MOLECULAR ASSESSMENT OF GENETIC DIVERSITY IN INDIAN ACCESSIONS OF ALOE VERA USING SSR MARKER

Objective: In this study Aloe vera (L.) Burm. f. collected from 12 states covering all the different agro-climatic zones of India were investigated for its genetic diversity analysis by using SSR marker assay.Methods: Total genomic DNA was isolated from young leaf samples using CTAB method. Twenty primers were selected which were used for Asparagus officinalis L a related species of A. vera and others were developed from available Aloe vera plant sequences with the help of primer 3 software. Similarity matrices and dendrogram were constructed by using NTSys software to show a phenetic representation of the genetic relationship. Polymorphic Information Content (PIC), the effective multiplex ratio (EMR) and Marker Index (MI) were calculated for the assessment of genetic diversity.Results: The neighbor-joining tree based on all SSR fragments of twelve Aloe vera germplasm accessions grouped into three major clusters. The similarity value ranged from 46 % to 100 %. The highest 100 % similarity was noted between Haryana and Uttar Pradesh accessions followed by 93% similarity between Haryana and Punjab accessions with Rajasthan. Minimum similarity was noted between Gujarat and Kerala accessions.Conclusion: This study revealed the rich genetic diversity among Aloe vera accessions from different agro-climatic zones of India. It is also concluded that SSR marker analysis can be a useful tool for the assessment of genetic diversity of the medicinal plants.Â

Screening of traditionally used medicinal plants for their antimicrobial efficacy against oral pathogens and GC-MS analysis of <em>Acacia nilotica </em>extract

162-168Oral diseases are one of the major public health issues. Due to acquisition of pathogenic resistance over conventional antimicrobials, the search for natural alternatives continues. In the present study, thirty two methanol and ethyl acetate extracts prepared from 14 different plant species were screened against oral pathogens. Principal Component Analysis indicated that methanol extract of Acacia nilotica twig was the most influential with highest F1 score and showed almost 2 fold higher antimicrobial activity in comparison to others. GC-MS analysis of Acacia nilotica twig revealed the presence of various bioactive such as limonene, stigmasterol, linoleic acid, ricinoleic acid, santalol, undecylenic acid. Evaluation of antimicrobial potential of medicinal plants may thrive a safe, inexpensive and efficient therapeutic in developing formulation for oral care products

ASSESSMENT OF GENETIC DIVERSITY IN TINOSPORA CORDIFOLIA BY INTER SIMPLE SEQUENCE REPEATS (ISSR) AND EXPRESSED SEQUENCE TAGGED- SIMPLE SEQUENCE REPEATS (EST-SSR)

Objective: In this study, assessment of genetic diversity was carried out using two kinds of molecular markers: Inter-Simple Sequence Repeats (ISSR) and Expressed Sequence Tag Simple Sequence Repeats (EST-SSR) in T. cordifolia. Methods: A total of 20 primers/primer pairs were tested for the detection of polymorphism. For genetic diversity assessment, certain parameters such as Polymorphic Information Content (PIC), Marker Index (MI), effective multiplex ratio (EMR) and DDI (Diversity detecting Index) were used. Results: The PIC, MI, EMR and DDI values ranges from 0.306-0.351, 0.76-1.18, 3.86-2.16 and 0.739-0.175 respectively. Cluster analysis based on Jaccard`s similarity coefficient using an Unweighted Pair Group Method with Arithmetic mean (UPGMA) classified all 24 accessions in to two major clusters respectively for both the marker system which demarcated the accessions according to different climatic zones. Similarity indices ranged from 0.68-1.0 for ISSR and 0.52-0.96 for EST-SSR. Conclusion: Both marker systems ISSR and EST-SSR separate out the accessions from different climatic zones in to different groups. In addition, both have shown a high genetic diversity and a good consistency among different genotypes of T. cordifolia. Out of these two, EST-SSR proves more efficient as it directly correlates with the geographical distribution of the plant

Screening of antimicrobial efficacy of traditionally used Indian plants against microorganisms associated with dandruff

Dandruff is a clinical condition affecting the scalp causing itching and relapsing inflammation. Staphylococcus epidermidis, Propionibacterium acne and Malassezia furfur pathogens are found more and less respectively in the scalps of dandruff subjects. In the present study, a total of 32 plant extracts were screened for their anti-dandruff activity by agar well diffusion method. Among the selected plant extracts of methanol and ethyl acetate, 25 showed significant activity while 7 extracts have not showed activity at a particular concentration. Among all the extracts ethylacetate extract of Cinnamomum zeylanicum and Glycyrrhiza glabra showed very high activity i.e. 18-20 mm while methanolic extract of Punica granatum and Syzygium aromaticum showed moderate activity against all the three pathogens. Thus the active plant extracts can be a potential source for the formulation of natural anti-dandruff agents

Molecular Detection of Hepatitis C Virus (HCV) by Conventional One-step RT-PCR Coupled with Nested PCR

Aims: HCV causes both acute and chronic infections and can be easily transmitted through contaminated blood or other body fluids. The present study deals with the molecular detection of HCV with help of one-step RT-PCR assay followed by nested PCR and agarose gel electrophoresis. Study Design: RNA extracted from the confirmed positive samples of HCV was utilized for the standardization of the one-step RT-PCR assay and nested PCR assay for diagnosis of HCV. Place and Duration of Study: Centre for Biotechnology, Maharshi Dayanand University, Rohtak Haryana, India, during period of one year (January-December 2015). Methodology: HCV positive samples were obtained from Department of Medicine, Maulana Azad Medical College (MAMC), New Delhi, India. Published primers from most conserved regions of HCV were taken and these primers were able to amplify all the strains of HCV. One-step RT-PCR kits, primers, extracted RNA from these positive samples were used for standardization of molecular diagnostic assays. The results were checked by 2% agarose gel electrophoresis. Results: Positive samples of HCV were detected by nested PCR. Positive samples showed sharp band of 405bp while there was no amplification in the negative control. Conclusion: Rapid tests have low sensitivity and specificity while molecular assays are rapid, sensitive and specific. Conventional one-step RT-PCR assay followed by nested PCR is rapid, specific, sensitive and it is also less costly than real-time RT-PCR. Cost of an assay is an important factor in controlling a disease in resource limited settings of developing countries

Data from: Spatial and seasonal influences on culturable endophytic mycobiota associated with different tissues of Eugenia jambolana Lam. and their antibacterial activity against MDR strains

Background: Present study focuses on diversity and distribution analysis of endophytic fungi associated with different tissues of Eugenia jambolana. The influence of season and geographical location on diversity and distribution of endophytic fungi has been analyzed. Antibacterial activity of isolated fungal species has also been investigated against MDR bacterial strains. Result: A total of 1896 endophytic fungal isolates were obtained from healthy, surface sterilized tissues of leaf, stem and petiole tissues during summer, monsoon and winter season. Out of 24 fungal species isolated, 20 species belong to class Ascomycetes, 2 to Basidiomycetes and 2 to Zygomycetes. Maximum species diversity was in rainy season whereas colonization frequency was in winter. All the diversity indices showed maximum species diversity at site 5 (Yamunanager), rainy among the seasons and leaf among the tissues studied. Aspergillus genus was most frequently isolated. Aspergillus niger and Alternaria alternata were most dominant species. Three way ANOVA results showed that effect of season was highly significant on species diversity in relation to sites and tissues. 60% endophytic fungal extracts showed significant antibacterial activity against one or more than one MDR bacterial strain. Conclusion: Different fungal species were recovered from different sites but the inter-site comparisons were not significant according to Jaccard similarity coefficient. Diversity of such fungal endophytes indicates that Eugenia jambolana plant acts as an ecosystem facilitating survival of many microbes with impressive antibacterial potential

Antiplasmodial potential and quantification of aloin and aloe-emodin in Aloe vera collected from different climatic regions of India

Abstract Background In this study, Aloe vera samples were collected from different climatic regions of India. Quantitative HPTLC (high performance thin layer chromatography) analysis of important anthraquinones aloin and aloe-emodin and antiplasmodial activity of crude aqueous extracts was done to estimate the effects of these constituents on antiplasmodial potential of the plant. Methods HPTLC system equipped with a sample applicator Linomat V with CAMAG sample syringe, twin rough plate development chamber (20 x 10 cm), TLC Scanner 3 and integration software WINCATS 1.4.8 was used for analysis of aloin and aloe-emodin amount. The antiplasmodial activity of plant extracts was assessed against a chloroquine (CQ) sensitive strain of P. falciparum (MRC-2). Minimum Inhibitory Concentration (MIC) of aqueous extracts of selected samples was determined according to the World Health Organization (WHO) recommended method that was based on assessing the inhibition of schizont maturation in a 96-well microtitre plate. EC (effective concentration) values of different samples were observed to predict antiplasmodial potential of the plant in terms of their climatic zones. Results A maximum quantity of aloin and aloe-emodin i.e. 0.45 and 0.27 mg/g respectively was observed from the 12 samples of Aloe vera. The inhibited parasite growth with EC50 values ranging from 0.289 to 1056 μg/ml. The antiplasmodial EC50 value of positive control Chloroquine was observed 0.034 μg/ml and EC50 values showed by aloin and aloe-emodin was 67 μg/ml and 22 μg/ml respectively. A positive correlation was reported between aloin and aloe-emodin. Antiplasmodial activity was increased with increase in the concentration of aloin and aloe-emodin. The quantity of aloin and aloe-emodin was decreased with rise in temperature hence it was negatively correlated with temperature. Conclusions The extracts of Aloe vera collected from colder climatic regions showed good antiplasmodial activity and also showed the presence of higher amount of aloin and aloe-emodin in comparison to collected from warmer climatic sites. Study showed significant correlation between quantities of both the anthraquinones used as marker compounds and EC50 values of the different Aloe vera extracts. Although, both the anthraquinones showed less antiplasmodial potential in comparison to crude extracts of different Aloe vera samples. Diverse climatic factors affect the quantity of tested compounds and antiplasmodial potential of the plant in different Aloe vera samples

Screening of antimicrobial efficacy of traditionally used Indian plants against microorganisms associated with dandruff

934-939Dandruff is a clinical condition affecting the scalp, causes itching and relapsing inflammation. In the present study, a total of 32 plant extracts of 16 plants were prepared using methanol and ethyl acetate and screened for their anti-dandruff activity by agar well diffusion method. Dandruff causing microorganisms including Staphylococcus epidermidis, Propionibacterium acne and Malassezia furfur were selected for the study. Ethyl acetate extract of Cinnamomum zeylanicum and Glycyrrhiza glabra exhibited maximum activity with zone of inhibition of 18-20 mm while methanol extract of Punica granatum and Syzygium aromaticum demonstrated moderate activity against the studied microbes. On the basis of ZOI, PCA and MIC the results revealed that the ethyl acetate extract of C. zeylinicum bark and G. glabra root are most influential extracts followed by methanol extract of P. granatum, G. glabra, S. aromaticum, and ethyl acetate extract of A. nilotica in term of antimicrobial activity against the studied dandruff causing microbes. Therefore the active plant extracts can act as a potential source for the formulation of natural anti-dandruff agents