022661   Madhavilatha L;Singh N V (Horticulture Dep, Institute of Agricultural Sciences, Banaras Hindu Univ, Varanasi-221 005) : In Vitro colonal propagation of herbal medicinal plant Ashwagandha (Withania somnifera (L.) Dunal). Adv Pl Sci 2008, 21(1), 11-13.

Study was undertaken with a view to develop a reproducible protocol for in vitro propagation of ashwagandha Withania somnifera. Plantlets (ashwagandha)were produced in vitro from the leaf and nodal segments. Axillary leaves gave better callus formation with a combination of 2, 4-D (2mg 1^-1) and Kin (Img I^-1). Shoots differentiated best from leaves calli on MS medium containing BAP (2 mg I^-1) and IAA (0.5 mg I^-1). Shoot proliferation was best from nodal explants on MS medium supplemented with BAP (2mg l^-1) and IAA (lmg l^-1). Regenerated shoots rooted best on half strength MS medium containing IBA (2.0 mg l^-1). Pantlets [ were transferred to pots containing sand and soil mixture (1:1) for hardening. The acclimatized plantlets under controlled environmental conditions were transferred to the field.

3 tables, 7 ref

022660   Kumar J;Ver5ma V;Quazi G N;Gupta P K (Division of Biotechnology, Regional Research Laboratory, , Canal Road, Jammu-180 001, Email: jitendra_kumar_2k1@rediffmail.com) : Genetic diversity in Cymbopogon species using PCR-based functional markers. J Pl Biochem Biotechnol 2007, 16(2), 119-22.

Genetic diversity among 25 accessions (involving 8 species, 2 interspecific hybrids and one hybrid mutant) of medicinally important genus Cymbopogon was assessed using 17 PCR-based functional markers, that were designed from members of three different multigene families. We developed 16 primer pairs from two multigene families, 8 primer pairs each from cytochrome P450 and UDP-glucosyltransferase (UGT); one primer pair was derived from 58 rRNA gene family. A total of 119 fragments were visualized, of which 108 (91%) were polymorphic. The level of diversity among different taxa/accessions observed during the present study was, however, low relative to the diversity level obtained due to RAPD markers in two earlier studies. The pattern of genetic diversity neither matched with the known taxonomic classification, nor did it always match with the distribution of chemical constituents of the essential oils available in these accessions. Thus, present investigation though revealed poor correlation between the molecular and chemical diversity, these gene-based markers may prove useful in the development of perfect markers for association mapping of genes involved in controlling agronomically important traits.

^ssc1 illus, 2 tables, 10 ref

022659   Khanh T D;Xuan T D;Linh L H;Chung I M (Biotechnology Dep, Ruykyu Univ, Okinawa 903-0213, Japan, Email: imcim@konkuk.ac.kr) : Allelopathic plants: 18. passion fruit (Passiflora spp.). Allelopathy J 2008, 21(2), 199-206.

Passion fruit (Passiflora incarnata and Passiflora edulis) is medicinal plant and widely grown in tropical and subtropical countries for edible fruits and medicines. In bioassay and greenhouse studies, application of plant materials significantly inhibited the germination and growth of indicator plants and major paddy weeds. In field trial, plant materials applied at 1-2 ton ha^-1 reduced the initial growth of paddy weeds by 70% and increased rice yield by 30%. The allelopathic properties of plant parts followed as the order: Leaf > stem> root. Numerous chemical substances belonging to alkaloids, phenolics, flavonoids, and volatiles were found in these plants, of which kavain, yagonin, dihydromethysticin and coumarin completely suppressed the barnyardgrass growth at 500 mg L^-. Hence Passion fruit may be used as natural herbicide to reduce the dependency on synthetic herbicides and other agrochemicals.

4 illus, 2 tables, 36 ref

022658   Khan J;Deka M (Biotechnology Dep, Gauhati Univ, Guwahati, Assam, Email: d_bhaity@rediffmail.com) : Effect of UV irradiation on cellulase production by A. Fumigatus isolated from agriculture wastes. Asian J Microbiol Biotechnol envir Sci 2008, 10(1), 123-6.

The production of cellulases and β-glucosidase from Aspergillus fumigatus was investigated in shake flask cultures. These species were isolated from different Agricultural wastes. The enzymatic sacchairfication of cellulase has been proposed as a means of producing glucose on an industrial scale. In UV irradiated species it was possible to increase the production of β-glucosidase up to 80 folds in 60 minutes, 73 fold increases in filter paper activity, 50 fold increases in Cl and 20 fold increases in CMCase activity. UV exposure was given for 30, 60 and 90 minutes. The effect of irradiation on cellulase production was examined in Mandel and Sternberg medium (1976) which revealed that the 60 minutes of UV irradiation can cause significant increase in the production of cellulase which in turn increase the production of glucose-a product of interest to industries.

4 illus, 5 tables, 7 ref

022657   Henning S B;Paula Santos P M;Lovaglio R;Costa Siddhartha G V A O;Jonas C;Loshchagin P E (Biological Sciences Institute, Sao Paulo State Univ-Unesp, Brazil, Email: jconti@rc.unesp.br ) : Factors affecting rhamnolipids production by Pseudomonas Aeruginosa LBI. Res J Biotechnol 2008, 3(1), 45-9.

This paper presents the results from study investigating the capacity of Pseudomonas aeruginosa LBI growing on several carbon (Hydrophilic substrate) and nitrogen sources. The carbon source (2%) studied were: glycerol, manitol, fructose, glucose, lactose and the nitrogen sources (2; 3; 4 and 5%) were: NaNO₃, NH₄NO₃, (NH₄)₄O₄ and (NH₄)₄CO. At the 96 h of fermentation, the medium with glycerol as a carbon source reached 7.9 g/L of rhamnolipids and 1.2 g/L cellular protein. The surface tension reduction was 38.46 % for glycerol as a carbon source. The NaNO₃ at 3% was the best concentration for rhamnolipids production (7.35 g/L) and cellular protein (1.12 g/L). The influences of metal ions [FeSO₄.7H₂O (0.001 g/L, 0.005 g/L and 0.1 g/L) and MgSO₄.7H₂O (0.001 g/L, 0.005 g/L and 0.1 g/L)] on ramnolipids production were studied. Fe²⁺ had a negative influence on the studied concentrations while Mg²⁺ had a positive influence when its concentration was increased.

5 illus, 29 ref

022656   Gurvinder Kaur;Mahal G S;Bains N S (Plant Breeding Dep, Genetics & Biotechnology Punjab Agricultural Univ, Ludhiana-141 004) : Improvement for productivity traits in durum wheat with introgression Triticum dicuccoides. Crop Improv 2007, 34(2), 124-7.

With the objective of enhancing productivity, introgression lines were generated in background of durum wheat cultivars PDW 233 and PDW 274 from crosses with five Triticum dicoccoides and five Triticum dicoccum accessions. During early generations single plant selection was performed for yield and its components followed by a replicated trial (BC₂F₄ stage). Selected high yielding progenies were purified and advanced to BC₂F₅ (two crosses) or BC₂F₇ (eight crosses) and a total of ninety six lines were evaluated for key productivity traits including days to flowering, grain filling period, plant height, tillers per plant, number of grains per spike, grain weight per spike, 1000-grain weight, grain yield per plot and harvest index. Forty-nine progenies showed superiority over recurrent parent for grain yield. Yield improvement could be achieved without loss of local adaptation as reflected by retention of recurrent parent's height and maturity in several hight yielding lines. Some advanced backcross progenies with improved yield attributes have been identified.

2 tables, 10 ref

022655   Gurnoor K;Verma N;Wheatley Denys N (Biosenor Development Laboratory, Biotechnology Dep, Punjabi Univ, Patiala-147 002, Email: gurnoorbiotech@yahoo.co.in) : Comparison of various cell disruption techniques for the extraction of Arginase from Neurospora crassa. Res J Biotechnol 2008, 3(1), 39-44.

The filamentous fungi, owing to their ability to secrete high levels of proteins, are attractive organisms for the secretion of heterologous proteins of commercial importance and medical value. Arginase is one such enzyme having immense therapeutic potential. The present investigation was carried out with the purpose of studying the effect of three different (physical and chemical) intracellular methods of cell disruption and enzyme extraction on the activity of the enzyme arginase using Neurospora crassa MTCC-858 [FGSC: 746{77:25C}] as the microbial source of the enzyme. In Neurospora crassa and other microbial organisms that do not have a urea cycle, the products of arginase, ornithine and urea allow utilization of arginine as a nitrogen source. Also in the present study, the growth pattern of the fungus was determined by employing two methods (dry weight method & absorbance method) and a comparison between both the methods of growth determination was made for the very first time.

6 illus, 1 table, 24 ref

022654   Goyali J C;Dhindsa G S;Sharma A (Plant Breeding Dep, Genetics and Biotechnology Punjab Agricultural Univ, Ludhiana-141 004) : Genetic parameters, genotype x environmental interactions, correlation and path coefficient analysis of yield components in hexaplod triticale. (X Triticosecale wittmack). Crop Improv 2007, 34(2), 133-41.

The estimates of heritability, genetic parameters, genotype x environment interactions, correlation and path coefficient analysis for yield and its components in twenty diverse genotypes of triticale ( X Triticosecale Wittmack) under six diverse environments were worked out. A high degree of variation was observed for all the characters. Most of the genotypes showed high genotypes x environments interactions for all the traits studied. The genetic parameters of variability showed that no character had high magnitude of all four parameters, genetic (GCV) and phenotypic (PCV) coefficients of variability, heritability (H) and genetic advance (GA) under all the environments. Only spike length showed moderate GCV, PCV coupled with high H and GA under all the environments as well as pooled over environments, followed by number of grains per spike and number of spikes per plant in at least four environments. The estimates of correlation coefficients indicated that plant height, number of spikes per plant, grain weight per spike, spike length, 1000 grain weight and harvest index were positively correlated with yield. Number of grains per spike and spike length influenced the grain yield indirectly through grain weight per spike. This suggests that selection for these component charcters can help in improvement of yield in triticale.

4 tables, 13 ref

022653   Govindaraj S;Selvankumar T;Hariprasad S; Alagesaboopathi C (P G Research Biotechnology Dep, Mahendra Arts and Science College, Kalippatti-637 501) : In Vitro regeneration and mass multiplication of Andrographis affinis Nees - an endemic medicinal species from India. Asian J Microbiol Biotechnol envir Sci 2008, 10(1), 173-7.

An efficient protocol has been developed for plant regeneration from nodal explants of in vitro grown Andrographis affinis Nees. Proliferation of shoots was achieved on Murashige and Skoog (MS) medium supplemented with various concentrations of BA, K.n either singly or in various combinations. The highest shoot regeneration frequency (100%) were obtained from nodal segment on MS medium fortified with 2 mg/1 BA and 1 mg/1 Kn. The regenerated shoots best on MS 2mg/l IBA. Regenerated plantlets with well-developed shoot and roots were hardened, successfully transferred to soil and maintained in green house. The present in vitro procedure can be used in conservation and mass multiplication of this endemic medicinal plant.

2 illus, 4 tables, 7 ref

022652   Gour V;Sharma S K;Emmanuel C J S K;Tarun Kant (Biotechnology Laboratory, Forest Genetics and Tree Breeding Division, Forest Research Institute, Jodhpur-342 005) : Rapid in vitro morphogenesis and acclimatization protocol for balanites aegyptiaca (L) del - a medicinally important xerophytic tree. J Pl Biochem Biotechnol 2007, 16(2), 151-3.

Cellus mediated regeneration system for Balanites aegyptiaca (L) Del is reported here. Different explants like apical buds young thorns and cotyledon pieces from mature tree and root segments from in vitro raised seedlings were used for callus induction on MS medium supplemented with 2.23 μM 2,4-Dichlorophenoxyacetic acid. Seven to eight weeks old calli were transferred on hormone free MS medium in order to get regeneration. Shoot morphogenesis was achieved only from cotyledon-derived callus. The shoots so produced rooted well, when cultured on 85 medium supplemented with 9.84 μM Indole-3-butyric acid. Plantlets have been transferred to the field after two-phase hardening and are performing Well.

^ssc1 illus, 1 table, 11 ref

022651   Gothi K K;Hire R S;Vijayalakshimi N;Dongre T K (Nuclear Agriculture and Biotechnology Div, Bhabha Atomic Research Centre Trombay, Mumbai-400 085) : Mating behaviour of radio-sterilized males of red palm weevil, Rhynchophorus ferrugineus (Oliv). J Nucl Agric Biol 2007, 36(1-2), 65-72.

Mating behaviour of normal and radio-sterilized males of red palm weevil, Rhynchophorus ferrugineus Oliv were carried out under laboratory conditions with 12:12 Light: Dark photoperiodic cycle. Results indicated that both sterilized as well as unsterilized males mated equally well in both the phases of photoperiodic cycle and majority of the males mated in middle (5 to 8 hrs.) part of the phases. There was no significant change in mating behaviour of sterilized males, except slight increase in mating frequency and period of copulation. The maximum number of males mate only once in a single day, however some males were observed to mate more than 4 times in control and 6 times in irradiated regimen. The average respective mating frequency of control and sterilized male were 16.4 and 19.2 in dark phase, 17.8 and 18.8 in light phase. The period of copulation varied from 15 to 45 minutes in control males and 15 to 60 minutes in sterilized males.

1 illus, 3 tables, 12 ref

022650   Gomes A;Giri B;Saha A;Mishra R;Dasgupta S C; Debnath A;Gomes A (Laboratory of Toxinology and Experimental Pharmacodynamics, Physiology Dep, Calcutta Univ, 92 A.P.C. Road, Kolkata-700 009, Email: agomesc_u@gmail.com) : Bioactive molecules from amphibian skin: their biological activities with reference to therapeutic potentials for possible drug development. Indian J expl Biol 2007, 45(7), 579-93.

The amphibian skin contains various bioactive molecules (peptides, proteins, steroids, alkaloids, opiods) that possess potent therapeutic activities like antibacterial, antifungal, antiprotozoal, antidiabetic, antineoplastic, analgesic and sleep inducing properties. Research on amphibian skin derived biomolecules can provide potential clue towards newer drug development to combat various pathophysiological conditions. An overview on the bioactive molecules of various amphibian skins has been discussed.

1 a, 1 table, 136 ref

022649   Goel M K;Malik V;Kumar S;Govil C M (Plant Tissu Culture Div, Central Institute of Medicinal and Aromatic Plants, Lucknow-226 015) : In Vitro Micropropagation in Ochna serrulata. Adv Pl Sci 2008, 21(1), 19-21.

Ochna serrulata L. is a very important plant species due to their medicinal, ornamental and several other properties. A micropropagation protocol was developed for this plant using axillary shoot buds. Of all the various cytokinis viz., 6-Benzyl amino purine, Kinetin and Methyl ammo purine, BAP was found most suitable for shoot multiplication followed by kinetin and MAP. Rapid in vitro shoot multiplication was most optimum on basal MS medium supplemented with BAP (2.0 mgl^-1) and IBA (0.2:5 mgl^-1). The shoots were successfully rooted on half strength semi solid basal MS medium with 2% sucrose containing IBA (1.0 mgl^-1).

1 illus, 3 tables, 15 ref

022648   Ghodake V P;Patil S A;Pattekari P P;Dhumal P A;Khambe S D (Biotechnology Engineering Dep, Tatyasaheb Kore Institute of Engineering and Technology, Warananagar-416 113, Email: ghodakevp2000@yahoo.com) : Detection and optimization of indole acetic acid (IAA) production from Rhizobium meliloti. Asian J Microbiol Biotechnol envir Sci 2008, 10(1), 147-50.

In this study Rhizobium sp. was isolated from root nodules of fenugreek (Trigonella foenum-graecum) and identified as Rhizobium meliloti following standard methods. This isolate was tested for the production of indole acetic acid (IAA). Detection of IAA was done with TLC where a specific spot from the extracted IAA preparation was found, corresponding with the standard spot of IAA with same Rf value. The addition of L. tryptophan (as a precursor for auxins) to the medium inoculated with Rhizobium meliloti, enhanced the production of IAA several fold compared to L-tryptophan non-amended medium. Moreover, the present results support the consensus that the tryptophan dependent pathway plays vital role to maintain very high IAA levels. The temperature 30°C, pH 7.0 and 0.4 % tryptophan concentration in the medium was found optimum for the production of IAA by Rhizobium meliloti. Production of IAA by Rhizobium meliloti increased with an increase in tryptophan concentration from 1 to 4- mg/mL.

4 illus, 29 ref

022647   Fan J;Wang D;Ranu R S (Laboratory of Plant Molecular Biology/Biotechnology, Bioagricultural S, Colorado State University, Fort Collins, CO 80523-1177, USA) : Characterization and expression pattern of 1-aminocyclopropane-1-carboxylate (ACC) synthases cDNA from geranium (Pelargonium x hortorum cv sincerity). J Pl Biochem Biotechnol 2007, 16(2), 87-96.

Enzyme 1-aminocyclopropane-1-caryboxylate (ACC) synthase catalyzes the rate limiting step in the regulation of ethylene bios thesis and is, primarily, regulated at the transcription level. To understand the role of ACC synthases in stress-induced ethylene bursts in geranium, degenerated oligonucleotide primers were used to amplify a DNA probe from cDNA; and used to screen a cDNA library prepared from an mRNA preparation from mature flowers. Six putative full-length ACC synthase clones were isolated; these clones were divided into two groups on the basis of deduced amino acid sequences and features contained in their mRNA. Clones (e.g. PHS-ACS41, 44, and 45) in group I contain a stop codon (TAG) just before the initiation codon (ATG). Clone PHS-ACS44 from this group contains an "extra" 780 nucleotide sequence at the untranslated 3' -end region (UTR) after the first polyadenylation signal and a short poly(A) tail. The "extra" UTR contains another polyadenylation signal and a poly(A) tail, otherwise the coding sequence is identical to PHS-ACS41. Clone PHS-ACS49 in group II contains stop codon (TAG) twenty one bases upstream from the initiation codon. All these genes contain eleven conserved amino acid residues, the substrate and pyridoxal 50-phosphate binding site that are characteristic of all ACC synthases. Using RT-PCR and gene specific primers, expression studies show that these genes are expressed differentially. PHS-ACS41 and PHS-ACS44 not only show differences in expression in vivo, but also exhibit differential translational efficiencies in vitro suggesting that the "extra" 780 UTR in vivo may serve as a regulatory element. Our finding of multiple ACC synthase genes and their expression in geranium is consistent with similar observations in other plant species in which ACC synthases are encoded by a multigene -family that are differentially regulated by internal and external factors.

7 illus, 1 table, 46 ref

022646   Dhandapani R;Prabuganesh G;Shanmugam K S; Mahendran M;RajaSekar S (NO, Thanthai Hans Roever College, Perambalur District-621 212) : Mass multiplication of Indian medicinal plant Salix tetrasperma Roxb & Cor.. Adv Pl Sci 2008, 21(1), 363-5.

Salix tetrasperma is an important Indian medicinal plant. In tamil it is called "NEER NOCHCHI" or "NEER VANJI" in Indian system of medicine. The plant is used for rheumatism, diuretic, anthelmintic, piles, epilepsy, venereal diseases, head ache and stones in bladder. It has not yet been brought under cultivation. Its taxanomy, morphology, ecology and medicinal uses were studied. Tissue culture technology is costly and requires a high-tech laboratory. Hence, a low cost mass multiplication technology has been invented through water culture experiment in order to make its sapling available to the interested herbal farmers in a large scale. The findings are reported in this paper, which will be immense help and use to the herbal farmers.

3 tables, 6 ref

022645   Dhandapani R;Mahendran M;Shanmugam K S; Prabuganesh G;Raja Sekar S (Medicinal Plants Research Unit, Post Graduate & Research Dep of Microb, Thanthai Hans Roever College, Perambalur District-621 212) : A low-cost mass multiplication technology for Justicia ganderussa Linn.. Adv Pl Sci 2008, 21(1), 353-4.

Justicia ganderussa Linn. is an important Indian medicinal plant. In Tamil it is called "KARU NOCHI". This plant is known to possess anti-inflammatory, antiseptic, stomach pain, diuretic, digestive and expectorant properties. The roots are useful in dysentery, wounds, ulcers, cough and malarial fever. Leaf powder, leaf and root decoctions are prepared and used for medicinal purposes. Its low-cost mass multiplication technology has been invented through IBA hormonal treatment stem cuttings experiment, in order to make its saplings available to the interested herbal farmers in large scale.

2 tables, 5 ref

022644   Dhandapani R;Jijo;Varghese G;Anwar Rahim A (Medicinal Plants Research Unit, Postgraduate and Research Dep of Micr, Thanthai Hans Roever College, Perambalur District-621 212, Email: paniroever2007@rediffmail.com) : Macropropagation of Vitex negundo L.. Adv Pl Sci 2008, 21(1), 15-16.

Vitex negundo is an important Indian medicinal plant. In Tamil is called "VELLAI-NOCHI". V. negundo is known to possess headache, catarrhal fever, swellings, tonic, expectorant and diuretic properties. For medicinal uses of V. negundo is collected only from their wild natural habitats. This plant has not yet been proper cultivation through IBA hormonal treatment. gives the macropropagation of V. negundo through IBA 100 ppm and 200 ppm hormonal treatment of stem cuttings. Numerous saplings can be preared efficiently by this method.

2 tables, 6 ref

022643   Dasgupta C N;Mukhopadhyay M J;Mukhopadhyay S (Centre of Advanced Study, Botany Department, University of Calcutta, 35, Ballygunge Circular Road, Calcutta-700 019, Email: sanmukh@vsnl.net) : Somatic embryogenesis in Asparagus densiflorus (Kunth) jessop cv sprengeri. J Pl Biochem Biotechnol 2007, 16(2), 145-9.

An efficient protocol has been developed for plant regeneration in Asparagus densiflorus (Kunth) Jessop cv Sprengeri (Asparagaceae) through somatic embryogenesis from spear sections. Callus culture was initiated on Murashige and Skoog (MS) medium formulation containing a-naphthaleneacetic acid (5.37 μM), indole-3-acetic acid (5.71 μM) and 6-benzylaminopurine (2.22 μM). The callus became embryogenic by transferring to 2, 4-dichlorophenoxyacetic acid (4.52 μM) containing MS medium. Somatic embryos developed and matured vigorously on MS medium with NAA (1.07 μM), 6-γ-γ-dimethylaminopurine (9.84 μM) and abscisic acid (1.89 μM). Mature bipolar embryos were converted efficiently into plants on MS medium in the presence of low level of kinetin (2.32 μM). Regenerated plants showed 80% survival after transfer to field. These plants were all diploid (2n=60). Peroxidase activity was maximum in the embryogenic callus as documented from the gel as well as spectrophotometric analysis.

^ssc1 illus, 1 table, 18 ref

022642   Cheema P S;Srivastava S K;Amutha R;Singh S; Singh H;Sandey M (Bacteriology and Mycology Div, Indian Veterinary Research Institute, Izatnagar-243 122, Email: sksrivastava@ivri.up.nic.in ) : Detection of pathogenic leptospires in animals by PCR based on lipL21 and lipL32 genes. Indian J expl Biol 2007, 45(6), 568-73.

Efficacy of primers capable of amplifying conserved outer membrane protein (OMP) genes i.e., lipL21 and HpL32 of Leptospira strains was tested for rapid and early diagnosis of the leptospirosis using a polymerase chain reaction (PCR). These OMP genes were found to be conserved in various leptospiral serovars viz., Canicola, Pomona, Icterohaemorrhagiae, Pyrogenes, Sejroe, Grippotyphosa, Ballum and Tarassovi as PCR products of 561 bp and 756 bp were obtained by PCR employing lipL21 and lipL32 based primers, respectively, in all these serovars. Absence of such amplicons in DNA extracted from Pasteurella, Campylobacter and Brucella confirmed the specificity of the primers. Serum and tissue samples collected from cattle, buffaloes and experimentally infected guinea pigs and calves were subjected to PCR using above primers as well as conventionally used primers G1/G2. All the sera and tissue samples, whether field samples or collected from experimentally infected animals, found positive for G1/G2 specific PCR were also positive for lipL21 and lipL32 specific PCR. The study indicated that lipL21 and lipL32 based primers could be used for PCR based diagnosis of leptospirosis. Since G1/G2 primers are known not to amplify the DNA of Grippotyphosa, the use of primers employed in the present study could have an additional advantage in detection of cases of the disease.

8 illus, 2 tables, 16 ref

022641   Anuradha M;Rao K N (NO, Central Tobacco Research Institute, Rajahmundry-533 105, Email: radhama2k2@yahoo.co.in) : Nitrogen dose-dependent metabolic shift from nitrate reduction to starch accumulation in flue-cured tobacco varieties. J Pl Biol 2007, 34(1), 1-4.

A sand culture experiment was conducted with three tobacco varieties and three levels of nitrogen (4.54, 9.08 and 18.16 g/plant) "using McMutrey's nutrient solution. Nitrate reduction measured as leaf in vivo nitrate reductase activity increased with increased level of applied nitrogen in all the three varieties. Leaf photosynthetic pigments, total nitrogen and nitrate nitrogen were higher at 45 days after planting and decreased to lower levels when leaves were at the stage of harvest, irrespective of nitrogen dose and vari-ety. Specific leaf weight was lower during plant growth period but reached its maximum when the leaves were at harvest. Depend-ing on nitrogen dose leaf pigments and total nitrogen increased in K-326 as compared to Gouthami and Hema. Specific leaf weight decreased with increased level of applied nitrogen in all the three varieties during plant growth period but it was higher at the time of harvest. Specific leaf weight was higher in K-326 followed by Gouthami and Hema. Leaf starch content was higher when nitro-gen dose was lower and it decreased with increased level of N. Higher nitrogen levels led to delay in shift from nitrogen utilization to starch accumulation resulting in lower starch, sugars and higher nitrogen and nicotine. Metabolic shift from nitrogen utilization to starch accumulation is of prime significance in flue-cured tobacco from the stand point of its quality as sugar-nicotine ratio de-termines the utility of the leaf as a smoking product.

13 ref

022640   Amin S B;Chellapandi P (Biogas Research Centre, Gujarat Vidyapith, Sadra-382 320, Email: pchellapandi@gmail.com) : Production of extra cellular proteinase by Asergillus foetidus NCIM 505 on milk powder and soya bean meal based media. Res J Biotechnol 2008, 3(1), 33-38.

The main interest in proteases in biotechnology is their potential to be used to hydrolyze protein substances in leather, food and dairy industry. Though most of the Aspergillus species are considered as alkaline type protease producers, some of the species are exceptionally suited for mild alkaline and metallo-protease production. We report here on a novel application of protease as a soya bean-milk protein hydrolyzing activity in Aspergillus foetidus NCIM 505. It was cultivated in solid-state and broth cultures on soya bean and milk protein based media. The cultures were characterized for growth, consumption of the carbon source, nitrogen source and protease production. Growth conditions have optimized for protease production and the best yield was achieved at 60-84 h, 30°C, with the initial pH of 5.7 in the production media. On solid surface of substrates, yeast extract and sucrose were favored for the maximum enzyme synthesis. Protease peptone was found to be the best inducer in broth culture of A.foetidus. The optimum moisture content for maximum enzyme activity was 50-55% in solid-state cultivation. The crude fraction of protease showed more activity at 40°C in O.I M Tris-HCl buffer (pH 8.0). Growth characteristics and enzyme activities were examined for potential correlations. Such correlations would allow the use of this strain for food processing industry.

3 illus, 2 tables, 24 ref

022639   Amala Balu P;Ibrahim S M;Muthuramu S (Cotton Dep, Tamil Nadu Agriculture Univ, Coimbatore, Tamil Nadu) : In Vitro response for callus induction in sunflower (Helianthus annuus L.). Adv Pl Sci 2008, 21(1), 23-6.

In sunflower the in vitro response of four CMS lines, .5 restorers and their 20 hybrids were studied by using MS medium with three different growth hormone compositions for callus induction. Hypocotyl was the explants used for this study. Among the three media studied the MS medium containing 2,4-D 5mg/L + Kinetin 0.5mg/L was found to be good irrespective of the genotypes studied. Among the parents 234 A (93.23%) and RHA 299 (9.66 days) and among the hybrids 852A x RHA 299 (93.36%) and 234A x RHA 29!) (9.61 days) recorded highest percentage of callus induction and minimum number of days taken for callus induction respectively in medium containing 2,4-D 5mg/L + Kinetin 0.5mg/L. The parents viz., 234A, and RHA 2.99 and the hybrids viz., 821 A x RHA 299, 852A x RHA 299, 852 A x Morden and 234 A x RHA 29 responded well for in vitro studies. These parents and hybrids can be successfully utilized in biotechnological approaches [ for the improvement of sunflower crop.

2 tables, 8 ref

022638   Ahmad S;Jamal A;Fazli I S;Alam T;Ali Khan M; Kamaluddin;Abdin M Z (Program in Molecular Medicine, 373 Plantation Street, Biotech II Suite 309, Univ of Massachusetts Medical School (UMMS), Worcester, MA-01605, USA, Email: asaif77@yahoo.com) : Estimation of psoralen content by HPLC method in different organs of Psoralea corylifolia L., as influenced by sulphur & nitrogen nutrition. J med aromatic Pl Sci 2007, 29(4), 197-203.

Psoralen is a naturally occurring furocoumarin which has been exploited for its photosensitizing properties since ancient time. Psoralen and its derivatives (linear furocoumarins) are used in dermatological disorders (vitiligo and psoriasis) and are also very promising in the treatment of other diseases. A simple randomized field experiment was conducted to estimate the concentration of psoralen by HPLC (High Pressure Liquid Chromatography) in different parts (root, stem, leaf and seed) of Psoralea corylifolia L. at various phenological stages as influenced by sulphur and nitrogen nutrition. Six different treatments that included three levels of sulphur (0, 20 and 20+20 kg ha^-1) in combination with two levels of nitrogen (20 and 20+20 kg ha^-1) were tested (T₁ = control -without manure and fertilizers, T₁ = manure @ 9 kg plot<6>-1 -10 t ha^-1, T₃ = S₀N₂₀ K₄₀ P₄₀, T₄, = S₂₀ N₂₀ K₄₀ P₄₀l, T₅ = S₂₀₊₂₀ N₂₀ K₄₀. P₄₀, T₆ = S₂₀₊₂₀ N₂₀₊₂₀ K₄₀ P₄₀). Maximum psoralen concentration was found in seed at mature unripe stage as compared to other plant parts. An increment of 15-16 % was observed in psoralen concentration with treatment T₅ as compared to control (T₁). Leaf area index and leaf area duration along with chlorophyll content were also reported to be higher in T₅ treatment. Our results may be beneficial for pharmaceutical Industry in view of large demand of psoralen.

4 illus, 5 tables, 38 ref

022637   Abasaheb R N;Shitole Mahadev G (Botany Dep, Annasaheb Awate College, Manchar, Dist. Pune-410 503, Email: abasahebnalwade_botany@yahoo.co.in) : Enzyme activity as a method to characterize salt-tolerance in calli and plantlets of grape (Vitis vinifera L.) cv. Tas-A-Ganesh. Res J Biotechnol 2008, 3(1), 27-32.

A relation between antioxidant defense system and salt tolerance in two types of grape (Vitis vinifera L.) cv. Tas-A-Ganesh calli and plantlets differing in salt tolerance was studied. Callus culture of this cultivar was obtained from axillary bud explants on Murashige and Skoog (MS) medium with 2.2 mM BAP and 10.72 mM NAA, which were adapted to grow in the medium with 128.25 mM NaCl, were termed as salt-tolerant calli. Both the unselected and salt-tolerant calli were grown on MS medium with increasing concentrations (0.0 to 171 mM) of NaCl. Unselected and salt-tolerant calli survived up to 85.5 mM and 128.25 mM NaCl respectively. Both the calli exhibited linear increase in the activities of peroxidase, catalase and superoxide dismutase with increase in the salt concentrations. The plantlets generated from salt-tolerant calli exhibited higher activities of these enzymes than the plantlets generated from unstressed calli. The activity of IAA oxidase enzyme was increased in the unselected calli and decreased in the salt-tolerant calli with increasing NaCl stress; its activity was less in the plantlets generated from salt-tolerant calli than plantlets generated from unstressed calli. Thus, more availability of IAA in the cells of salt-tolerant calli and plantlets must have helped to maintain more growth under saline conditions. Superior antioxidant system (through peroxidase, catalase and superoxide dismutase) and more availability of IAA helped salt-tolerant calli and plantlets generated from them to maintain more growth and to survive under wider range of salinity.

1 illus, 2 tables, 31 ref

021622   Yadav A;Yadav B R (Biotechnology Dep, Kurukshetra University, Kurukshetra-136 119) : DNA fingerprint: genetic relationship in six Indian goat breeds. Indian J Biotechnol 2008, 7(4), 487-90.

The DNA fingerprint and genetic relationship of 6 Indian goat breeds were studied by RAPD technique. Intra- and inter-breed divergence was estimated in terms of band sharing frequency (BSF) and mean average percentage difference (MAPD). The interbreed BSF was lower than intrabreed BSF due to more homogeneity within breed than between breeds. Inter reed BSF showed more similarity between Marwari and Sirohi (0.916±0.032) and less between Barbari - Black Bengal (0.855±0.087). MAPD provides a measure of genetic divergence in terms of genomic DNA fingerprints between breeds. The interbreed divergence was lower between Marwari and Sirohi (9.08±4.37) and higher between Black Bengal and Jhakrana (13.23±06.85).

021621   Vasantharaj David B (Sun Agro Biotech Res Centre, , Madanandapuram, Porur, Chennai-600 116, Email: drdavid@md4.vsnl.net.in) : Biotechnological approaches in IPM and their impact on environment. J Biopestic 2008, 1(1), 1-5.

Use of chemical pesticide and other agro chemicals are getting reduced /being banned globally because of their toxic effects on human beings and his live stock, residual toxicity, environmental problems, pest out-breaks and drastic effects on beneficial insects. Therefore, now it is imperative to develop a holistic system of tackling pests to make it more eco-friendly, economically viable and socially acceptable for the farmers. In the WTO regime, it is absolutely necessary to limit the usage of chemicals, to remain in the world market and sustain the competition. In this regard to tackle the major pests and diseases of major crops biotechnological approaches are gaining momentum. Compared with usage of chemical pesticides biopesticides constitute around 2% in the country. The biotechnological approaches of pest control such as use of botanical pesticides, use of microbial pesticides, augmentative biocontrol by inundative releases, pheromones and attractants in pest management and plant incorporated protectants (PIPs) / GM crops which are discussed in detail. Advantages and limitations of biopesticides have been outlined and the future approaches are highlighted.

10 ref

021620   Vadivel V;Pugalenthi M (Biotechnol Dep, Karpagam Arts and Sci Coll, Coimbatore-641 021, Email: pugalmuthiah@redifrmail.com) : Analysis of variability in seed storage proteins of Mucuna prurients var. utilis by using SDS-PAGE. Seed Res 2007, 35(2), 210-14.

The influence of agro-ecological conditions of different growing locations on the seed proteins of a tropical under-utilized legume, Mucuna pruriens var. utilis (Wall ex Wight) Baker ex Burck (velvet bean) was investigated. A wide variation was observed in the total seed protein content (22.8-26.1 g/l00 g seed flour) and different protein fractions with relation to different growing locations. However, the major seed protein fractions, the albumins and globulins of all the accessions exhibited similar kind of banding pattern and same molecular weight protein sub units when subjected to SDS-PAGE, indicating that the environmental conditions of the growing locations influence only the quantity of the proteins not their molecular characteristics in velvet bean seeds.

2 tables, 21 ref

021619   Tao X;Jang M S;Kim K S;Yu Z;Lee Y C (Biotechnology Dep, College of Natural Resources and Life Science, Dong-A University, Busan 604-714, South Korea, Email: yclee@dau.ac.kr) : Molecular cloning, expression and characterization of α-amylase gene from a marine bacterium Pseudoalteromonas sp. MY-1. Indian J Biochem Biophys 2008, 45(5), 305-9.

A gene (amyA) encoding an extracellular α-amylase from a marine bacterium Pseudoalteromonas sp. MY-1 was cloned and expressed in Escherichia coli. It comprised an open-reading-frame of 2,007 base pairs and encoded a protein of 669 amino acids with a predicted molecular weight of 73,770 daltons and a pI of 5.15. The entire amino acid sequence of amyA gene showed 86% similarity to the a-amylase preproprotein from Pseudoalteromonas haloplanktis. It consisted of a signal peptide, α-amylase catalytic domain and an amy C domain. The recombinant amylase was purified to homogeneity and biochemically characterized. The enzyme revealed maximum activity at pH 7.0 and 40°C. The enzyme hydrolyzed soluble starch and some maltooligosaccharides to several oligosaccharides, and maltose was the common product from different substrates.

021618   Swarna Piria R;Rajmohan K;Suresh S (Horticultural Research Station, , Pechiparai-629 161) : In vitro production of protocorms and protocorm like bodies in orchids. Agric Rev 2008, 29(1), 40-7.

In vitro multiplication is the most important biotechnological tool being used in orchids, especially for large scale commercial propagation. Usually in vitro methods would be useful in a crop where in the existing conventional methods are too-slow, non-profitable and not successful. As orchid seeds lack a functional endosperm, the germination of seeds require a long period and any disturbance to the soil or physical environment destroy the whole population. In this context the in vitro multiplication provide an appropriate solution to this problem. So development of tissue culture protocol is an essential pre-requisite for any other biotechnologicai manipulations at cellular and molecular level.

88 ref

021617   Srikanth A;Halami P M (Food Microbiology Dep, Central Food Technological Research Institute, Mysore-570 020) : Cloning of pediocin PA-1 and its immunity genes from Pediococcus acidilactici K7 using pAMJ shuttle vector into Lactococcus lactis MG1363. Indian J Biotechnol 2008, 7(4), 550-3.

The matured pediocin PA-1 encoding gene, pedA and its immunity counterpart, pedB from Pediococcus acidilactici K7 were cloned by PCR technique and ligated into the E. coli-lactic shuttle, protein expression vector pAMJ2008. The recombinant pAMJAB was constructed and sequenced to confirm the intactness of reading frame of the deduced pediocin PA-1 fusion protein. Lactococcus lactis MG1363 was electroporated with the plasmids and the transformation efficiency of <126> 2-3 x 10³ μg^-1 was observed. This was further confirmed by plasmid DNA isolation and analysis.

021616   Sreekumar J;Jose K K (NO, Central Tuber Crops Research Institute, Sreekariyam, Thiruvananthapuram-695 017) : Statistical tests for identification of differentially expressed genes in cDNA microarray experiments. Indian J Biotechnol 2008, 7(4), 423-36.

Microarrays experiments are becoming a common laboratory tool for monitoring expression level in cells for thousand of genes simultaneously. The new data promise to enhance fundamental understanding of life on a molecular level and may prove useful in medical diagnosis, treatment and drug design. The greatest challenge to array technology lies in the analysis of gene expression data to identify which genes are differentially expressed across tissue samples or experimental conditions. A simple fold change was used to test the differential expression of genes. Ordinary t-test and t-test approaches with minor variations are usually used in finding differentially expressed genes under two conditions. Analysis of variance (ANOVA) and mixed model ANOVA proved to be powerful under multiple conditions or several sources of variation. Since thousands of hypotheses are tested simultaneously there is increased chance of false positives and it becomes necessary to adjust for multiple testing when assessing statistical significance of findings. Bayesian variable selection and empirical Bayesian approaches offer yet another avenue.

021615   Sita Kumari K;Sarat Babu;Rao G H (Pharmacy Dep, MRPG College, Phool Baugh, Vizianagaram-535 002) : Process optimization for citric acid production from raw glycerol using response surface methodology. Indian J Biotechnol 2008, 7(4), 496-501.

Statistical experimental design was applied for the optimization of medium constituents for citric acid production by Yarrowia lipolytica NCIM 3589 in submerged fermentation using raw glycerol as the carbon source. Response surface methodology (RSM) involving central composite design (CCD) was adopted to evaluate the amount of citric acid produced by most important factors, such as carbon concentration, nitrogen concentration, and salt solution concentration. A second order polynomial regression model was fitted and was found adequate with R² of 0.9485. The optimum conditions were found to be carbon concentration 38.77 g/L, nitrogen concentration 0.401 g/L, and salt solution concentration 12.3% (v/v). Citric acid production at these optimum conditions was 13.41 g/L.

021614   Saxena H (Crop Protection Div, Indian Inst of Pulses Res, Kanpur-208 024, Email: hem_saxena@yahoo.com) : Microbial management of crop - pest. J Biopestic 2008, 1(1), 32-7.

In India most of the farmers depends upon synthetic pesticides for protecting their crops form pest attack. These pesticides not only caused environmental pollution, but also causing health hazardous to human being and domestic animals. This could be prevented by using bio-intensive integrated pest management (BIPM) were microbial insecticides place an important role. Here I have discussed about the important bacterial, fungal, viral, protozoan and nematode - based insecticides.

1 table, 47 ref

021613   Savita;Kusumakar A L;Malik Y P S;Minakshi; Prasad G (Veterinary Microbiology Dep, College of Veterinary Science & A H, JNKVV, Jabalpur-482 001) : Detection and characterization of group A and D avian rotaviruses in India. Indian J Biotechnol 2008, 7(4), 554-6.

Determines the association of rotaviruses with diarrhea in poultry. Avian rotaviruses were detected on the basis of migration pattern of discrete 11 segments of dsRNA in the RNA-polyacrylamide gel electrophoresis (RNA-PAGE). The prevalence of rotavirus infection in diarrhoeic (n=46) and environmental (n=31) samples was 17.39% (8/46) and 3.22% (1/31), respectively. Electrophoretic migration pattern characteristic of group A (5:1:3:2) and group D (5:2:2:2) avian rotaviruses was observed. Overall the prevalence of group D avian rotavirus was higher (77.8%; 7/9) than group A rotavirus (22.22%; 2/9) in central India. Occurrence of different electrophoretic migration patterns suggested existence and circulation of genetically diverse strains of avian rotaviruses. This study establishes the presence of group D avian rotavirus in India and need for expansion of such work to cover more geographical areas.

021612   Ruma Devi;Dhaliwal M S;Ajinder Kaur;Gosal S S (Vegetable Crops Dep, Punjab Agricultural University, Ludhiana-141 004) : Effect of growth regulators on in vitro morphogenic response of tomato. Indian J Biotechnol 2008, 7(4), 526-30.

Series of experiments were conducted to explore the in vitro morphogenic response of tomato genotypes viz., Castle Rock, Punjab Upma, VFN-8 and IPA-3, under different concentrations and combinations of growth regulators. The analysis of variance at 5% level of significance indicated that the differences among different genotypes, hormonal regimes as well as their interactions were statistically significant. The MS medium supplemented with BAP @ 3.0 mg L^-1 and IAA 2.5 mg L^-1 was optimum for callus induction, plant regeneration and number of shoots per explant. The maximum per cent callus induction and plant regeneration in the genotypes Castle Rock, Punjab Upma, VFN-8 and IPA-3. was 81.23, 76.69, 68.13 and 65.12%; and 46.91, 48.02, 57.14 and 60.23%, respectively, The respective average number of shoots per culture were 7.03, 6.92, 8.19 and 9.19. At higher and lower levels of hormones, a considerable decline was recorded in per cent callus induction, plant regeneration and number of shoots per explant. The best rooting was found to be in the 1/2 MS medium supplemented with 0.2 mg L^-1 IBA. Among the four soil mixtures studied viz., vermiculite, perlite, coco-peat and mixture of three (vermiculite, perlite and cocopeat in the ratio of 1:1:1), maximum plantlet survival rate was recorded in the mixture of three for genotypes Castle Rock and Punjab Upma and in vermiculite for genotype VFN-8.

021611   Roy B;Mandal A B (Biotechnology Section, Central Agricultural Research Institute, P O Box No. 181, Port Blair- 744 105) : Development of synthetic seeds involving androgenic and pro-embryos in elite indica rice. Indian J Biotechnol 2008, 7(4), 515-9.

Synthetic seeds were produced from anther-derived mass-multiplied embryos and pro-embryos of rice (Oryza sativa L.) var. IR 72. A high dose (4-6 mg L^-1) of BAP was found to produce a large number of dormant embryos, pro-embryos and embryo-like structures in about 45 d. These were encapsulated in sodium alginate (2.5% w/v) matrix. Germination and plantlet regeneration capacity of the encapsulated seeds were tested by culturing them on MS fortified with different combinations and concentrations of BAP, Kn and NAA. The result indicated that BAP in combination with lower concentrations of NAA increased germination of beaded embryos over control (MS without hormones). High percent of germination (55-87.5%) was observed when MS was supplemented with BAP and lower concentration of NAA; whereas, addition of Kn in MS reduced the germination percentage. The germination of unbeaded pro-embryos was 92.5% on MS basal medium. The reduced rate of germination of artificial seeds may be attributed to the damage incurred while separating the embryos from clusters and/or owing to adverse effects of chemicals used for encapsulation. Moderate germination (40.0%) was seen on sterile sand. Synthetic seeds may be used for in vitro propagation as well as genetic transformation experiments, especially involving biolistics.

021610   Rajput H J;Patil R B;Shinde S M (Diorectorate of Research, Dr.P.D.K.V., Akola-144 104) : Biochemical and molecular characterization of cotton hybrids. Ann Pl Physiol 2007, 21(1), 51-4.

Seven cotton hybrids i.e. PKVHY-2, CAHH-468, H-10, JKHY-I, PHH-3I6, RHH-1394 and RHB-0387 and their parents were characterized for proteins, oil and gossypol content. The parents and hybrids revealed that the oil content in various genotypes ranged from 17.95 to 23.05 per cent, protein from 36.00 to 44.25 per cent while gossypol range from 0.344 to 0.888 per cent. Molecular characterization by RAPD, using 15 random primers showed that only three primers OPD-15, OPD-05 and OPA-10 showed polymorphism while other primers showed monomorphic amplification or did not show any amplification of the genomic DNA. Primers OPD-15 could be used to test the hybridity of CAHH-468.

1 table, 18 ref

021609   Rajeswari V;Paliwal K (School of Biological Sciences, Madurai Kamraj University, Madurai-625 021) : In vitro plant regeneration of red sanders (Pterocarpus santalinus L.f.) from cotyledonary nodes. Indian J Biotechnol 2008, 7(4), 541-6.

Reliable and efficient micropropagation protocol was developed through axillary shoot proliferation from cotyledonary nodes of Pterocarpus santalinus. Cotyledonary nodes showed significantly (P<60>0.05) higher shoot multiplication rate and shoot length than leaf nodes on MS medium with 2.5 μM BAP and 2 μM 2-iP after first subculture (i.e. in the second harvest). For rooting, dipping of microshoots in 5 μM IAA solution proved superior to other in vitro methods. Of the various hardening media used for the acclimatization of rooted plants, a mixture of coarse sand, clay and farmyard manure (1:1:1) (v/v) supported the maximum percentage of survival (95%). There were no significant differences between the in vitro regenerated plants and seedlings of the same age for all the growth parameters measured (i.e. mean plant height, number of leaves per plant, fresh weight and dry weight of leaves, shoot and root per plant) four months after transfer to ex vitro conditions.

021608   Raja P;Balachandar D;Sundaram S P (Agricultural Microbiology Dep, Tamil Nadu Agricultural University, Coimbatore-641 003) : PCR fingerprinting for identification and discrimination of plant-associated facultative Methylobacteria. Indian J Biotechnol 2008, 7(4), 508-14.

Pink-pigmented facultative methylotrophic bacteria (PPFMs) belonging to the genus Methylobacterium, often associated with phyllosphere and roots and symbiotically benefit the plant species. For identification and discrimination of common plant-associated Methylobacterium species, different PCR-based DNA fingerprinting techniques were evaluated along with metabolic divergence. Standard/type strains of 10 species of plant-associated Methylobacterium were fingerprinted by means of RAPD, ARDRA, RISA, BOX and ERIC markers. These species showed divergence in carbon-substrates utilization pattern. Among the above PCR techniques evaluated, RAPD, RISA and ERIC fingerprinting had high discriminatory power than ARDRA and BOX-PCR. Being simple, rapid and repeatable, RAPD, RISA and ERIC-PCR techniques could be used for species identification and diversity analysis of plant-associated PPFMs belonging to Methylobacterium.

021607   Rai R K;Pushpa Singh;Shrivastava A K;Shukla S P;Namita Arya;Singh R K;Srivastava M K;Tewari P;Sawnani A;Khare A K; Ram Darash;Ram Kishore (Plant Physiol and Biochem Div, Indian Inst of Sugarcane Res, Lucknow-226 002, Email: ramakantrai@rediffmail.com) : Biochemical alterations in bud and root band zone of sugarcane setts due to chilling temperature. Indian J Pl Physiol 2007, 12(4), 344-52.

Effect of chilling temperatures (0, 5 and 10°C) on the sugarcane bud sprouting were compared with 25°C (normal). At the chilling temperatures, in the buds and the root band zones of the single bud sugarcane sett, the reducing sugars, specific activities of acid invertase (AI), IAA (indole acetic acid) oxidase and ATPase were decreased whereas the sucrose and IAA contents were increased as compared with 25°C. However, there was no change in sucrose content at the chilling temperatures but it markedly decreased at 25°C. Sucrose immobilization caused by the suppression of the acid invertase, hence decreased content of reducing sugars and accumulation of IAA might affected the sprouting at chilling temperatures.

4 illus, 42 ref

021606   Prabhu S;Kumar S;Subramaniam S;Sudha A (Nematol Dep, Tamil Nadu Agric Univ, Coimbatore-641 003, Email: somuprabhu@yahoo.com) : In vitro mass production of Pasteuria penetrans (Mankau) Sayre and Starr for the management of Meloidogyne incognita (Kofoid and White) chitwood. J Biopestic 2008, 1(1), 47-8.

Plant parasitic nematodes are parasitized and preyed by a variety of soil organisms which includes predatory nematodes, fungi, bacteria, viruses, protozoans, turbellarians, tardigrades, mites and other microarthropods. One such spore forming and an obligate parasite is Pasteuria penetrans. The major limitation in the use of Pasteuria spp. is their inability to grow in the absence of host nematodes. In vivo systems although proven are unlikely to be adopted for large scale application in most farming systems. Attempts were made to culture the organism in in vitro. Among the symbiotic bacteria used for mass production co-culturing with Enterobacter proved to be successful in culturing this obligate organism in a specially developed medium for mass multiplication.

1 table, 8 ref

021605   Pavon-Meza E L;Sarma S S S;Nandini S (Research Div, Postgraduate Studies, Universidad Nacional Autonoma de Mexico, Campus Iztazala, Av. de Los Barrios No. 1, Los Reyes, AP 314, CP 54090, Tlalnep, Email: sarma@servidor.unam.mx) : Combined effects of temperature, food availability and predator's (Asplanchna girodi) allelochemicals on the demography and population growth of Brachionus havanaensis (Rotifera). Allelopathy J 2008, 21(1), 95-106.

Studies the influence of asplanchnin (a non-toxic kairomone from the predator Asplanchna girodi) on the selected populations of prey Brachionus havanaensis cultured at two algal densities (0.5 x 10⁶ cells mL^-1 and 2 x 10⁶ cells mL^-1 of Chlorella vulgaris) and at two temperatures (15° and 25°C). At 15°C, the allelochemicals decreased the survival and offspring production of B. havanaensis at same food levels. Both gross and net reproductive rates were about 50% lower at 15°C than at 25°C. In the absence of asplanchnin, the rate of population increase (r) was highest (0.52 per day) at 25°C under 2 x 10⁶ cells mL^-1 of Chlorella. While in the presence of asplanchnin, the r was negative (-0.09 per day) at 15°C, at 0.5 x 10⁶ cells mL^-1. At higher temperature and higher food density, life history variables of B. havanaensis did not show adverse impact of asplanchnin. At 15°C under two food levels, B. havanaensis populations decreased in the presence of asplanchnin. However, at 25°C, B. havanaensis grown at low food level showed similar population densities (ca. 100 ind. mL^-1) regardless of the presence of asplanchnin. At 25°C and high food level, B. havanaensis had very low densities in the presence of Asplanchna than in controls. Thus, show the importance of food density and temperature as variables to evaluate the impact of predators' allelochemicals on the prey demographic characteristics.

4 illus, 1 table, 40 ref

021604   Nema R K;Dass S;Mathur M;Ramawat K G (Laboratory of Bio-molecular Technology, Botany Dep, M L Sukhadia University, Udaipur-313 001) : Morphactin and cytokinin promote high frequency bulbil formation from leaf explants of Curculigo orchioides grown in shake flask cultures. Indian J Biotechnol 2008, 7(4), 520-5.

High frequency bulbil formation from leaf explants of an endangered monocot herb, Curculigo orchioides, grown in shake flask cultures has been achieved. The leaf explants were obtained from in vitro grown plantlets maintained on MS solid medium containing BA (0.1 mg/L) and IBA (0.1 mg/L) for the past 8 years. Inoculum density of one leaf explant (<126>1 cm²) per two mL liquid medium (50 explants 100 mL^-1) was recorded optimal for bulbil formation. Increase in bulbil number per explant was recorded with increase in IBA concentration (from 0.1 mg/L to 1 mg/L) in the medium; level of substituted urea in the medium did not alter this response. Maximal percent explant response, number per explant and yield of bulbils was recorded in the medium supplemented with 1.0 mg/L IBA with 0.1 mg/L substituted urea. The number of bulbils per explant doubled during 4 to 8 wks growth while total yield (fresh biomass) increased several-folds. In IBA+2iP combination, optimal number of bulbils with maximum yield per litre was recorded in the medium supplemented with 1.0 mg/L IBA and 0.1 mg/L 2iP, at 8 wks growth. However, maximum fresh weight of bulbils (1027 g) was recorded in the medium containing 0.1 mg/L IBA and 1.0 mg/L 2iP. In morphactin and BA interaction, optimal number (38/explant), yield (19165/L) and FB (1450 g/L) of bulbils was recorded in the medium containing 1.0 mg/L BA and 0.1 mg/L morphactin followed by 0.01 mg/L and 1 mg/L of morphactin with the same concentration of BA. The use of this synthetic growth regulator in bulbil formation was new and very significant over other treatments. Formation of plantlets from bulbils was 80% and their survival was <126>80% in the field conditions. These results will be helpful in developing technology for micropropagation of the plant using bioreactor.

021603   Nagarajan G;Ashok Kumar C;Dechamma H J;Reddy G R;Ganesh K;Suryanarayana V V S (NO, National Research Centre on Camel, Post Bag No 7, Jorbeer, Bikaner-334 001) : Cloning and expression of FMDV-VP1 immunoreactive peptide in trivalent form and its application as immunogen. Indian J Biotechnol 2008, 7(4), 472-7.

A search for alternatives to conventional inactivated virus vaccine for FMD with an aim to control and eradicate the disease globally, is a continuous process till a promising one is identified. Development of such vaccines underlines necessity of avoiding the use of active virus, and to have broader antigenic coverage so as to make them suitable even for disease free countries. Subunit or peptide vaccines have been shown to elicit neutralizing antibody response. However, the titres are low as compared to sera from animals vaccinated with conventional vaccine and fail to protect animals against virus challenge. This is probably due to the inclusion of only limited epitopes. Under such conditions, mixing heterologous epitopes from the various serotypes may be a better approach for elicitation of high titred antibody response. Keeping this in view, we have linked C-terminal half of VP1 carrying two B cell and one T cell epitope of three FMDV serotypes (O, A and Asia 1), which are presently in use as vaccine strains in India. The linked polyvalent gene was expressed in Escherichia coli and the 59 kDa fusion protein was studied for its immunogenicity in guinea pigs in comparison with the specific epitopes of type 'O' produced as a similar fusion protein of 30 kDa. The trivalent protein showed better neutralizing antibody response, even with single booster injection, as compared to monovalent protein as observed in ELISA and SNT. These studies show future scope for the development of protein/DNA-based vaccine for FMD.

021602   Murugan K;Kishor Mohan T C (Plant Biochem and Molecular Biol Lab., Bot Dep, Univ Coll, Trivandrum, Kerala-695 034, Email: harimurukan@gmail.com) : Ameliorative mechanisms of sodium chloride stress in Sesbania sesban (L.) Merr. Indian J Pl Physiol 2007, 12(4), 373-7.

Study was to obtain a better understanding on salt-tolerance mechanism of Sesbania sesban to Na I. Rate of germination was not affected by the NaCI treatment with 50, 100, 150 and 200 mM. Fresh and dry weights were decreased with increasing NaCI concentration. Greater reduction of chlorophyll b content than chlorophyll a suggested that chlorophyll b is more susceptible to salt stress than chlorophyll a. Accumulation of Na⁺ and CI^- in the old leaves appears to be part of the mechanism to alleviate the salt toxicity. The higher accumulation of proline with increasing NaCI was found in the roots and shoots. Sesbania was found to have a greater tolerance to NaCI that could resist NaCI up to 200 mM is probably related with its ability to restrict Na⁺ and CI ªcontent in roots and translocate higher amount of the ions to shoots. The young and old leaves had a higher constitutive level of SOD, APX, CAT and GR activities with increasing NaCI concentrations. These findings indicated that S. sesban has higher levels of anti oxidative enzymes both constitutive and induced resulting in greater resistance to oxidative damage caused by NaCI stress.

2 illus, 4 tables, 14 ref

021601   Muraleedharan D;Gayathri Elayidam U (Centre for Arthropod Bioresources and Biotechnol, Univ of Kerala, Trivandrum, Email: drmurl@dataone.in) : Prospects and promises of endocrine biopesticides. J Biopestic 2008, 1(1), 6-11.

Environment hazards resulting from intensive use of synthetic organic crop protection agents, demands that the pest management studies should be biointensive. Environment friendly, safe and compatible approaches paved the way to develop biopesticides. The multiple functional capacities of insect neuropeptides, based on intervention on this system at any level provide opportunities for new insect control strategies. Potential biochemical for insecticidal action like Juvenile hormone, Juvenile hormone esterase, ecdysone, acetyl choline, ion channels and new peptides are discussed in detail.

47 ref

021600   Masud Hossain S;Anantharaman N (Chemical Engineering Dep, Mohamed Sathak Engineering College, Kilakarai-623 806) : Effect of wheat straw powder on enhancement of ligninolytic enzyme activity using Phanerochate chrysosporium. Indian J Biotechnol 2008, 7(4), 502-7.

A comparison made between the suspension culture of white-rot fungus, Phanerochate chrysosporium supplemented with wheat straw powder and control conditions (without wheat straw), revealed an enhancement in the ligninolytic enzymes activity in liquid shake flask in the former. Addition of 2% w/v of wheat straw powder improved greater stability to the lignin peroxidase (Li P) and manganese peroxidase (Mn P) enzymes when compared with the control. At optimum pH (4.5) and optimum temperature (40°C), maximum enzymatic activity was observed. The optimum shaking speed was found to be 60 rpm. The maximum activity of 538 and 512 μ mol/min mL was obtained for Li P and Mn P with wheat straw powder by the white-rot fungus at optimum conditions, respectively. However, maximum activity of 239 and 215 m mol/min mL, respectively was obtained for Li P and Mn P at similar conditions in the control.

021599   Maity B;Trivedi R;Kashyap V K (NO, Central Forensic Science Laboratory, 30 Gorachand Road, Kolkata-700 014) : Development of a macroarray based on 16S-23S rDNA probe hybridization for rapid diagnosis of human pathogenic bacteria. Indian J Biotechnol 2008, 7(4), 448-55.

The rapid identification of bacteria in biological specimens is important for selection of a suitable antimicrobial therapy. We have designed a rapid (<60>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.

021598   Mahantappa A;Banik A;Shivshankar J;Rao S G A (NO, Cryo Stem Cell Karnataka Pvt Ltd, Bangalore-560 024) : Stem cell in the umbilical cord blood is not related to volume and nucleated cell count. Indian J Biotechnol 2008, 7(4), 547-9.

The percent CD 34⁺ cells in umbilical cord tremendously varied from sample to sample. Study conducted on 100 cryopreserved samples for their volume, nucleated cell count and CD 34⁺ cell count showed no correlation between the three. Correlation study between volume and nucleated cell count showed poor correlation, where as correlation between volume and CD 34⁺, nucleated cell count and CD 34⁺ cell count showed no/negative correlation. Graphical representation of volume vs. nucleated cell count, volume vs. CD 34⁺ cell count, and nucleated cell count vs. CD 34⁺ cell count show the same results. These results have important bearing in umbilical cord blood banking, since sample acceptance/rejection at present is based only on volume of cord blood collected, which instead should be based on the number of CD 34⁺ stem cells.