019543   Goulart A J;Benedetti A C E P;Carmona E C; Rubens Monti (Biochemistry and Technological Chemistry Dep, Institute of Chemistry-Sao Paulo State Univ, UNESP, Araraquara, SP, Brazil, Email: montiru@fcfar.unesp.br) : Multipoint immobilization of invertase on agarose: stability and kinetic properties. Curr Trends Biotechnol Pharm 2008, 2(3), 462-70.

Enzyme immobilization is a specific method for restricting the enzyme freedom of movement. There are strategies for the proteins multipoint immobilization by amine-terminal residues, lysine residues and carboxylic groups. In the present work, a commercial invertase of Saccharomyces cerevisiae underwent multipoint immobilization on glyoxyl-agarose, amine-agarose and glutaraldehyde-agarose supports. Derivatives kinetic properties were determined and compared with the properties of the soluble enzyme. The copper influence on enzyme activity and its inhibition by fructose were also investigated. Amine-agarose exhibited activity closest to the soluble enzyme (93.3%). This same derivative maintained approximately 50% of initial activity when 800 mM of fructose were added to the reaction medium. However, glutaraldehyde-agarose exhibited the best stability to temperature and pH and none of the derivatives lost inhibition by copper. Glyoxyl derivative exhibited the lowest Km (0.023mM) and amine derivative achieved the highest maximum velocity (1666.7 U/mg prot.).

3 illus, 2 tables, 26 ref

019542   Dangi R;Gandhi J;Bhardwaj S (Chemistry Dep, M.P. Govt. P.G. College, Chittorgarh, Rajasthan, Email: sidsidsmart@yahoo.co.in) : Harvesting solar energy for treating dye stuff in industrial effluents-zns-cds used as photocatalyst. Rasayan J Chem 2008, 1(2), 335-61.

The removal of Crystal Violet was carried out through semiconductor ZnS-CdS. Visible light was used as the source of energy. Various parameters like amount of semiconductor, pH, light intensity, dye concentration etc. were varied. A tentative mechanism is proposed.

5 illus, 5 tables, 11 ref

019541   Chikkaswamy B K;Paramanik R C;Paramanik A (Biotechnology Dep, R.K. Institute of Management & Computer Science, Bellandur Gate, Sarjapura Main Road, Bangalore-560 037, Email: drchikkaswamy@rediffmail.com) : Study of peroxidase isozyme banding patterns in mulberry varieties. J phytol Res 2007, 20(2), 171-4.

Deals with peroxidase isozymes banding patterns in eight mulberry varieties.Of these Berhampur local is having five bands, whereas Sukasakwa, Jakkur local, Mutants, M.R. Midew, Coonoor-11, Selection -41 and Mysore local have revealed less bands. The significant changes were observed in banding patterns and in their Rt values. The possible relation between plant morphology and peroxidase is discussed, on the basis of similarity index.

1 illus, 3 tables, 31 ref

019540   Chakraborti S;Sinha S;Sinha R K (Cytogenetics & Plant Tissue Culture Laboratory, Life Science Dep, Tripura Univ, Suryamaninagar-799 130, Email: khsinhark@yahoo.co.in) : Biochemical changes during fruit ripening of Psidium guineense Swartz. (Myrtaceae) - an endemic wild guava of Tripura, India. J phytol Res 2007, 20(2), 183-7.

Biochemical analysis of the fruit of wild guava showed distinct variation in ascorbic acid and total soluble sugar contents during the course of fruit maturation and ripening. The acidity level along with total soluble protein and free phenolics were measured and analyzed as well. The fruit maturity was defined in terms of changes of colour from green to yellow. The result revealed that the ascorbic acid content increased from 1.15 ñ 0.05 mg/g to 2.30 ñ 0.12 mg/g fresh weight while the fruit titrable acidity decreased from 0.82 ± 0.01 to 0.60 ± 0.01 milli - equivalent of (N) Na₂CO₃/g in P. guineeme. Superiority in ascorbic acid and sugar content was recorded in P. guineense in comparison to P. guqjava while protein and phenol content was much higher in P. guajava.

4 illus, 3 tables, 16 ref

018506   Zeerak N A;Wani S A (Div of Plant Breeding and Genetics, Sher-e-Kashmir University of Agricultural Sciences and Technology of K, Shalimar, Srinagar-191 121) : Diversity of Irises from Kashmir Himalaya. J ornamental Hort 2007, 10(2), 115-18.

In Kashmir Himalaya, Irises form a important part of total ornamental plant diversity and grow abundantly in alpine and and sub-alpine grasslands, gardens, saffron fields, graveyards and stream banks. The genus is represented by several wild, cultivated and some endemic species. To assess the available Iris diversity, surveys were carried out in the diversity rich areas of the region and as a result the distribution and diversity of 14 species including cultivated varieties were studied. Most of the species studied grow wild, beautify their habitats and are put to several uses by local people. This important gene pool is however under threat due to urbanization, change in land use pattern and deforestation which has resulted in declining their frequencies. In view of thier dwindling populations, there is an urgent need to maintain their diversity through conventional and biotechnological approaches. Nonetheless this diversity is still being maintained only due to the fact that they have found their abode in sacred groves, graveyards and cemeteries where their genetic identities have remained preserved.

8 ref

018505   Yalcin S K;Ozbas Z Y (Food Engineering Dep, Faculty of Engineering, Hacettepe University, Beytepe 06532, Ankara, Turkey) : Effects of ammonium sulphate concentration on growth and glycerol production kinetics of two endogenic wine yeast strains. Indian J Biotechnol 2008, 7(1), 89-93.

Effects of initial ammonium sulphate concentration on growth and glycerol production kinetics of two endogenic wine yeast strains, Saccharomyces cerevisiae Kalecik 1 and Narince 3, were investigated in a batch system. Maximum specific growth rates for Kalecik 1 and Narince 3 were obtained at initial concentrations of 1.0 g/L, and 0.8 g/L ammonium sulphate, respectively. Further, maximum specific glycerol production rates and glycerol concentrations, for both of the strains, were obtained in the medium containing 0.3 g/L ammonium sulphate. In this medium, maximum glycerol concentrations were 6.4 g/L for the strain Kalecik 1, and 6.5 g/L for Narince 3.

018504   Vyas R V;Patel B;Maghodia A;Patel D J (Nematology Dep, B A College of Agriculture, Anand Agricultural University, Anand-388 110) : Significance of metabolites of native Xenorhabdus, a bacterial symbiont of Steinernema, for suppression of collar rot and root knot diseases of groundnut. Indian J Biotechnol 2008, 7(3), 371-7.

Native isolated Xenorhabdus from Steinernema spp. manifest wide variety of secretary proteins, mainly in three clusters having mol wt in the range of 20-21, 46-51 and 60-66 kDa. The proteins of high mol wt, 76-90 kDa, apart from regular proteins are produced only in Xenorhabdus isolates of S. riobrave (SrM & A), but not in the isolates S. carpocapsae (Sc) and S. thermophilum (St). Under pot culture efficacy, average galls/root of groundnut (root-knot index) was significantly low with higher shoot and root wts in exo- and endo-toxic factor (2% v/w) treatments of different Xenorhabdus isolates over control. Effect of Xenorhabdus metabolites against collar rot disease of groundnut revealed that soil application of exo- and endo-toxic metabolites (1:10 diluted) in root zone of plants were effective in suppressing fungus Aspergillus niger, giving better plant height and survival of groundnut at harvest. PCR amplification of genomic DNA showed multiple amplified products of Xenorhabdus isolates (Sr A & M, St).

018503   Vijaya Ch;Mallikarjuna Reddy R (Biotechnology Dep, Jawahar Bharati Degree College, Kavali) : Impact of soil composting using municipal solid waste on biodegradation of plastics. Indian J Biotechnol 2008, 7(2), 235-9.

Plastic wastes accumulating in the environment are posing an ever increasing ecological threat. In this context, an attempt was made to study the biodegradation of polyethylene films in the natural environment. The method suggested by ASTM standards, i.e. composting of polythene films with municipal solid waste, was adopted to examine the biodegradation of plastics in the natural environment. The samples were collected and tested for weight loss and reduction in tensile strength at 2 months interval for 12 months of composting. Loss of weight and reduction in tensile strength of polythene films were taken as the criteria indicating biodegradation of these materials. Composting of polythene films for 4 months did not show any degradation. After 4 months of composting, the loss in weight was 2.9-4.5% for HDPE films and 10.5-11.6% for LDPE films. Similarly, the reduction in tensile strength ranges from 16-20% for HDPE and 12-13% for LDPE films. The study indicates that the biodegradation of polythene films occur in natural environment at a very slower rate.

018502   Vasane S R;Kothari R M (NO, Jain Hi-Tech Agri Research Institute, Jain Irrigation Systems Ltd, Jalgaon-425 001) : Integrated approach to primary and secondary hardening of banana var. Grand Naine. Indian J Biotechnol 2008, 7(2), 240-5.

For improving banana productivity, virus-indexed Grand Naine plantlets were used for primary and secondary hardening. Among nine combinations of bio-fertilizers (including VAM) used, GNP₂V₁ medium comprising of peat fortified with nitrogen fixing and phosphate solubilizing microbes (each 1 g/plantlet) and VAM (2 g/plantlet) emerged as the optimal growth medium for primary hardening on the basis of statistically significant 98.9% survival, besides corroborating optimal morphological features (viz. root length, number of primary roots, % VAM colonization, height, girth, number of leaves, leaf area and chlorophyll content) at the end of primary hardening. For secondary hardening, among four combinations of bio-fertilizers and VAM used in conjunction with soil, press mud cake (PMC) and vermi-compost (VC), SNP₂V₁ medium comprising of soil, PMC and VC in a ratio of 1:1:1 (v/v/v) emerged as the optimal growth medium on the basis of statistically significant 97% survival, corroborated by optimal morphological features as above, along with uptake of macro- and micro-nutrients. The positive feedback by several monitoring parameters have left no doubt about the integrated outcome at the end of primary and secondary hardening.

018501   Vartak V;Shindikar M (Botany Dep, University of Pune, Pune-411 007) : Micropropagation of a rare mangrove Bruguiera cylindrica L. towards conservation. Indian J Biotechnol 2008, 7(2), 255-9.

In vitro propagation protocol has been developed for Bruguiera cylindrica L. (Rhizophoraceae), a rare tree mangrove found along the western coast of India. Hypocotyl segments of viviparous propagule were cultured in vitro on Murashige and Skoog's medium (MS). Shoot and root initiation was better on modified MS (MSM) than on MS. Shoots and roots were produced from the cut surface simultaneously in 74% explants on MSM devoid of growth regulators. The explants produced several visible shoot buds on their cut surfaces, though only 1-3 shoots were developed per explant. Multiple shoots were produced by 70% explants. When cultures were initiated in monsoon, 1.6-fold increase in shoot formation and 2.4-fold increase in root formation was observed over those initiated before monsoon. During acclimatization the survival was 87%. Considering the status of this species in terms of its distribution, threats and conservation values in the study area, its in vitro propagation will compliment and strengthen the large-scale plantation activities towards the conservation or restoration of mangrove forests.

018500   Vaishali;Khan S;Sharma V (Biotechnology Dep, Sardar Vallabh Bhai Patel University of Agriculture and Technology, Meerut-250 110) : RAPD based assessment of genetic diversity of Butea monosperma from different agro-ecological regions of India. Indian J Biotechnol 2008, 7(3), 320-7.

Butea monosperma (Lam.) Taub. (English. Flame of the forest) belonging to the family Fabaceae, is an anthropogenic tree of several castes and also a very useful tree for both local people and pharmaceutical industry. This valuable tree needs attention for the characterization of its genetic diversity, protection and cultivation. The present work describes randomly amplified polymorphic DNA (RAPD) analysis to assess the genetic divergence among 16 Butea accessions collected from four agro-ecological regions of India (nine agro-climatic sub zones), covering five states (Uttarakhand, Uttar Pradesh, Rajasthan, Madhya Pradesh and Karnataka). Out of the 30 ten mer random primers used for studying genetic divergence, 12 were polymorphic, generating a total of 145 amplification products with an average of 12 products per polymorphic primer and an estimated mean gene diversity of 0.43. Genetic relationships among accessions were evaluated by generating a similarity matrix based on Jaccard's coefficient ranging from 0.53 to 0.79. The phenetic dendrogram generated by UPGMA analysis grouped accessions into four clusters. Primer 5, 12 and 16 were found most informative based on their resolving power and their potential to differentiate all the accessions. The degree of genetic variation detected among the 16 accessions with RAPD analysis suggested that RAPD could be used for studying genetic diversity in Butea. The study also demonstrated that Butea germplasm collected from different agro-ecological regions showed no isolation based on sub-climatic zones as the accessions collected from different sub-climatic zones grouped together in the genetic tree.

018499   Suke S G;Pathak R;Ahmed R S;Tripathi A K; Banerjee B D (Environmental Biochemistry and Immunology Lab, Biochemistry Dep, University College of Medical Sciences and G.T.B. Hospital (University, Dilshad Garden, Delhi-110 095, Email: banerjeebd@hotmail.com) : Melatonin treatment prevents modulation of cell-mediated immune response induced by propoxur in rats. Indian J Biochem Biophys 2008, 45(4), 278-81.

The effect of melatonin, a major secretory product of the pineal gland, in attenuation of propoxur (2-isopropoxy phenyl N-methyl carbamate)-induced modulation of cell- mediated immune (CMI) response was studied in rats. Male Wistar albino rats were exposed to propoxur (a widely used pesticide) orally (10 mg/kg) and/or melatonin (10 mg/kg) orally for 4 weeks. CMI was measured by delayed-type hypersensitivity (DTH), leucocyte and macrophage migration inhibition (LMI and MMI) responses and estimation of cytokines TNF-α and IFN-γ levels. Rats exposed to propoxur for 4 weeks showed significant decrease in DTH, LMI and MMI responses. Propoxur also suppressed TNF-α and IFN-γ production significantly. Administration of melatonin alone caused a significant increase in DTH response. Although there were no changes in the LMI and MMI response, the cytokine levels were significantly increased, as compared to control. Co-administration of melatonin along with propoxur significantly nullified the effect of the pesticide on the CMI response, except DTH and reversed levels of cytokines to near control/normal values. Thus, melatonin treatment considerably attenuated immunomodulation caused by sub-chronic treatment of propoxur in experimental animals.

018498   Sudhir Kumar;Sangwan M L;Rupender (Animal Biotechnology Dep, College of Veterinary Sciences, CCS Haryana Agricultural University, Hisar-125 004) : Polymorphism in DRB3 exon 2 by PCR-RFLP and its association with mastitis in Nili-Ravi breed. Indian J Biotechnol 2008, 7(3), 398-400.

The investigation was undertaken to study the genetic polymorphism in exon 2 of DRB3 gene in Nili-Ravi (n=25) buffalo breed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and its association with mastitis. The gDNA was isolated from whole blood samples. When 304 bp PCR product of exon 2 of DRB3 gene was digested with Rsa1, 11 genotypes, viz., b/b, c/c, f/f, o/o, s/s, f/o, b/f, b/o, o/s, b/l and l/s, with frequency range 0.04-0.16 and 6 alleles, viz., b, c, f, l, o and s with frequency range 0.08-0.26 were observed. HaeIII detected 6 genotypes, viz., a/a, e/e, d/d, a/b, b/d and b/e with frequency range 0.04-0.28 and four alleles, viz., a, b, d and e with frequency range 0.08-0.6. However, Pst1 revealed 5 genotypes, viz., y/y, z/z, x/y, x/z, s/z and y/z with frequency range 0.08-0.32 and four alleles, viz., x, y, z, and s with frequency range 0.04-0.42. These results revealed that exon 2 of DRB3 gene was highly polymorphic in Nili-Ravi breed. Certain genotypes (c/c, f/l, b/f, o/o, l/s, a/a and e/e) were observed only in healthy animals, while others (b/f, b/o, f/o and y/z) in mastitis cases. Result to be tested on large sample size before its practical application.

018497   Srivastava R;Srivastava J;Behera S K;Khare P B (Pteridology Lab, National Botanical Research Institute, Lucknow-226 001) : In vitro studies on development of gametophyte, sex-ontogeny and reproductive biology of a threatened fern, Microsorium punctatum (L.) Copel.. Indian J Biotechnol 2008, 7(2), 266-9.

Studies on reproductive biology vis-a-vis conservation of threatened state of the species, Microsorium punctatum, is the major concern of the present study. The life cycle starting from spore germination, gametophyte growth and differentiation, sex ontogeny and success of sporophyte formation through intra-and inter-gametophytic selfing was scrutenised and reproductive biology of this species was completely studied. The result indicates that the species could moderately be a good colonizer, as considerable number of sporophytes was produced through selfing, but in contrast very few plants were observed in the area of its occurrence. The threatened state could be due to unfavourable conditions for gametophyte growth, mating, subsequent slow pace of sporophyte development, and both epiphytic and terrestrial habit of the species.

018496   Sreenivas Rao R;Jyothi C P;Venkateswar Rao L (Microbiology Dep, Osmania University, Hyderabad-500 007) : Biotechnological production of xylitol by mutant Candida tropicalis OMV5: Process optimization using statistical approach. Indian J Biotechnol 2008, 7(2), 218-24.

Orthogonal experimental design L₁₆ (2⁵) was used to investigate effects of key media components namely xylose, yeast extract, peptone, urea and inoculum size on the production of xylitol by a mutant strain of Candida tropicalis (CT-OMV5). Software for automatic design and analysis of the experiments, based on Taguchi approach was used. Optimal levels of key media components were also determined. Among the tested parameters, xylose and urea contributed higher influence and yeast extract concentration also played an important role in the conversion of xylose to xylitol. Application of Taguchi method helped in easy process optimization and higher xylitol yield. The increased yield was possible at lower levels of yeast extract and peptone. The yield of xylitol under these optimal conditions was 0.89g/g of xylose.

018495   Sood V, etal (Virology Lab, National Institute of Immunology, JNU Campus, New Delhi-110 067, Email: akhil@nii.res.in) : Host genes that affect progression of AIDS/HIV in India and novel gene therapeutic approaches against HIV. Indian J Biochem Biophys 2008, 45(3), 141-8.

A multitude of host and viral factors play critical role in susceptibility to HIV-1 infection and its subsequent progression to AIDS. Various host factors involved in HIV-1 infection include the chemokine receptors CCR5, CX3CR1, their ligands, RANTES, SDF-1 and cytokines like IL-10, IL-4, among others. The CCR5Δ32 allele is the most important genetic factor known to confer resistance to HIV-1 infection. However, other mutations in CCR5, CX3CR1 and SDF-1 have also been identified in Indian population. Polymorphisms in DC-SIGN, MHC class-I and II molecules are also known to affect HIV-1 progression. These polymorphisms can be utilized as genetic markers for evaluating disease progression and developing effective therapeutics. Also describes the development of anti-viral therapy, involving the use of catalytic nucleic acids like DNA-enzymes and ribozymes and the expression of ribozymes and si-RNA using lentiviral vectors for stem cell based anti-HIV therapy.

018494   Sirisha V L, etal. (Genetics Dep, Osmania University, Hyderabad-500 007) : Direct shoot organogenesis and plant regeneration from hypocotyl explants in selected genotypes of Leucaena leucocephala-a leguminous pulpwood tree. Indian J Biotechnol 2008, 7(3), 388-93.

Efficient in vitro plant regeneration system in subabul (Leucaena leucocephala), a leguminous pulp wood tree species, was established. The induction of shoots was achieved from selected elite clones of subabul K-8, K-636 and also wild type on MS medium supplemented with 2% sucrose and different concentrations (0.88 to 24.6 μM) of plant growth regulators (BA, Kn, 2iP & TDZ). The best medium for shoot regeneration was MS with 22.2 μM BA (5 shoots per explant), followed by 22.7 μM TDZ (4.6 shoots per explant). Addition of putriscine (9.3 μM) to MS medium containing 22.2 μM BA enhanced the number of multiple shoots to 7-8 but not the frequency of response. Shoot initials (measuring 1 cm) when separated and transferred on to MS medium containing 1.4 μM GA3 elongated to 2-5 cm in 15-20 d with 80% frequency. The per cent frequency of shoot differentiation was almost identical in the genotypes K-8 and K-636 but it differed significantly from the wild type. Leaf yellowing and abscission in all the genotypes was curtailed by supplementing the medium with 685 μM glutamine or 540 μM adenine. The excised shoots were transferred to root regeneration media containing 2.46 and 4.98 μM IBA or 2.6 and 5.3 μM NAA. Root regeneration was noticed with 100% frequency in all the three genotypes in presence of IBA or NAA. Plantlets were transferred successfully to the pots with 70% survival rate with no visible morphological variations. The protocol can be utilized for mass propagation and genetic transformation studies of this important pulpwood species.

018493   Singh M P;Srivastava A K;Viswakarma S K; Pandey V K;Pandey A K;Singh S K (Dep of Biotechnology, V.B.S. Purvanchal University, Jaunpur-222 001) : Extracellular enzymatic activities by Pleurotus species on vegetable wastes. Mushroom Res 2007, 16(2), 93-7.

Extracellular enzymes produced by three species of Pleurotus i.e. P. fossulatus, P. roseus and P. sapidus on two vegetable wastes - cauliflower and radish wastes revealed that laccase and polyphenol oxidase (PPO) appeared and peaked earlier than cellulases and xylanase. Cauliflower wastes supported maximum production of cellulases, xylanase, laccase and PPO activity was exhibited by P. sapidus.

3 tables, 24 ref

018492   Singh A K;Sharma D R;Singh R K (Dep of Biotechnology, Dr. Y S Pharmar University of Horticulture and Forestry, Nauni, solan-173 230) : In vitro shoot regeneration from water stress tolerant callus culture of tomato (Lycopersicon esculentum Mill.). Pl Cell Biotechnol molec Biol 2007, 8(1-2), 79-84.

Protocol was developed for in vitro shoot regeneration from water stress tolerant callus culture of tomato cv. solan vajar on MS medium supplemented with different concentrations and combinations of various auxins and cytokinins along with 100g/l polyethylene glycol (PEG-6000). Callus cultures of tomato were initiated from hypocotyl segment on MS medium supplemented with BAP (3mg/l) and NAA (0.5mg/l). cell clumps of about 1mm in diameter were exposed to increasing concentration of polyethylene glycol ranging from 10g/l to 100g/l for water stress tolerance. Upon incubation for 40 days, the cells, which could tolerate this concentration of PEG, grew to form cell clones. Selected clones were successfully subcultured on the 100g/l PEG selected medium for 6 weeks and then transferred to the normal MS medium. The selected calli when transferred from normal to the selective medium, were capable of gowing on it. The above selected calli were used for shoot morphogenesis. Frequency of shoot bud regeneration varied with hormonal concentrations and combinations in the medium. Among the combinations used, BAP (2mg/l) and IAA (1mg/l) combination was found to be best for higher frequency of shoot regeneration. Rooting of the regenerated shoot was found to be higher in MS medium supplemented with (0.1mg/l) combination was found to be best for higher frequency of shoot regeneration. Rooting of the regenerated shoot was found to be higher in MS medium supplemented with (0.1mg/l) NAA.

2 tables, 17 ref

018491   Sikka P;Sethi R K (NO, Central Institute for Research on Buffaloes, Sirsa Road, Hisar-125 001) : Genetic variability in production performance of Murrah buffaloes (Bubalus bubalis) using microsatellite polymorphism. Indian J Biotechnol 2008, 7(1), 103-7.

Sequence specific primers for microsattelite ETH131 from cattle were used to polymerise DNA using buffaloes' genomic DNA as the template. In light of the reported sequence similarity of 78-95% between cattle and buffalo, this anonymous repeat sequence, isolated from cattle cDNA library and associated to milk, was analysed in Murrah buffaloes. The extent of polymorphism revealed in buffaloes was less than that previously reported in cattle, in terms of number of alleles. Cluster analysis performed on the basis of sharing of specific lengths of repeated nucleotides (NTSysPC) showed DNA similarities of the order of over 91% in animals of alike performance as per production data records. A 17>37> DNA dissimilarity was observed in buffalo's DNA band length patterns of low and high production sub-groups in this study. Attempts have been made to associate the distinctive amplification profile identified genotypic variation with the high and low milk producing Murrah buffaloes. Inferences suggest that ETH131 amplification mediated genotyping has scope of differentiating characteristics of individual animals and utilization of this genomic sequence for early selection of better performing buffaloes.

018490   Shivaraj A;Madhusudan S;Ashok Kumar C;Isloor S;Gowda V;Dechamma H J;Reddy G R;Suryanarayana V V S (Molecular Virology Lab, Indian Veterinary Research Institute, Hebbal, Bangalore-560 024) : Amplification, cloning and sequencing of Enterococcus feacalis enolase gene. Indian J Biotechnol 2008, 7(3), 307-12.

α-Enolase, a key glycolytic enzyme, belongs to a novel class of surface proteins, which do not possess classical machinery for surface transport and transported on the cell surface through an unknown mechanism. It is a multifunctional protein and its ability to serve as a plasminogen receptor on the surface of a variety of hematopoetic, epithelial and endothelial cells suggest that it may play an important role in the intravascular and pericellular fibrinolytic system. Authors have amplified and cloned α-enolase gene of Enterococcus feacalis in a prokaryotic cloning vector, and then transferred it into Escherichia coli. The recombinant enolase vector (r-pBEnol) was isolated and sequenced. The sequence of the cloned enolase from E. feacalis was found identical to that of the E. feacalis V583. The sequence was submitted to NCBI nucleotide data bank and accession number (AM279410) was obtained.

018489   Sharma R K;Gupta V K;Jindal J;Dilawari V K (Insect Molecular Biology Lab, Entomology Dep, Punjab Agricultural University, Ludhiana-141 004) : Host associated genetic variations in whitefly, Bemisia tabaci (Genn.). Indian J Biotechnol 2008, 7(3), 366-70.

Genetic variability due to host plants was studied in whitefly, Bemisia tabaci (Genn.), populations that were collected from fields of different crops (cotton, brinjal, potato, tomato and soyabean) and a weed (Sida sp.), and maintained on their respective host plants for 12 generations. Comparative RAPD-PCR analysis of these populations led to identification of 85 different polymorphic bands or host specific markers. Of these, 39 markers were identified for single-host specificity; maximum markers were identified for tomato (14) and cotton (13); followed by brinjal (5), Sida sp. (4) and soyabean populations of whitefly (3). Similarly, of 23 two-host specific markers, maximum markers were identified in cotton (10) and tomato (11) whitefly. This is the first report establishing the existence of host-plant specific genotypes in B. tabaci that is likely to have broad impact on its pest status and vectoring ability for different geminiviral diseases. The genetic similarity dendrogram based upon the comparative RAPD profiles showed the existence of a high level of genetic relatedness (72-85%) amongst the investigated whitefly types and the lineage of their origin from a common type. This lineage suggests that the whitefly types holding specificity for different host plants under study have evolved as three distinct genetic groups formed by cotton, Sida and soyabean (Group 1); potato and brinjal (Group 2); and tomato (Group 3).

018488   Sharma N;Gautam N (Basic Science Dep, Dr YS Parmar University of Horticulture and Forestry, Nauni, Solan-173 230) : Antibacterial activity and characterization of bacteriocin of Bacillus mycoides isolated from whey. Indian J Biotechnol 2008, 7(1), 117-21.

Bacteriocin produced by Bacillus mycoides isolated from whey showed strong inhibition against food borne serious pathogens, Listeria monocytogenes and Leuconostoc mesenteroides. Antibacterial substance was partially purified by salt saturation method. Partially purified bacteriocin withstood temperature up to 100°C, found active at wider pH range (4 to 11) and was sensitive to trypsin. This inhibitory substance showed bactericidal effect against bacteriocin-sensitive indicators. Thus, on the basis of above mentioned characters, the antibacterial substance produced by B. mycoides is most probably a potent bacteriocin and can be used for biological preservation of food.

018487   Sharma M;Kumar V;Pundir C S (Biochemistry Research Lab, M D University, Rohtak-124 001) : Immobilization of porcine pancreas lipase onto free and affixed arylamine glass beads and its application in removal of oil stains. Indian J Biotechnol 2008, 7(3), 328-32.

Porcine pancreas lipase has been immobilized through diazotization onto free and affixed arylamine glass beads with 75.59% and 54.26% retention of the initial activity of free enzyme and conjugation yield of 16 mg g^-1 and 7 mg g^-1, respectively. Optimum pH of the enzyme was decreased on free glass beads but increased on affixed glass beads. Optimum temperature, energy of activation (Ea), time of incubation and Km for triolein were increased but Vmax remained almost unchanged after immobilization on both free and affixed arylamine glass beads. The utility of immobilized enzyme in the removal of oil stains from cotton cloth by various detergents was tested by chemical method. All the detergents gave better wash performance in the presence of immobilized lipase (both onto free as well affixed glass beads) than that by detergent alone. Furthermore, the washing by cheaper (non-enzymic) detergents in the presence of immobilized lipase was almost similar to that by expensive (enzymic) detergents. The free and affixed bead-bound enzyme could be used repeatedly about 100 times without any considerable loss of activity.

018486   Shah M R;Kamble S;Indurker S;Eapen S (Plant Biotechnology and Secondary Products Section, Nuclear Agricultur, Bhabha Atomic Research Centre, Trombay, Mumbai-400 085, Email: eapenhome@yahoo.com) : Advances in genetic transformation of leguminous crops. Pl Cell Biotechnol molec Biol 2007, 8(1-2), 1-18.

Crop legumes play a pivotal role as the major source of protein in the diet of the population of developing countries, while some crop legumes like peanut and soybean also serve as a source of high quality oil. Although several transgenic crops have been grown over several million hectares in the world, crop legumes are not yet commercially exploited using transgenic tools for improving the food security of the developing countries except soybean. Production of crop legumes is greatly hampered by several biotic and abiotic stresses and transgenic technology can be used for the improvement of these traits. This review summarizes the advances made in genetic transformation of "crop legumes" and the future prospects.

3 tables, 199 ref

018485   Sengupta P;Meena K;Mukherjee R;Jain S K; Maithal K (NO, Dabur Research Foundation, 22, Site IV, Sahibabad, Ghaziabad, Uttar Pradesh-201 010, Email: maithalk@rediffmail.com ) : Optimized conditions for high-level expression and purification of recombinant human interleukin-2 in E. coli. Indian J Biochem Biophys 2008, 45(2), 91-7.

Interleukin-2 (IL-2), a potent cytokine has been used in anti-cancer therapy for over a decade now. IL-2, originally identified as a growth factor for T lymphocytes is a 15 kDa hydrophobic glycoprotein that induces the activation, clonal proliferation and differentiation of T and B-lymphocytes and enhances the cytotoxicity of monocytes and natural killer (NK) cells. Here, we report a simple method for the cloning, high-level expression and purification of IL-2 protein, which can be easily extended to other bioactive therapeutic proteins. The IL-2 gene was amplified from human spleen cDNA and cloned in a prokaryotic (E. coli) expression system. An optimal expression of the IL-2 protein was determined by varying the expression conditions like temperature, inducer concentration and duration of induction. The protein was expressed as inclusion bodies and a panel of reagents including detergents, urea and guanidine hydrochloride were used to solubilize it. After solubilization, the protein was renatured and subjected to a single step gel-filtration chromatography to yield immuno-bioactive IL-2 protein with <62>99% purity.

018484   Sathyanarayana N;Rajesha R;Vikas P B;Bharath Kumar T N (Bio-Technology Dep, Sir M Visvesvaraya Institute of Technology, Bangalore-562 157) : Somatic embryogenesis and plant regeneration from stem explants of Leptadenia reticulata (Retz.) Wight. & Arn.. Indian J Biotechnol 2008, 7(2), 250-4.

Somatic embryogenesis and plant regeneration from stem explants of Leptadenia reticulata (Asclepeadaceae), an endangered medicinal plant possessing galactogogue property and rich alkaloid content, have been reported. Murashige and Skoog's medium supplemented with MS + 3% sucrose + NAA (2.68 μM) + BAP (4.40 μM) was the best for callus formation as well as pre-embryonic mass (PEM) induction. Of the two subculture conditions tested for proper embryo induction, MS liquid medium proved superior over solid culture medium in inducing healthy embryos. Presence of NAA in the induction medium was also found to be critical for PEM formation, as the callus raised without it could not be triggered for embryogenesis in any of the subsequent subcultures. The resulting embryoids on transferring to Murashige and Skoog's basal as well as other cytokinin containing medium attained maturation with varied frequencies, of which MS basal medium triggered maximum response compared to other cytokinin containing medium. The so-developed shoots on transferring to half strength MS + IBA (4. 90 μM) developed vigorous tap root system, which were later hardened on peat mass mixture with 75% survival.

018483   Saraswathy N;Nain V;Sushmita K;Ananda Kumar P (National Research Center on Plant Biotechnology, Indian Agricultural Research Institute, New Delhi-110 012) : Fusion gene encoding two different insecticidal proteins of Bacillus thuringiensis. Indian J Biotechnol 2008, 7(2), 204-9.

Chimeric fusion gene was constructed with the coding regions of insecticidal crystal protein (Cry1Ac) and vegetative insecticidal protein (Vip3Aa14) of Bacillus thuringiensis (Bt). Overexpression of the fusion gene in Escherichia coli resulted in the synthesis of a protein of <126>140 kDa size, as revealed by SDS-PAGE analysis. Western hybridization analysis using polyclonal antisera raised against Cry1Ac showed the presence of a band corresponding to <126>140 kDa. Stability of the fusion protein was studied by trypsin digestion. Insect bioassays of the fusion protein on three insect species viz., Helicoverpa armigera, Spodoptera litura and Plutella xylostella showed that the fusion protein retained the toxicity of Cry1Ac, but partially lost that of Vip3Aa14.

018482   Saisivam S;Bhikshapathi D V R N;Krishnaveni J;Kishan V (NO, K M College of Pharmacy, Madurai-62 5107) : Isolation of borrelidin from Streptomyces californicus-an Indian soil isolate. Indian J Biotechnol 2008, 7(3), 349-55.

In a screening programme for search of antibiotics active against clinical resistant strains of infectious bacteria, a potent antibiotic producer from soil capable of acting on vancomycin and methicillin resistant strains was isolated. The taxonomic and biochemical studies revealed that the culture was a strain of Streptomyces californicus. The fermentation schedule was determined in starch casein agar medium and was found to produce maximum antibiotic on 3rd d. The biological activities of the crude extract revealed that the active compound has both antibacterial and antifungal activity. The cytotoxic studies on HBL 100 cancer cells revealed weak activity of the compound in comparison to doxorubicin. The active compound was further found to inhibit the neoangiogenesis in fertile eggs. The active compound was extracted from culture filtrates by ethyl acetate and purified by column chromatography using silica gel. Mass and 1H and 13C NMR spectral information of the active compound were obtained. Finally, the active compound was identified as borrelidin, being reported as new compound from S. californicus.

018481   Sahoo S;Kashyap V K (National DNA Analysis Centre, Central Forensic Science Laboratory, 30 Gorachand Road, Kolkata) : Validation of a single tube fluorescent multiplex assay for simultaneous typing of 20 Y-STR loci. Indian J Biotechnol 2008, 7(1), 41-9.

Describes a single tube assay for 20 fluorescent labelled Y-STR loci, which is rapid and robust state-of-art multiplex system. Reaction conditions, including annealing temperature, concentration of primers, magnesium and template DNA, DNA polymerase and reaction volume, were optimised to yield robust amplification. The assay could withstand moderate fluctuations in reaction parameters with no affect on haplotype analysis. Sensitivity and robustness of the multiplex system is revealed from the examination of different matrices and environmental conditions wherein complete haplotypes could be obtained in all the samples with no failures except those exposed to severe environmental stresses. Genotyping of the 20 Y-STR loci showed that all loci included in the multiplex are highly polymorphic in the Indian populations and its inclusion increases the discriminatory power of the marker system. It could be summarized that the developed Y-STR multiplex assay is a simple, sensitive and robust system highly suitable for concurrent analysis of 20 polymorphic Y-STR loci.

018480   Roychoudhury J;Ali N (Infectious Diseases and Immunology Div, Indian Institute of Chemical Biology, 4 Raja S. C. Mullick Road, Kolkata-700 032, Email: nali@iicb.res.in) : Sodium Stibogluconate: Therapeutic use in the Management of Leishmaniasis. Indian J Biochem Biophys 2008, 45(1), 16-22.

Leishmaniasis causes significant morbidity and mortality worldwide. The disease is endemic in developing countries of tropical regions, and in recent years economic globalization and increased travel has also spread to people in developed countries. In the absence of effective vaccines and vector-control measures, the main line of defense against the disease is chemotherapy. Organic pentavalent antimonials, including sodium stibogluconate have been the first-line drug for the treatment of leishmaniasis for the last several decades, and clinical resistance to these drugs has emerged as a primary obstacle to successful treatment and control. The present review describes the structure, activity, mode of action of sodium stibogluconate and mechanism of resistance towards this drug in leishmaniasis.

018479   Roy U;Shalini R;Vanitha S V;Saha S K; Srivastava R C (Biological Sciences Group, Birla Institute of Technology and Science-Pilani, Goa Campus, Goa-403 726) : Preliminary study of smelling agents using electrical potential oscillations at liquid-liquid interface. Indian J Biotechnol 2008, 7(1), 73-82.

Aims to study the complex oscillations at liquid-liquid interface while mimicking sensing mechanisms of smell-oscillations of electrical potential differences across a bipolar liquid membrane induced by different classes of olfactory agents, e.g. amines, alcohols, acids, aldehydes and esters, in vitro. A preliminary attempt was made to classify and quantify various smelling agents and thereby developing a smell sensor. The bipolarity was induced by introducing a cationic (cetyl pyridinium chloride) and anionic surfactant (sodium lauryl sulphate) along with the electrolytes like sodium and potassium chloride in the experimental set-up. The data obtained indicate that olfactory agents of different groups exhibit characteristically different frequency and amplitude.

018478   Rathore P;Suthar R;Purohit S D (Plant Biotechnology Lab, Botany Dep, Mohanlal Sukhadia University, Udaipur-313 001) : Micropropagation of Terminalia bellerica Roxb. from juvenile explants. Indian J Biotechnol 2008, 7(2), 246-9.

In vitro micropropagation system has been developed for Terminalia bellerica Roxb., an important Indian medicinal plant. Nodal segments obtained from 15-d-old aseptically grown seedlings were used as explants. MS medium containing 1.5 mg L^-1 BAP was found most suitable for culture initiation. Although shoot multiplication was achieved on MS medium containing BAP and Kn, the maximum number of shoots was obtained with 1.5 mg L^-1 BAP. Best rooting response (60%) was observed on medium containing quarter strength MS salts, 0.6% agar and 0.1 mg L^-1 IBA. Plantlets were hardened initially in culture room conditions and then transferred to misthouse.

018477   Ramachandran G;Ananthanarayan L (Food Engineering and Technology Dep, Institute of Chemical Technology (Formerly UDCT), University of Mumbai, Matunga, Mumbai-400 019) : Optical resolution of (R, S)-ibuprofen in organic solvent by porcine pancreatic lipase catalyzed enantioselective esterification. Indian J Biotechnol 2008, 7(1), 94-8.

Porcine pancreatic lipase (PPL), a comparatively inexpensive enzyme, was used in the optical resolution of (R, S)-ibuprofen in methanol through enantioselective esterification. The different reaction parameters during the esterification of (R, S)-ibuprofen using PPL was studied with respect to temperature, time, enzyme concentration, substrate concentration and water content. The percentage conversion initially was 13%, which increased to 33% after optimization of the different reaction conditions. Surfactant coating of PPL resulted in a 2.5-fold increase in the hydrolytic activity of PPL. Surfactant coated PPL, when used in the esterification reaction slightly increased the percentage conversion from 21 to 22.5%. In all the reactions, PPL showed preference for catalyzing the esterification of S-(+)-ibuprofen.

018476   Rajendran C;Ray D D;Bansal G C (Parasitology Div, Indian Veterinary Research Institute, Izatnagar-243 122) : Expression of gene encoding immunodominant merozoite surface protein of Theileria annulata in Escherichia coli. Indian J Biotechnol 2008, 7(2), 200-3.

The asexual blood stage "merozoites" of an Indian strain of the tick-borne cattle haemoprotozoa, Theileria annulata, was generated in in vitro culture and the gene encoding the merozoite surface protein (Tams 1) was amplified from cDNA by using primers designed from T. annulata (Ankara strain). The amplified gene was cloned into pPROExHT b plasmid vector and expressed as fusion protein in Escherichia coli, DH5α strain. The recombinant protein was purified using NI-NTA affinity chromatography and used to detect antibody response in known anti-Tannulata bovine serum samples in a ELISA format.

018475   Raizunnisa K;Sai Krishna T;Schansker G; Strasser R J;Raghavendra A S;Mohanty P (Plant Sciences Dep, School of Life Sciences, University of Hyderabad, Hyderabad-500 046, Email: prasanna37@hotmail.com) : Application of fast chlorophyll a fluorescence transient (OJIP) analysis to monitor functional integrity of pea (Pisum sativum) mesophyll protoplasts during isolation. Indian J Biochem Biophys 2008, 45(1), 37-43.

Intact and metabolically very active mesophyll protoplasts were isolated rapidly from pea (Pisum sativum) leaves. The functional performance of protoplasts at various stages of their isolation was analyzed by using fast Chl fluorescence OJIP transients and compared with that of intact leaves. The results demonstrated that the OJIP transients could successfully be used to monitor the quality of mesophyll protoplasts at different isolation steps. The protoplasts maintained their integrity and photosynthetic status very well, and their performance was very similar to that of the intact leaves.

018474   Qader S A U;Aman A;Bano S;Syed N;Azhar A (NO, Pharmaceutical Research Center, PCSIR Laboratories Complex, Karachi, Pakistan) : Role of calcium ions and temperature on dextransucrase production. Indian J Biotechnol 2008, 7(3), 404-6.

Dextransucrase [E.C. 2.4.1.5] enzyme from newly isolated strain of Leuconostoc mesenteroides PCSIR-4 showed maximum production in the fermentation medium containing 0.005% CaCl₂. The production of dextransucrase was 2.5 times higher in the medium containing calcium ions as compared to the medium without calcium ions. Also, the stability of the enzyme increased up to120 d. Maximum dextransucrase production was obtained when culture was incubated at 25°C, while extracellular enzyme activity at 35°C. Thermal stability of extracellular enzyme was maximum at 30°C and at 50°C the total enzyme loss was in 240 min.

018473   Prema S;Sivaselvam S N;Karthickeyan S M K (Animal Genetics and Breeding Dep, Madras Veterinary College, Chennai-600 007) : Evaluation of genetic diversity in Mecheri sheep (Ovis aries) of Tamil Nadu using microsatellite markers. Indian J Biotechnol 2008, 7(3), 401-3.

Evaluation of genetic variation was carried out using microsatellite markers in 48 Mecheri sheep of Tamil Nadu as a part of genetic characterisation and conservation. The number of observed alleles ranged from 3 to 8 with a mean of 5 across all loci. The size of alleles ranged from 74 to 224 bp. The frequency of alleles ranged from 0.0208 to 0.6250. The polymorphism information content (PIC) values varied from 0.52 to 0.79 with a mean of 0.66. The population was not in Hardy-Weinberg equilibrium. The overall mean expected heterozygosity was 0.71. The inbreeding estimate within-population was 0.004, indicating excess of heterozygotes in the population of Mecheri sheep. The panel of microsatellites used was highly informative for molecular characterisation and could be used for exploitation of genetic diversity of the related breeds for conservation.

018472   Pramitha V S;Lipton A P;Thangaraj M (Marine Biotechnology Lab, Vizhinjam Research Centre of Central Marine Fisheries Research Institu, Vizhinjam-695 521) : Evaluation of α- and β-glucosidase inhibitory properties of macro-algae using intestinal extracts of marine snail, Thais rudolphi (Lamarck, 1822). Indian J Biotechnol 2008, 7(1), 61-5.

Glucosidase-inhibitory activity of marine macro-algae, Ulva fasciata and Hypnea musciformis in different solvents was detected using enzyme-agar plate method. Eenzyme-agar plate containing α- or β-D-glucosidase was used in combination with substrate-agar plate containing p-nitro phenyl α- and β-D-glucopyranoside (PNPG). Discs impregnated with algal extracts placed on the enzyme agar, followed by pre-incubation up to 2.5 h at 25°C, resulted in the characteristic inhibitory circle around the inhibitor. Extracts of red algae, H. musciformis had higher glucosidase-inhibition than the extracts of U. fasciata. The intestine of Thais rudolphi contained both α- and β-glucosidase enzymes and their inhibition was comparable with the commercial enzymes, suggesting the possible alternate method for assessing the glucosidase-inhibitory activity of the marine macro-algae.

018471   Ponnuvel K M;Ashok Kumar K;Velu D; Somasundaram P;Sinha R K;Kamble C K (Biotechnology Lab, Central Sericulture Germplasm Resources Centre, Thally Road, Hosur-635 109) : Characterization and genomic organization of esterase gene in silkworm, Bombyx mori L.. Indian J Biotechnol 2008, 7(2), 183-7.

N-terminal amino acid sequence of esterase gene was blast searched with Bombyx mori EST (expressed sequence tag) database. EST clone fbpv0006 showed 95% homology with N-terminal sequence of esterase protein. Further, the genomic organization of exons and introns were identified using the EST clone sequence. The results on genomic organization of esterase gene indicated that the blood esterase gene possesses two exons with varying length of 192 and 524 bp, and a long intron of 2124 bp length present between these two exons. The primers were designed to the intron and exon regions and the fragments were PCR amplified using genomic DNA from silk moth as template. The amplicon showed a mol wt 799 bp in the intron region and 547 bp covering the exon regions. When the esterase genes from two multivoltine (Pure Mysore & PMX) and bivoltine (NB4D2 & CSR-19) races, possessing contrasting characters to thermal tolerance, were PCR amplified and products were sequenced. The two sequences showed 97% homology with 3% mismatch through pair blast analysis. Furthermore, per cent identity per exon, number of gaps per exon, overall per cent identity, per cent coverage of the mRNA, number of splice donor sites, presence or absence of splice donor and acceptor sites for each exon were inferred through spidey programme. Also, the phylogenetic relationship of B. mori esterase gene sequence was compared with other organisms through Clustal W analysis and a dendogram was projected. The projection showed that majority of insect esterases formed a major separate cluster, while B. mori esterase clustered with aphid insect esterases.

018470   Pattnaik P;Sekhar K (NO, Defence Research and Development Establishment, DRDO, Jhansi Road, Gwalior-474 002) : Forensics for tracing microbial signatures: Biodefence perspective and preparedness for the unforeseen. Indian J Biotechnol 2008, 7(1), 23-31.

Biological weapons are assigned high priority in homeland security, defence, counterproliferation, nonproliferation, intelligence, and counterterrorist programmes. In order to strengthen active defense against development and use of these weapons, several comprehensive technological and forensic capabilities have been developed world over for investigative, intelligence, prosecutive, diplomatic and policy purposes. Microbial forensics is one of such strategies. It is a new discipline combining microbiology and forensic science. Microbial forensics associate the source of the causative agent with a specific individual or group by measuring molecular variations between related microbial strains. In this context, several advanced molecular techniques and practices including molecular phylogeny, whole genome sequencing, microarray analysis, DNA finger printing, etc., can be extremely valuable and effective analytical tools in a microbial forensic investigation. Results from such analyses may be related to the intentional use of microbial agents for bioterrorism or the accidental release of any offensive microorganisms or toxins of public health significance, specifically for the purpose of determining the origin. The new discipline of microbial forensics is an integration of an array of well established fields, such as microbial genomics, phylogenetics, forensic informatics and classical microbiology.

018469   Parab G V;Krishnan S (Botany Dep, Goa University, Goa-403 206) : Assessment of genetic variation among populations of Rhynchostylis retusa, an epiphytic orchid from Goa, India using ISSR and RAPD markers. Indian J Biotechnol 2008, 7(3), 313-9.

Rhynchostylis retusa (L.) Bl., a monopodial epiphytic orchid species with attractive flowers arranged in racemose inflorescence, ranks among the important Indian ornamental orchids. Comparative population studies using PCR based markers, RAPD and ISSR, were performed to assess the genetic diversity of the wild orchid. Among the 35 primers tested, 13 RAPD and 7 ISSR primers were selected for the analysis. In total, 74 RAPD and 30 ISSR fragments were generated. High level of polymorphism was recorded in RAPD (76.13%) than ISSR (62.6%). In case of RAPD, Nei's average genetic identities value for different populations of R. retusa ranged from 0.405 to 0.932. While for ISSR, it ranged from 0.733 to 0.933. The results of the present study can be seen as starting point for future research on the population and evolutionary genetics of this species.

018468   Pandey S;Malik S;Sharma S;Sharma M (Biotechnology Div, Institute of Himalayan Bioresource Technology, Palampur-176 061) : Studies on differential growth behaviour of two alpine herbs of Western Himalaya from different altitudes under in vitro conditions. Indian J Biotechnol 2008, 7(1), 137-40.

Inherent slow growth of alpine plant species as compared to temperate ones has consistently been observed even under in vitro conditions and considered as major bottleneck for rapid mass multiplication. Keeping this in view, different growth parameters including relative growth rate (RGR), specific leaf area (SLA), shoot mass fraction (SMF), leaf mass fraction (LMF), root mass fraction (RMF), respiration rate, carbohydrates, nitrogen content and nitrate reductase (NR) activity of different populations of Picrorhiza kurroa and Rheum emodi were studied under in vitro conditions. Gives an insight that regulation of growth in alpine plant species is genetic and it is strongly related with SLA and leaf thickness.

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018467   Nirmal N;Om Praba G;Velmurugan D (Center of Advanced Study in Crystallography and Biophysics, University of Madras, Guindy Campus, Chennai-600 025, Email: d_velu@yahoo.com) : Modeling studies on phospholipase A₂-inhibitor complexes. Indian J Biochem Biophys 2008, 45(4), 256-62.

Phospholipase A₂ (PLA₂) is a ubiquitous enzyme that specifically catalyzes hydrolysis of membrane phospholipids to produce lysophospholipids and free fatty acid, namely arachidonic acid, which provides substrate for eicosanoids biosynthesis. Thus, the compounds inhibiting PLA₂ have been implicated as potential therapeutic agents in treatment of inflammation related diseases. Plant and marine organisms serve as sources of compounds that act as potential therapeutic agents for treatment of various diseases. The present study reveals the relationship between the structure and function of the medicinally important herbal compounds (acalyphin, chlorogenic acid, stigmasterol, curcumin and tectoridin) and marine compounds (gracilin A and aplysulphurin A). To understand the binding mechanisms of these compounds, molecular modeling studies has been performed with Russell's viper and bovine pancreatic PLA2 as target molecules using molecular operating environment (MOE) software. These compounds show favorable interactions with the amino acid residues at the active site of Russell's viper and bovine pancreatic PLA₂, thereby substantiating their proven efficacy as anti-inflammatory compounds and antidotes.

018466   Nikam T D;Savant R S;Pagare R S (Botany Dep, University of Pune, Ganeshkhind, Pune-411 007) : Micropropagation of Ceropegia hirsute Wt. & Arn.- A starchy tuberous asclepid. Indian J Biotechnol 2008, 7(1), 129-32.

Protocol is described for micropropagation of the starchy, tuberous, herbaceous twinner, Ceropegia hirsute Wt. & Arm. by in vitro culture of nodal segments. Of the cytokinins (BAP and Kn) and auxins (IAA, NAA, 2,4-D) evaluated as supplements individually and in combinations to Murashige and Skoog (MS) medium, BAP (7.5 μM) was the most effective in inducing axillary multiple shoots (5.7±0.7 shoots/culture). Root were induced to in vitro derived shoots on half-strength MS medium supplemented with IAA (2 μM) and sucrose (5%). in vitro plants were acclimatized and were successfully established in the soil.

018465   Narayanan C;Dubey S;Wali S A;Shukla N;Randhir Kumar;Mandal A K;Ansari S A (Genetics and Plant Propagation Div, Tropical Forest Research Institute, Jabalpur-482 021) : Comparative efficacy of different DNA extraction methods for PCR-based assay in Tectona grandis L.f.. Indian J Biotechnol 2008, 7(1), 133-6.

Four extraction methods and sample types were evaluated for yield, quality and suitability of genomic DNA for ISSR (inter simple sequence repeats) marker amplification by PCR in teak (Tectona grandis). Both CTAB (hexadecyltrimetylammonium bromide) and SDS (sodium dodecyl sulfate) procedures extracted large amounts of high purity genomic DNA. Leaves of trees, bud grafts and seedlings yielded better DNA than seeds. Using identical PCR conditions, DNA extraction methods and sample types affected amplifications of ISSR markers with a pattern: seedling <62> graft <62> tree leaves. Genomic DNA of seed samples contained high amount of protein and showed no ISSR amplifications, which were overcome by inclusion of proteinase K in SDS based extraction method. In conclusion, DNA extraction method and sample types are very important for reproducible ISSR-based molecular marker analysis in teak.

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018464   Narayanamurthy G;Ramachandra Y L;Padmalatha Rai S;Manohara Y N;Kavitha B T (Biotechnology Dep, Kuvempu University, Shimoga-577 451) : Areca husk: An inexpensive substrate for citric acid production by. Indian J Biotechnol 2008, 7(1), 99-102.

Areca husk was used as a substrate for the production of citric acid under solid state fermentation (SSF) using a new local soil isolate of Aspergillus niger. A. niger produced 119.42±2.5 g citric acid/kg dry areca husk fermented in the presence of 3% w/w methanol at optimum pH 5.0, 50% moisture content and 30°C incubation temperature in 3 d. The citric acid yield was 66.7±1% based on the amount of fermentable sugars consumed during fermentation.

018463   Naik R M (NO, , , Email: rajeevnaik2@rediffmail.com) : Mitochondrial complex I impairment and differential carbon monoxide sensitivity of cytochrome c oxidase in wild type and CMS II mutants of Nicotiana sylvestris. Indian J Biochem Biophys 2008, 45(2), 126-9.

Plant mitochondria unlike their animal counterpart have some unique features with highly branched respiratory chain. The present work was undertaken in order to investigate the effect of loss/dysfunction of plant mitochondrial complex I on the relative flux of electrons through alternative oxidase (AOX) and cytochrome oxidase. Loss of a major subunit of mitochondrial complex I in cytoplasmic male sterile II (CMS II) mutant of Nicotiana sylvestris caused respiratory redox perturbations, as evident from the differential CO sensitivity of cytochrome oxidase. The leaf segments of CMS II mutant when exposed to CO under dark aerobic condition were insensitive to the inhibition of cytochrome oxidase, as against the wild type (WT). The differential CO response of WT and CMS II mutants appeared to be due to differences in the redox state of cytochrome a₃ (cyt a₃), the terminal electron acceptor during in situ respiration. Cyt a₃ appeared to be more in its oxidized form in CMS II and hence unable to form cyt a₃-CO complex. Pre-treatment of CMS II leaves with 2,4-dinitrophenol, an uncoupler of oxidative phosphorylation increased the CO response. The slight increase in rotenone-insensitive respiration of CMS II could be attributed partly to enhanced flux of electrons through cytochrome pathway to compensate for the loss of phosphorylation site and partly through AOX, which was induced by nitrate.

018462   Nagaraju V (National Research Centre for Orchids, , Pakyong-737 106) : Effect of sucrose on proliferation and conservation of Cymbidium hybrids in vitro. J ornamental Hort 2007, 10(2), 71-7.

Responses of protocorm like bodies (PLB's) of Cymbidium hybrids HXB, Lunavian Atlas (LA) and BXH cultured on Murashige and Sloog's (MS) medium with socrose were observed two weeks after culture. Proliferation of PLB's recorded early on medium supplemented with 20 g/l followed by 35 g/l sucrose in BXH, while in LA and HXB it was recorded at 40 g/l sucrose. Sucrose at 10 and 15 g/l in the medium completely suppressed the growth followed by gradual discoloration and ultimate death of the explants in 6-9 months period, while sucrose at 40 g/l in the media resulted in enhanced culture weight and bigger sized PLB's. Maximum per cent dry weight was recorded at 60 g/l sucrose. Among the hybrids, maximum number of protocorm like bodies and shoots was recorded in Cymbidium cross BXH. Plantlets regenerated on medium supplemented with 20 to 60 g/l sucrose were maintaind in vitro for upto 15 months.

3 illus, 5 tables, 13 ref

018461   Munshi A;Duvvuri V S (Genetics Dep, Shadan P G Centre for Biosciences, Hyderabad-500 004) : Nutrigenomics: looking to DNA for nutrition advice. Indian J Biotechnol 2008, 7(1), 32-40.

With the success of 'Human Genome Project' and the powerful tools of molecular biology, we have entered the era of genetic nutrition. The publication of the human 'blue print' has triggered an explosion in pharmaceutical research to utilize this knowledge in prescription of drugs to be tailored according to the genetic make up of susceptible individuals or in other words personalized medicine. Propelled by the recent unraveling of human genome; nutritional sciences are discovering the application of the so-called "omics" sciences. This has the potential of identifying and validating targets to improve personalized nutritional health and thus serves to define the added value for the next generation of foods and the crops. The first new term to emerge in this area was "Nutrigenomics." Today, the term nutrigenomics generally refers to the study of how dietary components interact with the genome and modify subsequent gene expression1. It is envisaged that nutrigenomics will lead to evidence-based dietary interventions for prevention of diet related common diseases.