Research Journal of Biotechnology

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Research Journal of Biotechnology

Antimicrobial and antioxidant activity of flower scape extracts of Agave salmiana: effect of the extraction solvent and development stage

María Ivette Medina-Galván, Aurea Bernardino-Nicanor, Javier Castro-Rosas, María de la Luz Xochilt Negrete-Rodríguez, Eloy Conde-Barajas and Leopoldo González-Cruz

Page No: 1-9

Abstract: The flower scape of Agave salmiana is traditionally used as anthelmintic agent, as insecticide against mosquitoes and as antimicrobial agent in some regions of Mexico. In this study, the antibacterial activity of the flower scape extracts against bacterial strains important in food production, the total phenolics content and antioxidant activity were evaluated. Five extracts were obtained using water, acetone, ethanol, methanol or hexane as solvents and the crude extract. All extracts were tested against Staphylococcus aureus, Listeria monocytogenes, Shigella sonnei, Escherichia coli and Salmonella typhimurium. Rifampicin resistance was induced in all strains. The extract yield, total phenolic content and antioxidant activity of the extracts were also tested. Gram-negative bacteria were more susceptible to growth inhibition than gram-positive bacteria.

This was especially true for aqueous extract and crude extract. A higher yield, antioxidant activity and total phenolic content was obtained with the water extract. Higher phenolic compound totals, antimicrobial activity and antioxidant activity were obtained using the middle development stage and middle section. The development stage and the tested section of the flower scape influenced the antimicrobial and antioxidant activities.

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Extraction of Diosgenin from Fenugreek and evaluation of its pharmacological role in alleviating Metabolic Syndrome in vitro

Myrene Dsouza, Rufina K. and Hana D.

Page No: 10-17

Abstract: Metabolic syndrome complications continue to rise with expanding urban lifestyles. Phytochemical components derived from botanicals have been used for decades in the treatment of metabolic diseases. One such component, diosgenin, a steroidal saponin is known to be present richly in fenugreek. The objective of the study was to evaluate whether diosgenin extracted from fenugreek by microwave assisted extraction (MAE) and ultrasound assisted extraction (UAE) methods, possesses in vitro anti-inflammatory and anti-diabetic properties.

In case of UAE, maximum diosgenin was obtained from fenugreek seeds with 80% ethanol for 5 min, whereas the maximum diosgenin in MAE was obtained with 80% ethanol for 1.3 min. UAE yielded greater amount diosgenin than MAE. The IC50 values for in vitro anti-inflammatory activities were evaluated by heat induced protein denaturation (IC50 crude=391.3 μg/ml, IC50 defat=25.2 μg/ml). The IC50 values for in vitro anti-diabetic activities were evaluated by the following assays: α-amylase inhibition (IC50 crude=371.7 μg/ml, IC50 defat=370.5 μg/ml) and pancreatic lipase (IC50 crude=550.0 μg/ml and IC50 defat=497.6 μg/ml). From the glucose diffusion assay, it is evident that diosgenin helps in delaying the glucose diffusion for up to 4h, thereby providing sufficient time for the system to eliminate glucose and prevent its uptake into the cells of the body. The results of these assays are promising for the use of diosgenin from fenugreek for the control of metabolic syndrome as it helps in the delay of fat and glucose absorption.

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Production and Characterization of Protease from TP2 isolate from Plant Swamp Silage

Ace Baehaki, Shanti Dwita Lestari, Agung Tirtayasa, Arif Hidayat and Nuni Gofar

Page No: 18-21

Abstract: The purpose of this research was production and characterization of protease from TP2 isolate of Plant Swam Silage. The optimum pH and temperature of protease from TP2 isolate were 11.0 and 45ºC respectively. Na+ and Mg2+ increased TP2 protease whereas K+, Fe2+ and Zn2+ inhibited protease from TP2 isolate inhibiting the enzyme. Study on the effect of metals ion indicated that protease from TP2 isolate was metaloenzyme. Moleculer weight of protease by using SDSPAGE from TP2 isolate was 34,75 kD to 185,51kDa.

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cpSSR marker based DNA fingerprinting and diversity assessment of mango genotypes

Ahmad Israr, Bajpai Anju and Rajan Shailendra

Page No: 22-26

Abstract: Germplasm characterization is a pre-requisite for their conservation and utilization in breeding programme. We used cpSSR markers for the first time to identify a set of polymorphic microsatellite loci and analysis of genetic diversity within mango genotypes. The present study was aimed to assess the intraspecific relationships among 28 diverse mango genotypes using uniparental and haploid chloroplast SSR (cpSSR) marker. A total of 27 alleles were identified with an average of 1.5 allele per locus. Seven out of 18 primer pairs showed polymorphic bands with a number of alleles ranging from one to three. The genotypes studied revealed significant levels of cpDNA genetic diversity. Shannon diversity index and gene diversity varied, from 0.07-0.32 and 0.34-0.95 respectively. Marker index and resolving power were ranged 0.04-2.14 and 0.07-1.71.

cpSSR markers are suitable for characterization and assessment across diverse mango genotypes and there is small difference in the genetic diversity with the species. Results indicated that two mango cultivars (Sukul and Himsagar) were found to be far distant of the genetic relationship from the other cultivars. Analysis of overall diversity of mango genotypes revealed the less intraspecies diversity (40%). This low intraspecific diversity is due to high degree of gene flow in population through random mating without barrier. The information of this work will be useful for selection and more efficient utilization of mango germplasm in breeding programs in the future.

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Effects of different nitrogen levels on photosynthetic characteristics, chlorophyll fluorescence and yield of allium sativumcv ‘Minlezipi’

Zhang Y.J., Gao H., Cheng H.Y. and Wang Y.W.

Page No: 27-31

Abstract: We studied the effects of different nitrogen levels on photosynthetic characteristics, chloropdy researchehyll fluorescence and yield of Allium sativumcv ‘Minlezipi’. The results showed that the application of nitrogen fertilizer could significantly increase the photosynthetic parameters. Improve photosynthetic performance and promote photosynthetic rate and alleviate the midday depression of photosynthesis to some extent. Meanwhile, proper application of nitrogen fertilizer is beneficial to the decline of F0 improving the values of Fm, Fv/Fm.

The results showed that increasing the amount of nitrogen fertilizer could improve the photochemical efficiency of photosystem II and play the potential of photosynthesis. When nitrogen application amount was, the bulb and appearance quality of garlic reached the maximum and the quality of fresh garlic, the diameter of garlic, the plant height and stem diameter increased 13.9%, 22%, 21.1% and 23.7% respectively. It showed under the condition of local soil fertility. Appropriate application of nitrogen fertilizer could increase photosynthetic rate of leaves. Alleviate photosynthetic midday depression. Improve the photochemical efficiency of photosystem and increase the garlic yield.

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Barrier effect of the M3 highway in Hungary on the genetic diversity of wild boar (Sus scrofa) population

Bendegúz Mihalik, Viktor Stéger, Krisztián Frank, László Szendrei and Szilvia Kusza

Page No: 32-38

Abstract: Highways are human-related barriers with multiple impacts on the environment. They reduce the habitat of the animals and plants, isolating parts of extant populations from each other and blocking migration routes. They also have a disturbing effect and increase mortality by animals being hit by vehicles. In Hungary, the M3 highway is one of the busiest motorways. In our hypothesis, it acts as a weak to medium barrier to gene flow between the populations on the two sides. To test this hypothesis and to establish the genetic profiles of the wild boars 40 samples (blood, meat, fur or hide) were collected from each side from which genomic DNA was isolated. 13 tetrameric STR markers described in pigs were used for genotyping.

Genetic diversity indices and structure were calculated with STRUCTURE v.2.3.4, PAST v.2.17c, Cervus v.3.0.6 and ARLEQUIN v.3 software. The results show that the populations on the two sides of M3 have only minor differences in allele frequencies and heterozygosity values. Nei’s and Fst distance values were both negligible (0.045 and 0.025 respectively), which means that the two groups are only slightly separated and still belong to the same population. This situation may be due to the recent building of M3, which started only 40 years ago, or to the functioning wildlife underpasses and the good mobility of wild boars. However, there exists a small effect, which can be intensified in time, therefore periodic monitoring is necessary to prevent stronger isolation.

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Nano-structure cathode catalysts for improving Microbial fuel cell performance

Mahmoud Rehab H., Hassan Rabeay Y.A., Samhan Farag A., Ali Gamila H. and Ibrahim Mohamed K.

Page No: 39-46

Abstract:Platinum-reduced graphene oxide- graphite nanocomposite (Pt/ RGO/gr) and Pt free catalysts, graphene nanosheets (GNSs), MnO2 and RGO/MnO2 were synthesized and investigated as cathode catalysts for oxygen reduction reaction (ORR) with the Platinum as a benchmark. Electrochemical and morphological characters of the synthesized catalysts were studied using cyclic voltammetry and transmission electron microscopy (TEM). Power densities produced from MFC closed circuit operation using catalysts on the cathodes were about 68 times more than those obtained using graphite electrode. Although, there was slightly reduction of 5.8% in MFC power density (from 170mW/m2 to 160 mW/m2) when Pt nanoparticles were loaded on RGO nanosheet compared to Pt/C cathode, by replacing the precious- Pt catalyst with other catalytic materials, a significant reduction in power density was observed.

For RGO based cathode, the MFC performance decreased by 31.1% (from 170mW/m2 to117 mW/m2) and the percent of reduction reached 48.5% (from 170mW/m2 to 88 mW/m2) for MnO2 based cathode MFC. However, in RGO/MnO2 nano-composite cathode MFC, the percent of reduction in power density was 26.4% (from 170 mW/m2 to 125 mW/m2). It can be concluded that, RGO nanosheet loaded with Pt nanoparticles could be a good start for finding an alternative economic and effective cathode.

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Classification of 5’ and 3’ Untranslated Regions in Human transcriptome by Machine Learning methods

Kumar Shailesh, Govil Sumit, Kumar Vikram, Kachhawah Sumita and Kothari S.L.

Page No: 47-53

Abstract: Untranslated regions (UTRs) are non-coding part present upstream and downstream of translation unit of mRNA and contribute to regulation of translational process, gene composition and expression. Here, 5’ and 3’ UTRs are classified by various machine learning methods on genomic features contributing to UTRs composition. Supervised Machine learning based classification algorithms such as Random forest, Logistic Regression and Naïve Bayesian are used to classify UTRs. Merged and resampled dataset were used for training of classifiers. Tree based method like Random forest was found best among all methods for classification of UTRs with average sensitivity on merged and resampled dataset 0.865 and 0.968 respectively.

The key finding is that 3’UTRs composition is different from 5’ UTRs and can be classified by every classifier with variable sensitivity and specificity. These models can be used to identify the diseased causing UTRs on its features.

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Antibacterial activity of fungal Sclerotium rolfsii lectin (SRL1) against Ralstonia solanacearum

Jayalakshmi K., Lingaraju S., Bhat R.S., Raju J., Raghu S., Bhagat Y. and Giri Madhu S.

Page No: 54-60

Abstract: Antibacterial activity against Ralstonia solanacearum was evaluated by using E.coli expressed Scleroitum rolfsii lectin (SRL1) which was collected from Department of Biotechnology, University of Agricultural Sciences, Dharwad. The expression of SRL1 was checked with total protein isolated from induced E. coli BL21(DE3)pLysS cells carrying respective recombinant expression vectors. Recombinant E. coli clones had a total protein content of 12 mg/ml compared to 10.00 mg/ml in control. SRL1 proteins showed 19.5 kDa band on SDS-PAGE. SRL1 agglutinated rabbit and human erythrocytes with a higher specificity for rabbit erythrocytes. Haemagglutination was not inhibited by the tested carbohydrates but glycoproteins exerted a strong inhibitory action. Minimum concentration of protein required for agglutination (MCA) 1.73 µg with total activity of 0.43 ×104. E. coli expressed SRL1 lectin was purified through His-tag purification kit and the purified lectin showed a single band on denaturing electrophoresis, with a molecular mass of 19.5 kDa.

SRL1 demonstrated a remarkable nonselective antibacterial activity. SRL1 strongly inhibited the growth of R. solanacearum with a MIC of 0.625 mg/ml with 1.86 mg/ml MBC which corresponded to the minimum concentration of the lectin capable of reducing the number of cfu for 0.1% of the initial inocula. Interaction effect of lectin and concentrations with an inhibition zone of 2.07 cm and gradually reduced the growth of R. solanacearum 0.865 OD.

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Proximate Composition of Marine Worm EEL (Myrophis Platyrhnchus, Breder, 1927) in Parangipettai Coastal Water, Tamil Nadu, Southeast Coast of India

Veeramani T., Ravi V. and Santhanam P.

Page No: 61-69

Abstract: Nutrients level of Worm EELs were estimated during one-year period in the species of M. platyrhynchus. Concentration of protein, lipid and carbohydrate was estimated. The higher concentration of protein 22.16%, lipid 1.66% and carbohydrate 1.68% found in male fish and moisture (79.55%) content predominantly occurred in female Worm EEL. Monthly variation of protein level between 19.08-24.75%, lipid 1.29-2.19%, carbohydrate 0.09-0.50% and moisture content 72.50-79.55% was estimated respectively.

Correlation coefficient value of p<0.05 was significantly observed. During the study period, higher level of proximate composition was estimated in male fish. Seasonal variations of nutrient level were observed. However, the higher concentration of proximate composition was found during monsoon season and lesser nutrient in summer season due to the availability of the food.

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Mapping of quantitative trait loci (QTL) for grain length in basmati rice (Oryza sativa L.) using Basmati-370 x Pusa Basmati 1 RIL population

Bhardwaj Rakhi, Salgotra R.K. and Bhat Javaid Akhter

Page No: 70-85

Abstract:Grain shape of basmati rice is an important quality trait determining consumer acceptance. In the present study, the mapping population comprised of 140 F7 recombinant inbred lines (RILs). Basmati 370 and Pusa Basmati 1 cross were used for identification and molecular mapping of simple sequence repeat (SSR) markers linked to grain length in basmati rice. The phenotypic data was recorded for various quality and yield contributing traits and for genotyping, 70 polymorphic SSR markers were used. Using both phenotypic and genotypic data, two QTLs linked to grain length were detected on chromosome 4 and 6 corresponding to marker intervals RM13838- RM3698 and RM13840- RM530 respectively. Earlier studies also reported the QTL on chromosome 4, however QTL on chromosome 6 has also been reported.

The QTL was identified on the chromosome number 4 with LOD score of 3.23 whereas the other QTL located on the chromosome number 6 with a LOD score of 2.7. Furthermore, the two identified QTLs were designated as qGRL-4.1 and qGRL-6.1 which can be further be used in rice breeding programme to enhance the grain length particularly in local basmati rice where grain length is very short. Moreover, these QTLs can also be used for the development of good genetic stock to be used in marker assisted backcross breeding. In the study, the identified major QTLs affecting basmati grain length can be used further for fine mapping and validation of specific genes to develop gene based perfect markers for use in rice breeding.

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Bioprospecting of indigenous Bacillus thuringinsis GBI-3 for extracellular biosynthesis of gold nanoparticles characterization and optimization

Thakur Rajni Kant and Shirkot Poonam

Page No: 86-96

Abstract:Nanoparticle research is currently an area of intense scientific interest as this plays an important role in a variety of potential applications in fields of biomedicine, optical, electronics, with newer potential roles in environmental sector for water remediation and biosensor sectors. Physical and chemical methods of nanoparticles synthesis are too expensive and toxic with requirements of high temperature and pressure condition. In contrast to chemical and physical methods, microbial processes for synthesizing nanomaterials can be achieved in aqueous phase under gentle and environmentally benign conditions.

Moreover, use of microorganisms for synthesis of functional nanoparticles is eco-friendly, comparatively inexpensive and faster. The ability of microorganisms to change oxidation state of metals and their microbial processes has opened up new opportunity to explore novel applications such as biosynthesis of metal nanomaterials. This approach has become attractive focus in current green biotechnology towards sustainable development. In present study an indigenous bacterial strain GBI-3 has been isolated from a local gold mine Khaltunala. It was characterized morphological, biochemically and also by using 16S rrna gene technology and was identified as Bacillus thuringensis. The phylogeny of Bacillus thuringensis was determined using various bioinformatics tools viz BLASTn, Cluster W. To achieve maximum gold nanoparticles synthesis various parameters such as pH, incubation temperature, incubation period, Wavelength were optimized. The incubation temperature of 37oC, incubation time of 36 hrs, 560 nm wavelength and pH of 6.8 was found to be optimized for this purpose.

GBI-3 bacterial strain has been explored towards its capability of gold nanoparticles synthesis. Stable, monodispersed, cubical gold nanoparticles (GNPs) formation with 18 - 53 nm dimensions were synthesized successfully under invitro condition upon exposure of gold chloride trihydrates HAuCl4 solution to supernatant of Bacillus thuringensis strain GBI-3. Gold nanoparticles synthesized have been characterized by Transmission electron microscope and Fourier transform infrared spectroscopy (FTIR). Successful synthesis of stable, cubical and monodispersed nanoparticles in the size range of 18-53 nm using this efficient Bacillus thuringensis GBI-3 has been reported in this study leading to the development of an easy bioprocess for synthesis of GNPs of desired size and shape. The study demonstrates that there is a possible protein present in the isolated strain GBI-3 which reduces Au3+ to Au0 through an electron shuttling mechanism leading to the synthesis of nearly monodispersed GNPs. This green route of biosynthesis of GNPs is a simple, economically viable and an eco-friendly process.

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Plant Defense against Pathogens: The Role of Salicylic Acid

Kumar Vinod and AlMomin Sabah

Page No: 97-103

Abstract:Increasing demand for the global food production in the agricultural sector is a crucial driving force for the development of new disease-management methods that are not only effective against known pathogens, but also to the ones that will evolve. Plants possess specialized structures, chemicals and sophisticated mechanisms to defend themselves from pathogens. Understanding these defense mechanisms and pathways are critical for developing innovative approaches to protect crop plants from diseases as pathogens are continually evolving intricate means of breaching plant defenses. Plant defense pathways involve a number of signaling compounds that regulate the production of defense-related chemicals. These pathways are strongly connected with salicylic acid (SA), ethylene (ET), jasmonic acid (JA) and abscisic acid (ABA). The compound that is in the center of interest in this review article is salicylic acid due to its involvement in a variety of functions including biotic and abiotic stress management in plants.

The adverse effects of overuse of pesticides have led to the development and adoption of genetically engineered crops including those expressing genes involved in SA mediated defense pathways for enhanced defense capabilities, higher yields under biotic stress with reduced use of harmful pesticides. This review article focuses on latest developments in the plant-pathogen interaction and in particular, on the functional role played by salicylic acid in plant defense. A better understanding of plant defense mechanisms will enable scientists to develop more efficient methods of protecting plants from pathogens for sustainable agriculture.

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