transition from stage 2 to stage 3. However, the levels of glycoproteins in the medium were markedly higher in stage 0 cultures compared to those in stage 1. The activities of a protein kinase, glycosidase, and xylanase increased markedly with progressing embryogenesis. Our observations suggest that in addition to being controlled at the level of stage-specific gene expression, somatic embryogenesis in sandalwood is also regulated at the level of controls on cell wall flexibility and posttranslational changes in the pool of preexisting proteins.

Direct somatic embroygenesis from immature embryos of rosewood (Dalbergia latifolia Roxb.)

Direct regeneration of somatic embryos was obtained from immature zygotic embryos of Dalbergia latifolia. Immature embryos dissected from green pods 90 d after flowering gave the highest frequency of somatic embryo formation. Preculture on high 2,4-D medium for 4 weeks induced direct somatic embryogenesis, which was expressed during the second culture phase in the presence of low 2,4-D along with a high sucrose concentration. Embryos were separated and transferred to the maturation medium containing MS + 0.5-1.0 mg/L BAP, where embryos developed into plantlets. Somatic embryos failed to convert into complete plants without BAP treatment. This method of direct regeneration of somatic embryos without a callus phase has direct application for genetic manipulation studies.

Biotransformation of cholesterol to diosgenin by freely suspended and immobilised cells of Dioscorea bulbifera L.

The cell suspension cultures, established from the friable callus which was risen from the nodal segments of Dioscorea bulbifera L. in Murashige-Skoog (MS) medium supplemented with indole-3-butryic acid (20 mg L- 1), was examined for cell growth in MS medium fed with cholesterol (50 mg L- 1 and 100 mg L- 1) after 8, 10, 12, 14, 16, and 18 days of culture. The growth index of the cell suspension culture on the 8th day was 1.2 and gradually inclined to 1.9 on the 16th day and remained the same at the 18th day. There is no marked difference in the cell growth of cholesterol-treated and control cell suspension culture. The maximum accumulation of diosgenin was noticed on the 14th day in control and cholesterol-treated cell suspension culture and immobilised cell cultures. The highest concentration of diosgenin, 2.94% and 2.14% dry weight, was obtained in immobilised cell culture and cell suspension culture treated with 100 mg L- 1 cholesterol, respectively.