Indian Journal of Biotechnology

 

Editorial Board

 


Prof. N S Punekar

Indian Institute of Technology Bombay

Mumbai 400 076

E-mail: nsp@iitb.ac.in

 

Prof. Sudhir Sopory

International Centre for Genetic

Engineering & Biotechnology

New Delhi 110 067

E-mail: sopory@icgeb.res.in

 

Prof. A Surolia

National Institute of Immunology

New Delhi 110067

E-mail: surolia@mbu.iisc.ernet.in

 

Dr Ashok Pandey

Regional Research Laboratory

Thiruvananthapuram 695 019

E-mail: ashokpandey56@yahoo.co.in

 

Prof. Suresh Chand

University of Lucknow

Lucknow 226 007

E-mail: sureshchand55@hotmail.com

 


Prof. Debabrata Das

Indian Institute of Technology

Kharagpur 721 302

E-mail: ddas52003@yahoo.com

 

 

Dr V K Kashyap

National Institute of Biologicals

NOIDA 201 307

E-mail:info@nib.gov.in

 

 

Prof. Rakesh Bhatnagar

Jawahar Lal Nehru University

New Delhi 110 067

E-mail rakbhat01@yahoo.com

 

 

Dr Rita Mulherkar

Advanced Centre for Treatment, Research & Education in Cancer

Navi Mumbai 410 208

E-mail: rmulherkar@actrec.gov.in

 

Dr Rakesh Tuli

National Botanical Research Institute

Lucknow 226 001

E-mail: rakeshtuli@hotmail.com


Prof. Jong M Park

Pohang University of Science & Technology

Pohang 790-784

Korea

E-mail: jmpark@postech.ac.kr

 

 

Prof. Christian Larroche

Laboratoire de Genie Chimique et

Biochemique

Aubiere Cedex

France

E-mail: christian.larroche@univ-bpclermont.fr

 

Prof. S D Pillai

Texas A&M University

Texas 77843-2472

USA

E-mail: spillai@poultry.tamu.edu

 

Prof. Indra K Vasil

University of Florida

Gainesville, FL

USA

E-mail: ivasil@ufl.edu


 

Director: NISCAIR (Ex-Officio)

Editor: Dr K Y Kavathekar  Associate Editor: Dr M A A Khan

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ă2008The Council of Scientific & Industrial Research, New Delhi.

 

Indian Journal of Biotechnology

 

http://www. niscair.res.in

 

VOLUME7

CODEN: IJBNAR 7(3) (2008) 277-414

NUMBER3

JULY2008

ISSN: 0972-5849

 

CONTENTS 

Review

 

Edible vaccines: A new approach to oral immunization

283

Neeraj Mishra, Prem N Gupta, Kapil Khatri, Amit K Goyal & Suresh P Vyas

 

 

 

Papers

 

Enhancement of NAPL bioavailability by induction of cell-surface hydrophobicity in Exiguobacterium aurantiacum and Burkholderia cepacia

295

Gita Mohanty & Suparna Mukherji

 

 

 

Amplification, cloning and sequencing of Enterococcus feacalis enolase gene

307

A Shivaraj, S Madhusudan, C Ashok Kumar, S Isloor, Veere Gowda & H J Dechamma

 

 

 

Assessment of genetic variation among populations of Rhynchostylis retusa, an epiphytic orchid from Goa, India using ISSR and RAPD markers

313

G V Parab & S Krishnan

 

 

 

RAPD based assessment of genetic diversity of Butea monosperma from different agro-ecological regions of India

320

Vaishali, Suphiya Khan & Vinay Sharma

 

 

 

Immobilization of porcine pancreas lipase onto free and affixed arylamine glass beads and its application in removal of oil stains

328

M Sharma, V Kumar & C S Pundir

 

 

 

Extracellular collagenase from Rhizoctonia solani: Production, purification and characterization

333

Hossam S Hamdy

 

 

 

Enhanced production of streptomycin and hydrolytic enzymes by Streptomyces griseus strains using different types of organic solvents and detergent compounds

341

Kamal Y I El-shahed, Ahmed I El-Diwany & Hassan M M Awad

 

 

 

Isolation of borrelidin from Streptomyces californicus—An Indian soil isolate

349

Srinivasan Saisivam, D V R N Bhikshapathi, J Krishnaveni & V Kishan

 

 

 

Production of alkaline protease by Bacillus circulans using agricultural residues: A statistical approach

356

R K Jaswal, G S Kocher & M S Virk

 

 

 

Optimization of malting conditions of Pearl millet [Pennisetum glaucum (L.)R.Br.] using
Box-Behnken design

361

P Mary Anupama, C Ayyanna & T Ramana

 

 

 

Host associated genetic variations in whitefly, Bemisia tabaci (Genn.)

366

R K Sharma, V K Gupta, J Jindal & V K Dilawari

 

 

 

Significance of metabolites of native Xenorhabdus, a bacterial symbiont of Steinernema, for suppression of collar rot and root knot diseases of groundnut

371

R V Vyas, Biren Patel, Ajay Maghodia & D J Patel

 

 

 

Increased isoflavonoids accumulation in cell suspension cultures of Pueraria tuberosa by elicitors

378

Shaily Goyal & K G Ramawat

 

 

 

In vitro propagation and quercetin quantification in callus cultures of Rasna (Pluchea lanceolata
Oliver & Hiern.)

383

Deepika Arya, Vidya Patni & Uma Kant

 

 

 

Direct shoot organogenesis and plant regeneration from hypocotyl explants in selected genotypes of Leucaena leucocephala—A leguminous pulpwood tree

388

V L Sirisha, S Prashant, D Ranadheer, P Ramprasad, N M Shaik, Manish Arha, S K Gupta, Sameer Srivastava, A K Yadav, P S Kulkarni, O U Abhilash, B M Khan, Shuban K Rawal &
P B Kavi Kishor

 

 

 

In vitro shoot proliferation in Emblica officinalis var. Balwant from nodal explants

394

Divya Goyal & Seema Bhadauria

 

 

 

Short Communications

         

Polymorphism in DRB3 exon 2 by PCR-RFLP and its association with mastitis in Nili-Ravi breed

398

Sudhir Kumar, M L Sangwan & Rupender

 

 

 

Evaluation of genetic diversity in Mecheri sheep (Ovis aries) of Tamil Nadu using microsatellite markers

401

S Prema, S N Sivaselvam & S M K Karthickeyan

 

 

 

Role of calcium ions and temperature on dextransucrase production

404

Shal Ali Ul Qader, Afsheen Aman, Saeeda Bano, Noman Syed & Abid Azhar

 

 

 

Efficient plant regeneration in small cardamom (Elettaria cardamomum Maton.) through somatic embryogenesis

407

C Mahonari, S Backiyarani, T Jebasingh, Archana Somanath & R Usha

 

 

 

Instructions to Contributors

411

 

 

AUTHOR INDEX


Abhilash O U

388

Aman A

404

Anupama P M

361

Arha M

388

Arya D

383

Awad H M M

341

Ayyanna C

361

Azhar A

404

 

 

Backiyarani S

407

Bano S

404

Bhadauria S

394

Bhikshapathi D V R

349

 

 

Dechamma H J

307

Dilawari V K

366

 

 

El-Diwany A I

341

El-shahed K Y I

341

 

 

Gowda V

307

Goyal A K

283

Goyal D

394

Goyal S

378

Gupta P N

283

Gupta S K

388

Gupta V K

366

 

 

Hamdy H S

333

 

 

Isloor S

307

 

Jaswal R K

356

Jebasingh T

407

Jindal J

366

 

 

Kant U

383

Karthickeyan S M K

401

Kavi K P B

388

Khan B M

388

Khan S

320

Khatri K

283

Kishan V

349

Kocher G S

356

Krishnan S

313

Krishnaveni J

349

Kulkarni P S

388

Kumar C A

307

Kumar S

398

Kumar V

328

 

 

Madhusudan S

307

Maghodia A

371

Mahonari C

407

Mishra N

283

Mohanty G

295

Mukherji S

295

 

 

Parab G V

313

Patel B

371

Patel D J

371

Patni V

383

Prashant S

388

Prema S

401

 

Pundir C S

328

Qader S A U

404

Ramana T

361

 

 

Ramawat K G

378

Ramprasad P

388

Ranadheer D

388

Rawal S K

388

Rupender

398

 

 

Saisivam S

349

Sangwan M L

398

Shaik N M

388

Sharma M

328

Sharma R K

366

Sharma V

320

Shivaraj A

307

Sirisha V L

388

Sivaselvam S N

401

Somanath A

407

Srivastava S

388

Syed N

404

 

 

Usha R

407

 

 

Vaishali

320

Virk M S

356

Vyas R V

371

Vyas S P

283

 

 

Yadav A K

388


 


 

Indian Journal of Biotechnology

Vol 7, July 2008, pp 283-294

 

Edible vaccines: A new approach to oral immunization

 

Neeraj Mishra, Prem N Gupta, Kapil Khatri, Amit K Goyal and Suresh P Vyas*

Drug Delivery Research Laboratory, Department of Pharmaceutical Sciences

Dr Harisingh Gour Vishwavidyalaya, Sagar 470 003, India

Received 13 March 2007; revised 5 December 2007; accepted 10 February 2008

Edible vaccines offer exciting possibilities for significantly reducing the burden of diseases like hepatitis and diarrhoea, particularly in developing world where storing and administering vaccines are the major problems. Edible vaccines are prepared by molecular farming using the science of genetic engineering. Selected genes are introduced into the plants. The transgenic plant is then induced to manufacture the encoded protein. Owing to its low cost, it will be suitable for developing countries like India. Edible vaccines are mucosal-targeted vaccines, which cause stimulation of both systematic and mucosal immune response. Edible vaccines are being developed for various diseases, such as measles, cholera and hepatitis B, and many more are in the process of development. Thus, they may also help to suppress autoimmune disorders such as Type-I diabetes, diarrhoea, multiple sclerosis, rheumatoid arthritis, etc. Human trials conducted by the National Institute of Allergy and Infectious Diseases (NIAID), US Department of Health and Human Services, USA show that edible vaccines are feasible. ProdiGene, a biotech company, has a patent for vaccine against, viral diseases of hepatitis and transmissible gastroenteritis virus. This review comprises methods of preparation, mechanism of action, recent developments, clinical trials and therapeutic applications of edible vaccines.

Keywords: Transgenic plant, edible vaccine, oral immunization, mucosal immunity, autoimmunity

 

 

Indian Journal of Biotechnology

Vol 7, July 2008, pp 295-306

 

Enhancement of NAPL bioavailability by induction of cell-surface hydrophobicity in Exiguobacterium aurantiacum and Burkholderia cepacia

 

Gita Mohanty and Suparna Mukherji*

Centre for Environmental Science and Engineering (CESE), Indian Institute of Technology Bombay, Powai, Mumbai 400 076, India

Received 26 February 2007; revised 10 December2007; accepted 25 February 2008

Induction of cell surface hydrophobicity in bacterial cultures can facilitate the direct interfacial uptake of non-aqueous phase liquids (NAPLs). This study explores bioavailability of NAPLs for Exiguobacterium aurantiacum and Burkholderia cepacia isolated from oil-contaminated soil and sediments. Surface tension measurements and emulsification activity tests did not provide evidence for release of extracellular biosurfactants/bioemulsifiers. Contact angle measurement on cell layers and bacterial adhesion to hydrocarbon (BATH) assay was conducted for determining the cell surface hydrophobicity. While the surfaces of cultures grown on soluble substrate, dextrose, were not hydrophobic, higher water contact angle and greater adherence to n-hexadecane/diesel revealed the highly hydrophobic nature of the cell surfaces for cultures grown on NAPLs (n-hexadecane and diesel), thus providing evidence for induction in cell surface hydrophobicity. Positive results in the carbocyanine assay indicating release of lipopolysaccharides/extracellular polysaccharides was observed over the log growth phase only for the NAPL-grown cells. Transmission electron microscopy (TEM) revealed an abundance of intracellular electron transparent globules within the NAPL-grown cells. Light microscopy and TEM images together revealed differences in cell surface characteristics of E. aurantiacum and B. cepacia, which was also affected by the growth substrate.

Keywords: Aliphatic hydrocarbons, biodegradation, LPS, oil, TEM, uptake mechanism

 

 

Indian Journal of Biotechnology

Vol 7, July 2008, pp 307-312

 

Amplification, cloning and sequencing of Enterococcus feacalis enolase gene

 

A Shivaraj1,3, S Madhusudan1,2, C Ashok Kumar1, S Isloor3, Veere Gowda3, H J Dechamma1,
G R Reddy1 and V V S Suryanarayana1*

1Molecular Virology Laboratory, Indian Veterinary Research Institute, Hebbal, Bangalore 560 024, India

2Department of Biotechnology, Indian Academy Centre for Research & Post Graduate Studies, Hennur Cross
Kalyananagar, Bangalore 560 043, India

3Department of Microbiology, Veterinary College, Hebbal, Bangalore 560 024, India

Received 26 April 2007; revised 12 November 2007; accepted 25 January 2008

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

Keywords: Enolase, Enterococcus feacalis, cloning vector, sequence, nucleotide

 

 

Indian Journal of Biotechnology

Vol 7, July 2008, pp 313-319

 

Assessment of genetic variation among populations of Rhynchostylis retusa, an epiphytic orchid from Goa, India using ISSR and RAPD markers

 

G V Parab and S Krishnan*

1Department of Botany, Goa University, Goa 403 206, India

Received 29 December 2006; revised 11 February 2008 ; accepted 15 April 2008

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

Keywords: Rhynchostylis retusa, RAPD, ISSR, genetic variation

 

 

Indian Journal of Biotechnology

Vol 7, July 2008, pp 320-327

 

RAPD based assessment of genetic diversity of Butea monosperma from different agro-ecological regions of India

 

Vaishali1, Suphiya Khan2, Vinay Sharma*2

1Department of Biotechnology, Sardar Vallabh Bhai Patel University of Agriculture and Technology, Meerut 250 110, India

2Department of Bioscience and Biotechnology, Banasthali Vidhyapith 304 022, India

Received 18 September 2007; revised 11 February 2008; accepted 20 April 2008

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

Keywords: DNA, Butea monosperma, molecular profiling, RAPD, agro- ecological regions

 

 

Indian Journal of Biotechnology

Vol 7, July 2008, pp 328-332

 

Immobilization of porcine pancreas lipase onto free and affixed arylamine glass beads and its application in removal of oil stains

 

M Sharma, V Kumar and C S Pundir*

Biochemistry Research Laboratory, Department of Biochemistry and Genetics, M D University, Rohtak 124 001, India

Receivd 17 March 2006; revised 9 January 2008; accepted 10 March 2008

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

Keywords: Porcine pancreas lipase, arylamine glass beads, immobilization, detergent, cloth washing, oil stains

 

 

Indian Journal of Biotechnology

Vol 7, July 2008, pp 333-340

 

Extracellular collagenase from Rhizoctonia solani: Production, purification and characterization

 

Hossam S Hamdy*

Biology Department, Faculty of Education, Ain Shams University, Roxy 11757, Cairo, Egypt

Received 2 March 2007; revised 5 September 2007; accepted 10 December 2007

Potentiality of R. solani grown on Sabouraud-glucose-collagen medium to produce glycosylated metallo-proteinase with collagenolytic activity was optimized and maximum production (212.33 U/mL) was recorded after 108 h of submerged incubation (175 rpm) at pH 5.5 and 30°C of temperature.  Two-step column chromatography technique on DEAE-cellulose and Sephadex G150 was adopted to purify the partially purified enzyme produced by ammonium sulfate (40%, w/v) precipitation.  Yield of purification was 60.49% of the original activity with specific activity of 18064.7 × 103 U/mg protein and 18.72-folds of purification.  The purified enzyme showed maximum activity at 40°C and pH 5 ,which was stimulated by ions of Ca, Co, Cu, K, Mg, Na or Zn and inhibited by ions of Fe and Hg.  Metal composition of the purified enzyme revealed that it contains Ca2+ and Zn2+. Tm (midpoint of thermal inactivation) was recorded at 65°C and 55°C after 1 and 6 h of exposure, respectively and T1/2 was found to be 7 and 5 weeks at 15°C and 4°C, respectively.  Molecular mass, Kmg/mL, Vmax of the enzyme were found to be 66 ± 4 kDa, 0.033 kDa and 0.28 U/mg min-1, respectively.  Activities toward gelatin and casein were also detected.

Keywords: Collagenase, production, purification, characterization, Rhizoctonia solani.

 

 

Indian Journal of Biotechnology

Vol 7, July 2008, pp 341-348

 

Enhanced production of streptomycin and hydrolytic enzymes by Streptomyces griseus strains using different types of organic solvents and detergent compounds

 

Kamal Y I El-shahed*, Ahmed I El-Diwany and Hassan M M Awad

National Research Center, Chemistry of Natural and Microbial Products Department, Dokki-Cairo-Egypt

Received 25 February 2006; revised 11 June 2007; accepted 10 November 2007

Different organic solvents and detergents showed profound effects on the production of streptomycin (SM), amylases and proteases by Streptomyces griseus strains. The maximum SM production (1128 and 780 mg/L) was obtained using Streptomyces NRRL-strain when ethanol (1.0% v/v) and Tween-80 (0.1%v/v) were added to the medium. Production of amylases was also enhanced using the strain with the addition of benzene at decline phase, n-butanol at the trophophase and isopropanol at the idiophase with values, 10.66, 8.28 and 8.05 U/mL for a-amylase (EC-3.2.1.1) and 6.28, 4.68 and 4.55 for b-amylase (EC-3.2.1.2), respectively. While the use of other solvents and detergents reduced the production of amylase specially activities by S. griseus DSM-40759, characterized by higher productions of a- and b-amylases in different growth phases especially at the death phase. Worthily, proteases secretion was only induced to 1.02 U/mL at the idiophase by culturing DSM-strain with the addition of Triton X-100 and SDS to the fermentation medium. Moreover, the solvents shifted the pH values in the acidic range by the two strains at different phases compared to the control.

Keywords: Streptomyces griseus, streptomycin, amylases, proteases, organic solvents, detergent compounds

 

 

Indian Journal of Biotechnology

Vol 7, July 2008, pp 349-355

 

Isolation of borrelidin from Streptomyces californicus—An Indian soil isolate

 

Srinivasan Saisivam1, D V R N Bhikshapathi2, J Krishnaveni2 and V Kishan2*

1K M College of Pharmacy, Madurai 62 5107, India

2University College of Pharmaceutical Sciences, Kakatiya University, Warangal 506 009, India

Received 17 January 2007; revised 23 October 2007; accepted 4 January 2008

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

Keywords: Antibiotic, borrelidin, fermentation, resistant strains, Streptomyces californicus

 

 

Indian Journal of Biotechnology

Vol 7, July 2008, pp 356-360

 

Production of alkaline protease by Bacillus circulans using agricultural residues: A statistical approach

 

R K Jaswal, G S Kocher* and M S Virk1

Department of Microbiology and 1Department of Maths, Stat and Physics, Punjab Agricultural University, Ludhiana 141 004, India

Received 7 June 2007; revised 14 November 2007; accepted 2 February 2008

Bacillus circulans, an alkaline protease producer isolated from vegetable waste, showed a pH of 10.5, temperature between 25-30°C and agitation rate of 200 rpm as optimum physical conditions for enzyme production. Factorial experiments were designed and the best CN combination of glucose and soybean meal produced a maximum alkaline protease of 461.65 U/mL. Soybean meal replaced with cotton deoiled meal (CDM) from enzyme production medium as the maximum supporter of alkaline protease produced 589 U/mL of protease. Protease values with different concentrations of CDM fitted in a regression equation showed that B. circulans produces maximum alkaline protease of 808.68 U mL-1 in 100.9 h using 0.5789% CDM.

Keywords: Alkaline protease, Bacillus circulans, CN combination, cotton deoiled mea, regression analysis l, soybean meal

 

 

Indian Journal of Biotechnology

Vol 7, July 2008, pp 361-365

 

Optimization of malting conditions of Pearl millet
[Pennisetum glaucum (L.)R.Br.] using Box-Behnken design

 

P Mary Anupama1*, C Ayyanna2 and T Ramana3

1Department of Biotechnology, ANITS, Visakhapatnam, India

2Al-Ameer College of Engineering and Information Technology, Visakhapatnam, India

3Department of Biotechnology, College of Science and Technology, Andhra University, Visakhapatnam, India

Received 4 July 2007;revised 12 February 2008; accepted 15 April 2008

In the present study, malting conditions of Pearl millet [Pennisetum glaucum (L.)R.Br.] were optimized using Box-Behnken design. The three variables chosen were gibberlic acid (X1), temperature (X2) of malting and days of germination (X3) for optimal yield of α-amylases and proteases in the malt. A total of 15 runs by combination of each factor were carried out and contribution of each factor was established. The t-values and p-values obtained revealed that all the three variables significantly contribute to the formation of both enzymes during malting. Optimal conditions required for the formation of maximal expression of α-amylases (323.03 units/g of malt) as provided by the model are as follows: gibberlic acid
1.053 ppm, temperature to be maintained at 32.98oC and the malt must be obtained at the end of 3.68 d. The set of conditions maintained to get an optimal protease yield (32.968 units/g of malt) are: gibberlic acid, 1.0743 ppm; temperature, 33şC; and the malt must be obtained at the end of 3.4 d (78 h). Following these conditions, malt with α-amylase and protease activities of 320 units and 31.24 units per g, respectively were obtained.

Keywords: Pearl millet, malting, a-amylase, Protease, Box-Behnken, Pennisetum glaucum

 

 

Indian Journal of Biotechnology

Vol 7, June 2008, pp 366-370

 

Host associated genetic variations in whitefly, Bemisia tabaci (Genn.)

 

R K Sharma, V K Gupta*, J Jindal and V K Dilawari

Insect Molecular Biology Lab, Department of Entomology, Punjab Agricultural University, Ludhiana 141 004, India

Received 18 May 2007 ; revised 20 December 2007; accepted 1 March 2008

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

Keywords: Bemicia tabaci, genotype analysis, genetic similarity, RAPD markers

 

 

Indian Journal of Biotechnology

Vol 7, July 2008, pp 371-377

 

Significance of metabolites of native Xenorhabdus, a bacterial symbiont of Steinernema, for suppression of collar rot and root knot diseases of groundnut

 

R V Vyas*, Biren Patel, Ajay Maghodia and D J Patel

Department of Nematology, B A College of Agriculture, Anand Agricultural University, Anand 388 110, India

Received 8 March 2006; revised 26 June 2007; accepted 10 October 2007

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

Keywords: Aspergillus, collar rot, exo- and endo-metabolites, groundnut, Meloidogyne, PCR, root-knot nematode, SDS-PAGE, Steinernema, toxicity, Xenorhabdus

 

 

Indian Journal of Biotechnology

Vol 7, July 2008, pp 378-382

 

Increased isoflavonoids accumulation in cell suspension cultures of Pueraria tuberosa by elicitors

 

Shaily Goyal and K G Ramawat*

Laboratory of Biomolecular Technology, Department of Botany, M L Sukhadia University
Udaipur 313 001, India

Received 23 August 2007; revised 1 January 2008; accepted 4 March 2008

Cell cultures of Pueraria tuberosa were established in modified Murashige and Skoog medium and challenged with yeast extract (YE), methyl jasmonate (MeJA) and salicylic acid (SA). Maximum isoflavonoids production was recorded at 48 h of YA incorporation at the stationary phase. 20 mM of MeJA and SA was most effective in isoflavonoids induction. Higher concentrations of these elicitors were negatively correlated with the isoflavonoids production. YE at 150 mg L-1 was optimal for isoflavonoids production, yielding 10 mg L-1 isoflavonoids, which was ~20% higher over the yields at optimal concentrations of MeJA and SA. YA incorporation can be used as a trigger to induce high yield of isoflavonoids.

Keywords: Cell culture, fungal elicitor, isoflavonoids, methyl jasmonate, Pueraria tuberosa, salicylic acid

 

 

Indian Journal of Biotechnology

Vol 7, July 2008, pp 383-387

 

In vitro propagation and quercetin quantification in callus cultures of Rasna (Pluchea lanceolata Oliver & Hiern.)

Deepika Arya, Vidya Patni* and Uma Kant

Plant Pathology, Tissue Culture and Biotechnology Laboratory, Department of Botany
University of Rajasthan, Jaipur 302 004, India

Received 11 April 2007; revised 19 September 2007; accepted 15 December 2007

A protocol for micropropagation of Pluchea lanceolata, an important medicinal herb was developed. Leaf explants obtained from field grown plants when tested for callus induction on Murashige and Skoog’s (MS) medium, supplemented with NAA in combination with BAP, produced the best callus. Maximum number of multiple shoots from the callus (26.6±0.67) was obtained on MS medium supplemented with BAP (1.0 mg/L) and Kn (1.0 mg/L). More or less uniform elongation of multiple shoots was obtained on MS medium with lower concentrations of cytokinins, i.e., BAP (0.25 mg/L) and Kn (0.5 mg/L). Further elongation and profuse rooting were achieved when the well-grown shoots were cultured on half strength MS medium supplemented with IBA (1.0 mg/L). The regenerated plantlets were hardened and established at 70% survival rate in pots. The bioactive secondary metabolite, quercetin, was isolated from callus tissues of different age groups and its identification and confirmation was carried out by the colour reaction, TLC behaviour, IR spectrum and HPLC techniques. Maximum quercetin content (0.23 mg/g dry wt of tissue) was obtained in 6-wk-old callus tissues.

Keywords: In vitro propagation, quercetin; callus, Pluchea lanceolata

 

 

Indian Journal of Biotechnology

Vol 7, July 2008, pp 388-393

 

Direct shoot organogenesis and plant regeneration from hypocotyl explants in selected genotypes of Leucaena leucocephala—A leguminous pulpwood tree

V L Sirisha1, S Prashant1, D Ranadheer1, P Ramprasad1, N M Shaik2, Manish Arha2, S K Gupta2, Sameer Srivastava2,
A K Yadav2, P S Kulkarni2, O U Abhilash2, B M Khan2, Shuban K Rawal2 and P B Kavi Kishor1*

1Department of Genetics, Osmania University, Hyderabad 500 007, India

2Plant Tissue Culture Division, National Chemical Laboratory, Pune 411 008, India

Received 17 January 2007; revised 29 October 2007; accepted 5 January 2008

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

Keywords: Leucaena lucocephala, pulpwood leguminous tree, hypocotyl explants, shoot organogenesis

 

 

Indian Journal of Biotechnology

Vol 7 July 2008, pp 394-397

 

In vitro shoot proliferation in Emblica officinalis var. Balwant from nodal explants

 

Divya Goyal* and Seema Bhadauria

Department of Botany, Raja Balwant Singh College, Agra 282 002, India

Received 17 April 2006; revised 10 April 2007; accepted 2 July 2007

An efficient protocol for in vitro shoot proliferation of Emblica officinalis var. Balwant has been developed by using nodal explants. In vitro shoot proliferation from nodal explants have been observed by using MS, B5 and WPM media. Amongst them, MS medium has been observed the best for shoot proliferation (63.3 %), while minimum phenol leaching (63.3 %) was observed on WPM medium. To enhance the shoot regenerative potential, explants were cultured on MS medium supplemented with various cytokinins, viz., BAP, Kn and TDZ. MS medium supplemented with BAP (4.44 µM/L) was observed the best among all cytokinins. As shoot proliferation depended upon the balance of cytokinins and auxins, and BAP was found ideal for shoot proliferation, additive effect of auxins – IAA, IBA and NAA was assessed at 4.44 µM/L conc. of BAP. Explants when cultured on MS + BAP (4.44 µM/L) with IBA (2.46 µM/L)) showed maximum percentage of shoots (85 %) with minimum callus formation (15%). Higher concentration of auxins suppressed the caulogenesis and induced callus formation.

Keywords: Emblica officinalis, micropropagation, nodal shoots, amla

 

 

Indian Journal of Biotechnology

Vol 7 July 2008, pp 398-400

 

Polymorphism in DRB3 exon 2 by PCR-RFLP and its association with mastitis
in Nili-Ravi breed

 

Sudhir Kumar, M L Sangwan* and Rupender

Department of Animal Biotechnology, College of Veterinary Sciences, CCS Haryana Agricultural University, Hisar 125 004, India

Received 18 September 2007; revised 6 February 2008;
accepted 18 April 2008

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

Keywords: BuLA-DRB3.2, PCR-RFLP, Nili-Ravi buffalo, mastitis

 

 

Indian Journal of Biotechnology

Vol 7, July 2008, pp 401-403

 

Evaluation of genetic diversity in Mecheri sheep (Ovis aries) of Tamil Nadu using microsatellite markers

 

S Prema, S N Sivaselvam* and S M K Karthickeyan

Department of Animal Genetics and Breeding, Madras Veterinary College, Chennai 600 007, India

Received 20 March 2007; revised 8 October 2007; accepted 28 December 2007

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

Keywords: Genetic variation, mecheri sheep, microsatellite markers

 

 

Indian Journal of Biotechnology

Vol 7, July 2008, pp 404-406

 

Role of calcium ions and temperature on dextransucrase production

 

Shah Ali Ul Qader1, Afsheen Aman1, Saeeda Bano1,
Noman Syed1 & Abid Azhar2

1Pharmaceutical Research Center, PCSIR Laboratories Complex, Karachi, Pakistan

2Institute of Biotechnology and Genetic Engineering, University of Karachi. Karachi, Pakistan

Received; revised; accepted

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

Keywords: Dextran, CaCl2, Stability, Dextransucrase, Glucosyltransferase, Leuconostoc mesenteroides

 

 

Indian Journal of Biotechnology

Vol 7, July 2008, pp 407-409

 

Efficient plant regeneration in small cardamom (Elettaria cardamomum Maton.) through somatic embryogenesis

 

C Manohari 1, S Backiyarani 2, T Jebasingh1, Archana Somanath 1 and R Usha1*

1Department of Plant Biotechnology, School of Biotechnology, Madurai Kamaraj University, Madurai, 625 021

2Cardamom Research Station, Kerala Agricultural University, Pampadumpara, Kerala 685 556

Received 20 July 2007; revised 16 November 2007;
accepted 5 February 2008

An efficient protocol for the induction of somatic embryogenesis and plant regeneration in small cardamom (Elettaria cardamomum Maton.) from the inner core region of rhizome has been established. Calli developed profusely on Murashige and Skoog (MS) medium containing 9.0 μM 2,4 D and 2.3 μM Kn. When the friable calli were cultured in MS medium containing 4.4 or 8.8 μM BAP + 0.5 μM NAA, abundant embryogenic calli were obtained. The highest frequency of embryogenic calli (68%) and plantlets (86%) were obtained from MS medium containing 4.4 μM BAP and 0.5 μM NAA. Further shoot development was observed with 13.2 μM BAP + 0.5 μM NAA. In the same medium, roots also appeared, thereby eliminating an additional step of in vitro rooting. The well-developed plants were hardened and transferred to a mist chamber in a greenhouse with 90% survival frequency.

Keywords: Cardamom, somatic embryogenesis, regeneration, Elettaria cardamomum.

 

 

 

INDIAN JOURNAL OF BIOTECHNOLOGY

 

Instructions to Contributors

 

The Indian Journal of Biotechnology, a quarterly journal, publishes original research papers, reviews, digests (biotechnology highlights), news-scan, etc. The journal covers papers on Biotechnology in the following main areas: (i) Agriculture; (ii) Animal husbandry; (iii) Environment; (iv) Industry; (v) Microbiology; (vi) Medicine; (vii) Bio-informatics; and (viii) Socio-legal and ethical aspects.

Indian Journal of Biotechnology invites original research and review manuscripts not submitted for publication elsewhere. The review article will only be entertained if author(s) has included his own research work in it or has been an authority in that field. It is mandatory on the part of the corresponding author to furnish the following certificate at the time of submission of the manuscript:

This is to certify that the reported work in the paper entitled "                " submitted for publication is an original one and has not been submitted for publication elsewhere. I/we further certify that proper citations to the previously reported work have been given and no data/tables/figures have been quoted verbatim from other publications without giving due acknowledgement and without the permission of the author(s). The consent of all the authors of this paper has been obtained for submitting the paper to the "Indian J Biotechnology".

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The copyright of the paper will be transferred from the author to publisher. One original and two copies of the manuscript should be submitted to the editor. The manuscript, after referees’ acceptance, will be sent back to the author(s) along with referees’ comments. For re-submission, two copies of the revised version of the manuscript, and a copy on compact disc (CD) using word processing software such as MS Word (version 6 and onwards), or PDF files (version 4 and onwards), or as an attachment to e-mail should be submitted to the editor.

 

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Manuscripts should be typed in double space (11 pt, Times New Roman font preferred) on one side of the bond paper of 22×28 cm. All pages should be numbered consecutively. Use SI units, and give equivalent SI units in parenthesis when the use of other units is unavoidable. Symbols should conform to standard guidelines.

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Results—Results should contain data, which are essential for drawing main conclusion from the study. Wherever needed, the data should be statistically analyzed. Same data should not be presented in both table and figure form.

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References—References should be cited in the text by the consecutive numbers of their occurrence; the numbers are to be shown as superscript at the end of the statement related to that particular reference, e.g. It also inhibits the activity of endogenous DNA polymerase of HBV7.

Following the same sequence of the text, the list of references be appended under the References heading. Each reference should provide names and initials of all the authors, giving coma in between the authors and ‘&’ before the last author. In case, the authors are more than five, then use et al after the 5th author. It should be followed by title of the paper, abbreviated title of journal (in italics), volume number, year of publication (within circular bracket), and the starting and closing page numbers. Abbreviated titles should conform to the international guidelines, e.g. The Chemical Abstracts Service Source Index (CASSI) or BIOSIS

The style of references should be:

 

Research Papers

·     Ghosh A C & Basu P S, Extracellular polysaccharide production by Azorhizobium caulinodans from stem nodules of leguminous emergent hydrophyte Aeschynomene aspera, Indian J Exp Biol, 39 (2001) 155-159 [If accepted for publication, give (in press) in place of volume, year and pages].

·     Newell C A, Lowe J M, Merryweather A, Rooke L M & Hamilton W D O, Transformation of sweet potato (Ipomoea batatas (L) Lam) with Agrobactericum tumefaciens and regeneration of plants expressing cowpea trypsin inhibitor and snowdrop lectin, Plant Sci, 107 (1995) 215-227.

·     Hoffman M P, Zalom F G, Smilanick J M, Malyj L D, Kiser J et al, Field evaluation of transgenic tobacco containing genes encoding Bacillus thuringensis d-endotoxin or cowpea trypsin inhibitor: Efficacy against Helicoverpa zea (Lepidoptera: Noctuidae), J Econ Entomol, 85 (1991) 2516-2522

 

Books & Proceedings of Conferences

·     Truzuki T & Irukayama K, Minamata disease (Elsevier, Amsterdam) 1977, 30-45.

·     Roels O A & Mahadevan S, Vitamin A, in The vitamins: Chemistry, pathology and methods, 2nd edn, vol VI, edited by P Gyorgy & W N Pearson (Academic Press, New York) 1967, 139-210.

·     Allossp P G, Nutt K A, Geijsk R J & Smith G R, Transgenic Sugarcane with increased resistance to canegrub, in Sugarcane pest management in the new millennium, 4th Sugarcane Entomol Workshop, held on 7-10 Feb, 2000 (Int Soc Sugarcane Technol, Khon-khon, Thailand) 2000, 63-67.

·     Chaturvedi H C & Sharma A K, Citrus tissue culture, in Proc Natl Semin Plant Tissue Cult (ICAR, New Delhi) 1988, 36-46.

·     Kapoor B C, 2000. Managing in the face of not-so-developed and organized environment, paper presented in Natl Symp Manag Dev, Institute of Public Administration, Jaipur, India, 21-23 July, 2000.

 

Thesis & Dissertation

·     Chaturvedi H C, In vitro growth and controlled morphogenesis in callus tissue of Rauvolfia serpentina. Ph D Thesis, Agra University, Agra, 1968.

 

Patent

·     Trepaginer J H, New surface finishings and coatings, US Pat 1276323 (to DuPont Inc, USA). 27 June, 2000; Chem Abstr, 49 (2000) 27689.

Manuscript along with referees’ comments will be sent to the author identified for correspondence on the title page of the manuscript. It should be checked carefully and the modified manuscript should be returned within ten days of receipt. No page proofs will be sent to author(s).

 

Reprints—Twenty five reprints will be supplied gratis.