Genetic Diversity among Desmodium Gangeticum (L.) DL Accessions a Desmodin Synthesising Ethno-Medicinal Plant With SSR and Internal Transcribed Spacer Region for Species Conservation

In the present study simple sequence repeat (SSR) andintertranscribed spacer markers markers were used inDesmodium gangeticum to find out the genetic diversityanalysis for the biodiversity conservation of the Species as theplant is being used in medicinal preparations as well as havemany industrial applications. The study was aimed to find outthe relationships among different accessions. We find intrapopulationmean genetic value of (1.835) and Shannon (0.641)among the accessions. in AMOVA analysis, we noticed smallvariation accessions as (P = 1).The dendrogram generatedthrough distance matrix using the SIMQUAL-Dice Coefficientmodule of NTSYSpc ver matches to some extent with the PCAgraph. In the barcode ITS marker analysis the UPGMA showsthe accessions clustered into two groups. Blast results showedmaximum E-value in DG2 accession. In conclusion the abovebased markers can be used for the population conservationand authentication of the above medicinally important plant

Isolation and characterisation of plant growth-promoting bacterial and fungal endophytes from Himalayan Yew (Taxus wallichiana) - an economically imperative pant of Himalayas

It is a known fact that the bacterial and fungal endophytes inhabit the plant tissues besides aiding in the better growth and health of the plants. The bark and leaves of Taxus wallichiana have drawn a lot of interest in recent years since they are the richest source of taxol, an anticancer drug. As it is a slow-growing tree that can only be regenerated via vegetative propagation, it has been classified as a critical rare species due to its extensive collection for medicinal and other purposes. Nonetheless, the use of endophytes as plant growth promoters is gaining much importance among environmentalists and agronomists because of their imperative role in crop production. Even then, there is hardly any information available regarding the growth-promoting endophytes isolated from bark and leaves associated with T. wallichiana commonly known as Himalayan Yew. Therefore, the present study was undertaken to isolate fungal and bacterial endophytes from T. wallichiana and to classify the growth-promoting properties of these endophytes. In total, seven fungal and ten bacterial endophytes were obtained from different parts of T. wallichiana. All of the isolated fungal and bacterial endophytes produced indole acetic acid while most of them also produced ammonia. Besides, the fungal and bacterial endophytes were also screened for antimicrobial and various enzymatic activities. Based on the above results, the two fungal endophytes were selected for their possible ability to promote seed growth. The results showed that the fungal endophytes isolated from T. wallichiana played an active role in increasing growth in other plant species and therefore, can be used as potential plant growth promoters

Evaluation of alkali and thermotolerant lipase from an indigenous isolated Bacillus strain for detergent formulation

Background: Lipases are used in detergent industries to minimise the use of phosphate-based chemicals in detergent formulations. The use of lipase in household laundry reduces environmental pollution and enhances the ability of detergent to remove tough oil or grease stains. Results: A lipase-producing indigenous Bacillus subtilis strain [accession no. KT985358] was isolated from the foothills of Trikuta mountain in Jammu and Kashmir, India. The lipase (BSK-L) produced by this strain expressed alkali and thermotolerance. Lipase has an optimal activity at pH 8.0 and temperature 37\ub0C, whereas it is stable at pH 6.0\u20139.0 and showed active lipolytic activity at temperatures 30 to 60\ub0C. Furthermore, lipase activity was found to be stimulated in the presence of the metal ions Mn2+, K+, Zn2+, Fe2+ and Ca2+. This lipase was resistant to surfactants, oxidising agents and commercial detergents, suggesting it as a potential candidate for detergent formulation. BSK-L displayed noticeable capability to remove oil stains when used in different washing solutions containing buffer, lipase and commercial detergent. The maximum olive oil removal percentage obtained was 68% when the optimum detergent concentration (Fena) was 0.3%. The oil removal percentage from olive oil-soiled cotton fabric increased with 40 U/mL of lipase. Conclusions: This BSK-L enzyme has the potential for removing oil stains by developing a pre-soaked solution for detergent formulation and was compatible with surfactants, oxidising agents and commercial detergents

Application of a multiplex PCR assay for the detection of Shigella, Escherichia coli and Shiga toxin-producing Esch. coli in milk.

A multiplex PCR (mPCR) assay using previously known genetic markers of Shigella, Escherichia coli and Shiga-toxic Esch. coli was standardized. uidA gene was targeted for the common detection of Esch. coli and Shigella, whereas ipaH and stx1 genes were used as markers for the detection of Shigella and shiga-toxin producing strains, respectively. The standardized assays detected the target organism specifically and selectively. The mPCR developed by combining all the three reactions generated specific products. The inclusivity and exclusivity tests depicted the precise specificity of the mPCR assay. Results were interpreted on the basis of the pattern of amplicons generated: amplifications of the ipaH and uidA gene fragments indicated the presence of Shigella spp., amplification of uidA alone revealed the presence of Esch. coli and additional presence of verotoxin gene amplicon indicated verotoxinogenic nature of the strain. Specific patterns of bands were obtained when different strains of Esch. coli and Shigella spp. were subjected to this assay. The reactions, individually as well as in the mPCR, could detect approximately 1 cell per 20-microl PCR assay. The protocols were validated by analyzing the coded samples of full fat milk spiked with different pathogens. In naturally contaminated raw milk samples (n=100), Esch. coli were detected in all samples and verotoxinogenic Esch. coli in 15 samples. Shigella, however, was not detected in any of the samples. When DNA purified from the samples found positive for Shiga-toxic Esch. coli was directly used as template for the mPCR, the results showed agreement with the enrichment based detection. The mPCR assay, standardized in this study, may be used for rapid microbiological evaluation of milk samples. Further, the study emphasizes the need for better hygienic conditions in dairies

Evaluation of alkali and thermotolerant lipase from an indigenous isolated Bacillus strain for detergent formulation

Background: Lipases are used in detergent industries to minimise the use of phosphate-based chemicals in detergent formulations. The use of lipase in household laundry reduces environmental pollution and enhances the ability of detergent to remove tough oil or grease stains. Results: A lipase-producing indigenous Bacillus subtilis strain [accession no. KT985358] was isolated from the foothills of Trikuta mountain in Jammu and Kashmir, India. The lipase (BSK-L) produced by this strain expressed alkali and thermotolerance. Lipase has an optimal activity at pH 8.0 and temperature 37°C, whereas it is stable at pH 6.0–9.0 and showed active lipolytic activity at temperatures 30 to 60°C. Furthermore, lipase activity was found to be stimulated in the presence of the metal ions Mn2+, K+, Zn2+, Fe2+ and Ca2+. This lipase was resistant to surfactants, oxidising agents and commercial detergents, suggesting it as a potential candidate for detergent formulation. BSK-L displayed noticeable capability to remove oil stains when used in different washing solutions containing buffer, lipase and commercial detergent. The maximum olive oil removal percentage obtained was 68% when the optimum detergent concentration (Fena) was 0.3%. The oil removal percentage from olive oil-soiled cotton fabric increased with 40 U/mL of lipase. Conclusions: This BSK-L enzyme has the potential for removing oil stains by developing a pre-soaked solution for detergent formulation and was compatible with surfactants, oxidising agents and commercial detergents. Keywords: Activity, Bacillus subtilis, Environment pollution, Fabric, Gras, Lipolytic activity, Oil, Removal, Surfactant, Thermotoleran

Piperine, a Phytochemical Potentiator of Ciprofloxacin against Staphylococcus aureus

Piperine, a trans-trans isomer of 1-piperoyl-piperidine, in combination with ciprofloxacin markedly reduced the MICs and mutation prevention concentration of ciprofloxacin for Staphylococcus aureus, including methicillin-resistant S. aureus. The enhanced accumulation and decreased efflux of ethidium bromide in the wild-type and mutant (CIP(r)-1) strains in the presence of piperine suggest its involvement in the inhibition of bacterial efflux pumps

Functional cloning and predictive structural modeling of a novel esterase from Bacillus subtilis strain, RRL 1789.

We have recently reported the purification and characterization of a novel esterase from the Bacillus subtilis strain. In the present study we report the genomic DNA cloning and predictive structural modeling of this novel esterase. Tributyrin- and Rhodamine B-based functional screen of a Bacillus subtilis genomic library led to the identification of a potential lipolytic gene. DNA sequence analysis of the cloned gene showed that it encodes a protein of 489 amino acid residues. Sequence homology search and multiple sequence alignment showed that the protein was highly homologous to known esterases. Secondary structure-driven multiple sequence alignment with the homologous esterase of known three-dimensional structures was performed and a 3D structure model of this enzyme was constructed. Based on the topological organization of the secondary structures, this protein belongs to the alpha/beta hydrolase superfamily. Moreover, the presence of serine in the context of amino acid sequence G/A-X-S-X-G (with X an arbitrary amino acid residue) in the protein indicates that it belong to the class of serine hydrolases of this superfamily