Indian Journal of Biotechnology

 Total visitors:4,770 since 31-03-07

VOLUME 6

CODEN:IJBNAR 6(1) 1-132 (2007)

NUMBER 2

APRIL 2007

ISSN:0972-5849

CONTENTS

 Reviews

 

Lipase applications in food industry

141

IPC Code: Int. Cl.8 C12N9/20

 

R Aravindan, P Anbumathi and T Viruthagiri

 

 

 

Site directed drug delivery by non-viral mode

159

IPC Code: Int. Cl.8 A61K 48/00

 

Shiv Kumar Dubey, Archana Pandey, Roli Mishra, Neha Kapoor, Alka Tiwari and Krishna Misra

 

 

Pichia pastoris: A notable heterologous expression system for the production of foreign proteins—Vaccines

175

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

 

V Balamurugan, G R Reddy and V V S Suryanarayana

 

 Papers

 

Development of monoclonal antibodies against chicken IgM and its application in immunity studies

187

IPC Code: Int. Cl.8 C07K16/08

 

S Rathnapraba, G Dhinakar Raj and V Thiagarajan

 

 

 

Effect of substrates on the production of Monascus biopigments by solid-state fermentation and pigment extraction using different solvents

194

IPC Code: Int. Cl.8 C07C403/24, C12N1/12

 

Julio C Carvalho, Bruno O Oishi, Adenise L Woiciechowski, Ashok Pandey, Sumathy Babitha and Carlos R Soccol

 

 

 

Immobilization of tannase from Rhizopus oryzae and its efficiency to produce gallic acid from tannin rich agro-residues

200

IPC Code: Int. Cl.8 A01N63/04; C12N9/20; C12N11/04

 

Swetansu K Hota, Jayati Ray Dutta and Rintu Banerjee

 

 

 

Analysis of genetic relationship among three varieties of indigenous Kadaknath breed using 25 chicken microsatellite markers

205

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

 

S N S Parmar, T C Tolenkhomba, M S Thakur, C G Joshi, D N Rank, J V Solanki, P N Srivastava and P V A Pillai

 

 

 

Genetic diversity and molecular characterization of pathogenic Escherichia coli from different species of laboratory rodents

210

IPC Code: Int. Cl.8. C12N15/10

 

B Maity and P Y Guru

 

 

 

Molecular and morphological divergence studies in Indian potato varieties

216

IPC Code: Int. Cl.8 C12N15/10

 

V P Chimote, D Pattanayak and P S Naik

 

 

 

Genetic polymorphism in rice (Oryza sativa L.) through RAPD analysis

224

IPC Code: Int. Cl.8 C12N15/10

 

Shamsun Naima Rahman, Md Shahidul Islam, Md Samsul Alam and Khondoker MNasiruddin

 

 

 

Immobilization of amylase onto arylamine glass beads affixed inside a plastic beaker: Kinetic properties and application

230

IPC Code: Int. Cl. 8 G01N 33/535

 

Pinki Rani, Minakshi Sharma, Vijay Kumar and C S Pundir

 

 

 

Studies on treatment of low-strength effluents by UASB reactor and its application to dairy industry wash waters

234

IPC Code: Int. Cl.8 C08K5/18

 

P Sankar Ganesh, E V Ramasamy, S Gajalakshmi, R Sanjeevi and S A Abbasi

 

 

 

Micropropagation of mature tree of Citrus limon

239

IPC Code: Int. Cl.8 A01H4/00

 

J S Rathore, M S Rathore, M Singh, R P Singh and N S Shekhawat

 

 

 

In vitro micropropagation of Heliotropium indicum Linn.—An Ayurvedic herb

245

IPC Code: Int. Cl.8 A01H4/00

 

M Senthil Kumar and M V Rao

 

 

 

Plant regeneration through somatic embryogenesis from stem and petiole explants of Indian chicory (Cichorium intybus L.)

250

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

 

M Z Abdin and A Ilah

 

   

 

 

Effects of different culture media on seed germination and subsequent in vitro development of protocorms of Hygrochilus parishii (Veith & Rchb.f.) Pfitz (Orchidaceae)

256

IPC Code:  Int. Cl.8  A01H4/00

 

R Shadang, Padmanabh Dwivedi, S N Hegde and N Ahmed

 

 

          

Plantlet regeneration of Pinus kesiya Royle ex Gord. from mature embryos

262

IPC Code: Int. Cl.8 A01H4/00, 7/00

 

Pramod Tandon, Suman Kumaria and Hiranjit Choudhury

 

 

A reappraisal of the plasmid stability problem in chemostat cultures

267

IPC Code: Int. Cl.8 C12N9/20

 

P R Patnaik

 

 Short Communications

 

Molecular markers for screening salinity response in Sorghum

271

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

 

M V Subba Rao, P Kusuma Kumari, V Manga and N Sarada Mani

 

 

 

In vitro propagation of tikhur (Curcuma angustifolia Roxb.): A starch yielding plant

274  

IPC Code: Int. Cl.8 A01H4/00

 

S K Shukla, Susmita Shukla, Vijaya Koche and S K Mishra

 

 

 

Studies on flavonoid production using in vitro cultures of Momordica charantia L.

277

IPC Code: Int. Cl.8 A01H4/00; A61P3/06

 

Mala Agarwal and Raka Kamal

     

Micropropagation of Coptis teeta Wall.–Threatened medicinal plant of Arunachal Pradesh, India

280

IPC Code: Int. Cl.8 A01H4/00

Pramod Tandon, Tilok S Rathore and Suman Kumaria

 

Instructions to Contributors

 

 

 

 


Reviews

Indian Journal of Biotechnology

Vol 6, April 2007, pp 141-158

 

 

Lipase applications in food industry

R Aravindan*, P Anbumathi and T Viruthagiri

Lipases are the most pliable biocatalyst and bring about a wide range of bioconversion reactions, such as hydrolysis, interesterification, esterification, alcoholysis, acidolysis and aminolysis. Lipases can act on a variety of substrates including natural oils, synthetic triglycerides and esters of fatty acids. They are resistant to solvents and are exploited in a broad spectrum of biotechnological applications. Lipase catalyzed transesterification, hydrolysis and esterification are the important class of reactions for food technology applications in fats and oil industry, dairy industry, pharmaceuticals and bakery industry. Lipases are very peculiar as they hydrolyse fats into fatty acids and glycerol at the water-lipid interface and can reverse the reaction in non-aqueous media. Novel biotechnological applications, like biopolymer synthesis, biodiesel production, treatment of fat-containing waste effluents, enantiopure synthesis of pharmaceuticals and nutraceutical agents, have been established successfully. The present article extends the frontier of lipase technology towards food processing applications and discusses the important characteristics of lipases and its sources. Various methods of lipase immobilization for food technology applications, various assay methods for lipase, production of lipase by submerged and solid state fermentation strategies, and various purification methods available have been discussed in detail.

Keywords: ester, fatty acids, food applications, lipases, lipase sources, triglycerides

IPC Code: Int. Cl.8 C12N9/20

Indian Journal of Biotechnology

Vol 6, April 2007, pp 159-174

 

 

Site directed drug delivery by non-viral mode

Shiv Kumar Dubey, Archana Pandey, Roli Mishra, Neha Kapoor, Alka Tiwari and Krishna Misra*

Drug Delivery Technology (DDT) has achieved new heights during the last few years as an essential component of drug development. The delivery of drugs exclusively to the diseased site is the only plausible solution for increasing their efficacy and simultaneously reducing the toxic effects. Ever since the unraveling of human genome in the beginning of the present century and subsequent emergence of proteomics and genomics as new disciplines has given a new thrust to gene therapy which is aimed at treating the cause rather than the symptoms unlike the conventional small molecule drugs. The desired gene is transferred to the target cell through a vector so that the desired protein itself is expressed in the patient’s body. This approach is an appropriate remedy for the fast mutations occurring in pathogens. The vector is invariably a virus or may be a non-virus, therefore, the novel drug delivery systems are either via viral or non-viral modes. The non-viral delivery systems may be either synthetic vectors, drug loaded tumour cells (DLTCs) as vectors, folates (folic acid conjugates) or it may be by applying physical or chemical methods. All these approaches have been described in this paper giving the latest achievements of each. The prodrug concept has also been elaborated concisely.

Keywords: drug delivery, liposome, biodegradable polymers, prodrug, folates

IPC Code: Int. Cl.8 A61K 48/00

Indian Journal of Biotechnology

Vol 6, April 2007, pp 175-186

 

 

Pichia pastoris: A notable heterologous expression system for the production of foreign proteins—Vaccines

 

V Balamurugan, G R Reddy and V V S Suryanarayana*

Improvements in yeast expression systems, coupled with the development of more information on methylotrophic yeast are expanding the role of yeast in the process of understanding and engineering eukaryotic proteins. Pichia pastoris has become a highly popular expression host for the production of a wide variety of intracellular and extracellular recombinant proteins of interest. Initial success with this system was greatly facilitated by the development of versatile expression vectors that were almost exclusively based on the strong, tightly regulated promoter of the P. pastoris major alcohol oxidase gene. Recent advances in the development of yeast as a host for the production of heterologous proteins have provided a catalogue of new applications, methods and system components, which will help us to understand about more details and applications of the system even further. Characteristic features of Pichia species, which are the most advantageous and favourable, have resulted in an increasing number of biotechnological applications. During the past two decades, P. pastoris has been developed into a highly successful system, due to its increasing popularity, which can be attributed to several factors such as, the simplicity of techniques needed for the molecular genetic manipulation; the ability to produce foreign proteins at high level, the capability of performing many eukaryotic post-translational modifications (glycosylation, disulfide bond formation and proteolytic processing) and finally the availability of the expression system as a commercially available kit.

Keywords: Pichia pastoris, heterologous expression system, foreign proteins, vaccines

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

Papers

Indian Journal of Biotechnology

Vol 6, April 2007, pp 187-193

 

 

Development of monoclonal antibodies against chicken IgM and its
application in immunity studies

S Rathnapraba*, G Dhinakar Raj and V Thiagarajan

Immunoglobulin M (IgM) from specific pathogen free (SPF) chicken serum was purified and used for production of monoclonal antibodies (MAbs) for subsequent application in the detection of primary immune response to infectious agents. Chicken IgM was fractionated by gel filtration technique using Sephadex G-200. Purity of the fractionated IgM was confirmed by agar gel precipitation test (AGPT) and immunoelectrophoresis (IE) using rabbit anti-chicken serum. In polyacrylamide gel electrophoresis (PAGE), under non-reducing conditions, a single band was noticed and under reduced conditions, a band with molecular weight of 74 kDa was observed. Cell fusion was carried out with spleenocytes from immunized BALB/c mice and Sp2/0 cells, and the high positive clones were selected and characterized for isotype and cross-reactivity with IgY in enzyme linked immunosorbent assay (ELISA). Three MAbs chosen for characterization were of IgG1 isotype and they did not cross-react with standard IgY in ELISA. Concentrated IgM MAbs developed in this study were used in ELISA for the measurement of immune response in sequentially collected serum samples of birds, experimentally infected with egg drop syndrome (EDS-76) virus. The purified virus was coated onto the ELISA plates, followed by the addition of sera samples to be tested, MAbs against chicken IgM, anti-mouse peroxidase conjugate and substrate. The assay revealed an increase in IgM response in individual birds from 5-12 d with a peak on 9th d post-inoculation (PI), followed by an increase in IgG on 12-25 d PI. IgM antibodies against infectious bursal disease (IBD) virus were tested in 34 field serum samples. Comparison with AGPT revealed a marginal increase in sensitivity with IgM ELISA for detection of IBD virus specific IgM.

Key words: Chicken IgM, ELISA, immunoglobulins, monoclonal antibodies

IPC Code: Int. Cl.8 C07K16/08

Indian Journal of Biotechnology

Vol 6, April 2007, pp 194-199

 

 

Effect of substrates on the production of Monascus biopigments by solid-state fermentation and pigment extraction using different solvents

 

Julio C Carvalho, Bruno O Oishi, Adenise L Woiciechowski, Ashok Pandey, Sumathy Babitha and Carlos R Soccol*

The aim of the present work was to study the potential of natural substrates for biopigment production using a strain of Monascus sp, and to compare different solvents for extracting these pigments. Rice was the best substrate for cultivation of the culture in SSF. However, some of the other substrates used also presented good biopigment production, especially corn, wheat and cassava. Cassava bagasse gave a low pigment yield, but it is an agro-industrial residue whose low price might compensate for its low yield. However, efficient pigment production with this substrate requires supplementation of the culture medium with nutrients. Optimum fermentation time for SSF in cassava bagasse was 10-11 d. The production of yellow pigments was superior to that of red pigments, but the ratio red/yellow (ABS 500/ABS 400) grew during the course of the process. The comparison of several solvents for extraction showed that methanol was the best solvent, closely followed by DMSO and ethanol. The results also indicated that a mixture of ethanol-water with ca. 60% ethanol (w/w) was more efficient than other ethanol concentrations. The solvent to substrate ratio had little influence in the extraction. Finally, temperature in the range of 0ºC to 60ºC showed no difference in the extraction of the biopigments under static condition.

Keywords: pigment, substrate, extraction, solvent, solid-state fermentation, Monascus

IPC Code: Int. Cl.8 C07C403/24, C12N1/12

Indian Journal of Biotechnology

Vol 6, April 2007, pp 200-204

 

 

Immobilization of tannase from Rhizopus oryzae and its efficiency to produce gallic acid from tannin rich agro-residues

Swetansu K Hota, Jayati Ray Dutta and Rintu Banerjee*

Different tannin rich agro-residues, like sal seed (21% tannin), fruit of myrobalan (37.6% tannin) and tea-leaf (14.1% tannin) were selected for enzymatic conversion of their tannin content to gallic acid. These tannin rich substrates were used as carbon source in Czapek-dox medium for the production of tannase (E.C. 3.1.1.20, tannin acyl hydrolase) from Rhizopus oryzae (RO IIT RB-13, NRRL-21498). The maximum enzyme production of 17.7 U/mL was obtained in sal seed powder incubated for 48 h at 30oC. The enzymatic conversion of these agro-residues was carried out using immobilized tannase. Substrate concentrations of 8% sal seed, 7% myrobalan and 6% tea leaf (w/v) were found to be the optimum for maximum bioconversion. The maximum bioconversion (90 and 87%, respectively) was achieved with sal seed and tea leaf as substrate at 40°C and initial pH 4.5. In case of myrobalan, the maximum bioconversion was 90.2% at 50°C and initial pH 5.0. Moreover, optimization of the pH and temperature largely reduced the incubation time to 36 h. The immobilized tannase was stable for 7 cycles. The kinetic properties of immobilized enzyme revealed that there was a decrease in maximal reaction velocity (Vmax) and increase in Michaelis constant (Km) when compared to its free native counterpart.

Keywords: immobilization, Rhizopus oryzae, sal seed, tannase, tea-leaf, myrobalan

IPC Code: Int. Cl.8 A01N63/04; C12N9/20; C12N11/04

Indian Journal of Biotechnology

Vol 6, April 2007, pp 205-209

 

 

 

Analysis of genetic relationship among three varieties of indigenous Kadaknath breed using 25 chicken microsatellite markers

S N S Parmar*, T C Tolenkhomba, M S Thakur, C G Joshi, D N Rank, J V Solanki, P N Srivastava
 and P V A Pillai1

The genetic variability of three varieties of indigenous Kadaknath breed of poultry was evaluated using 25 microsatellite markers. All the 25 microsatellite markers were found to be polymorphic in all the three varieties. Number of alleles observed for all the markers varied from 3 to 10. Allele size of polymorphic loci ranged from 98-356 bp and showed very large variation across loci. Average heterozygosity values were found to be 0.721, 0.694 and 0.689 in Jet black, Golden and Pencilled varieties, respectively. Overall heterozygosity value for all the microsatellite loci among all the three varieties was found to be 0.701. Average polymorphic information content (PIC) values were found to be 0.671, 0.699, and 0.617 in Jet black, Golden and Pencilled varieties, respectively. Overall PIC values for all the microsatellite loci among all the three varieties were found to be 0.662. The Nei’s standard genetic distance (Ds) values between Jet black and Golden, Jet black and Pencilled, and Golden and Pencilled varieties were found to be 0.1678, 0.0951 and 0.1943 respectively. Phylogenetic consensus tree constructed using the boostrapped data and neighbour-joining method grouped all the three varieties into one cluster. Therefore, it is concluded that all the three varieties of Kadaknath breed had negligible genetic distance between each other.

Keywords: poultry. Kadaknath breed, microsatellite markers, genetic relationship

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

Indian Journal of Biotechnology

Vol 6, April 2007, pp 210-215

 

 

Genetic diversity and molecular characterization of pathogenic Escherichia coli from different species of laboratory rodents

 

B Maity* and P Y Guru

Escherichia coli has been intensively studied under various aspects in general bacteriology. In this study, 10 RAPD primers were examined in pathogenic E. coli isolated from laboratory rodents like Swiss, PS, BALB/c, C3H/J and SMA mice; SD, CF, DR, Wistar and Lew rats; Cotton rats, Mastomys and Mongolian gerbils suffering from diarrhoea to establish genetic diversity and molecular characterization in different rodent species. It has been found that the largest amplified products were 2000 bp and lowest 200 bp and the number of scorable bands for responding primers ranging from 1 to 8 with an average of four bands. A total of 6,724 bands were scored against 10 RAPD primers for E. coli in different rodent species. RBM 1 (RAPD primer) was amplified with 1000 bp bands in all rodent species. The dendogram revealed three groups among pathogenic E. coli strains of 13 rodent species which were almost similar except Wistar and SD rats due to their genetic diversity. The linkage distance between Wistar vs. SD was 0.06, and CF vs. Mastomys was 0.01. It is concluded that the pathogenic E. coli that may cause watery, non-bloody diarrhoea to watery-bloody diarrhoea and dysentery could be easily identifed using RBM 1 primer in laboratory rodents.

Keywords: Escherichia coli, genetic diversity, molecular characterization, RAPD, laboratory rodents

IPC Code: Int. Cl.8. C12N15/10

Indian Journal of Biotechnology

Vol 6, April 2007, pp 216-223

 

 

 

Molecular and morphological divergence studies in Indian potato varieties

V P Chimote*, D Pattanayak and P S Naik

Genetic diversity analysis of 32 commercial Indian potato varieties was done using four multi-loci SSR (123 alleles at 12 loci), 14 RAPD (168 alleles) and 21 morphological markers. 2D PCO analysis in general, gave a better picture of the genetic divergence as they had low influence of any specific allele/marker. Though the degree of divergence varied, overall placement patterns were similar to a large extent in all the three PCO studies, except that in RAPD studies this was defined by the second component of the axis. These results were also reflected in a consensus 2D-scatter plot derived from mean placement values of the three PCOs, which showed that Indian varieties were placed into three separate groups. Solanum demissum derived late blight resistant varieties formed a closely placed group and andigena type varieties released prior to late blight programme formed a diverse group. Third group composed of either traditional tuberosum varieties or recently released neotuberosum showed overlapping pattern.

Keywords: genetic diversity, Solanum tuberosum, SSR, RAPD, morphology

IPC Code: Int. Cl.8 C12N15/10

Indian Journal of Biotechnology

Vol 6, April 2007, pp 224-229

 

 

 

Genetic polymorphism in rice (Oryza sativa L.) through RAPD analysis

Shamsun Naima Rahman, Md Shahidul Islam*, Md Samsul Alam and Khondoker M Nasiruddin

Random Amplified Polymorphic DNA (RAPD) assay was performed to estimate genetic polymorphism in six different rice (Oryza sativa L.) cultivars viz. Basmati 370, DM 25, IRATOM 24, Binadhan 6, TNDB 100 and Y 1281. Three out of the 15 decamer random primers showed amplification of genomic DNA in 24 individuals. The primers produced a total of 26 bands of which 14 were polymorphic. Proportion of polymorphic bands and gene diversity estimates were 26.92% and 0.09 for Basmati 370, 11.54% and 0.04 for DM 25, 11.54% and 0.05 for IRATOM 24, 7.69% and 0.02 for Binadhan 6 and 23.08% and 0.11 for TNDB 100 whereas Y 1281 cultivar was monomorphic indicating the existence of high level of intra cultivar genetic variation in Basmati 370 and TNDB, respectively 100. High levels of population differentiation (GST = 0.75) and low levels of gene flow (Nm = 0.16) estimates across all the loci indicate sufficient existence of genetic variation among these six cultivars. Low intra cultivar variation and significant differentiation in different cultivar pairs was observed at a number of loci. Nei’s genetic distances estimated among the different pairs of cultivars were correlated with geographical distances. The UPGMA dendrogram based on Nei’s genetic distance clubbed the cultivars into three clusters. RAPD analysis showed promise as an effective tool in estimating genetic polymorphism in different rice cultivars.

Keywords: RAPD, polymorphism, similarity index, genetic distance, rice

IPC Code: Int. Cl.8 C12N15/10

Indian Journal of Biotechnology

Vol 6, April 2007, pp 230-233

 

 

Immobilization of amylase onto arylamine glass beads affixed inside a plastic beaker: Kinetic properties and application

Pinki Rani, Minakshi Sharma, Vijay Kumar and C S Pundir*

Commercial fungal amylase (diastase) has been immobilized through diazotization onto zirconia coated arylamine glass beads, affixed inside a plastic beaker. The immobilized enzyme retained 73.3% of the initial activity of free enzyme with a conjugation yield of 8 mg/g. Optimum pH of enzyme was decreased, while temperatures for maximum activity, energy of activation, time of incubation and Km for starch were increased after immobilization. The utility of immobilized enzymes in removal of starch stain from cotton cloth by various detergents was tested by chemical method. All the detergents gave better washing in presence of immobilized amylase than that by detergent alone. Further, the washing by cheaper (non-enzymic) detergents in presence of immobilized amylase was almost similar to that by expensive (enzymic) detergents. The immobilized enzyme was used about 100 times without any considerable loss of activity.

Keywords: amylase, arylamine glass, cloth washing, detergent, immobilization, starch stain.

IPC Code: Int. Cl. 8 G01N 33/535

Indian Journal of Biotechnology

Vol 6, April 2007, pp 234-238

 

 

Studies on treatment of low-strength effluents by UASB reactor and its application to dairy industry wash waters

P Sankar Ganesh, E V Ramasamy, S Gajalakshmi, R Sanjeevi and S A Abbasi*

In the backdrop of lack of success achieved by the past workers in using upflow anaerobic sludge blanket (UASB) reactor for treating low-strength wastewaters, the paper highlights the importance of R&D in this area. The two main reasons behind the importance of using UASB are: (1) generation of large volumes of low-strength wastewaters, which are often disposed untreated due to high costs, and (2) the potential of stabilizing the organic wastes by producing valuable energy as byproduct. Results are presented on the successful operation of UASB in treating low-strength dairy industry wash waters [Chemical oxygen demand (COD 1200-2000 mg/L]. The reactors achieved treatment efficiency of the order of 75-85% and were able to withstand shock-loads without adversely affecting the treatment efficiency. One of the reactors which was accidentally contaminated with acid, recovered quickly.

Keywords: UASB, low strength, dairy wastewater, biogas, methane, organic loading rate

IPC Code: Int. Cl.8 C08K5/18

Indian Journal of Biotechnology

Vol 6, April 2007, pp 239-244

 

 

Micropropagation of mature tree of Citrus limon

J S Rathore, M S Rathore, M Singh, R P Singh and N S Shekhawat*

A protocol for in vitro cloning of mature plants of Citrus limon using nodal shoot segments has been developed. Three to four shoots were induced per axillary meristems on MS medium amended with 9 µM BAP. Repeated transfer of the initial explants for up to six passages on MS medium [having 50% of the original concentrations of NH4NO3 and KNO3, and supplement of 250 mg L-1 (NH4)2SO4] with 0.22 µM BAP yielded crops of shoots. Three to four-fold shoot multiplication rate was achieved by subculturing of shoots. The shoots have been multiplied for more than 48 months without loss of vigour. One hundred percent of the cloned shoots rooted: (i) in vitro on half-strength MS medium+27 µM NAA+0.1% activated charcoal, and (ii) ex vitro on soilrite if pulse-treated with 1.07-2.68 mM NAA. Ninety five percent of rooted plantlets survived hardening and transplanting. Micropropagated plants are under filed trials in states of Rajasthan and Gujarat of India. These have flowered and produced fruits.

Keywords: cloning, ex vitro rooting, in vitro, lemon, mature explant, nodal shoot segment

IPC Code: Int. Cl.8 A01H4/00

Indian Journal of Biotechnology

Vol 6, April 2007, pp 245-249

 

 

In vitro micropropagation of Heliotropium indicum Linn.—An Ayurvedic herb

M Senthil Kumar and M V Rao*

An efficient micropropagation method was developed for Heliotropium indicum, an Ayurvedic medicinal herb, using apical and axillary bud explants. The highest number of shoots (32.6 shoots/apical bud and 20.2 shoots/axillary bud) was yielded after 30 d of culture in the MS medium supplemented with Kn (1.0 mg L-1), BA (0.5 mg L-1) and IAA (0.05 mg L-1). The cluster of proliferated shoots elongated simultaneously on the same medium. High frequency of rooting (85%) was obtained in both apical and axillary bud derived shoots when transferred to half-strength MS medium supplemented with IBA (0.1 mg L-1). Rooted plants were hardened and successfully established in field conditions.

Keywords: Heliotropium indicum, Boraginaceae, micropropagation, apical bud, axillary bud

IPC Code: Int. Cl.8 A01H4/00

Indian Journal of Biotechnology

Vol 6, April 2007, pp 250-255

 

 

Plant regeneration through somatic embryogenesis from stem and petiole explants of Indian chicory (Cichorium intybus L.)

M Z Abdin* and A Ilah

An efficient protocol has been developed for rapid propagation of Indian chicory (Cichorium intybus L.) through somatic embryo development. Indirect somatic embryogenesis was induced through nodal stem and petiole explants, cultured on Murashige and Skoog (MS) medium containing different concentrations of 2,4-D, NAA or IAA. Callus was induced from both the explants after 3 wk of inoculation. Subsequently, calli were sub-cultured on MS medium containing different auxin/cytokinin ratios that influenced the intensity of embryo formation, germination and ability to regenerate plants. Somatic embryogenesis was more intensive in medium with higher concentration of Kn (1.5 mg L-1) and lower concentration of IAA (0.5 mg L-1) supplemented with vitamin-free casein hydrolysate (500 mg L-1). After sub-culture, the embryoids were matured onto a fresh MS medium supplemented with Kn (1.5 mg L-1) + IAA (0.1 mg L-1) + IBA (1.0 mg L-1) + CH (500 mg L-1). The germination of embryos into shoots and less developed roots occurred on the same medium with slight change in IBB concentration (0.5 mg L-1). Further, root formation occurred in MS medium supplemented with 1.0 mg L-1 IBA.

Keywords: Cichorium intybus L., Indian chicory, micropropagation, somatic embryogenesis

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

Indian Journal of Biotechnology

Vol 6, April 2007, pp. 256-261

 

 

Effects of different culture media on seed germination and subsequent
in vitro development of protocorms of Hygrochilus parishii
(Veith & Rchb.f.) Pfitz (Orchidaceae)

R Shadang, Padmanabh Dwivedi*, S N Hegde and N Ahmed

Seeds of Hygrochilus parishii (Veith & Rchb.f.) Pfitz were inoculated on eight different culture media (basal) MS, MKC, V&W, I&Y and half strength of respective media. Seeds germinated within 15-20 d almost on all media with varying percentage and developed into greenish protocorm after 80-100 d. Best seed germination was observed in ½ strength MKC medium. Healthy protocorms were observed within 100 d in V&W media. Sub-cultured protocorms in all the above media supplemented with NAA, BAP, 2,4-D and IAA in a range of 0.5-2.0 mg L-1, both individually and in combination, and additives like banana pulp (BP, 10%) and coconut milk (CM, 15%) produced well developed callus. Multiplication of protocorms was best on ½ strength MS + NAA (2.0 mg L-1). Formation of root and leaf was early in V&W medium having CM (15%) and BP (10%) but without sucrose.

Keywords: culture media, Hygrochilus parishii, orchid, protocorms, seed germination

IPC Code:  Int. Cl.8  A01H4/00

Indian Journal of Biotechnology

Vol 6, April 2007, pp 262-266

 

 

Plantlet regeneration of Pinus kesiya Royle ex Gord. from mature embryos

Pramod Tandon*, Suman Kumaria and Hiranjit Choudhury

A high frequency (90.5%) of shoot bud induction was observed in mature zygotic embryos of Pinus kesiya Royleex Gord. that were pre-cultured in Quoirin and Lepoivre (LP) medium containing 23.15 mM Kn for 4 wk and then transferred to growth regulator-free medium. Multiplication and elongation of the shoot buds resulted in 1/2 LP medium containing 0.5 mM IBA and 4.5 mM BAP or Kn. Rooting was 72.3% in isolated shoots that were treated with 53.76 mM NAA for 24 h and then cultured on water-agar medium. Plantlets were hardened and successfully established on soil collected from pine forests with 70% survival. Biomass of the micropropagated plants was 3.2 times higher than seed-derived plants.

Keywords: growth characteristics, micropropagation, shoot buds induction, zygotic embryos, Pinus kesiya

IPC Code: Int. Cl.8 A01H4/00, 7/00

Indian Journal of Biotechnology

Vol 6, April 2007, pp 267-270

 

 

A reappraisal of the plasmid stability problem in chemostat cultures

P R Patnaik*

Segregational instability models of recombinant bacteria conventionally assume that ¾(a) the probability of loss of a unit of a plasmid is constant and (b) upon cell division, the progeny either inherit the mother’s plasmid content entirely or not at all. However, two previous studies have shown that neither assumption is correct. So, in this work, both assumptions have been relaxed to formulate a more general model of plasmid dynamics in a chemostat. Unlike earlier work, this model predicts a time-dependent dilution rate to maximize segregational stability. Some biological implications are discussed.

Keywords: chemostat, decay probability variation, Escherichia coli, plasmid stability

IPC Code: Int. Cl.8 C12N15/00

 

Short Communications

Indian Journal of Biotechnology

Vol 6, April 2007, pp 271-273

 

 

Molecular markers for screening salinity response in Sorghum

M V Subba Rao*, P Kusuma Kumari, V Manga and
N Sarada Mani

Genomic DNAs from two salinity tolerant (IS 23190-Zm 90 & IS 23217-Zm 138), one moderately tolerant (IS 3566) and one sensitive(IS 213253-Zm 321)  accessions of Sorghum were subjected to RFLP and RAPD analyses. A combination of digestion with Dra I and probing with BADH 1 distinguished the sensitive from the tolerant and moderately tolerant accessions in the RFLP patterns. A primer having CGGCTAGGT sequence provided distinct RAPD pattern for the tolerants (presence of specific amplified products a, b & c), moderately tolerant (presence of only c) and sensitive (lacking a, b & c) genotypes.

Keywords: RAPD, RFLP, salinity tolerance, Sorghum

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

Indian Journal of Biotechnology

Vol 6, April 2007, pp 274-276

 

 

In vitro propagation of tikhur (Curcuma angustifolia Roxb.): A starch yielding plant

S K Shukla*, Susmita Shukla, Vijaya Koche and S K Mishra

In vitro regeneration of Curcuma angustifolia Roxb. was achieved through shoot meristem culture. The shoot buds (2-3 cm long) from rhizome were inoculated on MS medium supplemented with 3.0 mg/L BAP for initiation and elongation of shoots. As a result, 1.87±0.28 shoots per explant were produced These shoots were transferred on MS medium supplemented with 3.0 mg/L BAP and 25 mg/L adenine sulfate for further shoot multiplication. About 6.9±0.69 micro-shoots per explant were produced with in 6 wk. The roots appeared from shoots on shoot establishment as well as multiplication media. The rooted plants were transferred to pots and acclimatized, which showed 83% survival with normal growth.

Keywords: Curcuma angustifolia, meristem culture, micropropagation, shoot bud

IPC Code: Int. Cl.8 A01H4/00

Indian Journal of Biotechnology

Vol 6, April 2007, pp 277-279

 

 

Studies on flavonoid production using in vitro cultures of Momordica charantia L.

Mala Agarwal* and Raka Kamal

Callus cultures and in vitro plant development of Momordica charantia L. were establishment on Murashige and Skoog’s (MS) medium using different concentrations and combinations of auxins and cytokinins. Further, qualitative and quantitative production of flavonoids in callus cultures and different in vitro developmental stages were studied by using thin layer chromatography and spectrophotometric analysis. Presence of three flavonoids was detected and the maximum amount of total flavonoid content was observed in 6-wkold callus cultures (2.90 mg/g dry wt). While in different in vitro morphogenetic stages, the maximum amount of total flavonoid content was observed in multiple shoots (2.96 mg/g dry wt).

Keywords:   flavonoid production, in vitro regeneration, Momor-dica charantia

IPC Code: Int. Cl.8 A01H4/00; A61P3/06

Indian Journal of Biotechnology

Vol 6, April 2007, pp 280-282

 

 

Micropropagation of Coptis teeta Wall.–Threatened medicinal plant of Arunachal Pradesh, India

Pramod Tandon*, Tilok S Rathore and Suman Kumaria

Multiple shoots were produced from axillary buds with rhizome parts from mature plants of Coptis teeta on ½ strength Murashige & Skoog’s (MS) nutrient salts medium containing 4.42 mM benzylamino purine+0.56 mM indole-3-acetic acid. Isolated shoots were rooted on ½ strength MS medium supplemented with 7.35 mM indole-3-butyric acid. Plantlets were also regenerated directly from rhizome segments cultured on Gamborg’s medium containing 2.31 mM kinetin+0.56 mM indole-3-acetic acid. Regenerated plantlets were hardened and established in the glass house.

Keywords: Coptis teeta, axillary buds, rhizome, multiple shoots, micropropagation

IPC Code: Int. Cl.8 A01H4/00

 

 

 

 


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. 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 floppy disk [3.5¢¢ (1.44 MB)] 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

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.