Indian Journal of Biotechnology

Six New World Camelidae microsatellite primer pairs were used to investigate the genetic polymorphism in Jaisalmeri camel. Polymerase chain reactions were carried out for 30 unrelated camels of Jaisalmeri breed. The amplification products were resolved in 6% (denaturing) urea PAGE and stained with silver nitrate. All six microsatellite primer pairs were found polymorphic in Jaisalmeri camel. The number of alleles ranged from 2 to 5. The expected heterozygosity ranged from 0.32 to 0.651 and the polymorphic information content ranged from 0.268 to 0.588. The results indicated the utility of these microsatellite loci for studying genetic polymorphism in dromedary breeds.
Surfactants are surface active chemical compounds, which are extensively used in various industrial and household formulations. Now-a-days, linear alkylbenzene sulphonate (LAS) is the most important anionic surfactant in use. The biodegradability of LAS is the main reason for its acceptance as a major chemical in industrial applications. But the huge amount discharged upsets the efficient removal by biodegradation and its acute exposure pose harm to the environment. Studies on indigenous isolates capable of efficient LAS degradation were conducted and two strains of the genus Pseudomonas showing about 81% of LAS degradation were selected. The biochemical and molecular characterization of the isolates were done in order to identify them. The selected isolates were identified as P. nitroreducens (L9) (MTCC 10463) and P. aeruginosa (L12) (MTCC 10462). The sequences generated in the 16S rDNA analyses were deposited in the NCBI GenBank under the accession numbers HQ271083 (L9) and HQ271084 (L12). The role of plasmid in biodegradation was checked and it was found that genomic DNA and plasmid together code for the degradation capacity of the selected strains. Though the colony count was reduced at high LAS concentrations, selected isolates were able to withstand very high concentrations of LAS (12000 ppm). In the presence of an alternate carbon source, such as, dextrose, the isolate showed diauxic growth. The selected strains were found to be two promising candidates for the bioremediation of the anionic surfactant LAS.
Aspergillus ochraceus (NCIM-1146) has ability to decolorize various xenobiotic dyes. Biodegradation of dyes was demonstrated by their decolorisation in the culture medium. The extent of biodegradation was determined by monitoring the decrease in absorbance of each dye. Malachite green decolorisation activity is affected by various conditions such as composition of media, concentration of dye, amount of mycelia and agitation. The durability of decolorisation activity under optimum conditions was investigated in repeated batch mode. An increase in the amount of mycelia positively affected the durability of decolorisation activity. The decrease in dye decolorisation capability of mycelia occurred with increasing dye concentration in repeated batch mode. Spectrophotometric data revealed that the process involved in decolorisation is through microbial metabolism but not biosorption. This study showed that fungal mycelia (A. ochraceus) could effectively be used as an alternative to the traditional physico-chemical process.
– Screening of agro-industrial residues for cellulase production  
– Optimization of initial moisture content  
– Optimization of incubation temperature and initial medium pH  
Cellulase production studies were carried out using the fungal culture Trichoderma reesei NRRL 11460 using four different lignocellulosic residues (both raw and pre-treated) by solid-state fermentation. The effect of basic fermentation parameters on enzyme production was studied. Maximal cellulase production obtained was 154.58 U/gds when pre-treated sugarcane bagasse (PSCB) was used as substrate. The optimal conditions for cellulase production using PSCB were found to be initial moisture content - 66%, initial medium pH-7.0, incubation temperature -28°C, NH4NO3 at 0.075 M, and 0.005 M cellobiose. The optimal incubation time for production was 72 h. Results indicate the scope for further optimization of the production conditions to obtain higher cellulase titres using the strain under SSF.
Vanillin (4-hydroxy-3-methoxybenzaldehyde), a pleasant smelling aromatic compound, occurs naturally in vanilla beans. It is widely used as a flavouring additive for beverages and aromatic additive for candles, fragrances, and perfumes. It is also noted as nutraceutical. Efforts have been made to obtain aromatic compounds produced by means of biological process, which employ microorganisms. White rot fungus, Phanerochaete chrysosporium NCIM 1197 was used to transform lignocellulosic substrates like rice husk, groundnut shell, etc. Further, detection and quantification of vanillin were carried out by TLC and spectrophotometrically. The maximum vanillin production (up to 55 μg/mL) was observed within 72 h using groundnut shell as a substrate. The use of various carbon sources, viz., glucose and starch, along with groundnut shell, affected vanillin production. Supplementation of glucose along with groundnut shell showed more increase in vanillin production (37 μg/mL within 96 h) in comparison to the supplementation of starch (up to 21 μg/mL within 96 h).
LasB protease, also known as the elastase enzyme was purified from an environmental isolate of Pseudomonas aeruginosa MCCB 123. The enzyme production was optimized by response surface methodology (RSM) which yielded 24754.17 U/mL protease activity and 654.14 U/ml elastase activity. The yields were 1.83 fold and 2.39 fold higher for protease and elastase activities, respectively. Scale up study of enzyme production was carried in a 5 L bioreactor. Bioprocess technology for enzyme production was successfully developed. © 2018 National Institute of Science Communication and Information Resources (NISCAIR). All rights reserved.
Melatonin, a neurohormone, is formulated as a thermoreversible pluronic gel for nasal administration as an 'overlap dosage form' for chronobiological treatment of sleep disorders. Aqueous PF gels containing drug (0.5 mg & 1 mg/0.1 ml), PEG 400 and PEG 15,000 were prepared by cold method. Pluronic gels were evaluated for gelation and gel melting. Gelation temperature (T 1) decreased with pluronic concentration while gel melting temperature (T2) increased. Melatonin shifted gelation range to higher temperature while PEG narrowed the gel range. Flux of diffusion decreased with PF concentration. Drug flux decreased in higher drug strength gels due to more partitioning in micellar phase. Pluronic gel (20%w/w, 1 mg/0.1 ml) showed bimodal pattern with a desired second peak flux (0.248μg/min/cm2) at 300 min. Flux pattern changed invariably with PEG. Bioadhesion time and strength to sheep nasal mucosa were more for gels containing melatonin and PEG 400. Nasal gels produced fast onset of action and induced sleep within fifteen minutes. The low intensity and rounded α-EEG wave pattern was observed for sleep duration of 5 hrs. Good correlation was observed in sleep pattern and low intensity α-EEG. The results are encouraging- and nasal melatonin gels have potential in the treatment of circadian cycle sleep disorders.
Biotechnological production of 2,3-butanediol (BD), a value added chemical, from wastes and with excessive biomass is a promising and attractive alternative to traditional chemical synthesis. The production of BD by Klebsiella oxytoca NRRL-13-199 from deproteinated whey was studied in batch fermentations at pH 6.5. Lactose present in the deproteinated whey was found to be effectively utilized by K. oxytoca, producing BD 0.259 g/g of lactose utilized after an incubation period of 96 h. Moreover, addition of acetate at a concentration of 50 mM was found to increase the yield by 1.5-fold, resulting in BD 0.365 g/g lactose utilized. During the process, COD (chemical oxygen demand) reduction of 87% was observed in addition to 91% BOD (biological oxygen demand) reduction. Thus, the present process of utilization of whey by K. oxytoca clearly shows that it not only helps in the production of value added chemical BD from the waste but also helps in reducing the environmental problems faced with the disposal of untreated or unfermented whey directly into the river.
Production parameters of extracellular alkaline protease employing our laboratory isolate Thermoactinomyces thalpophilus PEE 14 under solid-state fermentation (SSF) were optimized. Wheat bran, the best substrate among the 15 substrates used for optimization, showed the highest activity (1620 PU/g). The physical and chemical parameters were also optimized. The maximum enzyme activity under optimum conditions was obtained with incubation period 72 h. incubation temperature 55°C. initial pH 10. inoculum level 20%. level of salt solution 2:10 and initial moisture level 80%. Increase of enzyme activity by 60% (2576 PU/g) was observed when compared with the unoptimized conditions.
The rapid identification of bacteria in biological specimens is important for selection of a suitable antimicrobial therapy. We have designed a rapid (<8 h) diagnostic system that uses universal PCR primers to amplify a variable region of bacterial 16S & 23S ribosomal DNA, followed by reverse hybridization of the products to a panel of oligonucleotides, spotted on nylon membrane macroarray slides. Culture dependent assays were used as reference methods in the development and evaluation of this diagnostic platform. Broad range PCR primers were selected from the published literature to amplify 16S & 23S rDNA segments. Species-specific probe sequences were designed based on sequence alignment of eight different bacteria. These species included Helicobacter pylori, Salmonella typhimurium, Shigella dysenteriae, Vibrio choleriae, Neisseria gonorrhea, N. meningitidis, Corynebacterium diphtheriae and Haemophilus influenzae. To verify specificity, five to six initial oligonucleotide probe sequences per bacterial species were tested by hybridization on nylon membrane macroarray glass slides using culture collection strains as templates. Finally, three oligonucleotide probes per bacterial species were selected based on hybridization results. This procedure was successful in discriminating a range of bacteria in pure cultures. Adding further oligonucleotides to the panel without significantly increasing the cost the accuracy, range, and discriminatory power of the assay can be extended. This method is versatile and makes it possible to detect a large number of bacterial species in a single assay and discriminate different bacterial genera of medical importance. Our results provide a proof of concept for the diagnostic use of macroarray technology based on broad-range ribosomal RNA gene amplification, followed by hybridization and specific detection of bacterial species.
Spirulina/Arthrospira is a species of cyanobacteria used in health foods, animal feed, food additives and fine chemicals. The present study conducted a comparison of the 16S rRNA and cpcBA-intergenic spacer (cpcBA-IGS) gene sequences in Spirulina/Arthrospira strains from culture collection of CCUBGA, IARI, New Delhi. All the strains of Spirulina used in this study had shown nearly 99% similarity amongst them. About fifty sequences (cpcBA-IGS) of Spirulina strains taken from NCBI with ten from the present strains of Spirulina, a neighbour-joing (NJ) tree was constructed with the help of MEGA5.0. The tree showed 99% similarity. All the sequences were put to Multiple Sequence Alignment with the help of T-Coffee (version 7.38) and BioEdit (version 7.38) software. Similarity studies undertaken based upon 16S rRNA and cpcBA-IGS genes sequence analysis indicated similarity coefficient of 0.84. S. platensis and Arthrospira sp. showed 100 percent similarity. Therefore, the current study supports some previous conclusions based on 16S rRNA gene and cpcBA-IGS sequences, which found that Arthrospira taxa are monophyletic. However, compared to 16S rRNA sequences, cpcBA-IGS sequences might be better suited to resolve close relationships and interspecies variability.
Assessment of genomic DNA contamination in mtDNA isolated by the conventional procedure and by cellulose acetate membrane based method by using D. melanostictus IgM gene specific primers: Lane 1, PCR product from genomic DNA showing positive result; lanes 2 & 3, PCR products of mtDNA from conventional source and cellulose membrane source, respectively showing negative results meaning both the mtDNA source are free of genomic DNA contamination; & lane 4, 100 bp DNA ladder. 
Mitochondria isolated by a non-conventional method of membrane filtration were used for mtDNA extraction. This technique allows trapping of mitochondria on cellulose acetate membrane. The procedure does not involve sophisticated instruments and can be performed out of laboratory conditions. The advantages of this procedure are discussed in this paper.
Allelic pattern of M. coucha based on RAPD marker DNA fingerprinting [lane M1-M10=M. coucha samples: M=100 bp ladder; RG 3 ,RG 4 & RG 7].  
Phylogenetic analysis of M. coucha based on similarity indices.  
Allelic pattern of M. coucha based on microsatellite marker DNA fingerprinting [lane M1-M10= M. coucha samples: M=100bp ladder, MH 30, MH 105, MH 133 &MH 174.  
Ten RAPD primers and nine microsatellite markers of Mastomys huberti were cross tested to establish molecular characters as well as identify microsatellite markers in the genome of Mastomys (Praomys) coucha. A total of 484 bands were scored against 10 isolates of M. coucha. The per cent polymorphism was 1.66 in individual isolates but this polymorphism was not statistically significant among the 10 isolates (Mas1 to 10). Mean difference, Mas6 vs. Mas10 = 0.005, Mas5 vs. Mas3 = 0.485 and Mas7 vs. Mas9 = 0.260 were calculated by Newman-Keuls multiple comparison test using one way ANOVA among Mastomys strains. The dendogram also revealed Single linkage euclidean distances among the 10 isolates (Mas3 vs. Mas1 = 1.3, Mas8 vs. Mas5 = 1.5 and Mas9 vs. Mas5 = 1.1) but the Single linkage euclidean distances were statistically not significant (P>0.05). Identifying RG-1, 3, 4, 7 and 8 RAPD markers were amplified in all the isolates with 1200 bp, 1200 and 200 bp; 400 bp; 1000 and 700 bp and 300 bp, respectively along with other polymorphic alleles. Out of 9 microsatellite markers, 5 (MH028 - 434 bp; MH30 - 249 bp; MH105 - 271 bp; MH133 - 338 bp and MH174 - 316 bp, respectively) were exactly amplified in the genome of M. coucha. RG-1, 3, 4, 7 and 8 RAPD primers and 5 microsatellite markers may assist in the identification of M. coucha.
Uremia is a major feature of chronic kidney diseases (CKD). Studies were conducted with a standard urease producing probiotic strain, Streptococcus thermophilus (MTCC 1938) to explore the effectiveness of probiotic organism against uremia. The therapeutic potential of the probiotic bacterium was tested against acetaminophen (APAP) induced uremic rats. Supplementation of S. thermophilus in food resulted in improved uremic profile compared to positive control (PC) rats (no probiotic supplementation in food) during 15 d experimental period. Much lower concentration of plasma urea (77%), plasma creatinine (80%), urinary protein (68%) and urine glucose (68%) was found in the earlier. Significant increment of plasma glutathione (GSH) level (58%) in probiotic treated group was also noticed. Moreover, during the feeding with probiotic strain at a dose of 1× 109 CFU/mL bacteria, reduction of enterobacteria in faeces was observed. Notable biochemical changes were reflected in histopathological examination of kidney sections. The uremic profile of the probiotic induced rats was very much comparable with the negative control (NC), even found better for some parametric values. The present study suggests that S. thermophilus could be used as a novel alternative natural therapy for uremia, a major syndrome of CKD.
Shigella species are non-sporulating, Gram negative, facultative anaerobes. IS (insertion sequence) elements are the major cause of the dynamics of Sf301 chromosome and due to IS-mediated DNA rearrangements and formation of pseudogenes, the Shigella spp. became highly specific human pathogens with variable epidemiological and pathological features. Nucleotide sequence analysis of S. flexneri 1a genomic DNA was performed through Big dye terminator chemistry using ABI 3730 48 capillary DNA analyzer. In total, 60 kb data in form of contigs were analyzed by homology search using various bioinformatics tools. IS elements were identified using nucleotide blast at NCBI as well as the Is finder. Eight different IS elements were identified, which were present in different copy number. A new IS element ISEhe3 was identified, which belonged to family IS3. ISEhe3 was found absent in S. flexneri 2a 301, but it was present in 2457T strain. Amongst the IS elements identified, IS2, IS3 and IS600 elements showed identity with SHI2 Shigella pathogenicity island. The presence of large numbers of IS-elements in the Shigella genomes is likely the major cause of many of the genome rearrangements. Further investigations on IS elements will help to study genome dynamics and rearrangement of S. flexneri 1a strain.
Micropropagation offers an opportunity to propagate and preserve of important plants. A simple and efficient protocol for in vitro micropropagation of lisianthus (Eustoma grandiflorum), an ornamental plant, is reported. We produced full plants only during 50 days. Axillary buds explants dissected from mother plants grown in a greenhouse were cultured on Murashige and Skoog (MS) medium supplemented with 16 combinations of 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzylaminopurine (BAP) at concentrations of 0.01 - 5.00 mg l⁻¹. The optimum plant growth regulators (PGRs) combination for maximal shoot regeneration (73.76 per explant) and proliferation rate (222.53) was 0.10 mg l⁻¹ 2,4-D along with 5.00 mg l⁻¹ BAP. These huge numbers of shoots were produced from callus dedifferentiated from explant. The best medium for shoot length (4.36 cm per plantlet) was 0.10 mg l⁻¹ 2,4-D. The largest number (2.40 per plantlet) and highest length (3.86 cm per plantlet) of roots were obtained on medium without PGRs. Roots were produced at the end of shoots only in medium without PGRs. All shoots produced on media containing PGRs got rooted in soil during 10 days. Rooting in the soil reduces the charge of micropropagation. About 98% of the micropropagated plantlets were established successfully in 15 cm pots containing a mixture of peat and perlite (1:1 ratio) and formed new leaves. © 2018 National Institute of Science Communication and Information Resources (NISCAIR). All rights reserved.
The enzyme neuraminidase (NA), coded by H1N1 virus, catalyses the removal of terminal sialic acid from viral and cellular glycoconjugates. It cleaves the terminal sialic acid on the glycosylated NA during virus budding to facilitate virus release. The outbreak of Swine flu is suspected to be caused due to H274Y mutation in the neuraminidase enzyme of H1N1 2009 strain. The present study involves 3-D structure modeling of mutated neuraminidase of the strain A/Poland/274/2009 (H1N1) by MODELLER9v7 and comparative analysis of coding statistics among NA gene of 3 mutated H1N1 strains with GenBank accession numbers GU112751, GU371269 and CY053923. The analysis of 3-D model revealed that NAs have a common fold characterised by ß-pleated sheet flanked either side by helices. The amino terminal end of the molecule is occupied by ß-α-ß motif and carboxy terminal end by ß-hairpin motif. The molecule is characterised by 24 strands and 3 helices. The a1 helix is the longest among the three helices. The comparative analysis of coding statistics indicates that the statistical features f1, f5, f6 and f7 have the most discriminating power for the individual recognition of the mutated neuraminidase genes of H1N1 2009.
Four spiny lobster species, Panulirus versicolor, P. ornatus, P. homarus and P. polyphagus, and a mud lobster Thenus orientalis were collected from the Gulf of Mannar marine biosphere reserve. Partial 28S rRNA gene sequences of the spiny lobsters were examined for their nucleotide diversity, pairwise genetic distances, transition/transversion rate and phylogenetic relationships. The spiny lobster species recorded higher transition over transversion; GC content was also higher as reported in many groups of animals. Pair-wise genetic distance analysis shows that all the spiny lobsters were distinctly distant from the outgroup species and had minimum distance among the ingroup species. A molecular phylogeny based on 28S rDNA D2 sequences indicates that the four species of Panulirus form a monophyletic group.
Earlier, natural active substances (fish oocyte extract) were used to induce mouse fibroblast cells to express stem cell marker antigens. Further, in the present study, authors have tested the ability of these natural active substances in combination with IL-2 to induce 293T cells to express stem cell marker antigens. It was demonstrated that treatment with the natural substance in combination with IL-2 caused significant increase in the expression of Oct-3/4, c-Myc and Nanog marker antigens in 293T cells. The present approach was proved to be a rapid method to generate multipotent stem cells.
As atherosclerosis is a prevalent non-communicable disease, and yet, no definitive medical treatment found for it, Trapping MCP1 as a key factor in inflammation could be effective. Therefore, we decided to display rabbit MCP-1 (R-MCP1) on human embryonic kidney 293T cell line surface. Firstly, R-MCP1 plasmid (pR-MCP1) containing kappa chain signal sequence, R-MCP1 sequence and PDGFR intra membrane domain was constructed. The delivered pR-MCP1 was transformed in E. coli TOP10F', and the resulted clones were assessed by PCR and digestion. After linearizing pR-MCP1 by BglII, HEK cells were transfected by them. MCP1 gene integration and expression was confirmed at RNA and protein levels by real- time PCR and flow cytometry, respectively. PCR product gel electrophoresis on genomic DNA of transfected HEK cells showed a 737 bp band. Based on real- time PCR results, We observed R-MCP1 gene expression significantly increased in transfected cells (272.26 ± 37.32) compare to untransfected HEK 293T cells (2.67 ± 0.12) (p = 0.001). The results of flow cytometry showed that about 85% of transfected cells were positive and express R-MCP1. Therefore, cell surface display of R-MCP1 has successfully been performed and the produced cells can be used in future research to prepare diagnostic and therapeutic agents like aptamers.
δ-Endotoxin Cry1Ab22 is produced by Bacillus thuringiensis BtS2491Ab. The toxic spectrum of this protein is reported to span Lepidopteron and Dipteran. Here, we predict the theoretical structural model of newly reported Cry1Ab22 toxin by homology modeling method on the structure of the Cry1Aa toxin. Proposed model resembles the target by sharing common three dimensional, three domain structure. The main differences being located in the length of loops, absence of helixes (α7b, α10a, α10b, α11a) and presence of additional components (β21, α9b). Few of the components like α9a, α9b and α12a are positioned spatially at different locations. A better understanding of the 3D structure will be helpful in designing the domain swapping and mutagenesis experiments aimed at improving toxicity, and will lead to a deeper understanding of the common mechanism of toxins.
Plant MYB (myeloblastosis) transcription factors play an important role in various biotic and abiotic defense responses. The plant R2R3 MYB proteins are predominantly involved in plant-specific regulatory processes. Saccharum spontaneum is a promising biomass crop because of its high fiber, polymorphism and adaptation to different environmental stresses. Efforts to cross S. spontaneum with sugarcane to introduce its desirable genes into modern cultivars showed minimum conclusive results. In the present study, we exemplify the isolation, cloning and in silica characterization of SsMYB2R gene isolated from S. spontaneum belonging to subfamily of R2R3-MYB transcription factor. The protein showed noteworthy alignments with a SANT/MYB DNA binding domain in members of MYB family proteins from dicotyledons and monocotyledons. The nucleotide sequence of the cloned SsMYB2R revealed a single open reading frame of 1,074 bp coding for 357 amino acids. To comprehend its sequence-to-structure-to-function paradigm, the putative 3D structure of SsMYB2R was generated using I-TASSER server. Further, PROCHECK, PROMOTIF and ProSA programs were used to appraise the stereo chemical quality of the model to conclude scores within the recommended intervals. The secondary and 3D structures thus provided better insight of active sites regarding protein-DNA interacting domains, which help abiding different environmental stress conditions by docking studies.
Human immunodeficiency virus (HIV), the causative virus for acquired immunodeficiency syndrome (AIDS) has become the world's greatest dispute. Inhibition of nucleocapsid protein domain 7 NCp7 receptors have been sturdily pursued as a promising target for the treatment HIV AIDS. A set of 36 thioesters derivatives has been reported as HIV NCp7 inhibitor was analyzed by employing PHASE method to inspect the structural requirements for diverse analogues to inhibit NCp7 receptors and to obtain a highly predictive model used for designing of novel NCp7 receptors inhibitors. A united study of pharmacophore prediction, atom based 3D-quantitative structure-activity relationship (QSAR) and molecular docking approaches were carried out on pyridinioalkanoyl thiolesters derivatives to understand their structural requisites and binding mode of the best fitted ligand for NCp7 inhibitory activity. Five point pharmacophore hypothesis AADDR (two acceptors, two hydrogen donor, one aromatic ring) yielded a statistically significant 3D-QSAR model with partial least square (PLS) factors 5, regression coefficient value of (R²) = 0.9625, cross validation coefficient value of (Q²) = 0.7775, root mean square error (RMSE) = 0.1358. The core structure of nucleocapsid NCp7 domain is docked with the compounds obtained from ZINC and NCI. Docking study also revealed the binding orientation of active ligands at active residues of NCp7. Using pharmacophore based database search, 3D-QSAR and docking studies, we identified ZINC29569253 as a stable inhibitor. The geometry and type of this pharmacophore model give emphasis to important binding features which will be useful for the design of selective HIV NCp7 inhibitors.
Molybdenum reduction to molybdenum blue by microbes is a potential bioremediation tool for molybdenum pollution. A previous work using cyanide as a respiratory inhibitor has shown that the site of molybdenum reduction in Enterobacter cloacae strain 48 is at the electron transport pathway. In the present work, several respiratory inhibitors together with cyanide were used to reinvestigate the site of molybdenum reduction in E. cloacae strain 48. All the respiratory inhibitors tested showed no inhibition to the molybdenum-reducing capability of the bacterium. It was also discovered that cyanide caused a large increase in the pH of the enzymatic reaction mixture and, therefore, the inhibition previously seen was likely due to unfavourable pH for the enzyme activity. Based on these results, the site of molybdenum reduction in this bacterium is suggested not to be the component of electron transport pathway.
A plasmid encoded pediocin CP2 sequence was analyzed using various bioinformatics tools. Multiple sequence alignment of pediocin CP2 depicted 100% sequence similarity with pediocin PA-1/pediocin AcH. Since no detailed 3D structure or structure-function analysis is available in literature, the 3D structure of pediocin CP2 was obtained based on the known structure of homologous sakacin P. It was analyzed and deduced that YGNGV motif contained in N-terminal β-sheet in pediocin CP2 makes it antilisterial. It was followed by a well defined central amphiphillic α-helix (residues 20-30), and this in turn was followed by the C-terminal tail (residues 31-44), which folds back onto the central α-helix, thereby creating a hairpin-like structure. This hairpin-like structure was stabilized by the presence of a second disulphide bridge in pediocin CP2 (because of extra cysteine residue C44) which was absent in the reference peptide sakacin P. High thermostability and broad antimicrobial range of the pediocin CP2 in comparison to sakacin P is attributed to the presence of this second disulphide bond that can further be verified by experimentation.
The objective of this study was to provide molecular evidences for enterocin biosynthesis by Enterococcus faecium MTCC 5153. The culture filtrate (CF) of E. faecium MTCC 5153 exhibited a broad inhibitory spectrum against several enterococci, food-spoilage lactic acid bacteria (LAB) as well as pathogenic Gram-positive bacteria. The antimicrobial compound present in the CF showed the properties of class IIa bacteriocin. PCR was employed for taxonomic identification and detection of genes coding for virulence factors and different enterocins. Based on amplification of enterocin A (entA) gene by PCR, additional characterization of enterocin A operon was carried out. Southern hybridization confirmed the presence of chromosomally encoded enterocin A (entA) gene. These results suggest distribution of conserved enterocin A operon among different strains of E. faecium. Tricine SDS-PAGE activity gel assay in combination with mass spectral analysis indicated the production of single enterocin of 4.8 kDa in size. Production of enterocin was carried out using different sugars in tryptone, yeast extract (TYE) medium, under microaerophilic condition and the antimicrobial activity in the range of 10-50 × 10 3 arbitrary units per milliliter (AU mL -1) was observed. This study indicates the potentiality of native isolate to produce broad spectrum bacteriocin which can be used to deal with spoilage bacteria and food-borne pathogens like listeria.
The present study reports the effects of aeration and agitation on growth and production of the cyclosporin A (CyA) in batch fermentor cultures of Tolypocladium inflatum MTCC 557. Dissolved oxygen concentration, pH, dry cell wt, CyA titer and sugar utilization were continuously measured and determined. In the first step, the effect of agitation was evaluated by changing the agitation speed in the range of 250 to 450 rpm at 1 vvm. Further, after selecting the optimum agitation speed, the aeration rate was varied between 0.5 vvm to 1.0 vvm. Efficiency of aeration and agitation was evaluated through oxygen mass transfer coefficient (kLa). Maximum CyA production of 1274 mg/L with specific productivity of 0.152 mg/g.h was obtained at 350 rpm agitation and 0.75 vvm aeration. The kLa of the fermentation system supporting maximum production (350 rpm, 0.75 vvm) was 0.113 h-1. The oxygen transfer rate (OTR), oxygen utilization rate (OUR), and specific oxygen uptake rates (qO2) in the fermentation broth were also determined.
The sugarcane variety CoC 671 is an early maturing, high sugar content variety in India. Recently, it has become very susceptible to red rot caused by Colletotrichum falcatum Went. To detect the pathogen at an early stage, a new serological technique, enzyme-linked immunosorbent assay was developed based on antiserum developed against the protein of host pathogen. Direct antigen coating enzyme-linked immunosorbent assay (Dac-Elisa) for the early detection of red rot infection has been standardized. Further, this technique was found reliable to screened planting material of sugarcane variety CoC 671 for red rot infection at an earlier stage. Besides, ill vitro red rot treated as well as healthy calli were also screened for red rot infection.
Two kb, BamHI fragment of duck embryo propagated Indian isolate (VN 1) of egg drop syndrome-76 (EDS-76) was cloned into pBluescript II KS + vector and the fragment was labelled with α 33p dATP label using random primer extension method. The labelled DNA was used as a probe for detecting viral DNA in faecal samples of experimentally infected birds. Proteinase K treated, phenol extracted, alkali denatured samples exhibited strong signals as compared to proteinase K digested samples alone. The probe could detect up to 1.25 pg of viral DNA. However, it did not show any positive signal with another adenovirus, i.e., hydropericardium syndrome viral DNA indicating its specificity for egg drop syndrome virus (EDSV).
cDNA library was constructed in λgt 11 expression vector from mRNA of immature seeds of pigeonpea. This cDNA was screened with specific non-radioactive, DIG-labelled heterologous pea vicilin cDNA probe (pRC-758). The positive vicilin (7S) encoding cDNA clones were isolated. One of the vicilin cDNA clones showed insert size of ∼1.3 Kb, when analyzed by PCR using λgt 11 forward and reverse primers. This PCR product was subcloned in pUC-18 vector for confirmation of gene and product by Southern and Western hybridizations, respectively. The clone was named as pSL-1 and sequenced with M13 universal forward and reverse sequencing primers. The partial nucleotide sequence (1341 bp) has been indexed in NCBI gene bank with accession number AF348366. The complete gene has 1417 base pairs with 972 bp coding sequence. Hence, the predicted polypeptide chain of this gene was determined, which contained 323 amino acids. After post translation modification, the predicted polypeptide has 310 amino acids long with approximately mol wt of 34.1 kDa. This gene encoding vicilin (7S) protein provides the basic understanding of the gene structure of pigeonpea storage proteins.
Equine influenza virus, A/Equi-2/Ludhiana/87 H3N8 isolated from influenza epizootic in India in 1987 was characterized for its uniqueness by sequencing of its neuraminidase gene as the vaccine prepared with this isolate is still in use and has not been updated for more than a decade. Comparison of the nucleotide and amino acid sequences of this gene with other H3 N8 isolates revealed that Indian isolate had maximum differences with avian like equine influenza isolate (Jilin/89) and had resemblance with equine influenza H3N8 isolates namely Alaska/91, Tennessee/86, Newmarket/79 and Kent/87 which further indicated that it belongs to the lineage of currently circulating H3N8 equine viruses in equine population.
Selection of the best nutrients is one of the most critical stage in media optimization for polygalacturonase production. Plackett-Burman design was used to screen various pectin substrates, nitrogen sources and mineral nutrients for polygalacturonase production by Aspergillus awamori MTCC 9166. Fifteen different pectin sources like crude pectin, polygalacturonic acid, orange peel, citrus peel, jackfruit peel, etc. were selected for polygalacturonase production using 16 experimental design of Plackett-Burman. Similarly, eleven nitrogen sources like yeast extract, tryptone, casein hydrolysate, sodium nitrate, ammonium chloride, etc. and eleven mineral nutrients like NaCl, MgSO4, KH2PO4, CaCl2, etc. were screened for polygalacturonase production using 12 experimental design of Plackett-Burman. The enzyme production was studied for 5 d, where the maximum production was observed on 3rd d and so this data was analyzed using Indostat software to obtain regression coefficients and t-values. Based on these values significant nutrients like seven pectin sources (orange peel, jack fruit rind, apple peel, pine apple peel, mango peel, banana peel & tomato pulp), four nitrogen sources (urea, yeast extract, casein hydrolysate & potassium nitrate) and four mineral nutrients (NaCl, KH2PO4, CaCl2 & KH2PO4) were selected for second level screening of efficient nutrients for polygalacturonase production using 16 experimental design of Plackett-Burman. Orange peel as pectin source, casein hydrolysate as nitrogen source and NaCI showed maximum enzyme production and so were selected for further quantitative optimization.
An inducible keratinase is produced by Bacillus sp. JB 99 in a feather medium. Under submerged fermentation condition with continuous agitation (180 rpm), high level of keratinase production occurred at 45°C after 36 h at pH 10. The presence of carbon source in feather medium suppressed the enzyme production, while 0.1% yeast extract enhanced the production. The purified enzyme showed maximum keratinase activity at temperature 65°C and pH 10. The enzyme was monomeric and has a mol wt of approximately 66 kDa (SDS-PAGE). The enzyme may belong to serine protease group as completely inhibited by PMSF. Presence of metal ions, such as, Ca2+, Mg2+, Co 2+ and Ba2+ stimulated the enzyme activity, while Hg 2+, Pb2+, Zn2+ and Fe2+ decreased the activity. Present results indicate that Bacillus sp. JB 99 can be a highly useful organism for feather meal production and in leather industry.
In the present investigation, authors have reported novel calcium ion independent α-amylase enzyme from less extensively studied marine Streptomyces strain A3. The isolated strain was identified on the basis of 16S rDNA sequencing and scanning electron microscopy. The α-amylase from strain A3 was purified to homogeneity with the aid of ammonium sulfate precipitation and gel filtration chromatography by using Sephadex G-75, insoluble corn starch and sephacryl S-100 column, with a 43.92-fold increase in specific activity. SDS-PAGE and zymogram activity staining showed a single band equal to molecular mass of 45 kDa. The activity and stability of enzyme didn't increase in presence of calcium ion, indicating calcium ion independent nature of amylase. Enzyme retained considerable activity in presence of oxidants and commercial detergents. Glucose, maltose and maltotriose were the main end product of starch hydrolysis, indicating that enzyme is an α-amylase. Therefore, it can be concluded that the surfactant, detergent stable and calcium ion independent α-amylase from strain A3 has widespread applications for detergent and pharmaceutical industry where higher salt concentration inhibits enzymatic conversion. In addition to that, application of this amylase may eliminate the use of calcium in starch liquefaction and subsequent removal by ion exchange.
Somatic embryos of banana cv. Rasthali were subjected to 0-8 h desiccation and abscisic acid (ABA) treatment for improving plant conversion. Somatic embryos exposed to 10 μM ABA followed by 2 h desiccation exhibited an increase in plant conversion (66%) compared to control (56%). Embryos treated with ABA followed by 2 h desiccation survived up to 5 weeks with 56% plant conversion upon storage at 10°C. This study suggests that combined treatment of banana somatic embryos with ABA and desiccation can improve conversion frequency.
Arsenite is considered to be more toxic than arsenate (an average of 100 times). It can be oxidized to arsenate by chemical reaction or microbiological interactions. The aim of the present study was to investigate the arsenite-oxidizing microbes that exist in iron ore mine in India. We cultured 13 morphologically distinct bacterial strains, among which 6 strains could grow in high concentrations and shows the arsenite transforming abilities. Analysis of the amplified 16S rDNA gene sequences of the isolates revealed them to belong to alpha-, gamma- proteobacteria and firmicutes particularly in genera Paenibacillus, Pseudomonas, Ochrobactrum, Enterobacter and Bacillus. Further, qualitative silver nitrate screening assay and quantification by HPLC-ICP-MS analysis of these strains indicated the transformation of arsenite to arsenate. Moreover, the isolates were genetically analyzed for presence of arsenic arsenite transporter gene (arsB) that indicates the genetic ability of bacteria to tolerate the most toxic arsenic species. © 2019 National Institute of Science Communication and Information Resources (NISCAIR). All rights reserved.
Abiotic environmental stresses, which limit the plant distribution and productivity, include low and high temperature, salinity and water deficit. Over the last century human activities have increased the level of environmental stress in the form of pollutants such as ozone and heavy metals, levels of UV light reaching the biosphere and salinity in irrigated areas. Plants are sessile and have, therefore, developed mechanisms to survive under extreme environments sometimes in vegetative stages of their life cycle. The recent surge of information on regulation of gene expression under stress conditions as well as the biochemical function of individual proteins in conferring tolerance to stress will help in isolating the genes of interest to produce desired transgenics. The understanding of molecular basis of these survival mechanisms discussed in this review may ultimately help enhance plant productivity in current marginal areas. The review deals with effect of drought and salt stress on plants and the regulatory mechanisms.
Molecular characterization of biotic (Ascochyta blight, Fusarium wilt & dry root rot) and abiotic (drought & salinity) resistant genotypes of chickpea was carried out using a specific set of 20 polymorphic STMS (sequence tagged microsatellite site) markers. The number of alleles ranged from 1 to 3 alleles per locus. The PIC (polymorphism information content) value ranged from 0.0 to 0.656 with an average of 0.268, indicating the considerable efficiency of markers for studying the polymorphism level. The primer GA-33 showed maximum PIC value (0.656), while the ten primers had the 0.0 value. The dendogram derived from the analysis had six clusters. Many pairs showed the maximum similarity (0.983), whereas accessions ICC-1392 and ICC-2065 showed the minimum similarity (0.900). The conservation of genotypes ICC-2580, ICC-1392 and ICC-2065 was recommended for their utilization in Middle-east Asia region chickpea improvement programme.
A majority of environmental signals of varied types as well as varied intensities are perceived at the membrane level in a cell. In case of multicellular organisms like plants or animals, this 'perception' not only needs to be transferred to the actual centre of controlling and responding unit i.e. the nucleus but sometimes from one cell to another cell which may just be lying close enough or even at an appreciable distance e.g. 'root-shoot communications'. In contrast to processes of cell-to-cell communication in a plant system, which is just beginning to be elucidated, the mechanism of 'transfer' of this information from outer surface to the core controlling units has been an active area of research since past few decades. Abundant reports do exit in literature, which support a kind of 'cascading mechanism' for this purpose. It is now a well-established fact that a divalent cation i.e. Ca 2+ plays an extremely important role in this process. The fluctuations in the level of Ca2+ at a given time in a given cell organelle is the crucial factor determining the activation stage of some special proteins, which have been proposed to have a high affinity for Ca 2+. Such proteins are known as Ca2+-binding proteins (CaBPs). Under environmental abuses, the level and activation of CaBPs play an important role in bringing about the 'ignition' of 'protective' as well as 'defensive' mechanisms, which ultimately are reflected in the form of the physiological adaptations in the plant as a whole system. The present review is an attempt to highlight the importance of CaBPs in plants under abiotic stress conditions.
Brucella abortus is a facultative intracellular bacterial pathogen infecting animals and humans. This preliminary study was designed to express two immunogenic genes of Brucella abortus as recombinant fusion proteins in mammalian cells for being studied as a vaccine candidate in mammalian hosts, especially mice and cattle, in our next study. The complete open reading frame sequences of two immune dominant genes of Brucella abortus namely, Omp16 and L7/L12 were fused with an intermediate spacer along with N-terminal fusion with secretory signal sequence from immunoglobulin M. The complete fusion gene sequence was codon optimized for expression in mammalian cells. For expression analysis, the codon optimized synthetic gene was cloned in pDsRed-Express-N1 mammalian expression vector, with C-terminal fusion with red fluorescent protein sequence. On transfection in MDBK and HEK-293 cells, appearance of red fluorescence in transfected cells indicated expression of Omp16-L7/L12 fusion proteins along with RFP. The Omp16-L7/L12 fusion construct without RFP sequence was also expressed in mammalian cells. The expressed Omp16-L7/L12 fusion proteins were confirmed through both indirect fluorescence antibody test and western blot. This preliminary study suggested that the codon optimized Omp16-L7/L12 fusion construct is ready to be studied in hosts like mice and cattle for its vaccine efficacy.
A. precatorius L.: Field grown plant (A); Pods of A. precatorius (B); & Seeds of three genotypes-Red (C), Black (D) & White (E).
RAPD and ISSR profiles of A. precatorius genotypes showing polymorphic banding patterns using selected primers (a. RUF202, b. RUF203, c. RUF210, d. RUF215, e. RUF216, f. IUF016, g. RUF219, h. IUF019, & i. IUF022). [M, Mol wt marker 1 Kb ladder; lane 1, White seeded; 2, Red seeded; & 3, Black seeded]
RAPD and ISSR profiles of A. precatorius genotypes showing monomorphic banding patterns using selected primers (a. RUF205, b. RUF207, c. RUF211, d. RUF217, e. RUF218, f. RUF220, g. IUF017, & h. IUF021). [M, Mol wt marker 1 Kb ladder; lane 1, White seeded; 2, Red seeded; & 3, Black seeded).
Three genotypes of a medicinal, climbing herb Abrus precatorius L. (Family: Fabaceae), having different seed coat colour (red, black and white), were subjected to molecular analysis using PCR based RAPD and ISSR markers. Twenty RAPD (decamers) and fourteen ISSR primers were screened for their amplification potential. Of which 12 RAPD and 5 ISSR primers produced clear and reproducible amplified products. These primers yielded a total of 149 amplified fragments with an average of 8.76 bands per primer including 14 (9.39%) polymorphic fragments. In the study, ISSR fingerprinting (14.28%) detected more polymorphic loci as compare to RAPDs (7.89%). The data analyses based on Jaccard's similarity coefficient and UPGMA cluster analysis revealed that genotypes with black and white seed coats were more diverse as compared to genotype with red seed coat. Combined data of RAPD and ISSR further revealed that Abrus with white seed coat was more closely related to those having red seed coat.
SSRs among 35 accessions of niger amplified by primer 2911. [Lane L, Mol wt standard (100 bp); Lanes 1-35, Corresponds to the codes in Table 1.] 
-Geographic origin and range of oil (%) and oil parameters in 35 accessions of niger based on five parameters
Association between 35 accessions of niger as revealed by bootstrap analysis based on the ISSR data. Table 3-Jaccard's similarity coefficient among niger genotypes of various origin Similarity coefficient Origin No. of genotypes Paired combination Mean Range 
Molecular markers ISSR were used to assess the genetic diversity in 35 niger [Guizotia abyssinica (L.f.) Cass.] germplasm accessions significantly differing for oil parameters. Of the 200 ISSR primers screened, 25 primers producing clear and polymorphic bands were selected. The efficiency of individual ISSR primers was also compared for the resolving power ranges (Rp; 0.16 to 7.5); number of banding patterns (Nb; 3 to 27); heterozygosity (H; 0.245 to 0.497); polymorphic information content (PIC; 0.215 to 0.374); and discrimination power (Dj; 0.138 to 0.992). The range of mean genetic similarities between accessions based on Jaccard's similarity coefficient was significantly high (0.1 to 0.95) with a mean value of 0.40. The UPGMA clustering based on bootstrap analyses grouped the accessions into three major clusters. Although a significant genetic diversity was observed based on ISSR and biochemical markers, but only partial correlation between diversity based on these two parameters was observed. On the basis of present findings, it is evident that significant genetic as well as biochemical diversity exists among the niger accessions. The genotypes, particularly, accessions IC259393, IC211080 and IC211078 with highest linoleic acid (>53%), highest stearic acid (<8%), highest palmitic acid (>9%) and least oleic acid (<29%) would be helpful in pre-breeding for hybridization to create a wide spectrum of variability in subsequent segregating generations for various uses of fatty acids. The finding would further elucidate the genetics of oil parameters in niger.
-In vitro initiation of shoot in different media: (a) abnormal shoot in MS + 9.08 µM TDZ, (b) shoot induction in MS + 6.66 µM BAP, (c) shoot induction in MS + 6.66 µM BAP + 0.465 µM Kinetin, (d) callusing at shoot base and poor quality shoot in MS + 6.66 µM BAP + 2.63 µM Kinetin + 0.268 µM NAA.
-Shoot proliferation of A. mangium at different subculture in MS + 6.66 µM BAP + 0.465 µM Kinetin: (a) 1 st subculture, (b) 2 nd sub-culture, (c) 3 rd sub-culture, (d) 4 th sub-culture.
-Rooting and acclimatization of in vitro grown A. mangium: (a) rooting in ½ MS + 9.80 µM IBA, (b) length of root in ½ MS + 9.80 µM IBA length,(c) hardening of plantlets in different growing media, (d) plantlets ready for transplanting.
-Genetic stability of regenerated plantlets (Lane 1,7,13,19-Molecular weight marker (500 bp ladder); Lane 2,8,14 and 20-mother plant; lane 3,9,15 and 21-plants from 1 st subculture, lane 4,10,16 and 22-plants from 2 nd subculture; Lane 5,11,17,23-plants from 3 rd subculture; Lane 6,12,18 and 24-plants from 4 th subculture)
The study describes an efficient, reproducible and stable in vitro propagation protocol for Acacia mangium from nine years old phenotypically superior plus tree. For effective control of contamination explants were dipped in absolute alcohol for 1 minute followed by 0.1% mercuric chloride solution for 6 minutes. Maximum establishment (87.8%) and shoot proliferation was achieved with an average value of 5.7 shoots per explants and shoot length of 2.8 cm in Murashige & Skoog (MS) media supplemented with 6.66 μM 6-benzylaminopurine (BAP) and 0.465 μM kinetin. The addition of 9.80 μM indolebutyric acid (IBA) in half MS media gave maximum rooting (88.3%) with average 2.7 roots per microshoots with longest root of 18 cm. Rooted plants were hardened and successfully established in the soil. Random amplified polymorphic DNA (RAPD) profile of micropropagated plants shown no change in genetic fidelity up to four growth cycles.
-DNA banding profiles obtained using OPB-12 RAPD primer (1 -45: Peach genotypes and M: 100 bp ladder).
-Dendrogram obtained after pooled RAPD and ISSR analysis in peach germplasm.
List of peach germplasm used in molecular characterization studies
Primer sequences, annealing temperature, size of amplicons, number of amplified bands, percent polymorphism and polymorphic information content values of ISSR markers studied in peach genotypes
Molecular characterization of 45 peach (Prunus persica) accessions was carried out using 48 RAPD and 46 ISSR molecular markers to assess the value and magnitude of genetic divergence. The RAPD primers revealed 84.20% polymorphism and ISSR markers generated 89.00% polymorphism. Pooled RAPD and ISSR along with UPGMA clustering based on Jaccard's coefficient were estimated with a view to assess efficiency of the marker system in Prunus persica. Polymorphic information conten (PIC) values varied from 0.13 to 0.50 in RAPD and 0.12 to 0.49 in ISSR with the mean values for all loci were 0.33 and 0.35, respectively. Jaccard's similarity coefficient among peach accessions with respect to RAPD and ISSR markers ranged from 0.37 to 0.95 and 0.43 to 0.95 which indicated a broad genetic base. Pooled analysis of both molecular markers concluded that genotypes 'Darli' and 'IC-2' are most distantly related to each other. The use of arbitrary oligonucleotide primers in the amplification reaction facilitated the study of uncharacterized genomes. In the present study, high level of polymorphism indicates their applicability in framing more extensive studies in development of superior progenies, quantitative trait loci (QTL) mapping, molecular breeding, investigation of population genetic diversity, comparative mapping, selection of the parents etc. among various peach crop improvement programmes. © 2018 National Institute of Science Communication and Information Resources (NISCAIR). All rights reserved.
-Analysis of variance for callus induction frequency 
Effect of accessions and explants on callus growth, colour, terxture and globular shaped somatic embryo induction. 
Effect of somaclones on callus induction frequency. 
Effect of somaclones on callus growth, colour and texture.
Plant regeneration in lucerne (Medicago sativa) is highly genotype dependent and inspite of considerable information on in vitro regeneration in this species the Indian genotypes have not been investigated so far. A system of recurrent somatic embryogenesis (RSE) was established for the first time in Indian accessions of lucerne and utilized for Agrobacterium mediated genetic transformation. Seeds, hypocotyls and cotyledons of LLC-3, C-10, A-3 and IL-75 accessions and ovary explants of 5 somaclones of LLC-3 were used and globular shaped somatic embryos were observed in all of the cultures. All the developmental stages of somatic embryogenesis were observed in ovary culture of one somaclone only. These somatic embryos have been undergoing cycles of RSE and the system has perpetuated for 24 months. For Agrobacterium mediated genetic transformation, this system of RSE required creation of injury in individual somatic embryos for enhanced transformation efficiency that limited its utilization for large-scale transformation experiments. An efficient system to overcome the requirement of creation of injury was developed in which pre-culture of somatic embryos on 2,4-D supplemented medium prior to their co-cultivation with Agrobacterium and supplementation of 2,4-D in co-cultivation medium could significantly improve the transformation efficiency.
Poly (β-hydroxyalkanoates) (PHAs) are natural polyesters produced by a variety of bacteria. They are represented most commonly by poly (β-hydroxybutyrate) (PHB), an intracellular storage biodegradable polymer material. The production costs of PHB are quite high compared with those of synthetic non-degradable plastics, hence search for potential strains with high PHB accumulating ability. Hundreds of indigenous bacterial strains were screened for the accumulation of PHB by Nile red, fluorescence microscopy (Nile blue A) and PCR. Three degenerate primers were used as PCR primers to detect PHA synthase genes. Among the tested isolates, 35 strains yielded a specific amplicon of 496 bp and 406 bp in colony PCR and seminested PCR, respectively. Among the 35 short chain length positive strains, only 2 isolates yielded a specific amplicon of 540 bp PCR product in medium chain length PCR, representing partial coding sequences of phaC1/phaC2 genes. The mcl-PCR positive Pseudomonas indigenous isolates (LDC-5 and LDC-25) could be potential candidates for bioplastic production.
Chitinase gene (Pr-3) specific transcript accumulation was increased by 5.57% in mature leaf compared to the other in vitro grown tissues in tea (Camellia sinensis) genotype (T383) during induced systemic resistance by methyl jasmonate. Chitinase (Pr-protein) gene specific two primers were used in RT-PCR reaction to measure the differential transcript accumulation in different tissue systems (young leaf, mature leaf, callus and shoot regenerated from somatic embryo). The time-course expression patterns resulted in differential expression of two transcripts (254 & 366 bp) in response to induction. The 366 bp transcript was sequenced bidirectionally and deposited into the GenBank of NCBI ( EU373553).
Time course of yeast extract (a) 20 mg L-1 (b) 200 mg L-1 induced accumulation of puerarin, genistin, daidzein and genistein (µg g-1 dry wt) in cell suspension cultures of P. tuberosa (• treated cultures, ■ control cultures). Data are the averages of three experiments, each in triplicate. 
-Effect of different concentrations of methyl jasmonate treatment on isoflavonoids content (µg g -1 dry wt) in cell cultures of P. tuberosa grown in modified MS medium
Total isoflavonoids yield (µg g-1 dry wt) in cell suspension of P. tuberosa. (a) 20 mg L-1 (b) 200 mg L-1 yeast extract (• treated cultures, ■ control cultures). Data are the averages of three experiments, each in triplicate. 
Cell cultures of Pueraria tuberosa were established in modified Murashige and Skoog medium and challenged with yeast extract (YE), methyl jasmonate (MeJA) and salicylic acid (SA). Maximum isoflavonoids production was recorded at 48 h of YA incorporation at the stationary phase. 20 μM of MeJA and SA was most effective in isoflavonoids induction. Higher concentrations of these elicitors were negatively correlated with the isoflavonoids production. YE at 150 mg L-1 was optimal for isoflavonoids production, yielding 10 mg L-1 isoflavonoids, which was ∼20% higher over the yields at optimal concentrations of MeJA and SA. YA incorporation can be used as a trigger to induce high yield of isoflavonoids.
Top-cited authors
Nilanjana Das
  • VIT University
Vimala R
  • VIT University
Mukesh Doble
  • Distinguished Prof Saveetha Dental College Chennai; Director Theevanam Additives & Neutraceuts p ltd(IIT M research park) Madras
Kakasaheb R Mahadik
  • Bharati Vidyapeeth Deemed University
Viruthagiri Thangavelu
  • Annamalai University