In vitro Floral Induction of Cuscuta reflexa.

Floral initiation and development in the angiosperms is the essential step on which the yield of the plant depends. Sometimes external climate or any abiotic stress hinders the floral initiation and ultimately affect the plant yield. Hence, in vitro floral induction and development can help to overcome the external climatic factor. Furthermore, the protocol for in vitro floral induction in the parasitic angiosperm like Cuscuta reflexa has not been reported yet. We have standardized the protocol for floral induction in the parasitic plant Cuscuta reflexa. In this study it is established that MMS (modified Murashige Skoog) media supplemented with 2 mg L-1 NAA (naphthalene acetic acid, plant growth regulator) and 2 mg L-1 2,4-D (2,4-dichlorophenoxy acetic acid, plant growth regulator) supported floral induction after shooting in vitro. Furthermore, we found that MMS media supplemented with 2 mg L-1 2,4-D rapidly induced floral buds directly from the nodal explants without any shoot elongation. This protocol will help the researcher to induce flower in vitro in the other angiosperm plants along with Cuscuta reflexa.

Characterization of biotechnologically relevant extracellular lipase produced by Aspergillus terreus NCFT 4269.10

Abstract Enzyme production by Aspergillus terreus NCFT 4269.10 was studied under liquid static surface and solid-state fermentation using mustard oil cake as a substrate. The maximum lipase biosynthesis was observed after incubation at 30 °C for 96 h. Among the domestic oils tested, the maximum lipase biosynthesis was achieved using palm oil. The crude lipase was purified 2.56-fold to electrophoretic homogeneity, with a yield of 8.44%, and the protein had a molecular weight of 46.3 kDa as determined by SDS-PAGE. Enzyme characterization confirmed that the purified lipase was most active at pH 6.0, temperature of 50 °C, and substrate concentration of 1.5%. The enzyme was thermostable at 60 °C for 1 h, and the optimum enzyme–substrate reaction time was 30 min. Sodium dodecyl sulfate and commercial detergents did not significantly affect lipase activity during 30-min incubation at 30 °C. Among the metal ions tested, the maximum lipase activity was attained in the presence of Zn2+, followed by Mg2+ and Fe2+. Lipase activity was not significantly affected in the presence of ethylenediaminetetraacetic acid, sodium lauryl sulfate and Triton X-100. Phenylmethylsulfonyl fluoride (1 mM) and the reducing, β-mercaptoethanol significantly inhibited lipase activity. The remarkable stability in the presence of detergents, additives, inhibitors and metal ions makes this lipase unique and a potential candidate for significant biotechnological exploitation.

Characterization of biotechnologically relevant extracellular lipase produced by Aspergillus terreus NCFT 4269.10.

Enzyme production by Aspergillus terreus NCFT 4269.10 was studied under liquid static surface and solid-state fermentation using mustard oil cake as a substrate. The maximum lipase biosynthesis was observed after incubation at 30°C for 96h. Among the domestic oils tested, the maximum lipase biosynthesis was achieved using palm oil. The crude lipase was purified 2.56-fold to electrophoretic homogeneity, with a yield of 8.44%, and the protein had a molecular weight of 46.3kDa as determined by SDS-PAGE. Enzyme characterization confirmed that the purified lipase was most active at pH 6.0, temperature of 50°C, and substrate concentration of 1.5%. The enzyme was thermostable at 60°C for 1h, and the optimum enzyme-substrate reaction time was 30min. Sodium dodecyl sulfate and commercial detergents did not significantly affect lipase activity during 30-min incubation at 30°C. Among the metal ions tested, the maximum lipase activity was attained in the presence of Zn(2+), followed by Mg(2+) and Fe(2+). Lipase activity was not significantly affected in the presence of ethylenediaminetetraacetic acid, sodium lauryl sulfate and Triton X-100. Phenylmethylsulfonyl fluoride (1mM) and the reducing, ß-mercaptoethanol significantly inhibited lipase activity. The remarkable stability in the presence of detergents, additives, inhibitors and metal ions makes this lipase unique and a potential candidate for significant biotechnological exploitation.

Enhanced production of pectinase by Aspergillus terreus NCFT 4269.10 using banana peels as substrate.

Aspergillus terreus NCFT4269.10 was implemented in solid-state (SSF) and liquid static surface fermentation (LSSF) for biosynthesis of pectinase. Amongst various substrates, like, mustard oil cake, neem oil cake, groundnut oil cake, black gram peels, green gram peels, chickling vetch peels/grass pea peels wheat bran, pearl millet residues, finger millet waste, broken rice, banana peels (BP), apple pomace (AP) and orange peels, banana peel (Musa paradisiaca L.; Family: Musaceae) was most suitable for pectinase biosynthesis (LSSF: 400 ± 21.45 Uml-1; SSF: 6500 ± 1116.21 Ug-1). Optimization of process parameters using one-variable-at-a-time method revealed that an initial medium pH of 5.0 at 30 °C and 96 h of incubation along with mannitol, urea, ammonium persulfate and isoleucine have positive influence on pectinase production. Further, K+ (1 mM), Riboflavin (10 mg 100 ml-1) and gibberellic acid (0.025 %, w/v) supported in enhanced pectinase production. Banana peels and AP at a ratio of 9:1, moisture content of 90 % with 2 % inoculum size were suitable combinations for production of pectinase. Similarly, 96 h of soaking time with 0.1 M phosphate buffer (pH 6.5) is essential for pectinase recovery. Purification to electrophoretic homogeneity revealed 1.42 fold purification with 8.08 % yield and a molecular weight of 24.6 kDa. Scaling up of various fermentation parameters and supplementing BP as the substrate for pectinase production with better recovery could make it promising for different industrial exploitation.

Physico-chemical assessment of paper mill effluent and its heavy metal remediation using aquatic macrophytes--a case study at JK Paper mill, Rayagada, India.

The present investigation aims to assess the phytoremediation potential of six aquatic macrophytes, viz. Eichhornia crassipes, Hydrilla verticillata, Jussiaea repens, Lemna minor, Pistia stratiotes and Trapa natans grown in paper mill effluent of JK Paper mill of Rayagada, Orissa, for remediation of heavy metals. The experiment was designed in pot culture experiments. Assessment of physico-chemical parameters of paper mill effluent showed significant decrease in pH, conductivity, total dissolved solids, total suspended solids, chlorine, sulphur, biological and chemical oxygen demand after growth of macrophytes for 20 days. Phytoremediation ability of these aquatic macrophytic species for copper (Cu) and mercury (Hg) was indicated by assessing the decrease in the levels of heavy metals from effluent water. Maximum reduction (66.5 %) in Hg content of untreated paper mill effluent was observed using L. minor followed by T. natans (64.8 %). L. minor showed highest reduction (71.4 %) of Cu content from effluent water followed by E. crassipes (63.6 %). Phytoextraction potential of L. minor was remarkable for Hg and Cu, and bioaccumulation was evident from bioconcentration factor values, i.e. 0.59 and 0.70, respectively. The present phytoremediation approach was considered more effective than conventional chemical treatment method for removing toxic contaminants from paper mill effluent.