Research Journal of Biotechnology

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Micropropagation of an Important Medicinal Plant Aloe barbadensis Mill (Aloe vera L) for Field Plantation

Parmar Vilas R. and Jasrai Yogesh T.

A rapid micropropagation of Aloe barbadensis Mill was achieved from shoot tip explants cultured on MS medium supplemented with BAP (1 mg/l), NAA (0.5 mg/l) and citric acid (10 mg/l). After 3rd subculture, almost 10-15 multiple shoots were obtained within four weeks. However, proliferation in subsequent subcultures declined. The developed shoots also demonstrated root induction on medium containing 0.7 mg/l NAA.. The plantlets so generated were processed through hardening procedure for acclimatization and transfer to the soil. Through this procedure about 42 plantlets were generated from one explant over three subcultures (total 04 months). The in vitro raised plants were similar to the mother plant with general morphology.

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Decision Making Plasticity of Swarming Honeybees (Apis cerana cerana)

Huang Qiang, Zeng Zhi-Jiang*, Yan Wei-Yu and Huang Kang

Honeybee which is a very important and useful model insect of social behavior language and pheromone study has been widely studied around the world from many aspects. With the development of the colony, honeybees make many decisions to flourish the colony and breed it stronger. During the swarming season, honeybees will decide when to swarm, who joins the swarming, who stays in the colony and where the new home is. In this paper, we focused on whether workers can consciously decide to leave or to stay in the colony during the swarming, and if they can, what can be the underline impetus. The experiment was conducted with three colony Apis cerana cerana under natural swarming conditions. Genetic relatedness and subfamily composition were analyzed by using four Microsatellites. The results showed that subfamily distribution in swarms and the workers staying in the colony were significantly different from random decision making. Workers prefer to stay with their super sister virgin queens. Our data first indicates genetic relatedness may affect workers making non random decisions on whether to leave or to stay in the colony during swarming. We presume non random decision making set the evolutionary success for honeybees.

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Modulation of Glycolytic and Gluconeogenic Enzymes by Treatment with Hemidesmus Indicus R.Br. Ethanolic Root Extract in Alloxan Induced Diabetic Rats

Sowmia C.*, Kokilavani R. and Gurusamy K.

Diabetes mellitus is a metabolic disorder in which the body does not produce or properly use insulin. Hemidesmus indicus R.Br (Asclepiadaceae) is a traditional plant that is used for the conventional therapy of several diseases such as diabetes mellitus and for blood purification. Our aim in the present study is to investigate the effect of Hemidesmus indicus R. Br. root extract on glycolytic and gluconeogenic enzymes to demonstrate their possible therapeutic effects. The root extract of Hemidesmus indicus R. Br. (400 mg/kg) body weight was administered orally to control and alloxan induced diabetic rats for 30 days. Applied doses did not cause any acute toxicity or behavioral changes. The glycolytic and gluco-neogenic enzymes were estimated after administra-tion of the extracts. The glycolytic enzyme activities of hexokinase, aldolase, phospho-glucoisomerase and malate dehydrogenase in both liver and kidney showed minimum effect in toxic rats but toxic rats treated with the plant extract showed elevated effect and their values were near normal. The gluconeogenic enzyme activities of glucose – 6 – phosphatase and fructose 1,6 diphosphatase in both liver and kidney showed higher level in diabetic rats but diabetic treated rats with the plant extract showed lower level to the near normal values. Glibenclamide was used as a reference drug for the experiment.

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Effect of Growth Regulators on Micropropagation of Rauvolfia Serpentina (L.) Benth

Harisaranraj R., Suresh K. and Babu S. Saravana

Rauvolfia serpentina (L.) Benth. called Sarpagandha (the snake root) in Hindi, belongs to family Apocynaceae. This plant is listed in earliest Ayurvedic medicinal text the Charaka Samhita (c. 700 B.C.) and has been used since in olden days for the treatment of mental illness and insomnia. The roots of R. serpentina contain numerous alkaloids. Its indiscriminate use and poor method of conventional propagation have led this species to be included in the list of endangered plants. Thus a need arises to generate an efficient protocol for cloning of R. serpentina in order to regenerate propagules to replenish depleting forests and meeting the demand of commercial cultivation. The present study reveals the development of an efficient and simple process for large-scale clonal micropropagation of selected plants of R. serpentina and their successful field establishment. In vitro shoot multiplication from nodal explants of Rauvolfia serpentina was standardized by using MS (Murashige & Skoog) medium supplemented with a cytokinin (BA or Kin). Inclusion of NAA in the culture medium along with BA promoted a higher rate of shoot multiplication. The mean number of shoots per explant after 4 weeks of culture was maximum (5.21) on the MS medium supplemented with 2.0 mg l-1 BA and 0.25 mg l-1 NAA. The elongated shoots rooted within 7-8 days in ½ strength MS medium supplemented with 0.25 mg l-1 of NAA and 2% (w/v) sucrose. About 85% of the rooted plantlets survived acclimatization and transfer to the greenhouse. The present study paves a way for conservation and propagation of this important medicinal plant..

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Phytoremediation for Arsenic Contamination: Arsenate Reductase

Shipley Sheena, Nordin Andrew B., Connie G. Tang1 and Kim Sung-Kun1, 2*

Arsenic is a toxic pollutant present in the environment that causes serious health issues. Phytoremediation would be a solution to mitigate arsenic contamination in soil or water. Two forms of arsenic mainly exist in the environment, arsenite, [As(III)] and arsenate, [As(V)]. In plant mechanisms, arsenate reductase reduces arsenate to arsenite with the help of thioredoxins or glutaredoxins, which serve as electron donors. Subsequently, arsenite can be stored in the plant vacuole. For example, the Arabidopsis thaliana transgenic plant contains bacterial arsenate reductase and glutamylcysteine synthase, which enhance its ability to store two to three times more arsenic per gram of tissue than the non-transgenic plant. Further investigation of the mechanism of arsenic uptake in plants is needed to provide a foundation to develop better transgenic arsenic hyperaccumulators.

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Bioconversion of 3-cyanopyridine to Nicotinic Acid by a Thermotable Nitrilase

Nigam V. K.*, Agarwal A., Sharma M., Ghosh P.1 and Choudhury B.2

A thermostable nitrilase producing isolate identified as Bacillus sp. MTCC 7545 has been studied towards production of nicotinic acid from 3-cyanopyridine. The benzonitrile was observed as inducer for the enzyme production. Maximum enzyme production was observed at 24 h of incubation and the activity of nitrilase was found to be optimum at 650C. The enzyme was stable for ~6 h at 500C. Besides pyridine-3 carbonitrile, benzonitrile was also a preferred substrate for the enzyme.

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Phylogenetic Analysis of Lamellidens corrianus obtained from Konkan Region of Maharashtra by 28s rRNA and 18s rRNA sequences

Jadhav B. L. and Jamkhedkar Suruchi *

The freshwater pearl producing bivalve Lamellidens corrianus was collected from the various parts of Konkan region of Maharashtra and the phylogenetic studies were carried out using 28s rRNA and 18s rRNA gene sequences. The 28s rRNA gene sequence exhibited 99.2% similarity with freshwater bivalves. The 18s rRNA sequence when aligned with homologus region of Pinctada albina showed 88% - 89% similarity indicating phylogenetic nearness to marine pearl producing bivalve. The dendrogram genetrated by Clustal W for both 28 s rRNA and 18s rRNA indicated these populations are sub species of L. corrianus. The accession numbers of the sequences submitted to NCBI GenBank are EU394233, EU394234, EU394235, EU394236, EU394237 and EU394238.

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Cytotoxic Consequence of D2O-Induced Chemical Transformation of Thymine and Uracil

Salih Fadhil M. 1*, Al-Hanshi Maysa S. S. 1 and Pillay Avin E.2

Chemical transformation of thymine and uracil was observed in the presence of D2O. The products of the relevant reactions were confirmed spectrophotometrically. Pure samples were dissolved in solutions of the D2O, corresponding to concentrations of typically 10-2 mM. The deuterated solvent caused chemical transformation in the two chemical compounds tested and produced intense characteristic absorbance maxima between 200–400 nm. The biological effects of such transforma-tions were investigated using Bacillus pumilis. Bacterial cells were suspended in D2O at 0oC and 37oC. Samples were taken after different periods of contact and numbers of survivors were scored. Dramatic reduction in cellular viability was observed in the first few minutes. Cells kept in ice continued losing viability up to about 90% in 80 minutes. However, when cells were maintained at 37oC, they managed to recover fully in 60 minutes after a lag period of 10 minutes. Cells exposed to D2O at 0oC for 20 minutes and then maintained at 37oC started recovering at a rate similar to that seen with cells kept continuously at 37oC. Potentially lethal damage is thought to be induced by D2O and this type of damage can be repaired if injured cells were kept for some time in an environment free of growth supporting materials.

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Detection of Alpha - Amylase Producing Bacteria (Bacillus sp. 5250) from soil: Isolation and Optimization of Various Conditions of Growth

Chandramouli Meenakshi*, Kumar Narender, Ambekar Vikrant1, Suresh Karupothula2, Bhide Shobhana3 and Sabharwal Sushma4

Starch and maltose are the only carbon sources that have effectively induced the bacteria to produce amylase whereas other sugars such as glucose, sucrose, lactose were found to suppress the production of the same. Media containing yeast extract and peptone together with carbon source was found to be optimal for production of amylase by the bacteria. The study of the nitrogen source such as ammonium chloride, potassium nitrate on production of amylase showed varied response on the growth of bacteria. The pure culture of the bacteria has been deposited at the NCIM depository and strain number has been obtained. The characterization of the pure culture was done and the bacterium was found to be Bacillus sp. Also effect of antibiotics and drugs on the bacteria was assessed.

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Impact of Lead ion on the Stability of Lipid-protein Organization of Photosynthetic Organell

Panda Sunakar* and Panda Sumita

Lead in its various ionic and molecular forms causes serious environmental hazards. The lead ion (Pb2+) influences the stability of lipid-protein organisation of isolated chloroplasts. Although the metal ion induces thylakoid membrane lipid peroxidation, it prevents the loss of pigments (chlorophylls and carotenoids) and loss of protein. Such changes bring about a drastic modification of the structural organisation and photochemical reactions of photosynth-etic assembly. The cation induced enhancement in membrane lipid peroxidation is attributed to formation of cation – calmodulin complex which activates the enzyme phospholipase – A2 leading to deesterification of membrane lipids. The liberated polyunsaturated fatty acids are readily exposed to reactive oxygen species and get peroxidised. The cation induced retardation in pigment and protein loss is explained in terms of metal ion binding to lipid protein complex through carboxylic acid group of lipids and sulfhydryl group of protein. The metal binding is evident from changes in absorption characteristics of organell.

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IPNM Practices to study Agronomical Characteristics of Vertisol Soil

Menaria K.* and Tenguria R.

The present study deals with the influence of IPNM treatments to vertisol soil in wheat-soybean cropping sequence system. The results observed for agronomical properties like plant height, number of tillers, yield parameters like seed index, production of grain and straw, test weight and harvest index in vertisol soil indicated that these properties were significantly enhanced for the crop on the application of different IPNM treatments. The best results were observed with T6 treatment (25% NPK+FYM). The incorporation of different IPNM treatments claimed that IPNM treatments have significant potential to improve agricultural properties of soil.

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Protective Effect of Apigenin against the Genotoxic Damage induced by Estradiol-17 b

Siddique Yasir Hasan* and Afzal Mohammad

The use of traditional natural preparations from plant materials for the treatment of various diseases by the majority of people have led to assess the modulating action of plants extract/ products when associated with other substances. Estradiol-17b is a well known steroid hormone (estrogen) and has been reported to induce genotoxic damage and tumor formation in various experimental models both in vitro as well as in vivo. Estradiol-17b is also used in oral contraceptives formulation in combination with synthetic progestins. The prolonged users of oral contraceptives have been reported to develop various types of cancers. Natural plant products play an important role in reducing the genotoxic damage by the steroids. In this context the effect of apigenin was studied against the genotoxic damage induced by estradiol-17b. The treatment of apigenin at concentration of 5, 10, 15 and 20 mM was given along with 10 and 20 mM of estradiol-17b separately and respectively. The treatment of apigenin results in the reduction of the sister chromatid exchanges (SCEs), thereby suggesting a protective role of apigenin against the estadiol-17b genotoxic damage.

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Premna obtusifolia: Membrane Stabilizing Effect

Chitra M.

Ethanolic extract of Premna obtusifolia was evaluated for the free radical scavenging and membrane stabilizing properties.The parameters studied were superoxide dismutase, catalase, glutathione reductase, reduced glutathione, gluta-thione peroxidase, ascorbic acid, Na+K+ATPase, Mg2+ATPase and total ATPase activities. Results of biochemical studies of blood samples of CCl4 treated animals showed significant increase in the above levels of marker enzymes which reflect liver injury caused by CCl4 whereas samples from animals treated with ethanol extract of Premna obtusifolia showed significant decrease in the levels of markers indicating the protection of hepatic cells. The results revealed the ethanolic leaf extract followed by the ethanolic root extract of Premna obtusifolia could afford significant free radical scavenging and membrane stabilizing effect against CCl4 induced hepatocellular injury.

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Mini Review: Aspergillus niger and Heavy Metal Removal : A Perception

Vala Anjana K.

Heavy metal pollution has been a pressing problem. Application of biosorbents can be a promising alternative to combat the problem. In this review, Aspergillus niger as a biosorbent has been discussed. An outline of advantages of using A. niger, types of metals removed and percentage removal of different metals by A. niger has been presented. A comparison of A. niger with a few other bio materials has been made. Analytical techniques used and mechanisms of metal removal have also been briefly discussed.

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