Total visitors:7,020 since 27-07-04

Indian Journal of Biotechnology

 

 

 

ISSN:0972-5849

 

CODEN:IJBNAR 3(3) 323-464 (2004)

VOLUME 3

NUMBER 3

JULY 2004

 

 

 

CONTENTS

 

Reviews

 

Microsatellites (SSRs): Puzzles within puzzle

331

IPC Code: Int. Cl.7 C 12 N 15/09, 15/10, 15/11

 

Seema Trivedi

 

 

 

Genetic conservation of plantation crops and spices using cryopreservation

348

IPC Code: Int. Cl.7 A 23 B 7/024, 7/055, 9/10

 

Rekha Chaudhury & S K Malik

 

 

 

Genetic management for increased tolerance to aluminium and iron toxicities in rice—A review

 

359

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00; C 12 N 15/10

 

Asit B Mandal, Asit K Basu, Bidhan Roy, T E Sheeja & Tarak Roy

 

 

 

Papers

 

Nasal melatonin gels using pluronic PF-127 for chronobiological treatment of sleep disorder

 

369

IPC Code: Int. Cl.7 A 61 K 38/00, 47/36

 

S S Pisal, P Reddy, A R Paradkar, K R Mahadik & S S Kadam

 

 

 

Isolation and characterization of proteins involved in cryoprotection from cold resistant mutants of Pseudomonas fluorescens ATCC13525

 

378

IPC Code: Int. Cl.7 A 01 N 63/00

 

Vandana Katiyar, D P Mishra, Satish Kumar & Reeta Goel

 

 

 

Role of phosphorylase and amylase in accelerated ripening of a mutant banana ‘Basrai-10Gy’

 

382

IPC Code: Int. Cl.7 C 12 N 15/01

 

K K Surendranathan, N K Ramaswamy & M B Pendharkar

 

 

 

Characterization of cotton (Gossypium hirsutum) germplasm by ISSR, RAPD markers and agronomic values

 

388

IPC Code: Int. Cl.7 C 12 N 15/10

 

Ashok Dongre, Vilas Parkhi & Santosh Gahukar

 

 

 

Kinetic studies of penicillin production during batch and repeated batch in fluidized bed bioreactor with agar immobilized P. chrysogenum cells

 

394

IPC Code: Int. Cl.7 A 01 N 63/00; C 12 P 37/00

 

A Swaroopa Rani, Annapurna Jetty & S V Ramakrishna

 

 

 

Generation and characterization of pea (Pisum sativum) somaclones for resistance to Ascochyta blight and powdery mildew

 

400

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00; C 12 N 15/01

 

Rajan Sharma & R P Kaushal

 

 

 

Banana streak virus from India and its detection by polymerase chain reaction

409

IPC Code: Int. Cl.7 C 12 N 15/10

 

Anita K Cherian, V K Baranwal, V G Malathi, R P Pant & Y S Ahlawat

 

 

 

Agrobacterium rhizogenes mediated hairy root induction in two medicinally important members of family Solanaceae

 

414

IPC Code: Int. Cl.7 A 01 H 5/06; C 12 N 15/03

 

Pankaj K Pawar & Vijay L Maheshwari

 

 

 

Silver nitrate induced rooting and flowering in vitro on rare rhoeophytic woody medicinal plant, Rotula aquatica Lour.

 

418

IPC Code: Int. Cl.7 A 01 H 4/00, 5/02, 5/06

 

M Chithra, K P Martin, C Sunandakumari & P V Madhusoodanan

 

 

 

In vitro propagation of Pterocarpus marsupium Roxb.: An endangered medicinal tree

422

PC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

Sharad Tiwari, Pankaj Shah & Kanchan Singh

 

 

 

In vitro clonal propagation of Momordica charantia L.

426

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

Mala Agarwal & Raka Kamal

 

 

 

Selection of transformants of Escherichia coli containing cellulase gene from Ruminococcus albus isolated from rumen of crossbred steers

 

431

IPC Code: Int. Cl.7 C 12 N 15/10, 15/11

 

M Chandrasekharaiah, A Thulasi & K T Sampath

 

 

 

Reconstruction of in vitro matured goat oocytes by using synchronized skin fibroblast cells as donor nuclei

 

435

IPC Code: Int. Cl.7 C 12 N 5/06, 15/06

 

S K Das & A C Majumdar

 

 

 

Effects of genotype and culture medium on in vitro androgenesis in soybean (Glycine max Merr.)

 

441

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

Sharad Tiwari, Prem Shankar & Manoj Tripathi

 

 

 

Establishment of an embryogenic suspension culture of Pinus kesiya (Khasi pine) from various explants

 

445

IPC Code: Int. Cl.7 A 01 H 4/00, 7/00

 

Chitta Ranjan Deb & Pramod Tandon

 

 

Short Communications

 

Direct organogenesis in Datura metel L. from in vitro and in vivo nodal explants

449

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

B Muthukumar, D I Arockiasamy & E Natarajan

 

 

 

Production of camptothecines from callus cultures of Nothapodytes foetida (Wight) Sleumer

 

452

IPC Code: Int. Cl.7 A 01 H 4/00; A 01 N 63/00

 

Sundravelan R, B Desireddy & Veeresham Ciddi

 

 

Plant regeneration from callus cultures in Suaeda nudiflora (Wild.) Moq.

454

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

Aneesha Singh, J Chikara & J B Pandya

 

 

 

Genetic characterisation of Jaisalmeri camel using microsatellite markers

457

IPC Code: Int. Cl.7 C 12 N 15/10

 

L Gautam, S C Mehta, R S Gahlot & K Gautam

 

 

 

Instructions to Contributors

461

 

 

Author Index

 

 

 

Indian Journal of Biotechnology

 Vol 3, July 2004, pp 331-347

 

Microsatellites (SSRs): Puzzles within puzzle

Seema Trivedi*

 

SSRs are not only found in all genomes (big or small) Mycoplasma vs animals but also in the smallest and compact genome amongst vertebrates like fish and birds. These findings suggest that microsatellites cannot be totally eliminated from the genome. Studies on distribution, frequency and pattern of SSRs in coding and non-coding regions reveal species specific, length or pattern dependent features, which suggest that SSRs are common in intergenic regions of all organisms. CG dinucleotide repeats are scarce and trinucleotide tolerance is more in the coding sequences. However, there are features about microsatellites, which may not fall in any paradigm, as what may be true regarding motifs in one organism may not be so for others. This is evident from the studies that show rarity of (GT)n repeat in plants vis-à-vis mammals which have these repeats in abundance. Similarly, abundance of AAT repeats is found in introns of most organisms but rodents are exceptional in having AAG abundance. To highlight these features and to address the question regarding microsatellite abundance in relation to increasing genomic complexity, data from various sources have been tabulated for this review. Different schools of thought about the origin and evolution of microsatellites are also discussed.

 

Keywords: microsatellites, simple sequence repeats, motifs, mutation, SSR birth and evolution, density and percentage

IPC Code: Int. Cl.7 C 12 N 15/09, 15/10, 15/11

 

 

 

Indian Journal of Biotechnology

 Vol 3, July 2004, pp 348-358

 

Genetic conservation of plantation crops and spices using cryopreservation

Rekha Chaudhury* and S K Malik

 

Germplasm conservation in the form of seed is convenient and most cost-effective ex situ conservation method for plant species producing orthodox seeds. Several species of plantation crops and spices produce non-orthodox seeds, which exhibit intermediate or recalcitrant seed storage behaviour. Storage of these species requires the use of in vitro conservation techniques for short- to medium-term conservation and cryopreservation to achieve long-term conservation. Various cryotechniques applicable to 11 genera totaling 16 species [rubber (Hevea brasiliensis Muell.- Arg.), cocoa (Theobroma cacao L.), coconut (Cocos nucifera L.), arecanut (Areca catechu L.), oilpalm (Elaeis guineensis Jacq.), Coffea spp., tea (Camellia sinensis L.(O.) Kuntze), black pepper (Piper nigrum L.), cardamom (Elettaria cardamomum Maton), cashew (Anacardium occidentale L.) and nutmeg (Myristica fragrans Houtt)] are reviewed in the present paper. Physical and physiological factors determining the success or failure of cryopreservation are also discussed.

 

Key words: cryopreservation, seed, embryo, embryonic axes, shoot apices, pollen

IPC Code: Int. Cl.7 A 23 B 7/024, 7/055, 9/10

 

 

 

Indian Journal of Biotechnology

 Vol 3, July 2004, pp 359-368

 

Genetic management for increased tolerance to aluminium
and iron toxicities in rice—A review

Asit B Mandal1*, Asit K Basu2, Bidhan Roy1, T E Sheeja1 and Tarak Roy3

 

Several tolerant donors were identified from rice germplasms and significant strides were made in their genetic improvement through conventional breeding during the last 40 years in Indian subcontinent. In majority of the cases the physiological mechanism governing tolerance towards Fe and Al toxicities were unzipped. It is also worth mentioning that progress in breeding largely depends upon the efficiency and effectiveness of the screening technique to identify the true tolerants, availability of the tolerant gene(s) in the germplasms and selection of desirable lines from the segregating population. With increase in Fe-toxicity level, plant height, number of ear bearing tillers, panicle length and grain yield were found decreased in rice; whereas, vegetative growth period got extended with increased spikelet sterility. Toxic concentrations of Al generally inhibit root growth, restrict water intake and nutrients uptake that lead to poor growth and low yield per se. In vitro screening for developing tolerant genotypes is relatively a straightforward and amply demonstrative method. Toxicity tolerance was also found transferable to a desirable plant type. Further, marker aided selection (MAS) would facilitate reliable selection for abiotic stress tolerant genotypes. Quantitative trait loci (QTL) for toxicity tolerance towards Fe and Al were identified and expected to help in developing tolerant, high yielding varieties through MAS. The present review deals with the possible regulatory mechanisms of Fe and Al tolerance and improvement of rice through identification of the tolerant lines with the help of efficient molecular markers en route biotechnological approaches.

 

Keywords: Biotechnological tools, Fe and Al toxicities, rice, stress tolerance mechanisms

IPC Code: Int. Cl. 7 A 01 H 4/00, 5/00; C 12 N 15/10

 

 

 

Indian Journal of Biotechnology

 Vol 3, July 2004, pp 369-377

 

Nasal melatonin gels using pluronic PF-127 for chronobiological treatment of sleep disorder

S S Pisal, P Reddy, A R Paradkar, K R Mahadik* and S S Kadam

 

Melatonin, a neurohormone, is formulated as a thermoreversible pluronic gel for nasal administration as an ‘overlap dosage form’ for chronobiological treatment of sleep disorders. Aqueous PF gels containing drug (0.5 mg & 1 mg/0.1 ml), PEG 400 and PEG 15,000 were prepared by cold method. Pluronic gels were evaluated for gelation and gel melting. Gelation temperature (T1) decreased with pluronic concentration while gel melting temperature (T2) increased. Melatonin shifted gelation range to higher temperature while PEG narrowed the gel range. Flux of diffusion decreased with PF concentration. Drug flux decreased in higher drug strength gels due to more partitioning in micellar phase. Pluronic gel (20%w/w, 1 mg/0.1 ml) showed bimodal pattern with a desired second peak flux (0.248mg/min/cm2) at 300 min. Flux pattern changed invariably with PEG. Bioadhesion time and strength to sheep nasal mucosa were more for gels containing melatonin and PEG 400. Nasal gels produced fast onset of action and induced sleep within fifteen minutes. The low intensity and rounded α-EEG wave pattern was observed for sleep duration of 5 hrs. Good correlation was observed in sleep pattern and low intensity α-EEG. The results are encouraging and nasal melatonin gels have potential in the treatment of circadian cycle sleep disorders.

 

Keywords: pluronic gels, melatonin, nasal, chronobiological release, sleep pattern

IPC Code: Int. Cl.7 A 61 K 38/00, 47/36

 

 

 

Indian Journal of Biotechnology

 Vol 3, July 2004, pp 378-381

 

Isolation and characterization of proteins involved in cryoprotection from cold resistant mutants of Pseudomonas fluorescens ATCC13525

Vandana Katiyar1, D P Mishra2, Satish Kumar3 and Reeta Goel1*

 

The proteins isolated from cold resistant mutants of Pseudomonas fluorescens ATCC13525 were biologically characterized. These proteins, designated as CRP-1 (41.6 kDa, from CRPF9) and CRP-2 (14.1 kDa, from CRPF8), were able to protect freeze-labile alkaline phosphatase (ALP) against freeze denaturation up to approximately 84% activity. Addition of BSA could not yield similar cryoprotection of ALP. However, 60% loss of enzymatic activity was recorded in absence of the proteins. Furthermore, the two proteins in combination protected the enzyme activity up to 86%. The freeze-thaw challenge given to the mutants and wild type cells revealed the mutant to be more resistant to freezing (-20°C). The survival percentage of the mutant cells after repeated freeze thaw was significantly higher (55.6%) than wild type (9.8%). Nevertheless, incubation at 10°C, prior to freezing (-20°C) resulted in a further increase in survival percentage of mutant cells (79.6%). The proteins isolated from these mutants possessed the cryoprotective ability, which could be correlated with increased freezing resistance of the mutants at low temperature.

 

Keywords: cold resistant mutants, cryoprotection, freeze-labile enzyme, freeze survival, P. fluorescens

IPC Code: Int. Cl.7 A 01 N 63/00

 

 

 

Indian Journal of Biotechnology

 Vol 3, July 2004, pp 382-387

 

Role of phosphorylase and amylase in accelerated ripening of mutant banana ‘Basrai-10Gy’

K K Surendranathan*, N K Ramaswamy and M B Pendharkar

 

‘Basrai-10Gy’, a mutant developed by γ- irradiation of shoot-cultures of the ‘Basrai’ banana, ripened much faster compared to its parent. The biochemical mechanism underlying the faster ripening of the mutant was investigated. Ripening was assessed by determining textural changes in the whole fruit, the peel and the pulp, as well as from the physical changes of ripening. The activities of amylase (E.C.3.2.1.3) and starch phosphorylase (E.C. 2.4.1.21), the enzymes that initiate the catabolism of starch, were comparatively evaluated in the parent and mutant variety. They were correlated with the ripening behaviour of the fruit. An increase in the amylase activity, associated with the climacteric and normal ripening behaviour, was observed in ‘Basrai’ bananas; while, the phosphorylase activity was comparatively insignificant. The mutant variety showed low amylase and very high phosphorylase activities, suggesting the direct availability of sugar phosphate instead of free sugars to accelerate the ripening of ‘Basrai-10Gy’ fruit.

 

Keywords: amylase, banana ripening, mutant, starch metabolism, starch phosphorylase

IPC Code:Int. Cl.7 C 12 N 15/01

 

 

 

Indian Journal of Biotechnology

 Vol 3, July 2004, pp 388-393

 

Characterization of cotton (Gossypium hirsutum) germplasm
by ISSR, RAPD markers and agronomic values

Ashok Dongre*, Vilas Parkhi and Santosh Gahukar

 

Using ISSR, RAPD markers and agronomic values, the characterization of 25 cotton (G. hirsutum) germplasms was carried out. 45 ISSR primers and 40 RAPD primers were used to amplify the germplasms. 19 scorable ISSR primers generated 90 markers while 21 reproducible RAPD primers generated 150 markers; of which, 49 markers from ISSR and 76 markers from RAPD were scored as polymorphic. Dendrograms were developed for ISSR and RAPD analysis by using NTSYS-pc software. Dendrogram of ISSR analysis showed 3 clusters while that of RAPD showed 4. Six agronomic values were also used for the characterization of germplasms. Based on these agronomic values, non-hierarchical Euclidean cluster analysis was performed to group the 25 cotton germplasms into 6 clusters. Significant similarities were found in the clustering of ISSR and RAPD analysis, however, correlations were not made between the clustering of ISSR, RAPD analysis and agronomic data analysis.

 

Keywords: ISSR, RAPD, agronomic values, cotton, germplasm

IPC Code: Int. Cl.7 C 12 N 15/10

 

 

 

Indian Journal of Biotechnology

 Vol 3 July 2004, pp 394-399

 

Kinetic studies of penicillin production during batch and repeated batch in fluidized bed bioreactor with agar immobilized P. chrysogenum cells

A Swaroopa Rani1, Annapurna Jetty1* and S V Ramakrishna2

 

The fermentation kinetics of penicillin production by Penicillium chrysogenum was carried out at 27°C and pH 6.0. Batch and repeated batch fermentations using agar-immobilized cells in fluidized bed bioreactor were studied for their potential application in production of penicillin from lactose, a fermentable sugar. Kinetics of immobilized cell fermentation, showed the penicillin yield of ~0.0155 g/l, with highest penicillin concentration of ~57.09 mg/l and the high reactor productivity of ~23.8 mg/l h-1. Repeated batch fermentation experiments showed that the immobilized biocatalysts could be recycled effectively for 5 cycles. Penicillin yield was 4-5-fold greater by this method of immobilization, with high productivity as compared to free cells and other immobilization methods.

 

Keywords: Penicillium chrysogenum, agar, penicillin, batch, repeated batch, fluidized bed reactor

IPC Code: Int. Cl.7 A 01 N 63/00; C 12 P 37/00

 

 

 

Indian Journal of Biotechnology

 Vol 3 July 2004, pp 400-408

 
Generation and characterization of pea (Pisum sativum) somaclones
for resistance to Ascochyta blight and powdery mildew

Rajan Sharma and R P Kaushal*

 

Somaclones of two pea cultivars, Palam Priya and Lincoln were generated and evaluated for resistance to Ascochyta blight and powdery mildew. Among the hormonal combinations tested for callus induction in MS medium, the best combination observed was 2,4-D + NAA + BAP 0.5 mg/l each using leaf explants. Frequency of callus differentiation was maximum (10 and 23.5% for Palam Priya and Lincoln, respectively) on MS medium containing NAA 0.5 mg/l + BAP 6 mg/l but shoots formed withered and did not root either. However, MS medium containing NAA 0.5 mg/l + BAP 5 mg/l gave good differentiation. In vitro flowering as well as seed formation was also observed in 11 shoots. Vigorous rooting was obtained when in vitro grown shoots having node at cut end were cultured on half strength MS medium containing NAA 2 mg/l. Five of 71 Lincoln somaclones and 1 of 21 Palam Priya somaclones showed resistance to Ascochyta pinodes but none were resistant to Erysiphe pisi. However, one somaclone (SP6-1) of Palam Priya possessing resistance to Ascochyta blight also showed moderate resistance to powdery mildew. Increased PAL as well as peroxidase activity was observed in somaclones showing resistance to Ascochyta blight. Variation in the banding pattern of esterase was also observed in the native-PAGE profiles of Palam Priya and Lincoln, and somaclones derived from them.

 

Keywords: Ascochyta pinodes, Erysiphe pisi, in vitro, isozymes, pea, Pisum sativum, somaclones

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00; C 12 N 15/01

 

 

 

Indian Journal of Biotechnology

 Vol 3, July 2004, pp 409-413

 

Banana streak virus from India and its detection by polymerase chain reaction

Anita K Cherian2, V K Baranwal1* V G Malathi1, R P Pant1 and Y S Ahlawat1

 

Banana streak virus (BSV) is an important pathogen of bananas and plantains (Musa spp.). The authors cloned and sequenced part of the genomes of two isolates of BSV from Kerala, BSV-K1 and BSV-K3. Both the clones contained a sequence covering a part of open reading frame III. The multiple sequence alignment of amino acids showed that both the isolates had a very high degree of identity with each other and clustered with Nigerian isolates of BSV (BSV-Onne) but not with Australian isolate of BSV (BSV-Mys). The relationship of these two isolates with other badnaviruses and Rice tungro bacilliform virus was also analysed. BSV was detected by PCR amplification in samples, which were symptomatic but negative in electron microscopy.

 

Keywords: Banana streak virus, banana, PCR detection, badnavirus

IPC Code: Int. Cl.7 C 12 N 15/10

 

 

 

Indian Journal of Biotechnology

Vol 3 July 2004, pp 414-417

 

Agrobacterium rhizogenes mediated hairy root induction in
two medicinally important members of family Solanaceae

Pankaj K Pawar and Vijay L Maheshwari*

 

Withania somnifera (L) Dunal. and Solanum surattense Burm f., the two medicinally important members of family Solanaceae, were investigated for induction of hairy roots using soil borne bacterium, Agrobacterium rhizogenes. Explants like stem segments, hypocotyls and leaves with midrib were infected with the bacterium in vitro. In both plants, extensive hairy roots were induced from leaf explant containing midrib within 15 days of infection. These roots were then established on MS basal medium and their growth was observed to be independent of exogenous supply of phytohormones. The growth rate of transformed roots was 10-fold as compared to control. These roots can be grown and established in phytohormone free liquid MS medium and used as a promising source of secondary metabolites of medicinal significance.

 

Keywords: Agrobacterium rhizogenes, Withania somnifera, Solanum surattense, withanoloids, solasodine, hairy roots

IPC Code: Int. Cl.7 A 01 H 5/06; C 12 N 15/03

 

 

 

Indian Journal of Biotechnology

 Vol 3 July 2004, pp 418-421

 

Silver nitrate induced rooting and flowering in vitro on
rare rhoeophytic woody medicinal plant, Rotula aquatica Lour.

M Chithra1, K P Martin2*, C Sunandakumari1 and P V Madhusoodanan1

 

Silver nitrate induced enhanced rooting and flowering in vitro was achieved on Rotula aquatica L., a rare rhoeophytic woody aromatic medicinal plant. Solid and liquid nature of the medium, and growth regulators significantly influenced in vitro rooting and flowering. Half-strength MS liquid medium fortified with 2.69 mM a-naphthaleneacetic acid (NAA) was effective among different auxins, which induced a mean of 7.3 roots per shoot. Plantlets acclimatized after auxin alone induced rooting exhibited 80% survival in field conditions. Nevertheless, the established plantlets exhibited 50% loss within 3 months. Addition of 11.7 mM silver nitrate to half-strength MS liquid medium containing 2.69 mM NAA increased the number of roots to 16.8 per shoot. Plantlets established after rooting in silver nitrate supplemented medium facilitated 95% survival, and the established plantlets did not exhibit loss even after 3 months. Dipping of the basal end of shoots developed in vitro in NAA (2.69 mM) and silver nitrate (11.7 mM) solution (made in water) for 25 days induced a mean of 12.4 roots per shoot. These plantlets transferred to small pots facilitated 90% survival. Half-strength MS liquid medium containing 2.69 mM NAA and 11.7 mM silver nitrate induced flowering earlier. The flowers were morphologically and structurally similar to that on plants growing in field conditions.

 

Keywords: Rotula aquatica, liquid medium, plantlet survival, solid medium

IPC Code: Int. Cl.7 A 01 H 4/00, 5/02, 5/06

 

 

 

Indian Journal of Biotechnology

 Vol 3, July 2004, pp 422-425

 

In vitro propagation of Pterocarpus marsupium Roxb.:
An endangered medicinal tree

Sharad Tiwari*, Pankaj Shah and Kanchan Singh

 

Nodal segments of Pterocarpus marsupium Roxb. were inoculated on seven different media compositions, viz. MS, B5 and White’s without growth hormones (MS00 , B500  and WH00), each supplemented with 3.0 mg l-1 BAP and 0.5 mg l-1 NAA (MSBN, B5BN, WHBN) and MS media supplemented with 0.2 mg l-1 IBA (MSIB). Seed germination improved in all the media studied, however, MS combinations were the best (95-100%). Maximum number of shoot induction per explant was in MS00 (3.25) followed by MSIB (2.26). Maximum nodes per shootlet were observed in medium MSIB (4.95), while shoot length was maximum in MSIB (2.92 cm) followed by MS00 (2.41). Regenerated plants were acclimatized and successfully transferred under field conditions.

 

Keywords: in vitro culture, nodal segments, plant regeneration, Pterocarpus marsupium

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

 

 

Indian Journal of Biotechnology

 Vol 3, July 2004, pp 426-430

 

In vitro clonal propagation of Momordica charantia L.

Mala Agarwal1* and Raka Kamal2

 

The present investigation outlines the in vitro propagation of Momordica charantia L. The explants from in vitro grown seedling were cultured on modified MS medium. Shoot differentiation was obtained on MS medium supplemented with BAP. Root, callus were formed on IBA and 2,4-D, respectively. Shoot as well as root differentiation was obtained on medium containing BAP+IBA/NAA. Multiple shoots with roots were formed on MS medium without hormones (MSO). Rooting on shoot grown occurred on medium containing IBA and 40% of the plants survived successfully, when transferred to the field.

 

Keywords: clonal propagation, Momordica charantia, in vitro regeneration, explant

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

 

 

Indian Journal of Biotechnology

 Vol 3, July 2004, pp 431-434

 

Selection of transformants of Escherichia coli containing cellulase gene from Ruminococcus albus isolated from rumen of crossbred steers

M Chandrasekharaiah*, A Thulasi and K T Sampath

 

Ruminococcus albus, considered best fibrolytic bacterium was isolated and characterized from the rumen of crossbred steers. It was found as wrinkled white colonies, slightly elevated with a slightly undulated margin, no surface spreading with the absence of liquefaction and a zone of hydrolysis. The cells were gram positive single cocci or diplococci. R. albus DNA was a high molecular weight DNA and it had just moved out of the well during electrophoresis. A genomic library of the Hind III fragments of R. albus DNA in pBR322 was constructed in Escherichia coli. Four clones were obtained with cellulase activity.

 

Keywords: Ruminococcus albus, cellulase genes, transformants, Escherichia coli, rumen

IPC Codes: Int. Cl.7 C 12 N 15/10, 15/11

 

 

 

Indian Journal of Biotechnology

 Vol 3, July 2004, pp 435-440

 

Reconstruction of in vitro matured goat oocytes by using
synchronized skin fibroblast cells as donor nuclei

S K Das* and A C Majumdar

 

Cloning of embryos by nuclear transplantation has been developed in several species using foetal fibroblasts as donor cells. In the present study, the developmental potential of foetal fibroblasts, arrested in metaphase stage using cytochalasin-B, was evaluated using nuclear transfer. Studies were undertaken to find out suitable stage of the cell cycle of goatskin fibroblast cells with enucleated in vitro matured goat oocytes and their capability of development of embryo. Skin cells were isolated from fetus as well as from adult skin and cultured to monolayer using RPMI-1640 media with 10% FCS in 5% CO2 incubator at 38.5º C ± 1º C with 95% humidity. Immature oocytes were matured in vitro for 22-24 hrs and enucleated using a Leitz micromanipulator. The cytochalasin-B blocked synchronized cells were used as donor cells for transferring into the perivitelline space of the enucleated oocyte and electrofused with constant AC pulse at 7-10 volts, while DC pulses at 180-400 volts were applied for 5-20 µ sec followed by culturing in CO2 incubator to observe proper electrofusion and cleavage. The embryos after fusion were activated with cytochalasin-B for 2-4 hrs and then transferred to IVC media. Foetal fibroblast became confluence within 2 days whereas adult fibroblast cells took 7-8 days to become confluence and, by 1-2 hr of cytochalasin-B treatment, around 85% foetal fibroblast cells arrested in the metaphase stage and 4.3% reconstructed embryos reached to 2-cell and morula stage in each category. Thus, 3-5 passaged foetal fibroblast cells synchronized by cytochalasin-B and around 300 volts would be better for electrofusion for reconstruction of goat oocytes.

 

Keywords: goat, reconstructed embryo, nuclear transfer and fibroblasts

IPC Code: Int. Cl.7 C 12 N 5/06, 15/06

 

 

 

Indian Journal of Biotechnology

Vol 3, July 2004, pp 441-444

 

Effects of genotype and culture medium on in vitro androgenesis in soybean (Glycine max Merr.)

Sharad Tiwari*, Prem Shanker and Manoj Tripathi

 

Anthers of ten genotypes of Glycine max were cultured on four fortified B5 media supplemented with different levels of growth hormones, viz. B5 DBIG (2.0 mg l-1 2,4-D+0.5 mg l-1 IBA+100.0 mg l-1 myo-Inositol+360.0 mg l-1 L-glutamine), B5 DB (2.0 mg l-1 2,4-D+0.5 mg l-1 BA), B5DK (2.0 mg l-1 2,4-D+0.5 mg l-1 Kinetin) and B5BKN (0.5 mg l-1 BA+0.5 mg l-1 Kinetin+1.0 mg l-1 NAA). All the media were supplemented with 90.0 g l-1 sucrose and 7.0 g l-1 agar. Significant differences in the response of genotypes, culture medium and genotype x medium interactions were observed for callus initiation, formation of morphogenic calli and plantlet regeneration. Genotype JS 90-41was found superior for in vitro androgenesis. Culture medium B5DBIG exhibited higher response for androgenic callus formation and haploid plant regeneration.

 

Keywords: Glycine max, anther culture, androgenesis, in vitro, genotype x medium interactions, haploid plants

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

 

 

Indian Journal of Biotechnology

 Vol 3, July 2004, pp 445-448

 

Establishment of an embryogenic suspension culture of
Pinus kesiya (Khasi pine) from various explants

Chitta Ranjan Deb1* and Pramod Tandon2

 

The embryogenic cultures were obtained from mature zygotic embryos (79.6%) and apical dome sections (92.6%) (pretreated with 0.4% activated charcoal at 4oC for 24 hrs) on mMS and ½DCR media, respectively containing 5 mg l-1 2,4-D, NAA in combination along with 1 mg l-1 BAP, while, from secondary needles (88.6%) it was on MS medium containing 3 mg l-1 each of 2,4-D and NAA in conjunction with 1 mg l-1 BAP. Using 150-200 mg embryogenic tissues per 15 ml medium at 120 rpm raised embryogenic suspension cultures. But the embryogenic cultures from apical dome sections were cold treated at 4oC for 24 hrs before transferring to suspension culture. The cultures were subcultured at 6-7 days interval and were diluted at 1:4 ratio. The embryonal suspensor masses developed from the resulting elongated single cells within 3-4 passages in respective basal medium with 1/10th growth regulators of initiation medium. The proembryonal head formed in growth regulators free medium at 100 rpm. The embryonal head formed in basal medium containing 4% sucrose and 4 mg l -1 ABA in combination.

 

Keywords: activated charcoal pre-treatment, cold-treatment, embryogenic suspension culture, pinus kesiya, somatic embryogenesis

IPC Code:Int. Cl.7 A 01 H 4/00, 7/00

 

 

 

Indian Journal of Biotechnology

 Vol 3, July 2004, pp 449-451

 

Direct organogenesis in Datura metel L. from in vitro and in vivo nodal explants

B Muthukumar1*, D I Arockiasamy2 and E Natarajan3

 

In vitro plant regeneration was achieved in Datura metel from nodal explants collected from both in vitro germinated seedlings and field grown plants (in vivo). The explants were cultured on MS medium with BAP (0.5-3.0 mg/l) and NAA (0.5 mg/l). The nodal explants isolated from in vivo source exhibited a greater number of healthy multiple shoots than in vitro. BAP at 3 mg/l with NAA at 0.5 mg/l was found to be optimal for regeneration of shootlets.

 

Keywords: Datura metel, medicinal, in vitro regeneration, nodal explants, BAP, organogenesis

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

 

 

Indian Journal of Biotechnology

 Vol 3, July 2004, pp 452-453

 

Production of camptothecines from callus cultures of Nothapodytes foetida (Wight) Sleumer

R Sundravelan, B Desireddy and Veeresham Ciddi*

 

Murashige and Skoog (MS) medium supplemented with picloram (2 mg/l), N6- benzyladenine(BA) (1 mg/l) and gibberellic acid (GA3) (1 mg/l) was found to be suitable for the establishment of callus cultures from leaves of Nothapodytes foetida (Wight) Sleumer. The callus upon extraction and analysis revealed the presence of a cytotoxic quinoline alkaloid, camptothecine (CPT) (2.893±2.38 mg %) and 9-methoxy camptothecine (MCPT) (0.4±0.4 mg %).

 

Keywords: Nothapodytes foetida, camptothecine, 9-methoxy camptothecine

IPC Code:Int. Cl.7 A 01 H 4/00; A 01 N 63/00

 

 

 

Indian Journal of Biotechnology

 Vol 3, July 2004, pp 454-456

 

Plant regeneration from callus cultures in Suaeda nudiflora (Wild.) Moq.

Aneesha Singh, J Chikara* and J B Pandya

 

Suaeda nudiflora plants, showing relatively high salt tolerance, were used for micropropagation and generation  of plantlets from callus cultures. Induction of somaclonal variability in the existing germplasm was another aim. Callus was initiated from young stem on MS medium supplemented with different concentrations of BAP and 2,4-D. Organogenesis was achieved by transferring callus on MS medium supplemented with BAP, KN and adenine. Well-grown shoots rooted easily on MS-half supplemented with IBA, IPA and NAA and 80-90% rooting was achieved. Plantlets got hardened by keeping them in hardening unit for few days. Hardened plants established very well in the nursery. Though originated from callus, these plants did not show any morphological variations and were similar to their parent donor plant.

 

Keywords: Suaeda nudiflora, callus culture, organogenesis, benzylaminopurine, root primordial, hardened plantlets

IPC Code: Int. Cl.7 A 01 H 4/00, 5/00

 

 

 

Indian Journal of Biotechnology

 Vol 3, July 2004, pp 457-459

 

Genetic characterisation of Jaisalmeri camel using microsatellite markers

L Gautam2, S C Mehta1*, R S Gahlot1 and K Gautam1

 

Six New World Camelidae microsatellite primer pairs were used to investigate the genetic polymorphism in Jaisalmeri camel. Polymerase chain reactions were carried out for 30 unrelated camels of Jaisalmeri breed. The amplification products were resolved in 6% (denaturing) urea PAGE and stained with silver nitrate. All the 6 microsatellite primer pairs were found polymorphic in Jaisalmeri camel. The number of alleles ranged from 2 to 5. The expected heterozygosity ranged from 0.32 to 0.651 and the polymorphic information content ranged from 0.268 to 0.588. The results indicated the utility of these microsatellite loci for studying genetic polymorphism in dromedary breeds.

 

Keywords: breeds, genetic characterisation, Jaisalmeri camel, microsatellite

IPC Code: Int. Cl.7 C 12 N 15/10

 

 

   

EXPLANATORY NOTE ON IPC Int. Cl.7

 

The field of biotechnology has seen exponential growth in recent times. It attracted the attention of R&D institutes, who have made significant achievements/breakthroughs in the fields of agriculture, health and medicine, environment and so on, and consequently resulted in granting of hundreds of patents by patent authorities the world over. It has also thus aroused concerns in the above areas on ethical issues as well as profit making/sharing. In this context, patents, their classification, access to patent databases, prior art, etc. have assumed great significance. In biotechnology, these issues as such are more crucial since it is completely a field of applied nature.

 

Indian Journal of Biotechnology has been assigning IPC codes to each article for creating more awareness for its readers and facilitating the search of patent examiners as well as establishing prior art. The subject matter is briefly explained below.

 

The International Patent Classification, which is commonly referred to as the IPC, is based on an international multi-lateral treaty administered by World Intellectual Property Organization, Geneva, Switzerland.

 

The Classification is indispensable for the retrieval of patent documents in the search for "prior art." Such retrieval is needed by patent-issuing authorities, potential inventors, research and development units, and others concerned with the application or development of technology.

 

In order to keep the IPC up to date, it is continuously revised and a new edition is published every five years. The current (seventh) edition has entered into force on January 1, 2000.

The IPC is a hierarchical classification system comprising sections, classes, subclasses and groups (main groups and subgroups). The seventh edition of the IPC consists of 8 sections, 120 classes, 628 subclasses and approximately 69,000 groups.

 

Every subdivision of the IPC is indicated by a symbol and has a title. The IPC divides all technological fields into eight sections designated by one of the capital letters A through H. Each section is subdivided into classes. In turn, each class contains one or several subclasses, for example, A 01 B.

 

Each subclass is broken down into subdivisions referred to as "groups", which are either main groups or subgroups. Main group symbols consist of the subclass symbol followed by a one-to three-digit number, the oblique stroke and the number 00, for example, A 01 B 1/00. Subgroups form subdivisions under the main groups. Each subgroup symbol includes the subclass symbol followed by the one-to three-digit number of its main group, the oblique stroke and a number of at least two digits other than 00, for example, A 01 B 1/02.

For further information, please refer to: www.wipo.int

 

 

 

NISCAIR Policy on Plagiarism

 

The system of formal communication in science through publication in primary journals is based on originality and quality of information, being the only criteria for publication. However, there have been tendencies to misuse the system and vitiate the process of science communication for personal benefits. One of the ills, afflicting science communication, is plagiarism. Attempts at plagiarism may range from verbatim copying of extensive material of other authors, misappropriating results/data of others with minor changes in language/presentation without giving credit to original source, and to publishing essentially the same information more than once.

 

As the premier institution of publishing primary scientific journals in various disciplines of science and technology in India, NISCAIR strongly reiterates its policy of discouraging plagiarism of all kinds. All efforts are made to detect and frustrate attempts at plagiarism through editorial screening and rigorous peer review in respect of communications received for publication in the NISCAIR publications. Cooperation of the scientific community is sought in our efforts to frustrate all attempts at plagiarism.

 

In case, an attempt of plagiarism is brought to our attention accompanied with convincing evidence, following steps would be taken:

(a) After consulting the respective Editorial Board Members, authors guilty of plagiarism will be debarred from publishing their papers in NISCAIR journals.

(b) Heads of the Departments/Institutes of the offending authors will be intimated of such incidences of plagiarism.

(c) Such incidents of plagiarisms will be publicized through the concerned NISCAIR journals in consultation with the respective Editorial Board Members.

 

 

 

ERRATA

 

CONTENTS (on page 156):

Title, ‘Transgenic pants as bioreactors’ be read as ‘Transgenic plants as bioreactors’

Header (on page 203):

Vol 2 be read as Vol 3

Article Title (on page 241):

‘Cloning of adult trrees---’ be read as ‘Cloning of adult trees---’

 

 

 

AUTHOR INDEX

 

Agarwal M

426

Kadam S S

369

Pisal S S

369

Arockiasamy D I

449

Kamal R

426

 

 

Ahlawat Y S

409

Kaushal R P

400

Ramakrishna S V

394

 

 

Katiyar V

378

Ramaswamy N K

382

Baranwal V K

409

Kumar S

378

Reddy P

369

Basu A K

359

 

 

Roy B

359

 

 

Madhusoodanan P V

418

Roy T

359

Chandrasekharaiah M

431

Mahadik K R

369

 

 

Chaudhury R

348

Maheshwari V L

414

Sampath K T

431

Cherian A K

409

Majumdar A C

435

Shah P

422

Chikara J

454

Malathi V G

409

Shankar P

441

Chithra M

418

Malik S K

348

Sharma R

400

Ciddi V

452

Mandal A B

359

Sheeja T E

359

 

 

Martin K P

418

Singh A

454

Das S K

435

Mehta S C

457

Singh K

422

Deb C R

445

Mishra D P

378

Sunandakumari C

418

Desireddy B

452

Muthukumar B

449

Sundravelan R

452

Dongre A

388

 

 

Surendranathan K K

382

 

 

Natarajan E

449

Swaroopa Rani A

394

Gahlot R S

457

 

 

 

 

Gahukar S

388

Pandya J B

454

Tandon P

445

Gautam K

457

Pant R P

409

Tiwari S

422, 441

Gautam L

457

Paradkar A R

369

Thulasi A

431

Goel R

378

Parkhi V

388

Tripathi M

441

 

 

Pawar P K

414

Trivedi S

331

Jetty A

394

Pendharkar M B

382