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

Devi Ahilya University

Indore 452 017

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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ă 2008 The Council of Scientific & Industrial Research, New Delhi.

 

Indian Journal of Biotechnology

 

http://www. niscair.res.in

 

VOLUME 7

CODEN: IJBNAR 7(4) (2008) 415-578

NUMBER 4

OCTOBER 2008

ISSN: 0972-5849

 

CONTENTS

 

Review

 

Statistical tests for identification of differentially expressed genes in cDNA microarray experiments

423

J Sreekumar & K K Jose

 

 

 

Papers

 

Structural stabilization of bovine b-Lactoglobulin in presence of polyhydric alcohols

437

S N Anuradha & V Prakash

 

 

 

Development of a macroarray based on 16S-23S rDNA probe hybridization for rapid diagnosis of human pathogenic bacteria

448

B Maity, R Trivedi & V K Kashyap

 

 

 

Cloning and characterization of histone H3 and replacement histone H3.3 encoding genes of
Manila clam (Ruditapes philippinarum)

456

Sumin Lu, Zhenmin Bao, Xiaolong Wang, Jingjie Hu & Jianguang Fang

 

 

 

Overexpression of tobacco osmotin gene leads to salt stress tolerance in strawberry (Fragaria ´ ananassa Duch.) plants

465

Amjad Masood Husaini & Malik Zainul Abdin

 

 

 

Cloning and expression of FMDV-VP1 immunoreactive peptide in trivalent form and its application as immunogen

472

G Nagarajan, C Ashok Kumar, H J Dechamma, G R Reddy, K Ganesh & V V S Suryanarayana

 

 

 

The PCR amplification, sequencing and computer-aided analysis of ovine a S1-casein gene promoter

478

S K Bhure & B Sharma

 

 

 

Genetic variability among sheep breeds by random amplified polymorphic DNA-PCR

482

S Kumar, A P Kolte, B R Yadav, Sushil Kumar, A L Arora & V K Singh

 

 

 

DNA fingerprint: Genetic relationship in six Indian goat breeds

487

Anita Yadav & B R Yadav

 

 

 

Genetic diversity between Murrah and Bhadawari breeds of Indian buffalo using RAPD-PCR

491

Anurag Barwar, M L Sangwan, Sudhir Kumar & Sonika Ahlawat

 

 

 

Process optimization for citric acid production from raw glycerol using response surface methodology

496

K Sita Kumari, I Sarat Babu & G H Rao

 

 

 

Effect of wheat straw powder on enhancement of ligninolytic enzyme activity using Phanerochate chrysosporium

502

S Masud Hossain & N Anantharaman

 

 

 

PCR fingerprinting for identification and discrimination of plant-associated facultative methylobacteria

508

P Raja, D Balachandar & S P Sundaram

 

 

 

Development of synthetic seeds involving androgenic and pro-embryos in elite indica rice

515

Bidhan Roy & Asit B Mandal

 

 

 

Morphactin and cytokinin promote high frequency bulbil formation from leaf explants of Curculigo orchioides grown in shake flask cultures

520

Rajesh K Nema, Suchismita Dass, Meeta Mathur & K G Ramawat

 

 

 

Effect of growth regulators on in vitro morphogenic response of tomato

526

Ruma Devi, M S Dhaliwal, Ajinder Kaur & S S Gosal

 

 

 

Response of some Iranian wheat genotypes to anther culture system

531

Behnam Naserian Khiabani, Cirus Vedadi, Esfandiar Rahmani & Mir Ahmad Mosavi Shalmani

 

 

 

Induction of embryos and plantlets from anthers of Curculigo orchioides Gaertn.—An endangered medicinal herb

536

Alice Clara Augustine, Shashikiran Nivas & L D’Souza

 

 

 

In vitro plant regeneration of red sanders (Pterocarpus santalinus L.f.) from cotyledonary nodes

541

V Rajeswari & Kailash Paliwal

 

 

 

Short Communications

 

The stem cell in the umbilical cord blood is not related to volume and nucleated cell count

547

A Mahantappa, Avijit Banik, Jyothsna Shivshankar & S G A Rao

 

 

 

Cloning of pediocin PA-1 and its immunity genes from Pediococcus acidilactici K7 using pAMJ shuttle vector into Lactococcus lactis MG 1363

550

Anusha Srikanth & Prakash M Halami

 

 

 

Detection and characterization of group A and D avian rotaviruses in India

554

Savita, A L Kusumakar, Y P S Malik, Minakshi & G Prasad

 

 

 

Book Review

557

 

 

List of Referees

559

 

 

Annual Author Index

567

 

 

Annual Subject Index

571

 

 

Instructions to Contributors

575

 

 

 

 

               AUTHOR INDEX

 

 


Abdin M Z

465

Ahlawat S

491

Anantharaman N

502

Anuradha S N

437

Arora A L

482

Augustine A C

536

 

 

Babu I S

496

Balachandar D

508

Banik A

547

Bao Z

456

Barwar A

491

Bhure S K

478

 

 

D’Souza L

536

Dass S

520

Dechamma H J

472

Devi R

526

Dhaliwal M S

526

 

 

Fang J

456

 

 

Ganesh K

472

Gosal S S

526

 

 

Halami P M

550

Hossain S M

502

Hu J

456

Husaini A M

465

 

 

Jose K K

423

 

 

Kashyap V K

448

Kaur A

526

Khiabani B N

531

Kolte A P

482

Kumar C A

472

Kumar S

482

Kumar Sushil

482

Kumar Sudhir

491

Kumari K S

496

Kusumakar A L

554

 

 

Lu S

456

 

 

Mahantappa A

547

Maity B

448

Malik Y P S

554

Mandal A B

515

Mathur M

520

Minakshi

554

 

 

Nagarajan G

472

Nema R K

520

Nivas S

536

 

 

Paliwal K

541

Prakash V

437

Prasad G

554

 

 

Rahmani E

531

Raja P

508

Rajeswari V

541

Ramawat K G

520

Rao G H

496

Rao S G A

547

Reddy G R

472

Roy B

515

 

 

Sangwan M L

491

Savita

554

Shalmani M A M

531

Sharma B

478

Shivshankar J

547

Singh V K

482

Sreekumar J

423

Srikanth A

550

Sundaram S P

508

Suryanarayana V V S

472

 

 

Trivedi R

448

 

 

Vedadi C

531

 

 

Wang X

456

 

 

Yadav A

487

Yadav B R

482, 487


 

 

 

 


Review

 

Indian Journal of Biotechnology

Vol 7, October 2008, pp 423-436

 

Statistical tests for identification of differentially expressed genes in cDNA microarray experiments

J Sreekumar* and K K Jose1

Central Tuber Crops Research Institute, Sreekariyam, Thiruvananthapuram 695 017, India

1Department of Statistics, St Thomas College, Pala, Kottayam 686 574, India

Received 20 July 2006; revised 15 January 2008; accepted 16 March 2008

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.

Keywords: ANOVA, Bayesian inference, bioinformatics, differential gene expression, DNA microarrays, t-test

Papers

 

Indian Journal of Biotechnology

Vol 7, October 2008, pp 437-447

 

Structural stabilization of  bovine b-Lactoglobulin in presence of polyhydric alcohols

S N Anuradha and V Prakash*

Department of Protein Chemistry and Technology, Central Food Technological Research Institute,

Mysore 570 020, India

Received 14 February 2007; revised 13 March 2008; accepted 18 May 2008

b-Lactoglobulin (b-Lg) is the major protein present in bovine milk whey. Addition of hydrogen-bonded cosolvents to proteins is known to modify the thermodynamic properties of proteins. Preferential interaction parameters of b-Lg were determined in various concentrations of cosolvents like sorbitol, glycerol and sucrose using the precision densitymetry. The apparent partial specific volumes determined at 20°C under both isomolal and isopotential conditions in 0.02 M phosphate buffer (pH 7.9) were 0.743±0.001 and 0.744±0.001, respectively. From the partial specific volume data with cosolvents the preferential interaction parameter and other thermodynamic parameters were calculated at different cosolvent concentrations. The values increased with solvent concentration up to 40% and reached a maximum of -0.190±0.02 and -0.140±0.02 in sucrose and sorbitol, respectively. In glycerol, the value increased up to -0.213±0.03 in 20% glycerol and then decreased with the increase in the cosolvent concentration. There were no changes in secondary structure of the protein as reflected by far UV-CD spectra. The fluorescence spectra showed changes in the fluorescence intensity due to perturbations in the tryptophan moiety without changes in the emission wavelength.  The above data was further supported by the degree of hydrolysis in the presence of cosolvents with the enzyme a-chymotrypsin. The degree of hydrolysis reduced to 12% from the control value of 18% in the presence of sorbitol and glycerol and increased by 3% in the presence of sucrose. All the cosolvents also enhanced thermal stability to various extents.

Keywords:   b-Lactoglobulin (b-Lg), cosolvents, sorbitol, glycerol, sucrose, preferential hydration, apparent thermal transition temperature (Apparent Tm).

 

Indian Journal of Biotechnology

Vol 7, October 2008, pp 448-455

 

Development of a macroarray based on 16S-23S rDNA probe hybridization for rapid diagnosis of human pathogenic bacteria

B Maity 1, R Trivedi 1 and V K Kashyap 2*

1Central Forensic Science Laboratory, 30 Gorachand Road, Kolkata 700 014, India

2National Institute of Biologicals, NOIDA 201307, India

Received 16 May 2007; revised 20 March 2008; accepted 22 May 2008

The rapid identification of bacteria in biological specimens is important for selection of a suitable antimicrobial therapy. We have designed a rapid (<8 h) diagnostic system that uses universal PCR primers to amplify a variable region of bacterial 16S & 23S ribosomal DNA, followed by reverse hybridization of the products to a panel of oligonucleotides, spotted on nylon membrane macroarray slides. Culture dependent assays were used as reference methods in the development and evaluation of this diagnostic platform. Broad range PCR primers were selected from the published literature to amplify 16S & 23S rDNA segments. Species-specific probe sequences were designed based on sequence alignment of eight different bacteria. These species included Helicobacter pylori, Salmonella typhimurium, Shigella dysenteriae, Vibrio choleriae, Neisseria gonorrhea, N. meningitidis, Corynebacterium diphtheriae and Haemophilus influenzae. To verify specificity, five to six initial oligonucleotide probe sequences per bacterial species were tested by hybridization on nylon membrane macroarray glass slides using culture collection strains as templates. Finally, three oligonucleotide probes per bacterial species were selected based on hybridization results. This procedure was successful in discriminating a range of bacteria in pure cultures. Adding further oligonucleotides to the panel without significantly increasing the cost the accuracy, range, and discriminatory power of the assay can be extended. This method is versatile and makes it possible to detect a large number of bacterial species in a single assay and discriminate different bacterial genera of medical importance. Our results provide a proof of concept for the diagnostic use of macroarray technology based on broad-range ribosomal RNA gene amplification, followed by hybridization and specific detection of bacterial species.

Keywords: DNA macroarray; pathogenic bacteria, 16S-23S, rDNA probe, diagnostic tool

 

Indian Journal of Biotechnology

Vol 7, October 2008, pp 456-464

 

Cloning and characterization of histone H3 and replacement histone H3.3 encoding genes of Manila clam (Ruditapes philippinarum)

 

Sumin Lu1, Zhenmin Bao1*, Xiaolong Wang1, Jingjie Hu1, Jianguang Fang2

1Ocean University of China, 5 Rushan Road, Qingdao, Shandong 266 003, China

2Yellow Sea Fisheries Research Institute, 106 Nanjing Road, Qingdao, Shandong 266 071, China

Received 28 September 2006; revised 4 March 2008; accepted 10 May 2008

A full-length cDNA (rp-h3.3) encoding a replacement histone H3.3 had been identified in Manila clam (Ruditapes philippinarum). The cDNA consisted of 975 bp, including a ORF 411 bp in length, a 3′ untranslated regions 490 bp in length, a poly (A) tail and two polyadenylation signals. In addition, the cDNA encoded a peptide with the S.//.A.IG amino acid motif, which was identical to the replacement histone H3.3 variant. rpg-h3.3 and rp-h3s two sequences were obtained by amplifying the genomic DNA of R. philippinarum. The rpg-h3.3, which was the genomic DNA sequence of rp-h3.3, had one intron and the rp-h3s had no intron. It was predicted that h3s might be the h3l-like gene of R. philippinarum. Histone h3 (rp-h3) was also cloned from R. philippinarum genomic DNA. Therefore, the evolutionary origin between histone h3 and its replacement subtypes was analyzed. The study inclined to the viewpoint that h3l-like was the ancesteral gene of h3 and h3.3, which might be two different offsets from h3l-like. During histone evolution process, one copy of h3l-like genes became h3.3 by introns insertion and S-phase-coupled regulatory mechanisms lose, another copy became h3 by gene-duplication and accumulation of mutations in the S.//.A.IG motif.

Keywords: Histone, Manila clam (Ruditapes philippinarum), replication dependent histone, replication independent histone

 

Indian Journal of Biotechnology

Vol 7, October 2008, pp 465-471

 

Overexpression of tobacco osmotin gene leads to salt stress tolerance in strawberry (Fragaria × ananassa Duch.) plants

Amjad Masood Husaini and Malik Zainul Abdin*

Centre for Transgenic Plant Development, Department of Biotechnology, Jamia Hamdard, New Delhi 110 062, India

Received 26 June 2007; revised 26 February 2008; accepted 29 April 2008

Transgenic plants of strawberry tolerant to salt stress were produced using Agrobacterium mediated gene transfer technology. Leaf discs of in vitro grown plantlets of strawberry cv. Chandler were used as explants. Agrobacterium tumefaciens strain GV2260 harbouring osmotin gene under the control of CaMV 35S promoter in a binary vector system pBinAR was used in transformation experiments. Transgene integration and copy number were assessed by PCR and Southern hybridization confirming single copy as well as multiple copies of transgene integration in 10 different lines of transgenic strawberry. Expression of osmotin gene was confirmed in transgenic lines TL3, TL5, TL9 using Northern hybridization, while biochemical analysis revealed enhanced levels of proline, total soluble protein and chlorophyll content as compared to the wild plants. Leaf disc assays performed using both wild-type and transgenic plants had shown that these transgenic lines were tolerant to salt stress.

Keywords: Agrobacterium, proline, stress, strawberry, transgenic plants.

 

 

Indian Journal of Biotechnology

Vol. 7, October 2008, pp 472-477

 

Cloning and expression of FMDV-VP1 immunoreactive peptide in trivalent form and its application as immunogen

G Nagarajan1, C Ashok Kumar2, H J Dechamma2, G R Reddy2, K Ganesh2

and V V S Suryanarayana2*

1National Research Centre on Camel, Post Bag No 7, Jorbeer, Bikaner 334 001, India

2*Molecular Virology Lab, Indian Veterinary Research Institute, Bangalore Campus, Hebbal, Bangalore 560 024, India

Received 2 October 2007; revised 20 March 2008; accepted 25 May 2008

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.

Keywords: Trivalent immunogen; FMDV; immunoreactive; proteins; vaccine

 

Indian Journal of Biotechnology

Vol. 7, October 2008, pp 478-481

 

The PCR amplification, sequencing and computer-aided analysis of  vine aS1-casein gene promoter

S K Bhure* and B Sharma1

*Project Directorate on Animal Disease Monitoring And Aurveillance, IVRI Campus, Hebbal, Bangalore 560 024, India

1Indian Veterinary Research Institute, Izatnagar 243 122, India

Received 12 October 2006; revised 13 February 2008 ; accepted 15 April 2008

The paper reports 5'-flanking sequences of ovine aS1-CSNGP (casein gene promoter) of 2185 bp. It has shown many deletions, substitutions and a 12 bp addition compared to bovine sequence. The comparative study showed 2136 bp of  5˘-flanking region and 49 bp exon I sequence. The exon I sequence contained two ribosomal binding sites. The computational analysis showed presence of core promoter elements, viz., TATA box, CAAT box and initiator sequence. However, no typical GC box was found. Of five known mammary gland specific sequences, three sequences, viz., milk box, Groenen structure and Yu Lee 6, were found. The 220 bp Groenen structure contained other milk protein gene specific sequences (MGF, MPBF, Yu Lee 2, 4 and 5, and Oka box C) and hormone responsive elements (PRE, PRL-RE). Other HREs (GRE, CRE, GHRE and IRE) and ubiquitous transcription factor binding sites were also present. These milk protein gene specific regulatory sequences and HREs are responsible for tissue specific and multi-hormone regulation of the ovine aS1-CSNG.

Keywords: Ovine aS1-caseine gene promoter, gene regulation, transcription factor binding sites, hormone responsive elements

 

Indian Journal of Biotechnology

Vol 7, October 2008, pp. 482-486

 

Genetic variability among sheep breeds by random amplified
polymorphic DNA-PCR

S Kumar1*, A P Kolte1, B R Yadav3, Sushil Kumar2, A L Arora2 and V K Singh2

1Animal Biotechnology Section and 2Animal Genetics and Breeding Division, Central Sheep and Wool Research Institute
Avikanagar, Rajasthan 304 501, India

3Livestock Genome Analysis Laboratory, National Dairy Research Institute, Karnal, Haryana 132001, India

Received 19 December 2006; revised 22 February 2008; accepted 25 April 2008

Random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) was employed to assess the genetic variability and phylogenetic relationship among six breeds of sheep, viz., Malpura, Kheri, Chokla, Garole and two crossbreds Avikalin and Bharat Merino. Twenty-four individuals from each breed/crossbred were selected randomly. Initially, 40 primers were screened, of which 16 were found polymorphic and utilized for estimation of genetic variability and phylogenetic relationship among the breeds. The genetic distance was found highest between Malpura and Garole (D=0.1428), and the lowest (D=0.0612) between Avikalin and Chokla. However, the genetic identity was observed highest (I = 0.9406) between Avikalin and Chokla and the lowest (I = 0.8669) between Malpura and Garole. The Kheri sheep was found close with Chokla (D=0.0741), followed by Malpura sheep (D=0.0902) based on genetic distance. The phylogenetic tree also showed that Avikalin and Chokla are more close, whereas Malpura and Garole are distant to each other. The present study suggested that RAPD-PCR can be used successfully for analyzing genetic variation and phylogenetic relationship among breeds of sheep.

Keywords: Dendrogram, genetic distance, genetic identity, RAPD-PCR, sheep breeds

 

Indian Journal of Biotechnology

Vol 7, October 2008, pp 487-490

 

DNA fingerprint: Genetic relationship in six Indian goat breeds

Anita Yadav1* and B R Yadav2

*1Department of Biotechnology, Kurukshetra University, Kurukshetra 136 119, India

2Livestock Genome Analysis Laboratory, National Dairy Research Institute, Karnal 132 001, India

Received 30 October 2006; revised 14 February 2008; accepted 28 April 2008

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).

Keywords: Goat breed, DNA fingerprint, RAPD, BSF, MAPD

 

Indian Journal of Biotechnology

Vol 7, October 2008, pp. 491-495

 

Genetic Diversity between Murrah and Bhadawari Breeds of Indian buffalo
using RAPD-PCR

Anurag Barwar, M L Sangwan*, Sudhir Kumar and Sonika Ahlawat

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

Received 20 June 2007; revised 16 November 2007; accepted 10 February 2008

Random amplified polymorphic DNA (RAPD) technique was used to characterize Murrah and Bhadawari breeds of buffalo. From the 9 random primers, a total of 84 bands were amplified between breeds and 51 of these (about 60.72%) were found to be polymorphic. In Murrah, overall percentage polymorphism of 56.36 was observed, while in Bhadawari it was 57.14. Higher genetic similarity of 0.81 and 0.80 within Murrah and Bhadawari breeds, respectively, was observed as compared to between breed genetic similarity of 0.31. Genetic distance between the breeds was 1.20. MAPD values of 10.72 and 16.10 were observed within Murrah and Bhadawari breeds, respectively, while MAPD of 60.73 was observed between breeds. Six primers (OPU-01, OPU-02, OPU-05, OPU-07, OPU-14 and OPV-14) in Murrah and five primers (OPU-05, OPU-07, OPU-14, OPU-19 and OPV-14) in Bhadawari were found to be specific for these breeds.

Keywords: RAPD, buffalo, characterization, polymorphism, genetic distance, Murrah, Bhadawari

 

Indian Journal of Biotechnology

Vol 7, October 2008, pp 496-501

 

Process optimization for citric acid production from raw glycerol using response surface methodology

 

K Sita Kumari1*, I Sarat Babu2 and G H Rao3

1Departmant of Pharmacy, MRPG College, Phool Baugh, Vizianagaram 535 002, India

2Department of Biotechnology, ANITS, Sangivalasa, Bheemunipatnam, Visakhapatnam 531 162, India

3Center for Biotechnology, Department of Chemical Engineering, Andhra University, Visakhapatnam 530 003, India

Received 26 May 2006; revised 31 March 2008; accepted 28 May 2008

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.

Keywords: Citric acid, central composite design, raw glycerol, response surface methodology, Yarrowia lipolytica

 

Indian Journal of Biotechnology

Vol 7, October 2008, pp 502-507

 

Effect of wheat straw powder on enhancement of ligninolytic enzyme activity using Phanerochate chrysosporium

 

S Masud Hossain1* and N Anantharaman2

1*Department of Chemical Engineering, Mohamed Sathak Engineering College, Kilakarai 623 806, India

2Department of Chemical Engineering, National Institute of Technology, Tiruchirapalli 620 015, India

Received 20 January 2006; revised 1 April 2008; accepted 26 June 2008

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 m 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.

Keywords: Enhancement, ligninolytic enzymes, optimization, wheat straw, white-rot fungus, Phanerochate chrysosporium

 

Indian Journal of Biotechnology

Vol 7, October 2008, pp. 508-514

 

PCR fingerprinting for identification and discrimination of plant-associated facultative methylobacteria

P Raja, D Balachandar and S P Sundaram*

Department of Agricultural Microbiology, Tamil Nadu Agricultural University, Coimbatore 641 003, India

Received 19 September 2007; revised 1 April 2008; accepted 15 June 2008

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.

Keywords: ARDRA, BOX-PCR, ERIC-PCR, Methylobacterium, RAPD, RISA

 

Indian Journal of Biotechnology

Vol 7, October 2008, pp. 515-519

 

Development of synthetic seeds involving androgenic and pro-embryos
in elite indica rice

Bidhan Roy*1 and Asit B Mandal

Biotechnology Section, Central Agricultural Research Institute, P O Box No. 181, Port Blair 744 105, India

Received 12 February 2007; revised 19 February 2008; accepted 22 April 2008

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.

Keywords: Androgenic embryo, anther culture, artificial seeds, pro-embryo, rice, sodium alginate

 

Indian Journal of Biotechnology

Vol 7, October 2008, pp 520-525

 

Morphactin and cytokinin promote high frequency bulbil formation from leaf explants of Curculigo orchioides grown in shake flask cultures

Rajesh K Nema, Suchismita Dass, Meeta Mathur and K G Ramawat*

Laboratory of Bio-molecular Technology, Department of Botany, M L Sukhadia University, Udaipur 313 001, India

Received 22 January 2007; revised 10 March 2008; accepted 28 May 2008

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 (~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 ~80% in the field conditions. These results will be helpful in developing technology for micropropagation of the plant using bioreactor.

Keywords: Curculigo orchioides, bulbil formation, micropropagation, morphactin, cytokinin

 

Indian Journal of Biotechnology

Vol 7, October 2008, pp. 526-530

 

Effect of growth regulators on in vitro morphogenic response of tomato

Ruma Devi1, M S Dhaliwal1, Ajinder Kaur2 and S S Gosal2*

1Department of Vegetable Crops and 2Department of Plant Breeding, Genetics & Biotechnology, Punjab Agricultural University, Ludhiana 141 004, India

Received 31 August 2007; revised 26 February 2008; accepted 8 May 2008

A 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 ˝ 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.

Keywords: Lycopersicon esculentum, callus induction, plant regeneration, acclimatization; tomato; growth regulators

 

Indian Journal of Biotechnology

Vol 7, October 2008, pp 531-535

 

Response of some Iranian wheat genotypes to anther culture system

Behnam Naserian Khiabani*, Cirus Vedadi, Esfandiar Rahmani and Mir Ahmad Mosavi Shalmani

School of Agricultural, Medical and Industrial Research
Nuclear Science and Technology Research Institute, Mahmood Abad Road, Rajaee Shahr, Karaj, Iran

Received 1 May 2006; revised 27 June 2008; accepted 20 July 2008

The response of five Iranian wheat cultivars and four segregating F3 wheat lines was investigated in anther culture system for haploid plantlet regeneration. Anthers were plated onto P4 induction medium and cold and gamma irradiation were also applied to study their effect on initiation of calli and regeneration of plantlets. Significant differences were found between genotypes. Most of the segregating lines showed high response to anther culture than cultivars. The high calli and plantlets production in segregating lines may be attributed to heterosis effect. Gamma irradiation in wheat anther culture does not promote anther culture response. The results showed significant difference between genotypes and gamma irradiation. The line of F32005 with 4 Gy dose (without cold pretreatment) produced the highest amount of calli (52%) per 100 anthers inoculated, whereas Red-Bofgi genotype (with cold treatment and 4 Gy dose) produced the lowest percentage of calli (0.25%). The results indicated that both the androgenic response and regeneration ability were greatly genotype dependent. It could be concluded that genotypic response to anther culture is vital.

Keywords: Anther culture, cold treatment, low dose gamma ray, wheat

 

Indian Journal of Biotechnology

Vol 7, October 2008, pp. 536-540

 

Induction of embryos and plantlets from anthers of Curculigo orchioides Gaertn.An endangered medicinal herb

Alice Clara Augustine, Shashikiran Nivas* and L D’Souza

Laboratory of Applied Biology, St Aloysius College, Mangalore 575 003, India

Received 28 May 2007; revised 31 March 2008; accepted 5 May 2008

Curculigo orchioides Gaertn. is an endangered medicinal herb belonging to Amaryllidaceae. Anthers cultured in MS liquid medium supplemented with 0.5 mg/L BAP or 0.5-1.0 mg/L NAA formed multicellular pollen. However, they did not develop further into embryos. The multicellular pollen cultured with 0.2-1.0 mg/L 2,4-D developed into embryos when transferred to a hormone free medium. Continued culture of embryos on the medium free of growth substances resulted only in root formation. The embryos were, however, converted into plants when they were transferred to a medium with BAP (0.5 mg/L). The matured plants were successfully transferred to the soil.

Keywords: Amaryllidaceae, anther culture, Curculigo, embryogenesis

 

Indian Journal of Biotechnology

Vol 7, October 2008, pp 541-546

 

In vitro plant regeneration of red sanders (Pterocarpus santalinus L.f.) from cotyledonary nodes

 

V Rajeswari and Kailash Paliwal*

School of Biological Sciences, Madurai Kamraj University, Madurai 625 021, India

Received 8 May 2007; revised 20 February 2008; accepted 5 May 2008

A reliable and efficient micropropagation protocol was developed through axillary shoot proliferation from cotyledonary nodes of Pterocarpus santalinus. Cotyledonary nodes showed significantly (P<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.

Keywords: Cotyledonary node, ex-vitro rooting, endangered species, leaf nodes, Pterocarpus santalinus, Red sanders

 

Short Communications

 

Indian Journal of Biotechnology

Vol 7, October 2008, pp. 547-549

 

The stem cell in the umbilical cord blood is not related to volume and nucleated cell count

A Mahantappa1, Avijit Banik2, Jyothsna Shivshankar2
and S G A Rao1*

1Cryo Stem Cell Karnataka Pvt Ltd and 2Sri Raghavendra
Biotechnologies Pvt Ltd, 72, SBM
Colony, Anandnagar,

Bangalore 560 024, India

Received 6 March 2007; revised 14 February 2008;
accepted 20 April 2008

 

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.

 

Keywords: Autologous banking, CD 34+ cells, haematopoietic stem cells, umbilical cord blood

 

Indian Journal of Bioteechnology

Vol. 7, October2008, pp 550-553

 

Cloning of pediocin PA-1 and its immunity genes from Pediococcus acidilactici K7 using pAMJ shuttle vector into Lactococcus lactis MG1363

Anusha Srikanth and Prakash M Halami*

Department of Food Microbiology, Central Food Technological Research Institute, Mysore 570 020 India

Received 17 October 2007; revised 25 March 2008;
 accepted 20 June 2008

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 ~ 2-3 ´ 103 mg-1 was observed. This was further confirmed by plasmid DNA isolation and analysis.

Keywords: Pediococcus acidilactici, pediocin PA-1, electroporation, P170 expression system, Lactococcus lactis

 

Indian Journal of Biotechnology

Vol 7, October 2008, pp. 554-556

 

Detection and characterization of group A and D avian rotaviruses in India

Savita1, A L Kusumakar1, Y P S Malik1*, Minakshi2,
G Prasad2

1Department of Veterinary Microbiology, College of Veterinary Science & A H, JNKVV, Jabalpur 482 001, India

2Department of Animal Biotechnology, CCSHAU,
Hisar 125 001, India

Received 9 February 2007; revised 29 February 2008;
accepted 2 May 2008

The present study was undertaken to determine 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.

Keywords: Avian rotavirus, diarrhea, diversity, genomic pattern, group A and D, RNA-PAGE

 

 

 

 

 

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".

Signatures and names of all the authors

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.

 

Preparation of the Manuscript

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.

Title—It should be short & informative (15 pt), to be typed in only first letter of the first word capital; also, after colon or hyphen, first letter of the first word capital. Latin names are to be given in italics.

Short Running Title—Not in excess of 50 characters, to be all in capitals.

Keywords—Five or six keywords (in normal; 9 pt) indicating the contents of the manuscript.

Authors—Names of authors to be typed in first letters capital (10 pt).

Addresses of Authors—Addresses of the institution (s) where the work was carried out including telephone (office only), fax number and e-mail address (9 pt). Author for correspondence should be indicated with an asterisk (*)

Main Headings—Each manuscript should be divided into the following main headings (typed in bold, first letters capital, on the left hand side of the page; 11 pt): Abstract, Introduction, Materials and Methods, Results, Discussion, Acknowledgement, References.

Sub-Headings—Typed in flush left, bold, first letters capital (9 pt).

Sub-Sub Headings—Bold-Italics, first letters capital
(9 pt).

Abstract—Should be brief not exceeding 200 words, typed in normal (9 pt).

Introduction—A brief and precise literature review with objectives of the research undertaken and essential background be given.

Materials and Methods—Materials and Methods should include the source and nature of material, experimental design and the techniques employed. New methods should be described in sufficient details, and others can be referred to published work.

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.

Discussion—The discussion should deal the interpretation of the results. Wherever possible, results and discussion can be combined.

Tables—Tables should be typed in double space on separate sheets, numbered consecutively, and only contain horizontal cells. The table headings should be typed with the first letter capital.

Figures—The line drawings, illustrations, photographs, etc. will be accepted in JPEG/TIFF files with hard copy. For each figure, a glossy print or original drawing may be submitted. Photomicrographs should have a scale bar. Line drawings should be roughly twice the final printed single column size of 7.5 cm width. Text figures should be numbered in Arabic numerals. Lettering, numbering, symbols and lines in the graphs/illustrations should be sufficiently clear and large to withstand reduction up to 50%. Captions and legends to illustrations should be typed on a separate sheet of paper. Line drawings and photographs should contain figure number, author’s name and the orientation (top) on the reverse with a soft lead pencil. Photostat copies and dot matrix prints will not be accepted.

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.