Asif S. Tamboli

Co-plantation of aquatic macrophytes Typha angustifolia and Paspalum scrobiculatum for effective treatment of textile industry effluent

Field treatment of textile industry effluent was carried out in constructed drenches (91.4m×1.2m×0.6m; 65.8m3) planted independently with Typha angustifolia, Paspalum scrobiculatum and their co-plantation (consortium-TP). The in situ treatment of effluent by T. angustifolia, P. scrobiculatum and consortium-TP was found to decrease ADMI color value by 62, 59 and 76%, COD by 65, 63 and 70%, BOD by 68, 63 and 75%, TDS by 45, 39 and 57%, and TSS by 35, 31 and 47%, respectively within 96h. Heavy metals such as arsenic, cadmium, chromium and lead were also removed up to 28-77% after phytoremediation. T. angustifolia and P. scrobiculatum showed removal of Congo Red (100mg/L) up to 80 and 73%, respectively within 48h while consortium-TP achieved 94% decolorization. Root tissues of T. angustifolia and P. scrobiculatum revealed inductions in the activities of oxido-reductive enzymes such as lignin peroxidase (193 and 32%), veratryl alcohol oxidase (823 and 460%), laccase (492 and 182%) and azo reductase (248 and 83%), respectively during decolorization of Congo Red. Anatomical studies of roots, FTIR, HPLC, UV-vis Spectroscopy and GC-MS analysis verified the phytotransformation. Phytotoxicity studies confirmed reduced toxicity of the metabolites of Congo Red.

Phylogenetic analysis, genetic diversity and relationships between the recently segregated species of Corynandra and Cleoserrata from the genus Cleome using DNA barcoding and molecular markers

Cleome is the largest genus in the family Cleomaceae and it is known for its various medicinal properties. Recently, some species from the Cleome genus (Cleome viscosa, Cleome chelidonii, Cleome felina and Cleome speciosa) are split into genera Corynandra (Corynandra viscosa, Corynandra chelidonii, Corynandra felina), and Cleoserrata (Cleoserrata speciosa). The objective of this study was to obtain DNA barcodes for these species for their accurate identification and determining phylogenetic relationships. Out of 10 screened barcoding regions, rbcL, matK and ITS1 regions showed higher PCR efficiency and sequencing success. This study added matK, rbcL and ITS1 barcodes for the identification of Corynandra chelidonii, Corynandra felina, Cleome simplicifolia and Cleome aspera species in existing barcode data. Corynandra chelidonii and Corynandra felina species belong to the Corynandra genus, but they are not grouped with the Corynandra viscosa species, however clustered with the Cleome species. Molecular marker analysis showed 100% polymorphism among the studied plant samples. Diversity indices for molecular markers were ranged from He=0.1115-0.1714 and I=0.2268-0.2700, which indicates a significant amount of genetic diversity among studied species. Discrimination of the Cleome and Corynandra species from Cleoserrata speciosa was obtained by two RAPD primers (OPA-4 and RAPD-17) and two ISSR primers (ISSR-1 and ISSR-2). RAPD and ISSR markers are useful for the genetic characterization of these studied species. The present investigation will be helpful to understand the relationships of Cleome lineages with Corynandra and Cleoserrata species.

Resolving Generic Boundaries in Indian—Australasian Cleomaceae: Circumscription of Areocleome, Arivela, and Corynandra as Distinct Genera

Abstract Novel molecular data and morphological studies have provided support for the segregation of numerous genera from Cleome s. l. (Cleomaceae). Corynandra has been proposed as a segregate genus including Indian and Australian species based on floral and seed morphology. Contrasting seed coat micro-morphology between Indian and Australian species included in Corynandra raised questions over the monophyly of that genus concept. Relationships among the Indian and Australian species remain unclear due to limited sampling in previous molecular analyses. We expanded the sampling of taxa from India and Australia in order to clarify relationships between these species and the circumscription of Corynandra. Comprehensive sampling of Indian and Australian species formed three well-supported clades based on analyses of molecular sequence data and we conclude that Corynandra is not related to the Australian species. The genus Arivela Raf. is reinstated for the majority of Australian species. The new genus Areocleome is described to accommodate the unusual Australian C4 species Cleome oxalidea. The following new combinations are made: Areocleome oxalidea, Arivela arenitensis, Arivela bundeica, Arivela cleomoides, Arivela insolata, Arivela kenneallyi, Arivela limmenensis, Arivela linophylla, Arivela lophosperma, Arivela microaustralica, Arivela tetrandra, Arivela uncifera, Corynandra aspera, and Corynandra simplicifolia. Arivela microphylla is raised to species level and provided with a new combination.

Relative profile analysis of molecular markers for identification and genetic discrimination of loaches (Pisces, Nemacheilidae).

Genus Nemacheilus, Nemachilichthys and Schistura belong to the family Nemacheilidae of the order Cypriniformes. The present investigation was undertaken to observe genetic diversity, phylogenetic relationship and to develop a molecular-based tool for taxonomic identification. For this purpose, four different types of molecular markers were utilized in which 29 random amplified polymorphic DNA (RAPD), 25 inter-simple sequence repeat (ISSR) markers, and 10 amplified fragment length polymorphism (AFLP) marker sets were screened and mitochondrial COI gene was sequenced. This study added COI barcodes for the identification of Nemacheilus anguilla, Nemachilichthys rueppelli and Schistura denisoni. RAPD showed higher polymorphism (100%) than the ISSR (93.75-100%) and AFLP (93.86-98.96%). The polymorphic information content (PIC), heterozygosity, multiplex ratio, and gene diversity was observed highest for AFLP primers, whereas the major allele frequency was observed higher for RAPD (0.5556) and lowest for AFLP (0.1667). The COI region of all individuals was successfully amplified and sequenced, which gave a 100% species resolution.

Molecular phylogeny and genetic diversity of genus Capparis (Capparaceae) based on plastid DNA sequences and ISSR markers

AbstractCapparis (Capparaceae) has been used as a medicinal plant since ancient time. Capparis species were divided into Old World and New World taxa as described by the sectional division of Capparis. However, plastid DNA sequence data of Indian Capparis species were not analyzed in previous phylogenetic studies. Here, we have added Indian Capparis data in previous phylogeny and analyzed the relationship of Indian Capparis with Old World and New World taxa. The plastid phylogeny presented here includes Capparis taxa from its major distribution areas, New World and African capparoids. The presented phylogeny is used for the determination of biogeographic history of Capparis and recently segregated genera. Phylogenetic analyses of the combined plastid data revealed that the Indian Capparis are more closely related to Old World taxa and have connections with African, Australian and Eastern Asian species. Sectional classification of Old World and Indian Capparis considered in this study is reflected from the presented plastid phylogeny. The ancestral area reconstruction using Bayesian Binary Markov Chain Monte Carlo method strongly supports for the Africa as the ancestral region for both Old World and New World Capparis. Molecular marker-based genetic diversity studies on Indian Capparis are scarce. This work also includes the genetic diversity study of Indian Capparis species. Utility and efficacy of ISSR markers to study inter- and intraspecies variation in Capparis is evident from the AMOVA results.

Chebulinic acid and Boeravinone B act as anti-aging and anti-apoptosis phyto-molecules during oxidative stress

INTRODUCTION Aquatic pollutant Malachite green (MG) induces oxidative stress by producing intracellular H2O2 and associated hydroxyl, hydroxymethyl or hydroperoxide radicals in Saccharomyces cerevisiae. These radicals disturb cellular functions leading to early aging. Exogenous supply of natural antioxidants may play a crucial role as anti-aging by ensuring the cellular survival. METHODS Protective effect of Chebulinic acid (CA) and Boeravinone B (BB) was biochemically evaluated by measuring the expression levels of antioxidant enzymes. Intracellular oxidants generation, nuclear damage, necrosis, apoptosis, reduction in caspase 3/7 activity studied microscopically, spectrofluorometrically and biochemically along with growth dynamics and relative quantitation of Yap1, Sir2 and Bir1 expression using RT-PCR. RESULTS Malachite green (MG) showed adverse effect on S. cerevisiae showing 400.83% enhancement in accumulation of intracellular H2O2 and associated hydroxyl, hydroxymethyl or hydroperoxide radicals. Independent supplementation of CA (5 μg/ml) and BB (3 μg/ml) significantly reduced the accumulation by 385.78 and 372.68%, respectively. Presence of MG extended the lag phase of growth curve and also reduced colony forming units (CFUs)/ml to 3 × 108 from 15 × 108. Whereas, CA and BB maintained the normal growth curve, CFUs and proved as anti-aging. Elevation in the activities of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) by 241.35, 539.02 and 432.60% was observed after 2 h MG exposure. However, CA and BB significantly reduced the CAT, SOD and GPx activities. Microscopic observation of CA and BB augmented cells revealed protection from H2O2 and associated hydroxyl, hydroxymethyl or hydroperoxide radicals accumulation, nuclear disorganization, morphological distortion, apoptosis and necrosis contrary to MG exposed cells. An enhancement of 112.78% in caspase 3/7 activity was noted in MG exposed cells over control. Both CA and BB supplementation reduced the caspase 3/7 activity by 106.06 and 105.82%, respectively which was almost near normal. MG was found to induce the expression of yeast transcription factor Yap1; while presence of CA and BB restored expression of Yap1. Expression of longevity responsible gene Silent Information Regulator (Sir2) was also found to be reduced during MG exposure. However, CA and BB triggered the expression of Sir2. Similarly, MG lowered the expression of Baculoviral IAP repeat (Bir1) which is the inhibitor of apoptosis while CA and BB aided the over expression of Bir1. CONCLUSIONS CA and BB supplementation could significantly decrease oxidative stress, enhance cell viability and ultimately protected S. cerevisiae cells form aging.

Enzymatic analysis, structural study and molecular docking of laccase and catalase from B. subtilis SK1 after textile dye exposure

Abstract The textile dye decolorizing efficiency of Bacillus subtilis SK1 against 70 mg/L each of Malachite Green, Methyl Orange, Rubine GFL and Red HE3B was observed as 71.7, 73.6, 74.4 and 82.6%, respectively within 3 h. UV–Vis spectroscopy, GC–MS and HPTLC analysis confirmed mineralization of model dyes into its metabolites. Physico-chemical characterization confirmed acidic and hydrophilic nature of both laccase and catalase enzymes. Both enzymes contain dominant random coiled secondary structure (SOPMA tool) and intracellular location (CELLO_v.2.5), however, laccase alone contains two disulfide bridges (CYS_REC tool). The validation of constructed 3D structure (Modeller 9.19) of laccase and catalase enzymes revealed, RAMPAGE- 96.3 and 95.8% residues in favoured region, ProSA- Z score −8.2 and −9.6, respectively and ERRAT-Overall quality factor > 68. Potential energies −1.328 × 106 kJ/mol and −2.685 × 106 kJ/mol remained constant after 1395 and 1545 steps for laccase and catalase, respectively in energy minimization. Molecular docking results showed interaction of Methyl Orange with laccase (Thr260) and catalase (Lys 48), Rubine GFL with laccase (Thr 262) and catalase (His 176) and Red HE3B with laccase (Asn 264, Thr 418, Gly 321, Thr 262 and Gly 378) and catalase (Gln 258). This study provides dye degrading potential of Bacillus subtilis strain SK1 with structurally different textile dyes and vital role of the polar amino acids of laccase and catalase in these interactions.