SONICATION AND VACUUM INFILTRATION ENHANCED AGROBACTERIUM RHIZOGENES MEDIATED TRANSFORMATION IN SOYBEAN

AbstractObjective: The present study involved the formulation of protocol of Agrobacterium rhizogenes-mediated transformation for the detailed study of isoflavones metabolism in soybean.Methods: Cotyledons were separated from 4-day-old soybean seedlings and infected with three different A. rhizogenes strains under various time durations of sonication, vacuum infiltration and co-cultivated on MS medium supplemented with various concentrations of acetosyringone. The induced hairy roots were established as a culture with the selection agent hygromycin B. Transgenes integrated in hairy roots were analysed at molecular level by PCR assay.Results: A. rhizogenes strain R1000 harbouring pCAMBIA1301 resulted in better transformation efficiency when compared with other strains. The optimum duration of sonication (2 min) and vacuum infiltration (2 min) enhanced the transformation efficiency up to 76.47 %.  PCR analyses revealed the integration of transgene in hairy roots lines.Conclusion:  From the present study, we could conclude that, sonication and vacuum infiltration techniques could be employed to produce genotype independent transgenic soybean hairy root lines and which could be used to study for the improved production of potent anti-cancer compounds, isoflavones in soybean.Keywords: Agrobacterium, isoflavones, soybean, sonication, vacuum infiltratio

Analyzing The Metagenomics of Bacterial Diversity in The Backwater Mangrove Regions of Pichavaram and Parangipettai, Tamil Nadu, India, Reveals Insights into Sediment Microbial Communities

This metagenomic study delves into the microbial diversity and functional potential of sediment samples from the Pichavaram and Parangipettai mangrove ecosystems. The dataset, initially comprising millions of base pairs and sequences, undergoes meticulous quality control, revealing refined characteristics post-artificial duplicate read identification. Taxonomic analyses unveil a rich microbial landscape, dominated by bacteria, with intriguing presence of Eukaryota, Archaea, and unclassified sequences. The intricate community structures, highlighted through phylogenetic distributions, showcase the diversity and unexpected taxa, providing a nuanced understanding of microbial dynamics. Functional gene exploration uncovers key enzymes driving biosynthesis of secondary metabolites and metabolic pathways, shedding light on the microbial community's ecological roles and biotechnological potential. This comprehensive metagenomic analysis serves as a foundation for further ecological assessments, taxonomic refinements, and investigations into the adaptive strategies of these microbial communities in mangrove sediments

Phytochemical Analysis (LC-MS) of Azadirachta Indica Ethanolic Extract, Antioxidant, Anticancer, in Vitro Wound Healing Activity, and Immunomodulatory Effects of Azadirachta Indica Ethanolic Extract

This study presents a comprehensive investigation of the medicinal attributes of the ethanolic extract obtained from Azadirachta indica (A. indica) leaves collected from the Jamia Salafiya Pharmacy College campus in Malappuram District, Kerala, South India. The plant was meticulously identified by Dr. Samuel Thavamni B, a plant pharmacognosy specialist. The ethanolic extraction process was performed using the Soxhlet method. Phytochemical analysis was performed using liquid chromatography-LC-MS (liquid chromatography-mass and Mass spectroscopy). The in vitro antioxidant, MTT ((3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) assay, Apoptosis, and Immunomodulatory activity of ethanolic A. indica were evaluated. The extraction resulted in a semi-solid, dark-coloured extract with a yield of 8.67%. Phytochemical analysis using LC-MS identified key constituents such as octadecanoic acid, tetradecanoic acid, caryophyllene, 1,4-Eicosadiene, and triacontanoic acid, methyl ester, known for anti-inflammatory, antioxidant, and antimicrobial properties. In vitro antioxidant assays revealed a concentration-dependent inhibitory effect, with the A. indica ethanolic extract demonstrating substantial antioxidant capacity (77.81% at 100 µg), comparable to that of ascorbic acid (97.22%). However, the MTT assay indicated a concentration-dependent decrease in cell viability, emphasizing the need for caution owing to potential cytotoxic effects. Acridine orange/ethidium bromide staining provided insights into the induction of apoptosis in MCF-7 cells, highlighting the beneficial effects of the extract on cellular morphology. The scratch wound healing assay suggested the potential of the extract to promote fibroblast migration, implicating its role in enhancing wound healing. Immunomodulatory activity assessment revealed concentration-dependent modulation of immune responses, with the extract demonstrating a notable immunomodulatory effect of 77.81% at 100 µg. These findings shed light on the medicinal potential of A. indica, emphasizing the need for further research to elucidate the underlying mechanisms. The diverse chemical composition and multifaceted effects of the extract, from antioxidant and cytotoxic activities to immunomodulation and wound healing promotion, underscore its significance in pharmaceuticals and herbal remedies

meta-Topolin and β-cyclodextrin enhance multiple shoot and root production in black gram Vigna mungo (L.) Hepper

314-322The recalcitrant nature of black gram is the major constraint of in vitro regeneration and agrobacterium- mediated genetic transformation, to overcome this, a productive shoot regeneration protocol has been achieved in black gram cultivar T9 using 7-day old cotyledonary node explants excised from in vitro- raised black gram seedlings using meta-topolin. An aromatic cytokinin, meta-topolin along with BA (1.5+0.5 mg/L) in 0.50 strength MS medium with 1.5% (w/v) sucrose exhibited a maximum number of multiple shoots (32.0±0.37 shoots/explant) at the end of 6 weeks of culture. The shoots were elongated in (6.40±0.50 cm/shoot) in MS medium supplemented with GA3 (2.0 mg/L). A maximum number of roots (9.60±0.50/shoot) and root length (11.20±0.73 cm/shoot) were obtained in combination with β-cyclodextrin (a cyclic oligosaccharide; 1.5 mg/L) and IBA (1.5 mg/L). The rooted plantlets were hardened and acclimatized with least mortality rate of 2% in pot mixture consisting red soil:sand:farm yard manure (FYM) (2:1:1) and grown in green house with 85% relative humidity. Ploidy levels were analyzed using flow cytometry which confessed the chromosomal stability in invitro raised plants similar to parent plants. This protocol may be useful for producing transgenic black gram with desirable agro-traits in Indian cultivars

Influence of Dicyclohexylamlne Nitrite in Epoxy Primer

Epoxy resins have many advantages over other binders in the fonnulation of primers to protect steel structures from various corrosive environments. Recent studies showed that inhibitor incorporated epoxy primers protect structures for longer duration than plain epoxy primers. In the present investigation, different concentration of dicyclohexylamine nitrite (DCHAN) (0.5-2%) were incorporated in epoxy primers and coated on steel surfaces. The perfonnance of the primers were evalauted by salts spray and electrochemical evalaution tests in 3% sodium cWoride solution. The perfonnance of the primer in presence the inhibitor showed that the inhibitor protected the steel surface for a particular period and thereafter the inhibitor efficiency was in decreasing trend. In this study, the ratios of the DCHAN was optimized in the epoxy resin based primer and the performance of the optimized primer was evaluated by accelerated and electrochemical techniques

Flow distribution in spherical header of a liquid metal-cooled fast breeder reactor

432-439Liquid sodium from primary pump of a typical Liquid Metal -cooled Fast Breeder Reactor (LMFBR) reaches a spherical header through a vertical pipe and flows to the inlet plenum of the reactor through two discharge pipes, which connect the header to the plenum. The header is provided with a central cone and flow directing baffles for the purpose of having a smooth propagation of sodium flow and hence a reduced pressure drop. 3D hydraulic analyses of sodium flow in the header for various design configurations have been carried out to investigate the effectiveness of flow baffles and to assess the pressure drop in the header. It has been found that a configuration of 3D baffle combined with a central cone reduces the pressure drop in the header to the desirable level (1.7 mlc).</span

Corrosion behavior of metals and alloys in marine-industrial environment

This work deals with atmospheric corrosion to assess the degrading effects of air pollutants on ferrous and non-ferrous metals and alloys, which are mostly used as engineering materials. An exposure study was conducted in the Tuticorin port area located on the east coast of South India, in the Gulf of Mannar with Sri Lanka to the southeast. Common engineering materials, namely mild steel, galvanized iron, Zn, Al, Cu and Cu–Zn alloys (Cu–27Zn, Cu–30Zn and Cu–37Zn), were used in the investigation. The site was chosen where the metals are exposed to marine and industrial atmospheres. Seasonal 1 to 12 month corrosion losses of these metals and alloys were determined by a weight loss method. The weight losses showed strong corrosion of mild steel, galvanized iron, Cu and Zn and minor effect on Al and Cu–Zn alloys. Linear regression analysis was conducted to study the mechanism of corrosion. The composition of corrosion products formed on the metal surfaces was identified by x-ray diffraction and Fourier transform infrared spectroscopy

Enhanced Biosynthesis of Withanolides by Elicitation and Precursor Feeding in Cell Suspension Culture of <i>Withania somnifera</i> (L.) Dunal in Shake-Flask Culture and Bioreactor

<div><p>The present study investigated the biosynthesis of major and minor withanolides of <i>Withania somnifera</i> in cell suspension culture using shake-flask culture and bioreactor by exploiting elicitation and precursor feeding strategies. Elicitors like cadmium chloride, aluminium chloride and chitosan, precursors such as cholesterol, mevalonic acid and squalene were examined. Maximum total withanolides detected [withanolide A (7606.75 mg), withanolide B (4826.05 mg), withaferin A (3732.81 mg), withanone (6538.65 mg), 12 deoxy withanstramonolide (3176.63 mg), withanoside IV (2623.21 mg) and withanoside V (2861.18 mg)] were achieved in the combined treatment of chitosan (100 mg/l) and squalene (6 mM) along with 1 mg/l picloram, 0.5 mg/l KN, 200 mg/l L-glutamine and 5% sucrose in culture at 4 h and 48 h exposure times respectively on 28^th day of culture in bioreactor. We obtained higher concentrations of total withanolides in shake-flask culture (2.13-fold) as well as bioreactor (1.66-fold) when compared to control treatments. This optimized protocol can be utilized for commercial level production of withanolides from suspension culture using industrial bioreactors in a short culture period.</p></div