Mohammad Faisal

Phytotoxic hazards of NiO-nanoparticles in tomato: A study on mechanism of cell death

Nickel oxide nanoparticles (NiO-NPs) in the concentration range of 0.025-2.0mg/ml were examined for the induction of oxidative stress, mitochondrial dysfunction, apoptosis/necrosis in tomato seedling roots, as an in vivo model for nanotoxicity assessment in plants. Compared to the control, catalase (CAT), glutathione (GSH), superoxide dismutase (SOD) and lipid peroxidation (LPO) in 2.0mg/ml NiO-NPs treatments exhibited 6.8, 3.7, 1.7 and 2.6-fold higher activities of antioxidative enzymes. At 2.0mg/ml, 122% and 125.4% increase in intracellular reactive oxygen species (ROS) and mitochondrial membrane potential (ΔΨm) of seedling roots confirmed the oxidative stress and mitochondrial dysfunction. Comet assay exhibited a significant increase in the number of apoptotic (21.8%) and necrotic (24.0%) cells in 2.0mg/ml treatment groups vis-á-vis in control 7% apoptotic and 9.6% of necrotic cells were observed. Flow cytometric analysis revealed 65.7% of apoptotic/necrotic cell populations and 2.14-fold higher caspase-3 like protease activity were recorded in 2.0mg/ml treatment groups. Ultrastructure analysis revealed NiO-NPs translocation, nuclear condensation, abundance in peroxisomes and degenerated mitochondrial cristae. The dissolution of Ni ions from NiO-NPs signifies its potential to induce cell death presumably by Ni ions, triggering the mitochondrial dependent intrinsic apoptotic pathway.

Regeneration of plants from alginate-encapsulated shoots of Tylophora indica (Burm. f.) Merrill, an endangered medicinal plant

Summary A protocol has been developed for plant regeneration from encapsulated nodal segments of Tylophora indica. Nodal segments collected from a 2-year-old plant were encapsulated in calcium alginate beads. The best gel complex was achieved using 3% (w/v) sodium alginate and 100 mM CaCl2.2H2O. The maximum frequency (91%) of conversion of encapsulated beads into plantlets was achieved on Murashige and Skoog (MS) medium containing 2.5 µM 6-benzyladenine (BA) and 0.5 µM αnaphthalene acetic acid (NAA) after 6 weeks of culture. Encapsulated nodal segments stored at 4°C for 1 – 8 weeks also showed successful conversion, followed by development into complete plantlets when returned to regeneration medium. Conversion of encapsulated nodal segment into plantlets also occurred when the calcium alginate beads were sown directly into SoilriteTM moistened with ½2MS salts. Plants regenerated from encapsulated nodal segments could be hardened, acclimatised and established in soil, with a success rate of 90%. Encapsulation of vegetative propagules in calcium alginate beads could be useful for the exchange, storage and micropropagation of germplasm of this endangered medicinal plant.

Nano‐silicon dioxide mitigates the adverse effects of salt stress on Cucurbita pepo L

Research into nanotechnology, an emerging science, has advanced in almost all fields of technology. The aim of the present study was to evaluate the role of nano-silicon dioxide (nano-SiO2 ) in plant resistance to salt stress through improvement of the antioxidant system of squash (Cucurbita pepo L. cv. white bush marrow). Seeds treated with NaCl showed reduced germination percentage, vigor, length, and fresh and dry weights of the roots and shoots. However, nano-SiO2 improved seed germination and growth characteristics by reducing malondialdehyde and hydrogen peroxide levels as well as electrolyte leakage. In addition, application of nano-SiO2 reduced chlorophyll degradation and enhanced the net photosynthetic rate (Pn ), stomatal conductance (gs ), transpiration rate, and water use efficiency. The increase in plant germination and growth characteristics through application of nano-SiO2 might reflect a reduction in oxidative damage as a result of the expression of antioxidant enzymes, such as catalase, peroxidase, superoxide dismutase, glutathione reductase, and ascorbate peroxidase. These results indicate that nano-SiO2 may improve defense mechanisms of plants against salt stress toxicity by augmenting the Pn , gs , transpiration rate, water use efficiency, total chlorophyll, proline, and carbonic anhydrase activity in the leaves of plants.

Thidiazuron induced high frequency axillary shoot multiplication in Psoralea corylifolia

The effect of thidiazuron (TDZ) was studied on in vitro axillary shoot proliferation from nodal explant of Psoralea corylifolia - an endangered medicinal plant. Proliferation of shoots was achieved on Murashige and Skoog (MS) medium supplemented with 0.5, 1, 2, 3, 4 and 5 μM TDZ. The maximum number (13.6 ± 1.4) of shoots per explant were obtained from nodal segment cultured on 2 μM TDZ for 4 weeks and this increased to 29.7 ± 2.1 on hormone free MS medium after 8 weeks. The in vitro proliferated and elongated shoots were transferred individually on a root induction medium containing 0.5 μM indole-3-butyric acid (IBA) and within 4 weeks 4.5 ± 0.5 roots per shoot were produced. The regenerated plantlets were transferred to 1:1 soil and vermiculite mixture and acclimatized with 80 % survival rate. Fully acclimatized plants were grown in garden soil in greenhouse and their morphological and physiological parameters were comparable with seedlings.

AN EFFICIENT PLANT REGENERATION SYSTEM FOR MUCUNA PRURIENS L. (DC.) USING COTYLEDONARY NODE EXPLANTS

SummaryThe purpose of this study was to develop an efficient micropropagation system for Mucuna pruriens, an important medicinal plant in India. A range of cytokinins was investigated for multiple shoot regeneration with cotyledonary node explants from 7-d-old aseptic seedlings. Of all the cytokinins, 6-benzyladenine (BA), kinetin (KIN) and 2-isopentenyl adenine (2-iP) tested in Murashige and Skoog medium (MS), BA was the most effective and 5.0 μM was found to be optimum for inducing maximum shoots. Medium types, medium strength and pH were also investigated for induction and proliferation of shoots. The highest efficiency of shoot proliferation was observed in 5.0 μM BA and 0.5 μM α-naphthalene acetic acid (NAA) in half-strength MS medium at pH 5.8. The best condition for rooting was half-strength MS medium solidified with agar and with 2.0 μM indole-3-butyric acid (IBA). After rooting, the plantlets were transferred to plastic pots filled with sterile soilrite where 90% grew and all exhibited normal development.

Assessment of Genetic Fidelity in Rauvolfia serpentina Plantlets Grown from Synthetic (Encapsulated) Seeds Following in Vitro Storage at 4 °C

An efficient method was developed for plant regeneration and establishment from alginate encapsulated synthetic seeds of Rauvolfia serpentina. Synthetic seeds were produced using in vitro proliferated microshoots upon complexation of 3% sodium alginate prepared in Llyod and McCown woody plant medium (WPM) and 100 mM calcium chloride. Re-growth ability of encapsulated nodal segments was evaluated after storage at 4 °C for 0, 1, 2, 4, 6 and 8 weeks and compared with non-encapsulated buds. Effects of different media viz; Murashige and Skoog medium; Lloyd and McCown woody Plant medium, Gamborg’s B5 medium and Schenk and Hildebrandt medium was also investigated for conversion into plantlets. The maximum frequency of conversion into plantlets from encapsulated nodal segments stored at 4 °C for 4 weeks was achieved on woody plant medium supplement with 5.0 μM BA and 1.0 μM NAA. Rooting in plantlets was achieved in half-strength Murashige and Skoog liquid medium containing 0.5 μM indole-3-acetic acid (IAA) on filter paper bridges. Plantlets obtained from stored synseeds were hardened, established successfully ex vitro and were morphologically similar to each other as well as their mother plant. The genetic fidelity of Rauvolfia clones raised from synthetic seeds following four weeks of storage at 4 °C were assessed by using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers. All the RAPD and ISSR profiles from generated plantlets were monomorphic and comparable to the mother plant, which confirms the genetic stability among the clones. This synseed protocol could be useful for establishing a particular system for conservation, short-term storage and production of genetically identical and stable plants before it is released for commercial purposes.

IN VITRO REGENERATION AND PLANT ESTABLISHMENT OF TYLOPHORA INDICA (BURM. F.) MERRILL: PETIOLE CALLUS CULTURE

SummaryA protocol has been developed for high-frequency shoot regeneration and plant establishment of Tylophora indica from petiole-derived callus. Optimal callus was developed from petiole explants on Murashige and Skoog basal medium supplemented with 10μM2,4-dichlorophenoxyacetic acid +2,5μM thidiazuron (TDZ). Adventitious shoot induction was achieved from the surface of the callus after transferring onto shoot induction medium. The highest rate (90%) of shoot multiplication was achieved on MS medium containing 2.5μM TDZ. Individual elongated shoots were rooted best on halfstrength MS medium containing 0.5μM indole-3-butyric acid (IBA). When the basal cut ends of the in vitro-regenerated shoots were dipped in 150μM IBA for 30 min followed by transplantation in plastic pots containing sterile vermiculite, a mean of 4.1 roots per shoot developed. The in vitro-raised plantlets with well-developed shoot and roots were successfully established in earthen pots containing garden soil and grown in a greenhouse with 100% survival. Four months after transfer to pots, the performance of in vitro-propagated plants of T. indica was evaluated on the basis of selected physiological parameters and compared with ex vitro plants of the same age.

An efficient in vitro method for mass propagation of Tylophora indica

A protocol of high frequency shoot organogenesis and plant establishment from stem derived callus has been developed for Tylophora indica (Burm. f.) Merrill. - an endangered medicinal plant. Callus was developed on Murashige and Skoog (MS) medium supplemented with 10 μM 2,4,5-trichlorophenoxy acetic acid (2,4,5-T). Multiple shoot induction was achieved from the surface of the callus after transferring onto shoot induction medium. The highest rate (80 %) of shoot multiplication was achieved on MS medium containing 5.0 μM kinetin. The developed shoots rooted best on half-strength MS medium supplemented with 0.5 μM indole-3-butyric acid (IBA). The in vitro raised plantlets with well developed shoot and roots were acclimatized successfully and grown in greenhouse.

Antigenic role of the adaptive immune response to d-ribose glycated LDL in diabetes, atherosclerosis and diabetes atherosclerotic patients.

AIMS Glycation of proteins leads to the formation of advanced glycation end products (AGEs, which have significant role in the pathophysiology of diabetes complications. d-ribose appears to be the most reactive among the naturally occurring sugars and contribute significantly to the generation of AGEs. Glycation also results in the generation of free radicals causing structural modification which leads to the generation of neoantigenic epitopes. The aim of the present study was to investigate whether LDL modification results in auto-antibodies formation against its glycated conformer in diabetes and atherosclerosis patients. METHODS The binding characteristics of circulating auto-antibodies in patients against native and modified LDL were assessed. T2D (n=105), ATH (n=106) and T2D-ATH patients (n=72) were examined by direct binding ELISA as well as inhibition ELISA, compared with healthy age-matched controls (n=50). Furthermore, ketoamine moieties, HMF and carbonyl content were also estimated in these patients and healthy subjects. KEY FINDINGS High degree of specific binding was observed by 41.91% of T2D, 54.72% of ATH and 70.83% T2D-ATH patients sera towards d-ribose glycated LDL, in comparison to its native analog (P<0.05). Normal human sera showed negligible binding with either antigen. Competitive inhibition ELISA reiterates the direct binding results. The higher concentration of HMF, ketoamine and carbonyl content was observed in patients sera than healthy subjects. SIGNIFICANCE LDL glycation results in structural perturbation causing generation of neoantigenic epitopes that are better antigens for antibodies in T2D, ATH and T2D-ATH patients where T2D-ATH subjects showed higher prevalence in auto-antibodies against ribosylated LDL.

An efficient system for in vitro multiplication of Ocimum basilicum through node culture

An efficient in vitro micropropagation system was developed for direct shoot growth of Ocimum basilicum , an important medicinal plant, using nodal explants. The excised nodes were cultured on Murashige and Skoog (MS) medium containing two plant growth regulators (6-benzyladenine and 2isopentanyl adenine) with various combinations and concentrations for the study of shoot induction. Addition of L-glutamine was essential to induce sprouting of axillary buds. The best condition for shoot growth was with 6-benzyladenine (BA) 10.0 µM + L-glutamine 30 mg/L in MS medium. The optimum shoot formation frequency was 100% and about 13.4 ± 1.80 shoots were obtained from each explant after 8 weeks of culture. Shoots (more than 4 cm) were rooted most effectively in 5.0 µM indole-3-butyric acid (IBA) supplemented with half-strength MS medium. The plantlets thus obtained hardened off and were transferred to natural soil, where they grew well and attained sexual maturity.