Indian Journal of Biotechnology

 

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VOLUME 6

CODEN:IJBNAR 6(3) 287-426  (2007)

NUMBER 3

JULY 2007

ISSN:0972-5849

 

CONTENTS

 

Reviews

 

 

Functions and significance of nitric oxide in patho-physiological processes

293

 

IPC Code: Int. Cl.8 C01B21/24

 

 

Pankaj Gautam & S K Jain

 

 

 

 

 

Genetic transformation of arthropod vectors for control of vector borne diseases

305

 

        IPC Code: Int. Cl.8 C12N15/10

 

 

        P Azhahianambi & S Ghosh

 

 

 

 

 

Papers

 

 

 

 

 

A unique thermostable lichenase from Thermotoga maritima MSB8 with divergent substrate specificity

 

315

 

            IPC Code: Cl.8 C12N9/24

 

 

            Mohammed Abdul Sattar Khan, Mohammed Akbar, Motomitsu Kitaoka  & Kiyoshi Hayashi

 

 

 

 

 

Mutational drift prevails over translational efficiency in Frankia nif operons

321

 

IPC Code: Int. Cl.8 C12N15/10

 

 

Saubashya Sur, Arnab Sen & Asim K Bothra

 

 

 

 

 

Expression of 3AB protein of foot and mouth disease virus in Pichia pastoris

329

 

IPC Code: Int. Cl.8 C12N15/09, 15/11

 

 

            M Latha Priyadharshini, V Balamurugan, K Prabhudas, V V S Suryanarayana & G R Reddy

 

 

 

 

 

Microsatellite markers for genetic characterisation of Kachchhi camel

336

 

IPC Code: Int. Cl.8 C12N15/10, 15/16

 

 

S C Mehta, A Goyal & M S Sahani

     

 

       

 

 

Genetic analysis of CMS, restorer, hybrid and open-pollinated genotypes of Indian pearl millet [Pennisetum glaucum (L.) R. Br.] using ISSR markers

 

340

 

IPC Code: Int. Cl.8 C12N15/10

 

 

Satish Yadav, Sunita Jain, Veena Jain & Rajinder K Jain

 

 

 

 

 

Genetic diversity in tetraploid and diploid cotton (Gossypium spp.) using ISSR and microsatellite DNA markers

 

349

 

IPC Code: Int. Cl.8 C12N15/10

 

 

A B Dongre, Manoj Bhandarkar & Shubha Banerjee

 

 

RAPD markers for identifying oil palm (Elaeis guineensis Jacq.) parental varieties (dura & pisifera) and the hybrid tenera

 

354

 

IPC Code: Int. Cl.8 C12N15/10

 

 

D K Sathish & C Mohankumar

 

 

 

 

 

Biosorption of Cr(VI) from plating effluent using marine algal mass

359

 

IPC Code: Int. Cl.8 C02F11/02

 

 

E Thirunavukkarasu & K Palanivelu

 

 

 

 

 

Hapten synthesis, generation of polyclonal antibodies and development of ELISA for determination of thiamethoxam residues in food and environmental samples

 

365

 

        IPC Code: Int. Cl.8 C07K19/00; G01N33/53

 

 

Atmakuru Ramesh, Perumal Elumalai Thirugnanam & Prakhya Balakrishnamurthy

 

 

 

 

 

In vitro developmental ontogeny and life cycle of a rare fern species —Thelypteris confluens (Thunb.) Morton

 

372

 

IPC Code: Int. Cl.8 A01H4/00, 9/00

 

 

S Catharin Sara & V S Manickam

 

 

 

 

 

In vitro propagation of Dendrobium microbulbon A. Rich―A rare ethnomedicinal herb

381

 

IPC Code: Int. Cl.8 A01H4/00, 5/04

 

 

Urvashi Sharma, V Rama Rao, J S S Mohan & A S Reddy

 

 

 

 

 

A repeatable protocol for in vitro micropropagation of mulberry variety S54

385

 

IPC Code: Int. Cl.8 A01H4/00, 5/04

 

 

R Kavyashree

 

 

 

 

 

Improving decision making for drug candidates: A computational approach for benzthiazoles as antifungal

 

389

 

IPC Code: Int. Cl.8 G01N33/00

 

  

Abhay T Sangamwar, Uday D Deshpande, Sanjay S Pekamwar &
Sudhir M Vadvalkar

 

 

 

 

 

Nitrogen metabolism, artificial association study in two cyanobacterial isolates and assessment of their potential as biofertilizer

 

397

 

IPC Code: Int. Cl.8 C05F9/04

 

 

Mayashree Borthakur Syiem, Arvind Kumar Singh, Amar Nath Rai, Nonibala Khumanthem, Rajkumar Somendrajit Singh, Samrat Adhikari &Amrita Bhattacharjee

         

 

 

 

 

Short Communications

 

 

 

 

 

Isolation and molecular characterization of β-Tubulin gene from Arthrobotrys musiformisA nematode trapping fungus

 

404

 

        IPC Code: Int. Cl.8 C12N15/10

 

 

        J B Chauhan, R Swetha Priya, Bushra Jarullah & R B Subramanian

 

 

 

 

 

Isolation of Salmonella Typhimurium from poultry eggs and meat of Tarai region of Uttaranchal

 

407

 

        IPC Code: Int. Cl.8 C12N15/10

 

 

        K Nagappa, Shantanu Tamuly, Brajmadhuri, M K Saxena & S P Singh

 

 

Study of quality parameters in vermicomposting

410

 

IPC Code: Int. Cl.8 C05F3/00, 9/04

 

 

M C Borah, P Mahanta, S K Kakoty, U K Saha & A D Sahasrabudhe

 

 

 

 

 

Efficient plant regeneration from cotyledons of black gram [Vigna mungo (L.) Hepper]

414

 

        IPC Code: Int. Cl.8 A01H4/00, 5/10

 

 

Y Varalaxmi, A Vijayalakshmi, S K Yadav, B Venkateswarlu & M Maheswari

 

 

 

 

 

In vitro shoot regeneration from leaf and nodal explants of  Vernonia cinerea (L.) Less

418

 

IPC Code: Int. Cl.8 A01H4/00, 5/04, 5/12

 

 

Y N Seetharam, L N Rajanna, G Jyothishwaran, B Aravind, G Sharanabasappa & P B Mallikharjun

 

 

 

 

 

Sire effect on in vitro fertilizability of matured cattle oocytes

421

 

IPC Code: Int. Cl.8 C12N5/06, 15/06

 

 

K M Thara & S P Suresh Nair

 

 

 

 

 

Instructions to Contributors

423

 


 

 

AUTHOR INDEX

 

 


Adhikari S

397

Akbar M

315

Aravind B

418

Azhahianambi P

305

 

 

Balakrishnamurthy P

365

Balamurugan V

329

Banerjee S

349

Bhandarkar M

349

Bhattacharjee A

397

Borah M C

410

Bothra A K

321

Brajmadhuri

407

 

 

Chauhan J B

404

 

 

Deshpande U D

389

Dongre A B

349

 

 

Gautam P

293

Ghosh S

305

Goyal A

336

 

 

Hayashi K

315

 

 

Jain R K

340

Jain S

340

Jain S K

293

Jain V

340

Jarullah B

404

Jyothishwaran G

418

 

 

Kakoty S K

410

Kavyashree R

385

Khan M A S

315

Khumanthem N

397

Kitaoka M

315

 

 

Mahanta P

410

Maheswari M

414

Mallikharjun P B

418

Manickam V S

372

Mehta S C

336

Mohan J S S

381

Mohankumar C

354

 

 

Nagappa K

407

Nair S S P

421

 

 

Palanivelu K

359

Pekamwar S S

389

Prabhudas K

329

Priyadharshini M L

329

 

 

Rai A N

397

Rajanna L N

418

Rama Rao V

381

Ramesh A

365

Reddy A S

381

Reddy G R

329

 

 

Saha U K

410

Sahani M S

336

 

 

Sahsrabudhe A D

410

 

Sangamwar A T

389

 

Sara S C

372

Sathish D K

354

Saxena M K

407

Seetharam Y N

418

Sen A

321

Sharanabasappa G

418

Sharma U

381

Singh A K

397

Singh R S

397

Singh S P

407

Subramanian R B

404

Sur S

321

Suryanarayana V V S

329

Swetha Priya R

404

Syiem M B

397

 

 

Tamuly S

407

Thara K M

421

Thirugnanam P E

365

Thirunavukkarasu E

359

 

 

Vadvalkar S M

389

Varalaxmi Y

414

Venkateswarlu B

414

Vijayalakshmi A

414

 

 

Yadav S

340

Yadav S K

414


 

 


Reviews

Indian Journal of Biotechnology

Vol 6, July 2007, pp 293-304

 

 

 

Functions and significance of nitric oxide in patho-physiological processes

Pankaj Gautam and S K Jain*

Nitric oxide (NO) is an important signaling molecule that regulates a diverse range of physiological processes in many tissues. NO is enzymatically synthesized from L-arginine by three isoforms of nitric oxide synthase (NOS). It plays an important role in a large number of normal physiological (regulation of blood pressure, neurotransmission, wound repair and host defense mechanisms) as well as in patho-physiological (inflammation, infection, apoptosis, neoplastic diseases, liver cirrhosis, diabetes) processes. With an unpaired electron, NO is a strong pro-oxidant produced in conditions like sepsis. It also exerts its effect through other free radicals such as superoxide and hydroxyl ions and causes oxidative stress within the cell. Reactive nitrogen intermediates play a central role in cell death (apoptosis), which is mediated by the induction of pro-inflammatory cytokines such as tumour necrosis factor. NO mediated apoptosis occurs in various cell types such as macrophages, lymphocytes, thymocytes, endothelial cells. At lower concentration (10 nM-1 μM) NO has been shown to have antiapoptotic effect, which is cyclic GMP dependent. One of the most beneficial functions of NO is its implication in host defense against intracellular pathogens (Salmonella and Leishmania). Its derivatives such as per-oxynitrite are strong bactericidal in nature. Involvement of NO in inflammatory responses has been shown not only in experimental models but also in human inflammatory diseases. The antioxidant and antiapoptotic properties make this molecule of great therapeutic significance.

Keywords: NO, iNOS, apoptosis, TNF

IPC Code: Int. Cl.8 C01B21/24

Indian Journal of Biotechnology

Vol 6, July 2007, pp 305-314

 

 

 

Genetic transformation of arthropod vectors for control of vector borne diseases

P Azhahianambi and S Ghosh*

The vector borne diseases cause heavy loss to mankind and livestock industry throughout the world. Global climate changes reported to contribute to the recurrence and new epidemics of vector borne diseases. Unfortunately, the available strategies to control vector borne diseases are insufficient and public health burden of the major vector borne diseases is on increasing trend. Complete eradication of a vector population is not realistic and desirable for many arthropod vector species due to biological constraints. Genetic transformation of vectors offers the way to control the diseases transmitted by the arthropod vectors without killing them and is also an economically viable option. Genetic transformation of arthropod vectors is a new strategy in the genomic era in which the synthetic effector gene is introduced into the genome of vectors to block the developmental stages of parasite/pathogen inside the vector and subsequent driving of the effector gene into the wild vector population in a geographical area. However, the knowledge on the putative risk factors associated with the release of transgenic arthropod vectors in the field is lacking and considerable work is needed before the deployment of transgenic arthropod vectors in nature.

Keywords:   arthropod vectors, vector borne diseases, genetic transformation, effctor genes, transformation marker genes, viral vectors, transposons

IPC Code: Int. Cl.8 C12N 15/10

 

Papers

Indian Journal of Biotechnology

Vol 6, July 2007, pp 315-320

 

 

 

A unique thermostable lichenase from Thermotoga maritima MSB8 with divergent substrate specificity

Mohammed Abdul Sattar Khan1*, Mohammed Akbar2, Motomitsu Kitaoka1 and Kiyoshi Hayashi1

A putative endoglucanase gene corresponding to locus TM 1752 (AAD36817.1, Q9X274) of Thermotoga maritima MSB8 was cloned and expressed in Escherichia coli. The enzyme contains a NEP (Asn-Glu-Pro) motif but lacks a PCD (proposed catalytic domain) block. This endoglucanase is grouped under family 5 of the glycosyl hydrolases members of which predominantly hydrolyze β-1,4-linkages of carbohydrate polymers. The enzyme efficiently hydrolyzes the glycosidic bonds of mixed β-1,3-1,4 linkages in lichenan and barley β-glucan, but did not display any activity on crystalline cellulose or laminarin. However, negligible amount of hydrolysis was observed with CM-cellulose. This enzyme produced glucosyl β-1,3 glucosyl β-1,4 glucose as the major end product and glucosyl β-1,4 glucosyl β-1,3 glucose was not detected. The putative endoglucanase of T. maritima appears to be a unique endoglucanase being the first lichenase producing glucosyl β-1,3 glucosyl β-1,4 glucose to be placed into family 5 of the glucosyl hydrolases.

Keywords: Thermotoga maritima, endoglucanase, lichenase, family 5, glucosyl β-1,3 glucosyl β-1,4 glucose, glucosyl β-1,4
glucosyl β-1,3 glucose

IPC Code: Int. Cl.8 C12N9/24

Indian Journal of Biotechnology

Vol 6, July 2007, pp 321-328

 

 

 

Mutational drift prevails over translational efficiency in Frankia nif operons

Saubashya Sur1, Arnab Sen1* and Asim K Bothra2

Frankia is a Gram-positive actinomycete, fixing atmospheric nitrogen while making associations with a number of dicotyledonous plants. The most important operon of Frankia is the nif operon. Codon usage bias was investigated in three open reading frames and known genes of this organism from three different strains. Genes and ORFs show high biasness due to mutation pressure. Based on our results we conclude that mutational drift is more relevant over translational efficiency. The nif genes showed fairly high expression levels. Analysis of the result of codon indices and BLAST searches revealed that ORF3 might be a good candidate for expression.

Keywords: codon usage, Frankia, mutational bias, nitrogen fixation, nif , translational efficiency, ORF

IPC Code: Int. Cl.8 C12N15/10

Indian Journal of Biotechnology

Vol 6, July 2007, pp 329-335

 

 

 

Expression of 3AB protein of foot and mouth disease virus in Pichia pastoris

M Latha Priyadharshini1, V Balamurugan, K Prabhudas, V V S Suryanarayana and G R Reddy*

3AB non structural protein (nsp) was used to diagnose the vaccinated animals from those infected with foot and mouth disease virus (FMDV). In order to express the gene encoding 3AB protein of FMDV type A22 in Pichia pestoris, the gene was amplified and cloned into the yeast transfer vector (pPIC-9K) at EcoRI site. The cloned gene was further characterized by colony PCR, restriction enzyme digestion and sequence analysis. The recombinant plasmid was transferred into GS115 strain of P. pastoris cells by electroporation. The His+ Pichia transformants were analyzed for the presence of the insert in the yeast genome by PCR. PCR positive clones were grown and expression was induced with 0.5% methanol. The expressed gene products were then characterized by SDS-PAGE and Western blot analysis. This is the first report on the production of FMDV non structural proteins in yeast. The expressed protein will be of diagnostic importance.

Keywords: 3AB protein, expression, FMDV, Pichia pastoris

IPC Code: Int. Cl.8 C12N 15/09, 15/11

Indian Journal of Biotechnology

Vol 6, July 2007, pp 336-339

 

 

 

Microsatellite markers for genetic characterisation of Kachchhi camel

S C Mehta*, A Goyal and M S Sahani

Sixteen microsatellite loci were investigated for studying the genetic polymorphism in Kachchhi breed of camel. Polymerase chain reactions were carried out for 50 unrelated camels of Kachchhi breed. The amplification products were resolved in 6% (denaturing) urea PAGE and stained with silver nitrate. Out of the 16 microsatellite loci, 13 were found polymorphic. The number of alleles ranged from 2 to 6. The expected heterozygosity ranged from 0.332 to 0.796. The polymorphic information content ranged from 0.277 to 0.765. The results indicated existence of enough genetic variation among dromedary individuals and the potential use of microsatellite markers for further genetic investigations including genetic diversity analysis, individual identification, parentage testing and production enhancement.

Keywords: breeds, camel, genetic characterisation, Kachchhi, microsatellite

IPC Code: Int. Cl.8 C12N15/10, 15/16

Indian Journal of Biotechnology

Vol 6, July 2007, pp 340-348

 

 

 

Genetic analysis of CMS, restorer, hybrid and open-pollinated genotypes of Indian pearl millet [Pennisetum glaucum (L.) R. Br.] using ISSR markers

Satish Yadav1,  Sunita Jain1, Veena Jain1 and Rajinder K Jain2*

Genetic diversity and relationships among 20 Indian pearl millet [Pennisetum glaucum (L.) R. Br.] genotypes representing the commercially important hybrids, cytoplasmic male sterile (CMS) lines, restorer (res) lines and open pollinated varieties (opv) were assessed using inter simple sequence repeat (ISSR) markers. A total of 349 reproducible bands (73 monomorphic, 276 polymorphic) were detected using 30 UBC#9 ISSR primers with polymorphism percentage of 79.1%. Fortyfive of the polymorphic bands were in fact the unique bands that were present or absent exclusively in only one of the 20 genotypes. A set of two or any five ISSR-PCR assays were sufficient to clearly distinguish among the 20 genotypes. The salient features of ISSR marker data analyzed using clustering algorithms (NTSYS-Pc and Win Boot) and principal component analysis (two-dimensional, NTSYS-pc) are as given below: (i) all the four restorer lines, which have been widely used for hybrid production, were clustered in a single group, which is indicative of their narrow genetic base, (ii) the seven CMS lines, which are basically bred from Tift23A or Tift23A derived lines after crosses/backcrosses with some downy mildew resistance sources, displayed low to moderate levels of polymorphism, (iii) the three OPVs were quite diverse with HC-20 being more distant to HC-10 and HC-4, and (iv) hybrids such as HHB-60 and HHB-67, with same restorer parentage exhibited greater relatedness. The high resolution and reproducibility of ISSR-PCR assay shown in this study shall provide a simple, rapid and cost-effective system for the characterization of huge collection of germplasm and testing of purity of genetic stocks/hybrids. The study also demonstrates the need for selecting/generating genetically diverse restorer as well as CMS lines for the breeding of genetically diverse hybrids in pearl millet.

Keywords: genetic diversity, genotyping, ISSR, pearl millet, Pennisetum glaucum, CMS, restorer, OPV

IPC Code: Int. Cl.8 C12N15/10

Indian Journal of Biotechnology

Vol 6, July 2007, pp 349-353

 

 

 

Genetic diversity in tetraploid and diploid cotton (Gossypium spp.) using ISSR and microsatellite DNA markers

A B Dongre*, Manoj Bhandarkar and Shubha Banerjee

Two marker systems viz., microsatellite and ISSR were used to generate DNA profiles of 19 genotypes of cotton (diploid and tetraploid), and each major DNA fragment was recorded. The results of cluster analysis indicated not only the separation of genotypes of the two major species (Gossypium arboreum and G. hirsutum) into separate groups, but also the separation of the genotypes among each group. The similarity coefficient based on ISSR and microsatellite markers ranged from 0.59 to 0.90 and 0.59 to 0.93, respectively, thus suggesting considerable genetic variation between the cotton species studied.

Keywords: Gossypium, genetic diversity, microsatellite DNA marker, ISSR

IPC Code: Int. Cl.8 C12N15/10

Indian Journal of Biotechnology

Vol 6, July 2007, pp 354-358

 

 

 

RAPD markers for identifying oil palm (Elaeis guineensis Jacq.) parental varieties (dura & pisifera) and the hybrid tenera

D K Sathish and C Mohankumar*

Random amplification of polymorphic DNA (RAPD) analysis was done by arbitrary primers for determining the DNA polymorphism among the oil palm (Elaeis guineensis) varieties dura, pisifera and tenera, and monitoring the specificity of the primers for identifying each genotype. The three varieties were evaluated using thirty, 10-mer primers. Of the 30 primers, 26 yielded significant polymorphic DNA bands. A total of 185 bands were determined (300-1500 bp), of which 59 were representing dura, 65 for pisifera and 61 bands for tenera. The dendrogram obtained, using UPGMA, grouped all the three varieties into two distinct clusters. In one cluster, the shelled ones (dura and tenera) were grouped, while the other comprised the shell-less one (pisifera). The data also provide sufficient evidence for identifying each variety, dura, pisifera and tenera separately as well as the parentals dura and pisifera together. This is the first report of DNA based polymorphism assay to assess the level of variability in oil palm varieties.

Keywords: Elaeis guineensis, hybrid, parental varieties, random primers, RAPD

IPC Code: Int. Cl.8 C12N 15/10

Indian Journal of Biotechnology

Vol 6, July 2007, pp 359-364

 

 

 

Biosorption of Cr(VI) from plating effluent using marine algal mass

E Thirunavukkarasu and K Palanivelu*

Biosorption of heavy metals is emerging as an effective technology for the treatment of industrial wastewater. In this study, biosorption potential of Padina boergesenli–a brown marine alga was assessed in the removal of Cr(VI) from electroplating effluent. Batch experiments were conducted to determine the adsorption properties of the P. boergesenli and it was observed that the biosorption capacity strongly depends on pH. The maximum removal of Cr(VI) was around 49 mg/g of dry weight of biomass at pH 1.0 in 180 min with 100 mg/L of initial concentration. The experimental results were fitted to the Langmuir and Freundlich isotherm models and the Freundlich isotherm was found to well represent the measured sorption data implying P. boergesenli has heterogeneous surface. The kinetic study showed that the biosorption of Cr(VI) by P. boergesenli follows second order. The ability of P. boergesenli to biosorb Cr(VI) in a packed column was investigated, as well. The experiments were conducted to study the effect of important design parameters such as bed height and flow rate. The biosorption decreased as the flow rate increased. The bed depth service time model was used to analyze the experimental data.

Keywords: biosorption, packed column, Padina boergesenli, electroplating, Cr(VI)

IPC Code: Int. Cl.8 C02F11/02Indian Journal of Biotechnology

Vol 6, July 2007, pp 365-371

 

 

 

Hapten synthesis, generation of polyclonal antibodies and development of ELISA for determination of thiamethoxam residues in food and environmental samples

 

Atmakuru Ramesh*, Perumal Elumalai Thirugnanam and Prakhya Balakrishnamurthy

An ELISA (enzyme-linked immunosorbent assay) method for the determination of thiamethoxam residues has been developed. The method involves the synthesis of hapten and generation of polyclonal antibodies in rabbit using the hapten-protein (bovine serum albumin) conjugate carrier. Recovery experiments show the accuracy of estimation of thiamethoxam residues that was within the linear working range of 1 to 60 ng mL– 1 with the r2=0.992. Influence of different parameters, like detergent concentration, solvent, pH, ionic strength, has been studied. Cross reactivity was tested using different analogs. The method tolerates the solvent methanol concentration up to 15%. However, other polar solvents influence negatively on the binding. The detergent Tween 20 has not made any impact on the absorbance. The optimized ELISA for thiamethoxam used 1-60 ng/well and an antiserum dilution between 10000 and 200000. Recoveries were above 98%. The method was successfully applied for the analysis of food and environmental samples.

Keywords: ELISA, residues, thaimethoxime, water and food samples, hapten, polyclonal antibodies

IPC Code: Int. Cl.8 C07K19/00; G01N33/53

Indian Journal of Biotechnology

Vol 6, July 2007, pp 372-380

 

 

 

In vitro developmental ontogeny and life cycle of a rare fern species—
Thelypteris confluens (Thunb.) Morton

 

S Catharin Sara* and V S Manickam

The conservation of Thelypteris confluens, which was under the list of rare and endangered ferns of Western Ghats of South India, is the major concern of this work. The in vitro life cycle starting from spore germination, followed by micromorphological changes and the reproductive biology with regard to the monocultures of this species was thoroughly studied in simple medium. The fern showed specific germination and growth pattern of Vittaria and Drynaria type. Sex organs in male, female and hermaphroditic gametophytes underwent fertilization and the sporophytes developed with sprouting leaves within 120 d in the cultures. Harderning and acclimatization procedures responded very well to the sea level as well as to high altitudes of 1800-2400 m and proved to be having a very good propagating potential with regard to spore culture. The regeneration of miniature gametophytes from the old male prothalli as filamentous structures indicates the autotrophy and rejuvenating potential of this species. Results of micromorphology, gender analysis and reproductive potential justify the rarity of this species.

Keywords: conservation protocols, in vitro life cycle, reproductive studies, Thelypteris confluens (Thunb.) Morton

IPC Code: Int. Cl.8 A01H4/00, 9/00

Indian Journal of Biotechnology

Vol 6, July 2007, pp 381-384

 

 

 

In vitro propagation of Dendrobium microbulbon A. Rich―A rare
ethnomedicinal herb

Urvashi Sharma, V Rama Rao, J S S Mohan* and A S Reddy

 

Dendrobium microbulbon A. Rich (Orchidaceae) is a small epiphytic rare orchid used in stomachache by the tribal people of Gujarat. The present study deals with the development of a suitable protocol for its clonal propagation using pseudobulbs as explants. Murashige and Skoog (MS) basal medium supplemented with various concentrations of auxin and cytokinin was used either alone or in combination for induction, multiplication, elongation and rooting of in vitro shoots. MS medium fortified with 3% sucrose, 7.5 mg L-1 indole-3-acetic acid (IAA) and 20 mg L-1 benzyl aminopurine (BAP) showed axillary bud initiation. MS medium with 2.0 mg L-1 BAP favoured production of maximum number of shoots
(39 shoots), elongation of shoots as well as bulblets formation. In vitro raised shoots were rooted on Knop salts with MS vitamins and MS iron salts. During acclimatization, 60% of the plantlets survived after their removal from in vitro conditions.

Keywords: conservation, in vitro flowering, secondary explants, shoot multiplication, pseudobulbs, Dendrobium microbulbon

IPC code: Int. Cl.8 A01H4/00, 5/04

Indian Journal of Biotechnology

Vol 6, July 2007, pp 385-388

 

 

 

A repeatable protocol for in vitro micropropagation of mulberry variety S54

R Kavyashree*

 

Apical buds of mulberry (Morus indica L.) variety S54 were cultured on LSBM supplemented with different concentrations of BAP and TIBA to study their in vitro micropropagation potential. It was found that LSBM fortified with 8.88 µM BAP in combination with 2 µM TIBA was the most suitable medium for initiation (94% response) and multiplication of shoots (10.6) when compared to other concentrations tried. The well developed, elongated, healthy multiple shoots were subjected to ex vitro rooting technique to obtain complete plantlets. The statistical analysis of the data revealed significant difference between and within the treatments with regard to multiple shoots and root formation. The well developed axenic plants with similar genetic makeup were successfully established in the field with 90% survival frequency through sequential hardening process.

Keywords: apical bud, ex vitro rooting, micropropagation, multiple shoots, survival frequency

IPC Code: Int. Cl.8 A01H4/00, 5/04

Indian Journal of Biotechnology

Vol 6, July 2007, pp 389-396

 

 

 

Improving decision making for drug candidates: A computational approach for benzthiazoles as antifungal

 

Abhay T Sangamwar1*, Uday D Deshpande2, Sanjay S Pekamwar1and Sudhir M Vadvalkar1

To improve the decision making for selecting a molecule for synthesis from a set of virtually designed antifungal compounds, four in silico approaches are discussed and their optimization have been confirmed by in vitro methods. First, a set of compounds was designed over QSAR analysis and selected over predicted activities by best QSAR equation (r2>0.9) to increase the probability of finding new chemical entities. To reduce the burden of synthetic chemistry, Rule of Five was applied for screening druggable compounds. Second, OSIRIS was used to predict toxicity, mutagenicity and carcinogenicity. Third approach was prediction of biological activity spectra for substances (PASS) and fourth was molecular docking to analyze the actual interaction involved after binding with the target receptor. All compounds of the series passed rule of five. On the basis of OSIRIS prediction results, nine compounds were selected. PASS predictions have resulted into suitability of only one compound (test 30) as antifungal. Molecular docking at BioMed CAChe workstation was then performed for this compound. Comparison of active site residues at receptor binding and dock score with fluconazole has further confirmed the compound for synthesis. The synthesized compound has given MIC 16 µg/mL at antifungal assay against Candida albicans by potato dextrose agar method. It confirmed that in silico approaches are useful to find out a new drug with more accuracy.

Keywords: antifungal assay, benzthiazoles, BioMed CAChe, docking, OSIRIS, PASS, QSAR, Rule of Five

IPC Code: Int. Cl.8  G01N33/00

Indian Journal of Biotechnology

Vol 6, July 2007, pp 397-403

 

 

 

Nitrogen metabolism, artificial association study in two cyanobacterial isolates and assessment of their potential as biofertilizer

 

Mayashree Borthakur Syiem*, Arvind Kumar Singh, Amar Nath Rai1, Nonibala Khumanthem,
Rajkumar Somendrajit Singh, Samrat Adhikari and Amrita Bhattacharjee

Two strains of cyanobacteria, viz. Nostoc ANTH and Mastigocladus sp, were isolated from local separate temperature zones of Meghalaya, India. Both the strains showed preference for different temperatures for optimum growth [45°C for Mastigocladus sp.(thermophile) and 25°C for Nostoc ANTH (mesophile)]. The addition of nitrogen sources in the growth media (nitrate, ammonia and glutamine) supported their better growth but repressed heterocyst development and nitrogenase activity. Nitrate and nitrite uptake rates, NR and NiR activities increased by NO3Ż and decreased by NH4+ in Nostoc ANTH. However, such effects were only partial in Mastigocladus sp. The presence of fixed nitrogen sources in the media led to decreased GS activity and repressed methylammonium uptake in both the strains. Glutamine uptake was substrate inducible, energy-dependent and required de novo protein synthesis. Artificial association studies revealed successful establishment of association of rice roots with both cyanobacteria, including prolonged association of Mastigocladus sp. at high temperature (~45°C). Little modifications in growth temperature and growth media led to profuse akinete differentiation in target cyanobacteria. The replacement of normal cells by akinetes as field inoculants might have profound biotechnological implications in future biofertilizer programme.

Keywords: cyanobacteria, N2-fixation, mesophile, thermophile, biofertilizer

IPC Code: Int. Cl.8 C05F9/04

 

Short Communications

Indian Journal of Biotechnology

Vol 6, July 2007, pp 404-406

 

 

 

Isolation and molecular characterization of β-tubulin gene from Arthrobotrys musiformisľA nematode trapping fungus

J B Chauhan1, R Swetha Priya2, Bushra Jarullah2 and R B Subramanian2*

A new isolate of Arthrobotrys musiformis, a naturally occurring predacious hyphomycete, has been found to be effective against Haemonchus contortus, a roundworm, which seriously affects milk and meat production in ruminants. However, further studies on the suitability of this fungus for use in integrated pest management (IPM) showed that it is highly susceptible to benzimidazole derivatives, which are the most common antihelmintics currently used for deworming cattle. β-tubulin plays a major role in conferring resistance to benzimidazols. The β-tubulin gene of A. musiformis was isolated from genomic DNA through PCR and sequenced. BLAST analysis revealed 95% homology with β-tubulin gene of A. oligospora followed by other filamentous fungi. The isolated gene lacks mutation, which confers susceptibility to benzimidazols in the gene sequence of A. musiformis.

Keywords: Arthrobotrys musiformis, nemataphagous fungus, β-tubulin, PCR, pGEM-T-easy vector, benzimidazole

IPC Code: Int. Cl.8 C12N15/10

Indian Journal of Biotechnology

Vol 6, July 2007, pp 407-409

 

 

Isolation of Salmonella Typhimurium from poultry eggs and meat of Tarai region of Uttaranchal

K Nagappa1*, Shantanu Tamuly2, Brajmadhuri1, M K Saxena2 and S P Singh1

Presence of Salmonella in samples, 100 each of chicken eggs and meat, collected from various retail outlets of the Tarai region of Uttaranchal, was detected by polymerase chain reaction (PCR) using InvA gene primer, a rapid and sensitive DNA based method. The results were further validated by established conventional method of Salmonella isolation and biochemical tests, including slide agglutination test (poly “O” antiserum specific for Salmonella). Of 200 samples analyzed, 4 (one meat and 3 egg) were found to be contaminated with Salmonella enterica var Typhimurium. Further, the antibiogram of the isolates revealed sensitivity in the decreasing order to Chloromphenicol, Colistin, Polymixin, Enrofloxacin, and Ciprofloxacin.

Keywords:      antibiotic sensitivity, eggs, meat, PCR, poultry, Salmonella

IPC Code: Int. Cl.8 C12N15/10

Indian Journal of Biotechnology

Vol 6, July 2007, pp 410-413

 

 

 

Study of quality parameters in vermicomposting

M C Borah, P Mahanta, S K Kakoty*, U K Saha and A D Sahasrabudhe

In the present study, vermicompost samples were tested for chemical constituents. Effect of temperature and density of cocoons on the production of vermicompost was also studied. Samples collected from vermicompost units installed in the laboratory as well as in the field provided an opportunity to analyze quality assessment of the vermicompost and introduction of a three-pit system of vermicomposting.

Keywords: vermicompost, denitrification, cocoon, temperature, earthworm, Perionyx excavatus

IPC Code: Int. Cl.8 C05F3/00, 9/04

Indian Journal of Biotechnology

Vol 6, July 2007, pp 414-417

 

 

 

Efficient plant regeneration from cotyledons of black gram [Vigna mungo (L.) Hepper]

 

Y Varalaxmi, A Vijayalakshmi, S K Yadav, B Venkateswarlu and M Maheswari*

An efficient and rapid regeneration protocol was developed from cotyledons of black gram [Vigna mungo (L.) Hepper] variety T-9. Murashige and Skoog’s (MS) medium supplemented with Gamborg B5 vitamins containing BAP @ 4.0 mg/L was most effective in producing regenerative calli. The cultures expressed maximum plant regeneration potential with 12 shoots per calli on regeneration media with no exogenous amino acid or plant growth regulator supplementation. Green and robust shoots thus developed were successfully rooted within 15 d on 1/3 MS medium. Over 90% of rooted plants grew well and produced seeds normally when transferred to green house.

Keywords: black gram, grain legumes, in vitro culture, plant growth regulators, plant regeneration, Vigna mungo

IPC Code: Int. Cl.8 A01H4/00, 5/10

Indian Journal of Bioatechnology

Vol 6, July 2007, pp 418-420

 

 

 

In vitro shoot regeneration from leaf and nodal explants of Vernonia cinerea (L.) Less

 

Y N Seetharam*, L N Rajanna, G Jyothishwaran, B Aravind, G Sharanabasappa and P B Mallikharjun

Multiple shoots were obtained from leaf and nodal explants of Vernonia cinerea on MS media supplemented with different concentrations of BAP. Maximum numbers of shoots were obtained from leaf and nodal explants at 3.0 mg L-1 and 2.5 mg L-1 concentrations of BAP, respectively. Shoot multiplication and their development was better on MS medium supplemented with BAP (2.0 mg L-1) along with NAA (1.5 mg L-1) for both the explants. Rhizogenesis was observed when in vitro shoots were subcultured in half strength MS medium containing IAA (1.5 mg L-1). The in vitro raised plantlets were acclimatized and established successfully in the field.

Keywords: multiple shoot, proliferation, V. cinerea

IPC Code: Int. Cl.8 A01H4/00, 5/04, 5/12

Indian Journal of Biotechnology

Vol 6, July 2007, pp 421-422

 

 

 

Sire effect on in vitro fertilizability of matured cattle oocytes

 

K M Thara1* and S P Suresh Nair

Sire effect on in vitro fertilization of matured cattle oocytes were tested using frozen thawed semen. Oocytes collected from ovaries of slaughtered cattles were matured at in vitro conditions and fertilized with frozen thawed semen in the fertilization medium with different heparin concentrations. Fertilization rate for five different bulls was determined. Semen from different bulls was found to have different rates of fertilization on in vitro matured cattle oocytes.

Keywords: cattle, in vitro maturation, in vitro fertilization, oocytes

IPC Code: Int. Cl.8 C12N5/06, 15/06


 

 


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