Research Journal of Biotechnology

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Research Journal of Biotechnology





Cultivar specific response of callus induction and plant regeneration from mature embryos in different elite Indian wheat

Padaria Jasdeep, Tarafdar Avijit, Sharma Varsha, Vishwakarma Harinder, Singh G.P. and Singh Sanjay

Page No: 1-8

Abstract: To develop a successful cultivar-independent in vitro plant regeneration protocol in wheat, mature embryos of nine Indian elite wheat (Triticum aestivum) cultivars were taken to establish a reliable effective, reproducible callus culture and plant regeneration procedure. Two different auxins, naphtalenacetic acid (NAA) and dichlorophenoxyacetic acid (2,4-D) and two different cytokinins, 6-Benzylaminopurine (BAP) and kinetin (Kn) were assessed in several combinations for their effect on callus induction and plant regeneration from mature embryos in nine elite Indian wheat cultivars. The cultivar-dependent substantial variances were noted in per cent of regeneration response. The callus induction was assessed in terms of size of callus induced in seven days of initiation. Induction of callus did happen in all cultivars on all media combinations tried, though there was difference in size of callus induced in different cultivars cultured with different combination of growth regulators.

However, the induced callus, irrespective of cultivar when nurtured on MS media supplemented with 2,4-D (4.0 mg/L) and NAA (1.0 mg/L or 2.0 mg/L) showed prominent callus growth. There were significant differences among the nine cultivars in regeneration of shoots from embryonal callus on the various shooting media used. In spite of cultivar dependent response, the MS media supplemented with kn (2.0 mg/L), NAA (0.5 mg/L) and BAP (0.5 mg/L) was identified as the best media for shoot regeneration for some of the elite Indian cultivars.

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Detection and molecular diagnosis of pathogenic Escherichia coli in local hospitals

Aljohnai Nasser D., Almasaudi Saad B., Kabl Selah A., Al-Garni Saleh M., Abu-Zeid Mohamed, El Shafey Hatem M. and Ahmed Mohamed Morsi M.

Page No: 9-15

Abstract: Escherichia coli is an essential member of the healthy intestinal microflora of humans and different mammals. Pathogenic versions of E. coli pathotypes cause entire morbidity and mortality globally. E. coli is responsible for many outbreaks in advanced and developing countries and they every so often have deadly consequences. We used microbiological bacterium study such as gram stain, biochemical test and then used molecular diagnosed techniques the usage of ribosome gene sequence, additionally to detect toxin-producing genes in some E. coli bacterium.

Our result showed that forms of bacteria had been diagnosed and distinguished between E. coli, O157: H7 and non-E. coli, O157: H7 and the usage of molecular biology method inclusive of gene collection. Some toxin-producing microorganism were recognized. Also, interpretation of the use of molecular techniques on the gene sequencing may be very useful in figuring out organisms as well as toxicity genes.

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In vitro conservation and asymbiotic propagation of Dendrobium nobile Lindl.: A rare threatened orchid of Northeast India

Hemanta Laishram, Thokchom Rocky, Sharma Sachin, Hijam Lakshmi and Devi M. Preema

Page No: 16-20

Abstract: An efficient in-vitro propagation protocol was developed for Dendrobium nobile, an important threatened medicinal epiphytic orchid species of Northeast India using immature green seed pods. In the present investigation, full strength Murashige and Skoog medium supplemented with different concentrations and combinations of either α-naphthalene acetic acid (NAA) or 6-benzyladenine (BA) were used for seed germination and development of the seedlings. MS medium supplemented with 2.0 mg/l NAA was found the best for early seed germination (11.53 weeks), protocorm formation (14.20 weeks) as well as for seedling development (21.27 weeks) from the day of inoculation. MS basal medium in combination with equal amount of NAA and BA (1.0 mg/l each) produced the tallest shoots per culture (4.93 cm) and maximum number of leaves per shoot (7.33).

However, maximum number of roots per shoot (4.53) was observed in the MS basal medium supplemented with 1 mg/l NAA and 2.0 mg/l BA. The well developed in vitro rooted plantlets were hardened successfully in the potting mixture containing only vermiculite (91.25 %) followed by vermiculite + soilrite (91.25%) at 1:1 ratio.

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Biogenic Synthesis and Characterization of Gold Nanoparticles using Aqueous Leaf Extract of Scoparia dulcis L. and Assessment of their Antimicrobial property

Radhika P.

Page No: 21-31

Abstract: Biosynthesis of gold nanoparticles using aqueous leaf extract of Scoparia dulcis as a simple, green and economical raw material has been investigated. Gold nanoparticles synthesis was carried out using leaf extract of Scoparia dulcis and aqueous solution of 1 mM HAuCl4.3H2O at room temperature. Several factors which influenced the nanoparticles formation are pH, concentrations of extract and salt, temperature, reaction time and methods of extract preparation. The stability studies of gold nanoparticles reveal that they are more stable in phosphate buffer and with PVP. From the UV-Visible spectroscopic studies, the characteristic Surface plasmon resonance peak for 1:6 ratio is observed at 540 nm.

Further characterization was done with X-ray Diffraction (XRD), Fourier Transform Infrared (FTIR) Spectroscopy, Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM) and Dynamic Light Scattering (DLS) studies revealing that the spectra were characteristic for gold, crystalline, spherical and size ranging from 5-22 nm. In addition, the biosynthesized nanoparticles exhibited significant antimicrobial activity towards most of the tested bacterial and fungal cultures.

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In vitro mass multiplication and molecular validation of Ocimum gratissimum using DNA based markers

Chaudhary Priyanka and Sharma Vikas

Page No: 32-41

Abstract: The aim of the present investigation was to set up a proficient mass propagation system via nodal explants of Ocimum gratissimum; therapeutically imperative herbaceous plant belongs to Lamiaceae family. Surface sterilized nodal explants were inoculated on MS medium fortified with BAP and Kn (0.5, 1.0, 1.5, 2, 2.5,3 mg/l) alone as well as in combinations with IAA, NAA and IBA (0.25, 0.5, 1, 1.5, 2.0mg/l). Highest shoot proliferation was observed in MS medium containing 1mg/l BAP and 0.5mg/l IAA having 37.60 + 0.4 maximum shoot number and maximum shoot length of 9.08 + 0.03cm. For root induction, in vitro regenerated shoots were excised aseptically and then implanted on half strength MS medium fortified with various concentrations of IAA, NAA and IBA (0.25, 0.5, 1.0, 1.5, 2.0mg/l). MS medium fortified with 0.25 mg/l IAA proved to be most effective for the induction of healthy roots. Well-developed rooted plantlets were transferred to plastic pots containing a mixture of soil: sand: manure (1:1:1) and then plantlets were subsequently hardened, acclimatized and then successfully established in original habitat with 90% survival rate.

Molecular validation was carried out to check genetic stability of in vitro raised clones using RAPD primers. No genetic alterations were observed when compared in term of number of bands and their respective sizes with mother plant and samples of subsequent stages of the protocol thus ascertaining the true nature of the in vitro raised plants. Present study holds tremendous application in large scale production and successful establishment of genetically stable plants of this valuable herb before release for commercial purposes.

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The Effect of Cinnamon Bark on the Expression of Quorum Sensing System and Virulence Genes in Pseudomonas aeruginosa Strain PAO1

Batoul Kavyani, Mohammad Reza Arabestani, Amir Sasan Mozaffari Nejad, Shirin Moradkhani, Farideh Kamarehei, Mohammad Taheri and Mohammad Yousef Alikhani

Page No: 42-48

Abstract: Antibiotic resistance is widespread rapidly between pathogenic bacteria which causes main problems in clinical treatments. Since antimicrobial activities of plants have been proven, they could be suitable choice for antimicrobial agents. Also, recently it has been determined that one of the attractive targets for inhibiting the pathogenicity of bacteria is Quorum sensing (QS) system.

In this study, we investigated the ability of alcoholic and aqueous cinnamon bark extract on the expression of lasI/lasR QS system, elastase and exotoxin A genes in Pseudomonas aeruginosa strain PAO1 by Real-Time qPCR method. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were measured. Then, in order to investigate the gene expression of elastase and exotoxin A, quantitative Real-Time PCR was performed at sub-MBC concentrations. According to obtained results, aqueous extracts of cinnamon have strongest inhibition activity on the gene expression against P. aeruginosa PAO1 rather than the alcoholic extract. The results showed that aqueous extracts of cinnamon at the concentration of 0.125 mg/ml decreased the expression of lasB, toxA and lasI genes. The obtained results indicated that cinnamon extract can inhibit LasI/LasR QS system and also subsequent products such as elastase and exotoxin A.

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Study of the cell cycle length in Allium species

Kumar Sanjay

Page No: 49-53

Abstract:Aliium species (A. chinense, A. tuberosum, A. hookeri, A. ascalonicum and A. sativum) were collected from different parts of Nagaland. Mitotic cell cycle length of each species was studied and it was found that none of the species showed comparable cell cycle duration existing in Allium cepa.

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In silico and denaturing gradient gel electrophoresis- based evaluation of 16s rdna primers for metagenomic studies

Pasha Malik Ahmed, More Sachin A. and Krishnaraj P.U.

Page No: 54-60

Abstract: Microbial community structure and their composition in environmental sample are commonly analysed by amplification of 16S rDNA followed by fingerprinting or sequencing. The success of these studies is largely dependent on the primer pair used in analysis. Commonly used five pairs of 16S rDNA primers were analysed in silico and in DGGE in this study. E1052 | E1193 primer pair showed better coverage in silico using SILVA database but failed to produce similar results in DGGE. PRBA338 | PRUN518 produced good coverage in silico using RDP database and good diversity in DGGE analysis. Hence, critical analysis of primer is essential for metagenomic studies.

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Effects of Culture Medium on Growth and Glucosinolate Accumulation in the Hairy Root Cultures of Watercress (Nasturtium officinale)

Park Chang Ha, Kim Nam Su, Yeo Hyeon Ji, Bong Sun Ju, Park Jong Seok, Park Nam Il and Park Sang Un

Page No: 61-66

Abstract: Watercress (Nasturtium officinale) is considered to be one of the most promising plant sources of a variety of glucosinolate compounds. The present study investigated the effect of various types of media on growth and glucosinolate production in the hairy root cultures of watercress. Hairy root cultures are generated using watercress explants infected with Agrobacterium rhizogenes strain R1000. We used six types of media (B5, 1/2 B5, MS, 1/2 MS, SH and 1/2 SH media) in the establishment of hairy root cultures and quantified variations in the production of four glucosinolates (4-hydroxyglucobrassicin, glucobrassicin, 4-methoxyglucobrassicin and gluconasturtiin).

Of the six media conditions, MS basal medium increased the accumulation of glucosinolates wherein the contents of glucobrassicin, 4-methoxyglucobrassicin and 4-hydroxyglucobrassicin were 25, 44.88 and 217.33 times higher respectively than those in 1/2 SH medium. Moreover, gluconasturtiin content was 10.55 times higher than that in 1/2 B5 medium. Conversely, SH and 1/2 SH media were the most suitable for hairy root growth. These results suggest an alternative approach for glucosinolate production in watercress hairy root cultures.

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A reliable and efficient method to obtain high quality RNA from Centella asiatica (L.) Urb. without using liquid nitrogen for molecular analysis

Borthakur Ananya, Phukon Munmi, Deka Diganta, Modi Mahendra Kumar and Sen Priyabrata

Page No: 67-70

Abstract: Centella asiatica is a multipurpose medicinal plant which has been used traditionally in several systems of Asian medicine. The important role of C. asiatica in medicinal applications is attributed to its biologically active components of triterpene saponins. The isolation of good quality RNA is a pre-requisite for understanding the downstream molecular mechanisms leading to the synthesis of these secondary metabolites. The high cost of liquid nitrogen and commercially available kits is a major bottleneck for the developing countries. The present study describes a low cost, simple and reproducible RNA isolation method from leaves of C. asiatica without the use of liquid nitrogen or expensive RNA isolation kits.

The protocol produced a yield of 2.7-3.1 µg/µl of total RNA and recorded an A260/A280 ratio in the range of 1.8-2.0. The packaged cDNA library constructed from the isolated RNA showed an efficiency of 1.5 x 106 pfu/ml and the number of recombinant plaques comprised of 82%.

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Marine algae as a promising natural resource in treating Periodontitis: Current status and applications

Leena Sharan and Jannet Vennila

Page No: 71-82

Abstract:Periodontitis is a disease caused by microorganisms that affects the periodontium of the oral cavity which when left untreated leads to loss of tooth. Research reports have confirmed that Periodontitis has significant contribution in development of systemic diseases if unattended. Though there are adequate development in treatment, scientists and physicians are looking for new efficient natural components to combat the side effects.

Recently, there is huge thrust on Marine algae research to explore its medicinal properties. Sufficient literature has indicated that marine algae are rich source of natural compounds. The marine bromophenol has been widely studied for its antimicrobial activities. This review focuses on current and recent advancements in research that explores the possibility of using marine components including bromophenols as a potent natural substance to combat bacterial infections and various ailments. The significant application of marine bromophenols in treating the periodontal pathogens is also highlighted.

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Fuelling the future: Recent advancement in microbial Engineering for biofuel production – A Review

Aiswarya Rajan, Ranjitha J. and Vijayalakshmi S.

Page No: 83-91

Abstract:Number of reviews and reports show the capacity of microorganisms to create biofuels and microorganisms such as algae, yeasts, bacteria, and fungi. The recent approaches in microbial biofuel production are well explored and recognized and a possibility of micro algal cultivation strategies for direct energy conversion to produce biofuels has been recommended. For example, biofilm cultivation of microalgae or cyano-bacteria could be the new platform of biomass production pathways that ultimately explored for biofuel processing pathway.

The recent approaches in microbial biofuel production are well explored and recognized and a possibility of micro algal cultivation strategies for direct energy conversion to produce biofuels has been recommended. For example, biofilm cultivation of microalgae or cyano-bacteria could be the new platform of biomass production pathways that ultimately explored for biofuel processing pathway. Commonly alcohols are widely used as biofuels. Ethanol and methanol are the most commonly used fuels in internal combustion engines.

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Nanoparticles: their synthesis and their applications

Thakur Rajni Kant and Shirkot Poonam

Page No: 99-102

Abstract:Nanotechnology has become one of the most promising technologies applied in all areas of science. Metal nanoparticles produced by nanotechnology have received global attention due to their extensive applications in the biomedical and physiochemical fields. Recently, synthesizing metal nanoparticles using microorganisms and plants has been extensively studied and has been recognized as a green and efficient way for further exploiting microorganisms as convenient nanofactories. Here, we explore and detail the potential uses of various biological sources for nanoparticle synthesis and the application of those nanoparticles. Nanobiology is a new discipline that integrates fundamental research in biology, physics, engineering and the biomedical sciences.

The use of nanoparticles in the biomedical sciences opens the doors to the delivery of new, more potent, effective and better targeted agents to the cell, enabling the development of new strategies for improving human health. The peculiar nature and the novelty of nanoparticles’ chemistry demand the proper characterization of the nanoparticles functional building blocks and the common patterns relating chemically diverse moieties. Nanotechnology could produce a revolutionary wave of innovation in society. The form that such a revolution might take will depend upon many things but certainly upon the context, content and purposes of research projects.

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