Indian Journal of Biotechnology

 

Total visitors:3,006 since 16-10-06

VOLUME 5

CODEN:IJBNAR 5(4) 427-582 (2006)

NUMBER 4

OCTOBER 2006

ISSN:0972-5849

 

CONTENTS

 

Reviews

 

 

Single nucleotide polymorphism (SNP)ľMethods and applications in plant genetics:
A review

435

 

        IPC Code: Int. Cl.8 C12N15/00

 

 

        Tabassum Jehan & Suman Lakhanpaul

 

 

 

 

 

Double stranded RNAs as gene silencing tool: An overview

460

 

        IPC Code: Int. Cl.8 A61K31/713, 48/00

 

 

        Kapil Khatri, Amit Rawat, Sunil Mahor, Prem N Gupta & Suresh P Vyas

 

 

 

 

 

Antibiotics business: A glimpse

471

 

IPC Code: Int. Cl.8 A61K31/00

 

 

B K Bhattacharyya & S K Sen

 

 

 

 

 

Papers

 

 

 

 

 

A simple method for extraction of high molecular weight genomic DNA from buccal cells in mouthwash

477

 

IPC Code: Int. Cl.8 C12N15/10

 

 

Priya Koppikar & Rita Mulherkar

 

 

 

 

 

Twinning in Marwari and Bharat Merino ewes and its relationship with Booroola FecB mutation

482

 

IPC Code: Int. Cl.8 C12N15/01

 

 

S Kumar, Atul P Kolte & V K Singh

 

 

 

 

 

Ovine interferon-γ gene of indigenous sheep: Cloning, sequencing and expression studies in Escherichia coli

486

 

        IPC Code: Int. Cl.8 C12N15/09, 15/21

 

 

T J Rasool, M Hosamani, A Premraj, E Sreekumar & R K Singh

 

 

       

 

 

Vacuum foam drying for preservation of lasota virus: Screening of foaming agent and cycle optimization

491

 

IPC Code: Int. Cl8 A23B7/024; A61K39/155

 

 

S S Pisal, G M Wawde, M M Bandivadekar, A A Hajare, H N More & S S Kadam

 

 

 

 

 

Experimental studies and mathematical modeling of a semibatch bio-digester using municipal market waste as feed stock

498

 

IPC Code: Int. Cl.8 C02F3/20

 

 

        J Biswas, R Chowdhury & P Bhattacharya

 

 

Uptake of Cr(VI) in the presence of sulphate by Bacillus mycoides in aerobic environment

506

 

IPC Code: Int. Cl.8 C02F11/02; C12R1/07

 

 

        Asha Lata Singh, P Murali Krishna, T Nageswara Rao & P N Sarma

 

 

 

 

 

Detection of transgenes in genetically modified soybean and maize using polymerase chain reaction

510

 

IPC Code: Int. Cl.8 C12N15/10

 

 

        Gurinder Jit Randhawa & P K Firke

 

 

       

 

 

Isolation and characterization of L-asparaginase from marine actinomycetes

514

 

IPC Code: Int. Cl.8 C12N9/00; C12R1/01

 

 

        P Dhevagi & E Poorani

 

 

       

 

 

Desiccation and ABA treatment improves conversion of somatic embryos to plantlets in banana (Musa spp.) cv. Rasthali (AAB)

521

 

IPC Code: Int. Cl.8 A01H4/00

 

 

        L Srinivas, T R Ganapathi¸ P Suprasanna & V A Bapat

 

 

 

 

 

Assessment of variability in the regenerants from long-term cultures of ‘safed musli’ (Chlorophytum borivilianum)

527

 

IPC Code: Int. Cl.8 A01H4/00; C12N15/10

 

 

        Dilip K Arora, Sarabjeet S Suri & K G Ramawat

 

 

       

 

 

In vitro corm induction and genetic stability of regenerated plants in taro [Colocasia esculenta (L.) Schott]

535

 

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

 

 

Z Hussain & R K Tyagi

 

 

 

 

 

In vitro cloning of apple (Malus domestica Borkh) employing forced shoot tip cultures of M9 rootstock

543

  

IPC Code: Int. Cl.8 A01H4/00

 

 

        M Amin Dalal, B Das, A K Sharma, M Amin Mir & Amarjeet Singh Sounduri

 

 

 

 

 

Micropropagation of Embelia ribes Burm.f. using inflorescence segments

551

 

IPC Code: Int. Cl.8 A01H4/00

         

 

        K Shankarmurthy & V Krishna

 

 

 

 

 

Micropropagation of Capparis spinosa L. subsp. rupestris Sibth. & Sm. by nodal cuttings

555

 

IPC Code: Int. Cl.8 A01H4/00

 

 

        L Chalak & A Elbitar

 

 

 

 

 

Short Communications

 

 

 

 

 

Molecular analysis of bacterial population inhabiting coconut husk retting area

559  

 

IPC Code: Int. Cl.8 C12N15/10

 

 

Sabu Thomas, K Harikrishnan & Sandhya C Nair

 

 

 

 

 

In vitro multiplication of Asteracantha longifolia (L.) Nees–A medicinal herb

562 

 

        IPC Code: Int. Cl.8 A01H4/00

 

 

        J Panigrahi, R R Mishra & M Behera

 

 

In vitro propagation of Ceropegia bulbosa using nodal segments

565

 

        IPC Code: Int. Cl.8 A01H4/00

 

 

Divya Goyal & Seema Bhadauria

    

 

 

 

 

List of Referees

569

 

 

 

 

Annual Author Index

573

 

 

 

 

Annual Subject Index

575

 

 

 

 

Annual IPC Code Index

578

 

 

 

 

Instructions to Contributors

579

 

 

 

 


 

 

AUTHOR INDEX

 

 


Arora D K

527

 

 

Bandivadekar M M

491

Bapat V A

521

Behera M

562

Bhadauria S

565

Bhattacharya P

498

Bhattacharyya B K

471

Biswas J

498

 

 

Chalak L

555

Chowdhury R

498

 

 

Dalal M A

543

Das B

543

Dhevagi P

514

 

 

Elbitar A

555

 

 

Firke P K

510

 

 

Ganapathi T R

521

Goyal D 

565

Gupta P N

460

 

 

Hajare A A

491

Harikrishnan K

559

 


Hosamani M

486

Hussain Z

535

 

 

Jehan T

435

 

 

Kadam S S

491

Khatri K

460

Kolte A P

482

Koppikar P

477

Krishna P M

506

Krishna V

551

Kumar S

482

 

 

Lakhanpaul S

435

 

 

Mahor S

460

Mir M A

543

Mishra R R

562

More H N

491

Mulherkar R

477

 

 

Nair S C

559

 

 

Panigrahi J

562

Pisal S S

491

Poorani E

514

 


Premraj A

486

 

 

 

 

Ramawat K G

527

 

Randhawa G J

510

 

Rao T N

506

 

Rasool T J

486

 

Rawat A

460

 

 

 

 

Sarma P N

506

 

Sen S K

471

 

Shankarmurthy K

551

 

Sharma A K

543

 

Singh A L

506

 

Singh R K

486

 

Singh V K

482

 

Sounduri A S

543

Sreekumar E

486

Srinivas L

521

Suprasanna P

521

Suri S S

527

 

 

Thomas S

559

Tyagi R K

535

 

 

Vyas S P

460

 

 

Wawde G M

491

 


 


Indian Journal of Biotechnology
Vol 5 October 2006, pp 435-459

 

Single nucleotide polymorphism (SNP)–Methods and applications in
plant genetics: A review

 

Tabassum Jehan and Suman Lakhanpaul*

 

An array of genetic markers viz. morphological, biochemical and DNA based has been used in various fields including plant genetics and crop improvement. A novel class of DNA markers namely single nucleotide polymorphisms (SNPs) has recently become highly preferred in genomic studies. They are single nucleotide base polymorphism in genomic DNA and are the most abundant class of markers. In recent times, various SNP databases have been constructed to assess the SNP data available in humans, animals and plants. Seeing the huge potential of SNPs in pharmacogenomics and crop genetics, various assays for their genotyping have been described, which include direct sequencing, mining from EST databases, cleavage assays, molecular beacons, etc. Each of these assays are having their own merits and demerits and the choice of assay depends on the objective of the study and availability of the resources. These assays utilize detection platforms that range from the conventional gel based detection to high throughput systems like microarrays, mass spectrometry and flow cytometry. SNPs have tremendous applications and prospects in crop genetics. They can be used in association studies, tagging of economic important genes, genotyping, diversity analysis and evaluation among the plant species.

Keywords: single nucleotide polymorphism (SNP), genotyping assay, microarrays, application in plants, linkage disequilibrium (LD), diversity analysis

IPC Code: Int. Cl.8 C12N15/00

Indian Journal of Biotechnology
Vol 5, October 2006, pp 460-470

 

Double stranded RNAs as gene silencing tool: An overview

Kapil Khatri, Amit Rawat, Sunil Mahor, Prem N Gupta and Suresh P Vyas*

 

Gene silencing has evolved in a broad range of organisms probably as defense mechanism against invasive nucleic acids. RNA interference (RNAi) is a process of sequence specific post-transcriptional gene silencing that has been triggered by double stranded RNA ds(RNA) in a wide variety of organisms. Two major strategies are utilized. Transcriptional gene silencing (TGS) acts to prevent RNA synthesis and post-transcriptional gene silencing (PTGS) acts to degrade existing RNA. Introducing transgenes, RNA viruses, or DNA sequences that are homologous to expressed genes can activate silencing. RNA silencing is a potent means to counteract foreign sequences and its use had made it possible to ascribe certain biological functions to sequenced genes. It could play an important role in the development of functional genomics. This review elaborates the current progress on the understanding of the molecular basis of RNA silencing and further possibilities to use this as a technique to develop novel therapeutic molecules.

Keywords: gene silencing, TGS, PTGS, RNAi, ds RNA

IPC Code: Int. Cl.8 A61K31/713, 48/00

 

Indian Journal of Biotechnology
Vol 5, October 2006, pp 471-476

 

Antibiotics business: A glimpse

B K Bhattacharyya* and S K Sen

 

Early history of medicine is largely supported by the secondary metabolites from microorganisms. Before the introduction of antibiotics, between 1940s and 1950s, patients with bacteraemia suffered from low survival chances, and mortality from tuberculosis was well above 50%. The emergence and spread of antibiotic resistant pathogens has increased substantially over the past two decades. At the same time, the development of new antibiotics has decreased alarmingly, because pharmaceutical companies are pulling out of antibiotic research. Reasons include, the changes in regulations of elaborate drug trials, clinical preference for narrow spectrum compounds, prolonged post marketing surveillance, hence the cost of development.

Keywords: Streptomyces, antibiotic, pharmaceutical industry, business, broad spectrum

IPC Code: Int. Cl.8 A61K31/00

 

Indian Journal of Biotechnology
Vol 5, October 2006, pp 477-481

 

A simple method for extraction of high molecular weight genomic DNA from buccal cells in mouthwash

Priya Koppikar and Rita Mulherkar*

 

With tremendous advances made in the field of molecular biology techniques, the analysis of genomic DNA has become a matter of routine. However, there is a growing need for better, cost-effective, non-invasive methods for sample collection and DNA extraction. A simple protocol for sample collection and extraction of high molecular weight genomic DNA from buccal cells in mouthwash has been standardized. The method does not require elaborate instrumentation and can be applied to large population based studies. It has been observed that good quality high molecular weight genomic DNA can be obtained from exfoliated buccal cells in the early morning mouthwash samples, and that the DNA yield from similar samples decreases during the day, with very low yields obtained in the late evening. The amount of DNA obtained is enough for approximately 300 PCR amplifications using 100 ng DNA as template. DNA so obtained was used successfully for numerous amplifications by PCR with human gene specific primers and can be used for Restriction Fragment Length Polymorphism (RFLP) studies as well as for other molecular biology applications.

Keywords: mouthwash samples, genomic DNA extraction, PCR

IPC Code: Int. Cl.8 C12N15/10

Indian Journal of Biotechnology
Vol 5, October 2006, pp 482-485

 

Twinning in Marwari and Bharat Merino ewes and its relationship with Booroola FecB mutation

S Kumar*, Atul P Kolte and V K Singh

 

The Booroola fecundity (FecB) is a single autosomal gene, which increases ovulation rate and litter size in sheep. High prolificacy in Garole and Booroola Merino sheep is due to a mutation in FecB gene, and by its virtue they produce twin or more lambs. The FecB genotyping was carried out by Forced RFLP-PCR technique in prolific Garole, non-prolific Marwari and Bharat Merino ewes with the aim to confirm linkage of twinning with FecB mutation. Garole ewes were selected which had at least one record of twins, triplets or more lambs. Marwari and Bharat Merino ewes were selected into two groups, one group had at least one record of twin and other group had only single lamb. Results showed that FecB mutation was associated with twinning/high litter size in Garole ewes, while all Marwari and Bharat Merino ewes were found non-carriers (FecB++) for FecB mutation. The present study indicates that twinning in Marwari and Bharat Merino ewes is not linked with FecB mutation, and it may be due to various other extrinsic reasons, including environmental factors or other unknown genes.

Keywords: FecB mutation, sheep, twinning

IPC Code: Int. Cl.8 C12N15/01

Indian Journal of Biotechnology
Vol 5, October 2006, pp 486-490

 

Ovine interferon-γ gene of indigenous sheep: Cloning, sequencing
and expression studies in Escherichia coli

T J Rasool*, M Hosamani, A Premraj, E Sreekumar and R K Singh

 

Ovine interferon-gamma (IFN-γ) cDNA was isolated from total mRNA of peripheral blood mononuclear cells from indigenous sheep by reverse transcription PCR. The cDNA was successfully cloned, sequenced and expressed in Escherichia coli as thioredoxin fusion protein. Sequence comparison of Indian sheep IFN-γ showed high nucleotide homology with published IFN-γ sequences of sheep and other species of ruminants. The cDNA codes for a mature polypeptide with a putative molecular weight of 17 kDa. The recombinant ovine IFN-γ, expressed in BL21 E. coli cells as His-tagged protein, was purified using Ni-chelation chromatography. Up to 250 µg of recombinant ovine IFN-g could be obtained from 50 mL of bacterial culture. The production of this important cytokine has significant implications in therapy of major infectious diseases of small ruminants. It also has potential applications as adjuvant to enhance the protective efficacy of the existing vaccines.

Keywords: cDNA, cytokine, expression, interferon gamma, sheep

IPC Code: Int. Cl.8 C12N15/09, 15/21

 

 

Indian Journal of Biotechnology
Vol 5, October 2006, pp 491-497

 

Vacuum foam drying for preservation of lasota virus: Screening of foaming agent and cycle optimization

S S Pisal*, G M Wawde, M M Bandivadekar, A A Hajare, H N More and S S Kadam

 

Solid-state product of lasota virus was obtained by using vacuum foam drying (VFD). In the process, pluronic polymers, viz. F-108, F-68 and F-87 (plain and with sugars) were screened for their foaming ability. First, aqueous infrared (IR) spectrum of lasota virus harvest was obtained and pH and temperature stability of the harvest were evaluated. Then the temperature and vacuum of VFD cycle were optimized. The processed virus products were also evaluated for foam characteristics, moisture content and EID50 titre value. Aqueous solution of plain pluronic F-108 at 3.0% (w/v) strength showed the highest foaming ability as compared to other pluronics used in the study. The foaming ability of F-108 increased further in the presence of sugars. However, optimum foaming ability of pluronic F-108 during VFD was observed at 0.6% (w/v). Aqueous IR spectrum reveals symmetrical α-helix structure of lasota virus. The virus remained stable at pH 3 to 9, but denatured at temperatures above 20şC. Effective and uniform foam formation with low residual moisture and a virus titre of 6.24 was obtained in VFD cycle III; whereas, Lasota virus was completely denatured at lower vacuum with higher temperatures. Thus, the optimized cycle III is suitable for screening of protectants for dehydration-induced denaturation of lasota virus during VFD.

Keywords: cycle optimization, foamability, lasota, pluronic, vacuum foam drying

IPC Code: Int. Cl.8 A23B7/024; A61K39/155

 

Indian Journal of Biotechnology
Vol 5, October 2006, pp 498-505

 

Experimental studies and mathematical modeling of a semibatch bio-digester using municipal market waste as feed stock

J Biswas, R Chowdhury* and P Bhattacharya

 

An anaerobic digester of 10 dm3 capacity has been operated in batch mode at an optimum temperature of 40oC and pH 6.8 using vegetable market-waste as the feed material. The effect of slurry concentration and that of the concentration of carbohydrate, protein and fat in the slurry on the biogas production rate and methane concentration in the biogas have been studied. A maximum concentration of methane in the order of 80% has been obtained. The liquid medium of a running digester based on vegetable market waste has been used as seed culture. Both acidogenic and methanogenic bacterial strains have been isolated from the broth. Growth kinetics of acidogenic bacteria have been determined using carbohydrate, fat and protein individually as limiting substrates. Kinetics of exponential growth of methanogenic bacteria have been determined using different volatile fatty acids and long chain fatty acids as substrates. In all the cases, Monod type model has been observed to fit well with the experimental growth patterns. The performance of the digester has been studied using slurry concentration and concentration of carbohydrate, protein and fat in the slurry as parameters. A deterministic mathematical model has been developed using the suitable growth parameters of isolated bacterial strains to explain the transient behaviour of the digester. The model predictions have been compared successfully with the experimental findings.

Keywords: anaerobic digestion, biogas, concentration of slurry, mathematical modeling, municipal market waste

IPC Code: Int. Cl.8 C02F3/20

 

Indian Journal of Biotechnology
Vol 5, October 2006, pp 506-509

 

Uptake of Cr(VI) in the presence of sulphate by Bacillus mycoides in
aerobic environment

Asha Lata Singh, P Murali Krishna, T Nageswara Rao and P N Sarma*

 

Studies on the uptake of Cr(VI) were carried out using the bacterium Bacillus mycoides, isolated from industrial wastewater in the presence of sulphate. Cr(VI) uptake by B. mycoides was found to be concentration dependent. The uptake of Cr(VI) was maximum at pH 7. The study suggested that sulphate acted as a non-competitive inhibitor of Cr(VI) and it was transported inside the cell through the cell membrane by some different sites rather than sulphate uptake transport site of the B. mycoides system.

Keywords: Bacillus mycoides, Cr(VI) uptake, non- competitive inhibition, sulphate uptake

IPC Code: Int. Cl.8 C02F11/02; C12R1/07

 

Indian Journal of Biotechnology
Vol 5, October 2006, pp 510-513

 

Detection of transgenes in genetically modified soybean and maize using polymerase chain reaction

Gurinder Jit Randhawa* and P K Firke

 

Polymerase chain reaction (PCR) has been successfully employed to detect transgenes in genetically modified (GM) soybean and maize. Three pairs of primers for CaMV 35S, NOS and EPSPS genes were used in the detection. The results reveal that PCR could differentiate the GM plants from non-GM plants. Evidently, PCR is a highly reproducible and sensitive technique that can be successfully employed in detecting transgenes for screening GM soybeans and GM maize from their conventional plants.

Keywords: genetically modified soybean and maize, PCR, transgenes

IPC Code: Int. Cl.8 C12N15/10

 

 

Indian Journal of Biotechnology
Vol 5, October 2006, pp 514-520

 

Isolation and characterization of l-asparaginase from marine actinomycetes

P Dhevagi and E Poorani*

 

The marine environment is a potential source of novel actinomycetes, which are a potent source of antibiotics and novel bioactive compounds. Marine actinomycetes were enumerated in sediment samples from Parangipettai and Cochin coastal areas of South India (PDK7 and PDK2). The isolates showed potential L-asparaginase activity. The partially purified L-asparaginase showed a specific activity of 64.07 IU/mg protein, 83-fold pure and yielded 2.18 per cent of protein. The enzyme activity was maximum between pH 8 and 9 and maintained stability at pH 8. The L-asparaginase showed maximum activity at 60°C, and stable up to 80°C. The enzyme has a molecular weight of 140 kDa. It showed cytostatic effect on JURKAT cells (Acute T cell leukemia) and K562 cells (Chronic myelogenous leukemia) at 24 h and cytotoxic effects at 48 h.

Keywords: marine actinomycetes, therapeutic L-asparaginase, anti-lymphoma activity

IPC Code: Int. Cl.8 C12N9/00; C12R1/01

Indian Journal of Biotechnology
Vol 5, October 2006, pp 521-526

 

Desiccation and ABA treatment improves conversion of somatic embryos to plantlets in banana (Musa spp.) cv. Rasthali (AAB)

L Srinivas, T R Ganapathi¸ P Suprasanna and V A Bapat*

 

Somatic embryos of banana cv. Rasthali were subjected to 0-8 h desiccation and abscisic acid (ABA) treatment for improving plant conversion. Somatic embryos exposed to 10 mM ABA followed by 2 h desiccation exhibited an increase in plant conversion (66%) compared to control (56%). Embryos treated with ABA followed by 2 h desiccation survived up to 5 weeks with 56% plant conversion upon storage at 10°C. This study suggests that combined treatment of banana somatic embryos with ABA and desiccation can improve conversion frequency.

Keywords: banana, somatic embryos, abscisic acid, desiccation

IPC Code: Int. Cl.8 A01H4/00

Indian Journal of Biotechnology
Vol 5, October 2006, pp 527-534

 

Assessment of variability in the regenerants from long-term cultures of
‘safed musli’ (Chlorophytum borivilianum)

Dilip K Arora, Sarabjeet S Suri and K G Ramawat*

 

Somatic embryogenesis in long-term calluses of seedling and leaf explants, obtained from in vitro grown plants of Chlorophytum borivilianum—an endangered medicinal herb, has been achieved. Large number of plants were established in the field and evaluated for variability among the regenerants. The plantlets obtained through seedling-derived embryonic callus showed high level of morphological and cytological variations, which increased with the increase in age of cultures. Occasionally, variegated plants were also observed. Variation in leaf size, stomatal number, epidermal cell size and chromosomal number was observed in regenerants. Chromosomal variation was the least (3X-3 to 3X+3) in regenerants from 1 to 4-month-old cultures and increased (5X-1 to 7X) with the age in regenerants from cultures older than 6 months. Alternatively, regenerated plants from somatic embryos recurrently obtained from leaf explants showed very little variation (0.62%) in RAPD fingerprinting. However, further improvement in somatic embryogenesis is required for domestication of the plant using biotechnological method of propagation.

Keywords: Chlorohphytum borivilianum, RAPD, somaclonal variation, somatic embryos

IPC Code: Int. Cl.8 A01H4/00; C12N15/10

Indian Journal of Biotechnology
Vol 5 October 2006, pp 535-542

 

In vitro corm induction and genetic stability of regenerated plants in taro [Colocasia esculenta (L.) Schott]

Z Hussain and R K Tyagi*

 

In vitro corm formation was achieved on Murashige and Skoog’s (MS) medium, containing 8-10% sucrose, 22 µM N6-benzyl aminopurine (BAP), 0.6 µM α-naphthaleneacetic acid (NAA) and 0.8% agar, in taro (IC 420791). The corm forming cultures could be conserved up to 15 months at 25+2oC, whereas shoot-forming cultures could last for only 6 months on MS medium, containing 3% sucrose + 2.2 µM BAP + 0.6 µM NAA + 0.8% agar. Plantlets with in vitro-formed corms showed 100% survival in the field, and developed normal uniform corm-producing plants. Uniformity of tissue culture regenerated plants with corm (Ro) was determined on the basis of 12 qualitative and 10 quantitative morphological traits related to leaf, petiole, corm and root. Additionally, two PCR-based markers—RAPD and ISSR were also applied to determine the genetic stability of Ro plants. A total of 13 RAPD primers (of 35 tested initially) and 6 ISSR primers gave 111 distinct bands in RAPD and 43 in ISSR, and exhibited uniform RAPD and ISSR banding patterns for Ro plants tested. Our results suggested that present protocol used for in vitro corm formation may cost-effectively be applied for conservation of taro germplasm along with maintaining the genetic stability and functionality of plants. Further possibility may also be explored to use this protocol for production of disease-free planting materials. This will facilitate the national and international exchange of taro germplasms.

Keywords: conservation, genetic integrity, inter-simple sequence repeats, random amplified polymorphic DNA, taro, tissue culture

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

Indian Journal of Biotechnology
Vol 5, October 2006, pp 543-550

 

In vitro cloning of apple (Malus domestica Borkh) employing forced shoot tip cultures of M9 rootstock

M Amin Dalal*, B Das, A K Sharma, M Amin Mir and Amarjeet Singh Sounduri

 

For micropropagation of adult apple (Malus domestica Borkh) rootstock M9, explants were harvested from pre-chilled (4±3°C) dormant cuttings forced in growth chamber. Primary explants were established by using a slightly modified version of culture initiation procedure developed earlier. Multiple shoots, raised by axillary branching of surviving explants, were obtained from established cultures in two cultural pathways: (i) repeat transfer of primary explants on the MS supplemented with BAP (2.22 µM), IBA (0.49 µM) and Kn (2.2 µM), and (ii) repeat subculture of harvested microshoots and basal portion of sectored proliferating clumps on the same but suitably PGR amended medium, in which cytokinins concentration was reduced by half. A 4-fold shoot multiplication was achieved during each sub culture of 3±1 week’s duration that repeatedly produced crop of proliferated shoots without loss of vigour. Nodal segments (5-15 mm), obtained from in vitro raised microshoots were also used to initiate a new cycle of proliferating cultures. Isolated cloned microshoots (15-20 mm) with apical bud were cultured on MS basal medium supplemented with IBA (14.70 µM) for 8±3 d to initiate root. The microshoots were re-implanted in PGR free half strength basal MS medium with full complement of organics for
4±1 week for root development. The transferred in vitro hardened plantlets to polyvinyl cups or polybags, under carefully controlled descending RH regime of 95% to 70±5% over a period of 5±1 week, resulted in 80% ex vitro survival. The present protocol highlighted a novel strategy of micropropagation of apple rootstock M9 using three-step culture initiation procedure of forced primary explants.

Keywords: apple, dwarfing rootstocks, forced explants, hardening, in vitro propagation

IPC Code: Int. Cl.8 A01H4/00

 

Indian Journal of Biotechnology
Vol. 5, October 2006, pp 551-554

 

Micropropagation of Embelia ribes Burm.f. using inflorescence segments

K Shankarmurthy and V Krishna*

 

An efficient micropropagation protocol was developed for a threatened medicinal plant, Embelia ribes using inflorescence explants. The immature ovaries of the inflorescence segments proliferated into luxuriant mass of callus on Murashige and Skoog’s (MS) medium supplemented with IBA (3.5 mg/L) and Kn (0.5 mg/L). The combination of Kn and NAA in the range of 2.0 to 4.0 mg/L and 0.2 to 0.6 mg/L, respectively provoked the calli to differentiate into both shoot and root initials from the ovary callus. Highest number of regenerants (25±0.81per callus) were obtained at the concentration of 3.0 mg/L Kn and 0.4 mg/L NAA. The regenerants were transferred to the pots containing sterilized soil and hardened for a week. A mean of 21.82±1.02 regenerants per harvest were well acclimatized to the natural conditions exhibiting a normal development.

Keywords: Embelia ribes, inflorescence culture, micropropagation

IPC Code: Int. Cl.8 A01H4/00

Indian Journal of Biotechnology
Vol 5, October 2006, pp 555-558

 

Micropropagation of Capparis spinosa L. subsp. rupestris Sibth. &
Sm. by nodal cuttings

L Chalak* and A Elbitar

 

In vitro propagation of Capparis spinosa subsp. rupestris was conducted using single nodal cuttings. Multiple shoots were obtained on Murashige and Skoog (MS) medium supplemented with 1 mg/L zeatin. Shoot multiplication was optimized on the same medium by subculturing shoot segments with 2-3 nodes every 6 weeks. Aptitude to proliferation was successfully maintained during nine subcultures with a mean rate exceeding 20 new shoots per explant. High rooting response of shoots (92%) was obtained after 4 h pulse treatment period in darkness with 100 mg/L IAA solution, followed by culture on solid half-strength MS basal medium. These results indicate the enormous potential of C. spinosa subsp. rupestris that could be used for large-scale multiplication.

Keywords: caper, Capparis spinosa subsp. rupestris, tissue culture, micropropagation

IPC Code: Int. Cl.8 A01H4/00

 

Short Communications

 

Indian Journal of Biotechnology
Vol 5, October. 2006, pp 559-561

 

Molecular analysis of bacterial population inhabiting coconut husk retting area

Sabu Thomas*, K Harikrishnan and Sandhya C Nair

 

As part of the biodegradation of the phenolic compounds, a microbiological survey was conducted in the Kadinamkulam estuary during 2003. The aim of this study was to screen the bacterial population and to elucidate their plasmid profile in the context of coconut husk retting. Species of Pseudomonas, Escherichia, Enterobacter, Micrococcus, Shigella, Salmonella, and Klebsiella were found to be present in the area. Of these the most dominant species was Pseudomonas. The study was also planned to delineate the antimicrobial susceptibility pattern prevalent in the area. Of the various antibiotics tested, only gentamycin and ofloxacin were found to be sensitive to all the bacterial strains. During the present study nineteen isolates showed the presence of plasmids of varying sizes as their extrachromosomal genetic material.

Keywords: husk retting, estuary, bacteriology, antibiogram, plasmid profile

IPC Code: Int. Cl.8 C12N15/10

Indian Journal of Biotechnology
Vol 5, October 2006, pp 562-564

 

In vitro multiplication of Asteracantha longifolia (L.) Nees–A medicinal herb

J Panigrahi*, R R Mishra and M Behera

 

Plantlet regeneration in Asteracantha longifolia (L.) Nees (Acanthaceae), a medicinal herb, has been achieved from leaf segments of seedlings germinated on basal MS medium. Leaf explants were cultured on MS medium fortified with various concentrations of BA and NAA either alone or in combination. Among the combinations used, BA(2.0 mg/L) + NAA
(0.2 mg/L) was found to be the best for direct multiple shoot bud induction, while BA(2.0 mg/L) + NAA(0.5 mg/L) was suitable for high frequency shoot bud regeneration through an intervening callus phase. Well-developed shoots (>3 cm) were successfully rooted on MS medium containing NAA (0.1 mg/L). Eighty percent of the regenerated plantlets were successfully established in soil. The protocol described is simple, rapid, and efficient for in-vitro propagation of A. longifolia from leaf explants and soil establishment of plants.

Keywords: Asteracantha longifolia, leaf explant, organogenesis, shoot bud

IPC Code: Int. Cl.8 A01H4/00

 

Indian Journal of Biotechnology

Vol 5, October 2006, pp 565-567

 

In vitro propagation of Ceropegia bulbosa using nodal segments

Divya Goyal* and Seema Bhadauria

 

In vitro propagation of Ceropegia bulbosa, a threatened species has been achieved, which could be used to conserve it. Initially, nutrient media used were basal MS medium and MS media supplemented with cytokinins, BAP (2.22-8.87 µM/L) or kinetin (2.32-9.29 µM/L) or TDZ (2.27-9.08 µM/L) with combination of GA (0.58 µM/L) and NAA (0.27 µM/L) Nodal explants were used for establishing in vitro cultures. Mercuric chloride eliminated the contamination effectively from nodal explants after treating with Bavistin and Streptomycin with a good survival rate. One or two buds developed from a node after two weeks. BAP (4.44 µM/L) with GA (0.58 µM/L) and NAA (0.27 µM/L) showed highest frequency of formation of shoots (85%). Microshoots were transferred on MS medium supplemented with different concentrations of auxins IAA (2.85-11.42 µM/L) and NAA (2.69-10.74 µM/L) with BAP (3.55 µM/L) for rooting. IAA (11.42 µM/L) with BAP (3.55 µM/ L) showed highest rooting percentage. The rooted plantlets were acclimatized and hardened successfully within ten days of transferring them into plastic cups containing sterilized vermiculite for hardening.

Keywords:   in vitro propagation, Ceropegia bulbosa, threatened species, nodal explant

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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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.