Showing papers in "Indian Journal of Biotechnology in 2017"

Novel microorganisms for the treatment of Ni and V as spent catalysts

Abstract: Oil refining industry has used a great amount of catalysts, which once exhausted due to metal and hydrocarbon poisoning are disposed in special confining sites. Catalyst disposal represents a serious environmental problem due to the potential risk of metal leaching by natural events. The necessity to treat great amounts of catalysts and the existence of microorganisms that coexist with metals suggest that microorganisms can be used for the treatment of hazardous wastes such as spent catalysts. The aim of the present study was to isolate microorganisms from rich metal sites, mines, soils and water from rivers close to mines and then evaluate their ability to remove Ni and V from spent catalyst. Twenty-six isolates were obtained from samples using 9K liquid media and from them, only twelve isolates presented a minimum inhibitory concentration (MIC) higher than 200 ppm of Ni and V, then were evaluated for their ability to remove Ni and V in 9K liquid media added with 16% (w/v) pulp density of the catalyst. Results showed that isolates MNSH1-9K-1 and PRGSd-9K-4 had the highest removal for Ni and V corresponding to 149.5 mg Kg and 920.5 mg Kg, respectively and were identified by sequencing of 16S ribosomal RNA gene as Bacillus megaterium and Bacillus subtilis, respectively.

Sugar cane molasses as culture media component for microbial transglutaminase production

Abstract: Sugar cane molasses as culture media component for microbial transglutaminase production Oscar M Portilla*, Vicente Espinosa, Lorenzo Jarquin, Arturo Salinas, Gonzalo Velazquez, Manuel Vazquez Universidad Autónoma de San Luis Potosí CARHS, Km 5 Carretera TamazunchaleSan Martín.Tamazunchale, SLP-79960, México, Universidad Politécnica de Guanajuato, Av Universidad Norte, Cortazar, Guanajuato-38483, México, Instituto Politécnico Nacional, CICATA Unidad Querétaro, Cerro Blanco141, Colinas del Cimatario, Querétaro-76090, México, Facultad de Veterinaria, Universidad de Santiago de Compostela, Campus Lugo, Lugo-27002, España

Molecular markers in assessing genetic variation of Indian citron ( Citrus medica L.) cultivars collected from different parts of India

Abstract: Genetic diversity of forty six representative accessions of Citrus medica L. (Rutaceae) was estimated using 17 (RAPD) random amplification of polymorphic DNA and 11 inter simple sequence repeat (ISSR) markers. A total of 213 RAPD and 105 ISSR bands were generated, out of which, 175 (RAPD) and 77 (ISSR) bands were polymorphic. Polymorphism percentage of RAPD (82%) was comparatively higher than ISSR (73%). Genetic similarity value was found in the range of 0.49 to 0.96 (average 0.73), 0.38 to 0.99 (average 0.69) and 0.47 to 0.96 (average 0.72) in RAPD, ISSR and combined data analyses, respectively. Moderately high levels of polymorphism and genetic similarity within C. medica suggested that citron cultivars have a significant level of genetic diversity. A dendrogram generated based on unweighted pair group method of arithmeticaverage (UPGMA) separated all the accessions into two main clusters. The placement of C. medica accessions in various sub-clusters and groups in the dendrogram was based on molecular differentiation of individual accessions rather than their geographical origin. Both, RAPD and ISSR analyses were able to identify elite genotypes of C. medica with unique accession specific DNA fragments. Thus, both the markers can be utilized for inferring the extent and distribution of genetic diversity in C. medica.

Plant regeneration through isolated microspore culture in recalcitrant durum wheat genotypes.

Abstract: In the present study, a protocol of isolated microspore culture was optimized to regenerate green plants in Turkish durum wheat genotypes (Kiziltan-91, C-1252, Mirzabey 2000 & Kunduru-1149). The bread wheat cultivar (Gun-91) was used as control because of its high androgenic response in the microspore culture. First significant step was to treat the anthers with four pretreatments (cold, cold with mannitol, cold with sorbitol & mannitol at room temperature). Anther maceration was used as an isolation method and microspores were plated on induction culture medium supplemented with arabinogalactan-proteins (AGP) and ovary coculture. When the embryos reached the size of 2 mm, they were transferred to the differentiation medium having a combination of phenylacetic acid (PAA) and gibberellic acid (GA3). The best results were obtained with the pretreatment of mannitol (+4°C) for 7 d on providing embryos and regenerated green plants in four durum wheat genotypes. The cultivar Kiziltan-91 gave the best response for embryo (>2 mm) formation (20.63%) and cultivar Kunduru-1149 for green and total plant regeneration (6.25% and 18.75%). These results indicated that the present protocol performed well to increase the embryo formation and green plant regeneration in the recalcitrant durum wheat genotypes studied.

Comparative analysis of bioethanol production involving saccharification by mixed recombinant clostridial enzymes using sugarcane leaves and kans grass as sustainable feed stocks from North-east India.

Abstract: The present study involves bioethanol production from sugarcane (Saccharum officinarum) leaves (SL) and Kans grass (Saccharum spontaneum) (KG), under two separate approaches of fermentation, i) SHF (separate hydrolysis & fermentation) and ii) SSF (simultaneous saccharification & fermentation). The H3PO4-acetone pretreatment strategy was performed for effective lignin removal and enhanced porosity, confirmed by FT-IR (fourier transform infrared spectroscopy) and FESEM (field emission scanning electron microscopy). Saccharification was performed by involving recombinant cellulase GH5 (family 5 of glycoside hydrolase) and hemicellulase GH43 (family 43 of glycoside hydrolase) from Clostridium thermocellum, expressed and isolated from Escherichia coli. Successively, the bioethanol producers Saccharomyces cerevisiae (NCIM 3215) and Candida shehatae (NCIM 3500) were employed. Comparative SHF trials with 1% (w/v) pretreated KG and SL gave bioethanol titre of 0.52 and 0.64 g/L, respectively, whereas SSF experiments resulted in bioethanol titre of 0.8 and 1.0 g/L, respectively at 100 mL shake flask. An increased feedstock concentration of 5% (w/v) KG in shake flask resulted in increased bioethanol titre (4.2 g/L) in SSF. However, the increased substrate concentration of 5% (w/v) SL in individual shake flask resulted in 1.4 fold more bioethanol titre (5.7 g/L) in comparison to KG, and its scale up in bioreactor with 1 L working volume gave bioethanol titre of 11.4 g/L and yield of 0.267 g of bioethanol/g of pretreated SL.

Bioethanol production from watermelon rind by fermentation using Saccharomyces cerevisiae and Zymomonas mobilis.

Abstract: Bioethanol production potential of watermelon rind as feed stock was studied using Saccharomyces cerevisiae and Zymomonas mobilis. Significant production (5.86%) of bioethanol could be obtained at a five per cent substrate concentration of watermelon rind along with peptone as an additive by carrying out a single batch bioconversion and fermentation. The use of Zymomonas mobilis for saccharification and Saccharomyces cerevisiae for fermentation was found to be more effective compared to the use of single organisms for both the steps. The efficacy of bioconversion by Zymomonas mobilis was confirmed by the reduction in cellulose content of the biomass and increase in the total reducing sugar content (490 μg/ml) and glucose content (0.09%).

Revisited immune reactivity between native semi-purified protoplasmic (caprine) versus commercial purified protoplasmic (bovine) antigens for the screening of goatherds endemic for Johne’s disease

Abstract: The present study compared the immune-reactivity of 3 antigens of Mycobacterium avium subspecies paratuberculosis (MAP) sourced from different geographical regions and livestock species for the diagnosis of Johne’s disease (JD) in goats. Screening of 360 faecal and serum samples of goats by microscopy, i-ELISA, b-ELISA and r-ELISA gave 56.9, 40.0, 34.4 and 5.2% positive samples, respectively. Considering all the 4 tests (microscopy, i-ELISA, b-ELISA & r-ELISA kits), 3.0 and 35.2% goats were found common positive and negative, respectively. Of 3 ELISA tests, i-ELISA had the highest sensitivity, followed by b-ELISA and r-ELISA kit. ‘i-ELISA’ based on ‘indigenous antigen’ recovered from native (‘Indian Bison Type’) MAP strain of goat origin had superior immune reactivity as compared to imported commercial PPAs (protoplasmic antigens) of bovine origin (b-ELISA from Allied Monitor Inc., USA) and commercial ELISA kit for ruminant species (ID Vet, France). Lower immune-reactivity of commercial antigens as compared to ‘indigenous antigen’ indicated that search for universally acceptable ‘ELISA kit’ is not practically possible.

Effect of thidiazuron on in vitro regeneration potential of cotyledon and hypocotyl explants of cauliflower (Brassica oleracea L. var. botrytis)

Abstract: Plant regeneration studies were carried out with cotyledon and hypocotyl explants on four different combinations and concentrations of thidiazuron (TDZ), i.e., TDZ, TDZ+IAA (indole acetic acid), TDZ+NAA (naphthalene acetic acid) and TDZ+adenine, in cauliflower (Brassica oleracea var. botrytis cv. Pusa Snowball K1). Among the forty combinations tried, the maximum percent shoot regeneration was observed in hypocotyl explants (96%) on MS medium supplemented with 0.44 mg/L TDZ+0.08 mg/L IAA; whereas in case of cotyledon explants, only 64% shoot regeneration was achieved on MS medium supplemented with 0.77 mg/L TDZ+79.7 mg/L adenine. For root regeneration, the in vitro regenerated shoots were cultured on MS medium containing different concentration of IBA (indole butyric acid). About 100% root regeneration was observed on MS medium supplemented with 0.10 mg/L IBA. The regenerated plantlets were transferred to the pots containing cocopeat and acclimatized. Thus, an efficient and reproducible plant regeneration protocol has been standardized in cauliflower cv. Pusa Snowball K1.

Determination of genetic relationship among basmati and non-basmati rice (Oryza sativa L.) genotypes from North-West Himalayas using microsatellite markers.

Abstract: In the present study, 25 microsatellite markers were used to determine the genetic relatedness among the 51 basmati and 14 non-basmati rice (Oryza sativa L.) genotypes. A total of 82 alleles were detected by 25 markers, all of them (100%) were polymorphic. The polymorphic information content (PIC) among genotypes varied 0.253 (RM520) to 0.695 (RM206) with an average of 0.46. Pairwise genetic similarity coefficients between all genotypes ranged 0.1 to 0.6 with average of 0.39. Phylogenetic-based cluster analysis of the SSR data, based on distance, divided all genotypes into four groups (I, II, III & IV), consisting of 39, 7, 16 and 3 genotypes, respectively. Principal coordinate analyses (PCoA) confirmed the separation of basmati and non-basmati rice genotypes comparable to those from UPGMA analysis and were well in agreement. These results suggest that the microsatellite SSR markers are efficient for measuring genetic relatedness among the rice genotypes, and can be utilized effectively for the differentiation of basmati and non-basmati rice genotypes. Present study also indicated that genetically basmati rice is different from that of coarse rice type, and supports the concept of independent evolution of basmati rice. The low level of diversity in local basmati suggested the introduction of diverse germplasm in the basmati breeding programme.

A study on phosphate uptake by Acinetobacter sp. in presence of arsenate under aerobic condition

Abstract: Phosphate is the main hindrance for the removal of arsenic from the arsenic contaminated waste water. Therefore, phosphate removal from contaminated water has become imperative for the successful removal of arsenic. In the present study, an attempt was made to remove phosphate from the waste water by Acinetobacter sp. in the presence and absence of arsenate. When phosphate (25 ppm) containing synthetic solution was treated with Acinetobacter sp. at pН 6 at ambient temperature under aerobic condition, the bacterium was able to remove 71.88% (17.97 ppm) phosphate. However, in the presence of arsenate (5 ppm), only 54.24% (13.56 ppm) phosphate uptake was observed from the waste water by Acinetobacter sp. Thus the presence of arsenate (5 ppm) inhibited phosphate uptake by 17.64%. The phosphate uptake by Acinetobacter sp. follows the Michalis-Menten kinetics. In the presence and absence of arsenate, the maximum velocity (Vmax) of phosphate uptake was 1.07 and 1.03 μM mg−1 h−1; while the kinetic constant (Km) was 1.13 and 0.37 mM, respectively. Consequently, arsenate was observed as competitive inhibitor for the phosphate uptake. The data thus underlines the significance of Acinetobacter sp. for the removal of phosphate along with arsenate.

Evaluation of genetic diversity using random amplified polymorphic DNA (RAPD) markers in Melia dubia Cav.

Abstract: Random amplified polymorphic DNA (RAPD) molecular markers were used to evaluate the genetic diversity in populations of Melia dubia Cav. syn. M. composita Willd. (Family: Meliaceae) (Burma dek) from different agroclimatic regions of India. Of the 38 decamer primers used, 13 yielded polymorphic banding patterns. In total, 105 different bands were reproducibly obtained, of which 69 (65.7%) were polymorphic. The polymorphisms were scored and used in bandsharing analysis to identify genetic relationships. Cluster analysis based on Jaccard’s similarity coefficient using UPGMA grouped all the 24 populations into two major groups. Similarity indices ranged from 0.80 to 0.91, indicating that Burma dek germplasm within India constitutes considerably narrow genetic base.

Crustacean hyperglycemic hormone family gene silencing in Penaeus monodon mediated through dsRNA synthesized in vitro from genomic and cDNA

Abstract: S Vrinda1, C Reshmi1, Seena Jose1, P Reynold2, K K Vijayan 2, Rosamma Philip3 and I S Bright Singh1* 1National Centre for Aquatic Animal Health, Cochin University of Science and Technology, Cochin 682 016, India 2Central Marine Fisheries Research Institute, Cochin 682 018, India 3Department of Marine Biology, Microbiology and Biochemistry, Cochin University of Science and Technology Cochin 682 016, India