Research Journal of Biotechnology

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The Antidiabetic Activity of Brown Seaweed Sargassum confusum Polysaccharide Hydrolysates in Insulin Resistance HepG2 Cells in vitro

Chengfeng Yang, Yuqing Chen, Mingjun Chen, Ruibo Jia, Bin Liu and Chao Zhao

The purpose of this study was to investigate the antidiabetic activity of Sargassum confusum C. Agardh polysaccharide hydrolysates (SCO). In order to produce the highest yield of soluble polysaccharide hydrolysates, response surface methodology and Box-Behnken design were employed to optimize the temperature, time and pH on hydrolysis reaction. Effects of SCO on α-glucosidase activity and glucose metabolism in insulin-resistance HepG2 cells were evaluated in vitro. The yield of SCO was 5.9±0.3% which gave the optimal conditions for enzymatic hydrolysis at pH6.2 and 51°C for 3.9h. SCO showed strong inhibitory activity against α-glucosidase with an IC50 value of 9.9mg/mL. SCO could improve the glucose consumption and may ameliorate the insulin resistance in insulin-resistance HepG2 cells. Our results demonstrate that SCO exerts remarkable antidiabetic effect.

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Comparative Secretome Prediction and Analysis of Two Phytophthora spp.

- Kumar Arvind, Rahul C.U., Merin Babu, Tony Grace and Rajesh M.K.

Phytophthora spp. are widely distributed pathogens causing some of the most devastating diseases in plants. They accomplish parasitic colonization in hosts by virtue of an array of signaling proteins. Secretome analysis is one of the methods to attain molecular insights into species pathogenicity. In this study, we have analyzed the proteome of two species viz., P. ramorum and P. sojae and predicted extracellular secretory proteins particularly involved in host-pathogen interactions. The potential Phytophthora spp. secretome comprising of both the classical and non-classical secretory proteins was predicted with the aid of a stringent computational pipeline. Out of the 15,743 P. ramorum and 19,027 P. sojae proteins which were computationally analyzed in the current study, 1396 (8.86%) and 1666 (8.75%) proteins were categorized as classical secretory proteins (CSPs) while a total of 71 (0.45%) and 96 (0.50%) proteins were categorized as non-classical secretory proteins (NCSPs) in P. ramorum and P. sojae respectively. In addition, 235 and 399 effector proteins were also predicted from P. ramorum and P. sojae respectively. The functional annotation of the effector proteins revealed the occurrence of SSPs (small specific proteins), virulence and avirulence factors which could prove to be future target to control the pathogenicity and to decipher its role in host-parasite interactions.

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In vitro clonal propagation of Cyperus scariosus R. Br; An essential oil yielding medicinal plant

Kure S.R., Tungenwar A.G., Ekambe P.S. and Surwase B.S.

Cyperus scariosus R. Br. of family Cyperaceae is an essential oil yielding medicinal plant used for gonorrhea, syphilis, diarrhea, general weakness, intestinal disorders, fever and cholera. It is also being used in aroma therapy, cosmetics and perfumery. It shows various pharmacological activities like antidiarrheal, hepatoprotective, antimutagenic, antioxidant, anti-inflammatory, anti-arthritic and analgesic activity. High frequency in vitro clonal propagation protocol was standardized from the rhizome explants of Cyprus scariosus; an important oil yielding plant. Maximum shoot multiplication was obtained on MS medium containing 6 mg/l 6-benzyl amino purine alone. The number of shoots per explant were 15 with 90% response and 3.77±0.17 cm mean shoot length. The elongated healthy shoots were subcultured for successful root development on MS + 1.5 mg/l IBA. The number of roots per shoot were 4. The in vitro raised plantlets were hardened in green house and progressively shifted to natural conditions with 75% survival.

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Spatial distribution and correlation analysis of soil nutrients in tea gardens of Wuyi Mountain area

Hong Yongcong and Xin Wei

Although soil nutrients have direct impacts on the yield and quality of tea, the spatial distribution and correlation analysis of soil nutrients in tea gardens of Wuyi Mountain area were still in their infancy. Our results showed that the organic matter, available potassium and available magnesium in the tea garden soil of Wuyi Mountain area increased with the altitude. But the total nitrogen and available phosphorus were not correlated with the altitude. The organic matter, the available potassium and the available magnesium of tea garden soil with an altitude of 300-400 m, and the organic matter, the total nitrogen, the available phosphorus and the available potassium in the tea soil with a slope of 10o were higher than the others. And the available magnesium concentration in the tea gardens with a slope of 0o was the higher. The soil organic matter was correlated with total nitrogen, available phosphorus and available potassium. Although the available potassium and the available magnesium in some of the tea gardens were relative low, the organic matter and the total nitrogen were relatively rich. And the available phosphorus was at an appropriate level. Herein, the overall fertility level of the tea gardens of Wuyi Mountain area was suitable for tea production. Our investigation on the spatial distribution and correlation analysis of soil nutrients would certainly be helpful for the improvement for the yield and quality of Wuyi Mountain tea.

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Genetic diversity of Ascochyta rabiei causing blight of chickpea in India

Baite M.S., Dubey S.C. and Upadhyay B.K.

Ascochyta blight of chickpea is caused by Ascochyta rabiei (Pass.) Labr. which can infect all above-ground parts of the plant. Since Ascochyta is known to be genetically quite diverse, an attempt was made to analyse the genetic diversity of 25 A. rabiei isolates from northern India using universal rice primers (URP) and simple sequence repeats (SSR) markers. For identification, the ITS regions of the pathogen isolates were analysed varying from 524-550 bp revealing that the populations of A. rabiei have high genetic similarity (98.5-100%) amongst themselves. The genetic diversity assessed by URP assembled the isolates in two major clusters of which the first one contained 22 isolates whereas; the second major cluster had only three isolates. The SSR markers also classified the isolates into two major clusters. The first major cluster contained 18 isolates whereas the second major cluster had seven isolates. However, the groups generated by the molecular markers did not correspond to the geographic origin of the isolates barring few cases. The molecular markers showed high polymorphism and are valuable for analysis of the genetic diversity by revealing high genetic similarity amongst the A. rabiei isolates.

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Antioxidant activity of characterized silver nanoparticles synthesized using flower extracts of Chrysanthemum indicum

Rajeshkumar S.

Green synthesis of nanoparticles using biological materials exhibits low cost, simple, eco-friendly and less toxicity. In this study, silver nanoparticles were synthesized using flower extract of Chrysanthemum indicum. Flower extract treated with silver nitrate produced colour change within 10 minutes from yellow to yellowish brown indicating formation of silver nanoparticles. UV-Vis spectrum displayed a characteristic peak at 410 nm for synthesized silver nanoparticles. XRD and EDX confirm particles are crystalline in nature. Morphology of silver nanoparticles observed by Scanning Electron Microscope revealed spherical as well as unidentified shape of nanoparticles with aggregation. Fourier transform infra-red spectroscopy (FTIR) indicated the roles of different functional groups amine and alcohol in the synthesis mechanism. The antioxidant property of silver nanoparticles achieved using DPPH assay shows good free radicals scavenging properties.

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Characterization of Indian basmati rice (Oryza sativa L.) germplasm collected from NW Himalayas using agro-morphometric markers

Salgotra Romesh Kumar, Bhat Javaid Akhter, Gupta B.B. and Sharma Sandeep

The present study was undertaken for exploration, collection and morphological characterization of 141 basmati rice genotypes from different geographic regions of North Western Himalayas. Analysis of variance showed significant differences among the accessions for all traits. The narrow difference between PCV and GCV for the traits was studied indicating the low level of environment influence on their expression. High value of heritability and GA (%) together with GCV was observed for plant height, total tillers per plant and grain yield per plant revealing additive gene action for the expression of these traits. Grain yield per plant showed positive significant correlation with effective tillers per plant and 1000-grain weight. The UPGAMA cluster analysis based on ten agro-morphological traits divided all genotypes into six clusters (I, II, III, IV, V and VI) and in the same way PCA method also classified these genotypes into six groups (G1, G2, G3, G4, G5 and G6). The results from both the analyses are in well agreement with each other for grouping the genotypes on the basis of place of collection and geographic region and the corresponding groups in both cases are identical for the number and type of genotypes. The first four principal components contributed 68.80% of the variability. Results indicate considerable flow of basmati germplasm across different basmati growing regions of India. Both methods have proved to be effective in grouping basmati genotypes and helped in better understanding of the existing variability that will facilitate genotypic selection for crop improvement.

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Shoot formation efficiency of mature carob trees (Ceratonia siliqua L.) based on plant growth regulators pre-culture

Aguinaz Hayat, Qaddoury Ahmed and Anjarne Mohamed

An efficient protocol for in vitro propagation of mature carob tree (Ceratonia siliqua L.) was developed using nodal segments. It involves shoot initiation including a pre-culture period, multiple shoot formation and rooting. Half strength Murashigue and Skoog (MS/2) medium supplemented with various auxin and cytokinin used at different concentrations was tested for their bud initiation and shoot multiplication efficiency while different auxins were used to induce rooting of elongated shoots. Obtained results showed high percentage (>80%) of well-established and healthy cultures with shoot regeneration capacity that varied depending of BAP (6-Benzylaminopurine) and IBA (Indole acetic acid) concentrations. Hormonal pre-treatments of explants, whatever may be its duration and plant growth regulators, considerably increased bud initiation. Pre-culture in 5 mg/L TDZ (Thidiazuron) for one week induced the highest bud initiation (87%), 4 times higher than in non-pre-cultured explants. The maximum shoot (16.35) and leave (21.80) formation with best shoot length (2.42 cm) were observed when 0.5 mg/L BAP was combined with 0.5 mg/L IBA. Elongated shoots rooted more frequently produced multiple roots (5.89) with the highest root length (6.83 cm) on MS/2 medium supplemented with 1 mg/L IAA (Indole-3-acetic acid). Acclimatization of plantlets was successful; 66% survived after transfer to ex vitro conditions.

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Production and Characterization of Cellulase-free Xylanase by Aspergillus flavus ARC-12 and its application in pre-bleaching of Ethanol-Soda Pulp of Eulaliopsis binata

Gautam Archana, Kumar Amit and Dutt Dharm

Pre-treatment of pulp with cellulase-free xylanases can minimize the consumption of bleaching chemicals. A. flavus ARC-12 produced xylanase (1699.50 IU/gds) using pearl millet stover as the carbon source under SSF while cellulase activity was not detected in the crude xylanase. Xylanase production was enhanced up to 2219.85 IU/gds by the addition of beef extract (1.2%, w/v) as nitrogen sources. Xylanase production by A. flavus ARC-12 was further improved up to 2539.54 IU/gds by supplementing 0.10% (w/v) Tween-60 as surfactant. Partially purified xylanase showed optimum activity at pH 6.0 and temperature 50 °C. Xylanase was stable at 50 °C for a longer time (180 min) while declined activity drastically at 55 and 60°C at incubation time of 60 min. Crude xylanase was utilized effectively in the pre-bleaching of ethanol-soda pulp of E. binata. At an enzyme dose of 10 IU/g, kappa number was decreased by 18.51% while pulp brightness and viscosity were improved by 4.2% (ISO) and 4.96% respectively.

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Alcohol Hangover relieving effect of Hovenia dulcis Thunb associated with Antioxidant Activities through Ultra-High-Pressure Extraction Process

Lee Hyeon Yong

The ultra-high-pressure extraction (HPE) process at 400 MPa for 1 hour greatly increased the contents of total phenolic acids in Hovenia dulcis Thunb extracts to 112.7 mg/g compared with 75.64 mg/g from a conventional hot water extraction (WE) at 100C for 24 h. The HPE extract showed a good ability of decreasing residual alcohol concentrations in the blood of the rats to 0.06% after 6 hr treatment while the alcohol concentrations in rats fed with ethanol only increased from 0.004% at the starting point to about 0.139% after 6 h with 0.168% of the highest concentration after 2 hr. It was also confirmed that the HPE more effectively enhances both alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) activities, not one of each, than the WE such as 38.5% vs 29.0% and 43.1% vs 38.4% for ADH and ALDH respectively. Moreover, the HPE showed better glutathione-S-transferase activities (GST) at various concentrations than the WE. Thus, the HPE should play an important role in lowering the aldehyde concentrations in the blood which could result in relieving an alcohol hangover. This work also confirmed a close relationship between high antioxidant activities and hangover relief since the HPE extract contained higher amounts of polyphenols and also showed better hangover relieving activities that the WE or the control. Therefore, it can tell that the HPE process is effective for processing hard shelled plants which could result in enhancing their biological activities including hangover relief activities.

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Transcriptomics: A successful approach to unravel the molecular mechanism of plant-pathogen interaction in post-genomic era

Chakraborty Nibedita and Basak Jolly

Plants are constantly interacting with a wide variety of potential pathogens within their environment. During compatible interaction, the pathogen manipulates the immune system of the plant leading to susceptibility whereas in incompatible interaction, the plant prevents invasion of the pathogen by inducing various defense mechanisms. Transcriptome profiling plays a vital role in deciphering the mechanisms behind altered gene expression during plant-pathogen interactions. Till date hundreds of successful transcriptome profiling has been carried out in the area of plant-pathogen interactions, establishing transcriptomics as a mature platform to unravel the molecular mechanism of such interactions. Transcriptomics is an assembly of several techniques and each of these techniques has its own applications, utilities, inherent advantages and limitations. This review discusses major transcriptome profiling techniques along with their applications, utilities, advantages and disadvantages under one roof which will immensely help researchers for considering the choice of the technique during transcriptome profiling of plant-pathogen research.

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