Perumal Venkatachalam

Effect of lead on phytotoxicity, growth, biochemical alterations and its role on genomic template stability in Sesbania grandiflora: A potential plant for phytoremediation

The present study was aimed at evaluating phytotoxicity of various concentrations of lead nitrate (0, 100, 200, 400, 600, 800 and 1000mgL(-1)) in Sesbania grandiflora. The seedling growth was significantly affected (46%) at 1000mgL(-1) lead (Pb) treatment. Accumulation of Pb content was high in root (118mgg(-1) dry weight) than in shoot (23mgg(-1) dry weight). The level of photosynthetic pigment contents was gradually increased with increasing concentrations of Pb. Malondialdehyde (MDA) content increased in both the leaves as well as roots at 600mgL(-1) Pb treatment and decreased at higher concentrations. The activity of antioxidative enzymes such as superoxide dismutase and peroxidase were positively correlated with Pb treatment while catalase and ascorbate peroxidase activities increased up to 600mgL(-1) Pb treatment and then slightly decreased at higher concentrations. Isozyme banding pattern revealed the appearance of additional isoforms of superoxide dismutase and peroxidase in Pb treated leaf tissues. Isozyme band intensity was more consistent with the respective changes in antioxidative enzyme activities. Random amplified polymorphic DNA results indicated that genomic template stability (GTS) was significantly affected based on Pb concentrations. The present results suggest that higher concentrations of Pb enhanced the oxidative damage by over production of ROS in S. grandiflora that had potential tolerance mechanism to Pb as evidenced by increased level of photosynthetic pigments, MDA content, and the level of antioxidative enzymes. Retention of high levels of Pb in root indicated that S. grandiflora has potential for phytoextracting heavy metals by rhizofiltration.

Kinetic and thermodynamic evaluation of adsorption of Cu(II) by thiosemicarbazide chitosan.

A modified biomacromolecule, chitosan-thiosemicarbazide framework (TSCS) as an adsorbent for Cu(II) was prepared from dialdehyde chitosan through condensation reaction with thiosemicarbazide, stabilized by the reduction reaction with sodium borohydride. TSCS was characterized by means of FT-IR and XPS. Surface morphologies were studied by FESEM and BET, which revealed the highly macro porous structure. The thermal analyses was done through TGA showing much stable chemical configuration at about ≥400°C. The experimental equilibrium data was evaluated by Langmuir, Freundlich and Dubinin-Radushkevich isotherm models. The Langmuir adsorption model was best fitted with experimental value, suggests the existence of monolayer coverage of adsorbed molecules with a maxima of 142.85mgg-1. The kinetic data was analyzed using pseudo-first-order, pseudo-second-order and intraparticle diffusion models and the pseudo-second-order kinetics were found for all the concentrations. The calculated thermodynamic parameters such as ΔGo, ΔH and ΔS were -2.33kJmol-1, 570.40Jmol-1 and 9.75Jmol-1K-1 respectively signifies the adsorption of Cu(II) onto TSCS is endothermic, spontaneous and a process of physisorption. The regeneration efficiency of the TSCS as an adsorbent was found to be ≥90-95% using 0.1M EDTA.

Effect of auxins and cytokinins on efficient plant regeneration and multiple-shoot formation from cotyledons and cotyledonary-node expiants of groundnut (Arachis hypogaea L.) by in vitro culture technology

Tissue cultures were established from cotyledon and cotyledonarynode segments ofArachis hypogaea L. on Murashige and Skoog (MS) medium supplemented with different concentrations of auxins (IAA, NAA, IBA, and 2, 4-D) and cytokinins (KIN and BAP). For callus initiation, high concentration of auxins and low concentration of cytokinins were used, whereas high concentration of cytokinins and low concentration of auxins were used for shoot-bud differentiation. Callus induction and shoot-bud regeneration frequency, however, varied with genotype, expiant, and the different plant-growth regulators combination in the medium. The shoot-bud multiplication was also influenced by genotype, explant type, and growth regulators. The combination of BAP and NAA produced more shoots than other combinations. The maximum number of shoots was obtained from cotyledonary-node segments on a medium containing BAP (5.0 mg/L) and IBA (1.0 mg/L). Rooting of regenerated shoots was achieved on a medium augmented with NAA or IBA (2.0 mg/L) in combination with KIN (0.5 mg/L). Rooted plantlets were successfully established in the soil, where 95% of them survived.Tissue-culture studies of these expiants suggests the shoots to be ofde nova origin, which would make the system suitable for gene-transfer technology.

Molecular cloning and characterization of a 3-hydroxy-3-methylglutaryl-coenzyme A reductase 1 (hmgr1) gene from rubber tree (Hevea brasiliensis Muell. Arg.): A key gene involved in isoprenoid biosynthesis

Natural rubber (cis-1,4-polyisoprene) is a secondary metabolite produced in the laticiferous tissue of Hevea tree. Mevalonate synthesis, which is the first step in isoprenoid biosynthesis, is catalyzed by the enzyme 3-hydroxy-3-methylglutarylcoenzyme A reductase 1 (hmgr1). We have cloned and characterized a full-length cDNA as well as genomic DNA for hmgr1 gene from an elite Indian rubber clone (RRII 105). The nucleotide sequence of the genomic clone comprises 4 exons and 3 introns, giving a total length of 2440 bp. The sequences of 42 bp 5′ UTR and 69 bp of the 3′ UTR were also determined. The hmgr1 cDNA contained an open reading frame of 1838 bp coding for 575 amino acid protein with a theoretical pI value of 6.6 and the calculated protein MW was 61.6 kDa. The deduced amino acid sequence showed high identity with other plant hmgr1 sequences. The amino acid sequence of the Hevea hmgr1 revealed several motifs which are highly conserved and common to the other plant species. These sequence conservations suggest a strong evolutionary pressure to maintain amino acid residues at specific positions, indicating that the conserved motifs might play important roles in the structural and/or catalytic properties of the enzyme. Southern blot analysis of genomic DNA from Hevea probed with a genomic fragment indicated that there were at least three isoforms of hmgr in Hevea. This result reveals that hmgr1 is one of the members of a small gene family. (Northern blot analysis showed that hmgr1 mRNA transcripts were noticed in all tissues — latex, leaf, immature leaf, and seedlings), however, the abundance of transcript level was higher in latex cells. As one step towards a better understanding of the role that this enzyme plays in coordinating isoprenoid biosynthesis in plants, hmgr1 cDNA was over expressed in transgenic Arabidopsis plants. Transgenic plants were morphologically distinguishable from control wild-type plants and an increased expression level of hmgr1 mRNA was detected. These data suggest that hmgr1 gene expression is playing an important role in isoprenoid biosynthesis.

Biogeochemistry of uranium in the soil-plant and water-plant systems in an old uranium mine

The present study highlights the uranium (U) concentrations in water-soil-plant matrices and the efficiency considering a heterogeneous assemblage of terrestrial and aquatic native plant species to act as the biomonitor and phytoremediator for environmental U-contamination in the Sevilha mine (uraniferous region of Beiras, Central Portugal). A total of 53 plant species belonging to 22 families was collected from 24 study sites along with ambient soil and/or water samples. The concentration of U showed wide range of variations in the ambient medium: 7.5 to 557mgkg(-1) for soil and 0.4 to 113μgL(-1) for water. The maximum potential of U accumulation was recorded in roots of the following terrestrial plants: Juncus squarrosus (450mgkg(-1) DW), Carlina corymbosa (181mgkg(-1) DW) and Juncus bufonius (39.9mgkg(-1) DW), followed by the aquatic macrophytes, namely Callitriche stagnalis (55.6mgkg(-1) DW) Lemna minor (53.0mgkg(-1) DW) and Riccia fluitans (50.6mgkg(-1) DW). Accumulation of U in plant tissues exhibited the following decreasing trend: root>leaves>stem>flowers/fruits and this confirms the unique efficiency of roots in accumulating this radionuclide from host soil/sediment (phytostabilization). Overall, the accumulation pattern in the studied aquatic plants (L. minor, R. fluitans, C. stagnalis and Lythrum portula) dominated over most of the terrestrial counterpart. Among terrestrial plants, the higher mean bioconcentration factor (≈1 in roots/rhizomes of C. corymbosa and J. squarrosus) and translocation factor (31 in Andryala integrifolia) were encountered in the representing families Asteraceae and Juncaceae. Hence, these terrestrial plants can be treated as the promising candidates for the development of the phytostabilization or phytoextraction methodologies based on the accumulation, abundance and biomass production.

Agrobacterium-mediated genetic transformation of groundnut (arachis hypogaea L.): An assessment of factors affecting regeneration of transgenic plants

Transgenic groundnut (Arachis hypogaea L.) plants were produced efficiently by inoculating different explants withAgrobacterium tumefaciens strain LBA4404 harbouring a binary vector pBM21 containinguidA (GUS) andnptll (neomycin phosphotransferase) genes. Genetic transformation frequency was found to be high with cotyledonary node explants followed by 4 d cocultivation. This method required 3 days of precultivation period before cocultivation withAgrobacterium. A concentration of 75 mg/l kanamycin sulfate was added to regeneration medium in order to select transformed shoots. Shoot regeneration occurred within 4 weeks; excised shoots were rooted on MS medium containing 50 mg/I kanamycin sulfate before transferring to soil. The expression of GUS gene (uidA gene) in the regenerated plants was verified by histochemical and fluorimetric assays. The presence ofuidA andnptll genes in the putative transgenic lines was confirmed by PCR analysis. Insertion of thenptll gene in the nuclear genome of transgenic plants was verified by genomic Southern hybridization analysis. Factors affecting transformation efficiency are discussed.

Effects of concentration and gas flow rate on the removal of gas-phase toluene and xylene mixture in a compost biofilter

The aim of this work was to study the performance of a compost/ceramic bead biofilter (6:4 v/v) for the removal of gas-phase toluene and xylene at different inlet loading rates (ILR). The inlet toluene (or) xylene concentrations were varied from 0.1 to 1.5gm-3, at gas flow rates of 0.024, 0.048 and 0.072m3h-1, respectively, corresponding to total ILR varying between 7 and 213gm-3h-1. Although there was mutual inhibition, xylene removal was severely inhibited by the presence of toluene than toluene removal by the presence of xylene. The biofilter was also exposed to transient variations such as prolonged periods of shutdown (30days) and shock loads to envisage the response and recuperating ability of the biofilter. The maximum elimination capacity (EC) for toluene and xylene were 29.2 and 16.4gm-3h-1, respectively, at inlet loads of 53.8 and 43.7gm-3h-1.

Bioremoval of trace metals from rhizosediment by mangrove plants in Indian Sundarban Wetland

The study accentuated the trace metal accumulation and distribution pattern in individual organs of 13 native mangrove plants along with rhizosediments in the Indian Sundarban Wetland. Enrichment of the essential micronutrients (Mn, Fe, Zn, Cu, Co, Ni) was recorded in all plant organs in comparison to non-essential ones, such as Cr, As, Pb, Cd, Hg. Trunk bark and root/pneumatophore showed maximum metal accumulation efficiency. Rhizosediment recorded manifold increase for most of the trace metals than plant tissue, with the following descending order: Fe>Mn>Zn>Cu>Pb>Ni>Cr>Co>As>Cd>Hg. Concentrations of Cu, Ni, Pb and Hg were found to exceed prescribed sediment quality guidelines (SQGs) indicating adverse effect on adjacent biota. Both index of geoaccumulation (Igeo) and enrichment factor (EF) also indicated anthropogenic contamination. Based on high (>1) translocation factor (TF) and bioconcentration factor (BCF) values Sonneratiaapetala and Avicenniaofficinalis could be considered as potential accumulators, of trace metals.

Biomolecule-loaded chitosan nanoparticles induce apoptosis and molecular changes in cancer cell line (SiHa)

The present study reports on the synthesis of chitosan nanoparticles (CNPs) using methanol extracts of Gymnema sylvestre (GS) leaves and Cinnamomum zeylanicum (CZ) bark. Biomolecule-loaded nanoparticles induced apoptosis in a human cervical cancer (SiHa) cell line, and experiments were carried out to elucidate the underlying molecular mechanisms. FT-IR and XRD showed possible functional groups of the biomolecules and the crystalline nature of CNPs, respectively. Transmission electron microscopy images revealed that synthesized GSCNPs and CZCNPs had a smooth spherical shape with average sizes of about 58-80 and 60-120nm, respectively. Dynamic light scattering studies indicated that both GSCNPs and CZCNs were structurally stable with homogenous and heterogeneous natures, respectively. Furthermore, synthesized GSCNPs and CZCNPs exhibited dose-dependent cytotoxicity against the SiHa cancer cell line, with inhibitory concentration (IC50) values of 102.17μg/ml, 87.75μg/ml, 132.74μg/ml and 90.35μg/ml for GS leaf extract, GSCNPs, CZBE and CZCNPs, respectively.

Phytofabrication of bioactive molecules encapsulated metallic silver nanoparticles from cucumis sativus L. and its enhanced wound healing potential in rat model

The present study describes a rapid method for synthesis of metallic silver nanoparticles using callus (CAgNPs) and leaf extracts (LEAgNPs) of Cucumis sativus and evaluation of its wound healing activity in ratmodel. The prepared silver nanoparticles showed a peak at 350 nm corresponding to the surface plasmon resonance band. The FTIR spectroscopy measurements showed the presence of the possible biomolecules. X-ray diffraction analysis confirmed the crystalline structure of the synthesized silver nanoparticles. TEM images showed the size of the synthesized CAgNPs with diameter ranged from 21 nm to 23 nm with polygonal shape whereas, in the case of LEAgNPs, spherical shape was noticed with an average size between 11 nm and 19 nm. The EDX results indicated the chemical composition at specific locations on synthesized nanoparticles. Furthermore the topical application of ointment prepared using synthesized AgNPs was found to show enhanced wound healing activity in Wistar albino rat model. By the 21st day, the ointment base containing 5% (w/w) of silver nanoparticles showed 100% potential wound healing activity than the standard drug as well as control bases. Results strongly showed that the ointment base containing LEAgNPs was found to be very effective in wound repair mechanism in the experimental rats.