Indian Journal of Biotechnology

 

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VOLUME 8

CODEN: IJBNAR 8(3) (2009) 249-342

NUMBER 3

JULY 2009

ISSN: 0972-5849 (print); 0975-0967 (online)

 

  CONTENTS

 

 

Review

 

Catalytic antibodies as potential therapeutics

253

Murtuza Ali, A G Hariharan, Neeraj Mishra & Sanjay Jain

 

 

 

Papers

 

The saga of cytotoxin evolution — Switching of destructive role to a constructive role

259

Subhamay Panda, Santamay Panda & Jayanta Sinha

 

 

 

Isolation, purification and biochemical characterization of conotoxin from Conus figulinus Linnaeus (1758)

266

R Saravanan, S Sambasivam , A Shanmugam, D Satish Kumar, T Tamil Vanan & R A Nazeer

 

 

 

Isolation of symbiotic bacteria and bioactive proteins from the marine sponge, Callyspongia diffusa

272

S Boobathy , T T Ajith Kumar & K Kathiresan

 

 

 

Differentiation of Staphylococcus aureus strains based on 16S-23S ribosomal RNA intergenic space polymorphism

276

Ashok Dubey, S K Ghorui & S K Kashyap

 

 

 

Production of alkaline protease from Streptomyces gulbargensis and its application in removal of blood strains

280

N Vishalakshi, K Lingappa, S Amena , M Prabhakar & A Dayanand

 

 

 

Optimization of protease production from newly isolated strain of Bacillus sp. PCSIR EA-3

286

Shah Ali Ul Qadar, Erum Shireen, Samina Iqbal & Abida Anwar

 

 

 

Thermostable and alkaline tolerant xylanase production by Bacillus subtilis isolated form marine environment

291

N Annamalai, R Thavasi, S Jayalakshmi & T Balasubramanian

 

 

 

Biopolymer based intervention for Salmonella in water

298

Moushumi Ghosh & Santosh Pathak

 

 

 

Influence of Pediococcus acidilactici as a starter on the flavour of tempoyak (fermented durian)

304

Neti Yuliana & Vergilio V Garcia

 

 

 

In vitro propagation of Citrus indica Tanaka — An endangered progenitor species

311

M A Laskar, M Hynniewta & C S Rao

 

 

 

Influence of biotic and abiotic elicitors on accumulation of hyoscyamine and scopolamine in root cultures of Datura metal L.

317

L Ajungla, P P Patil, R B Barmukh & T D Nikam

 

 

 

Growth and shoot proliferation in Chlorophytum borivilianum Sant. et Fernand. in vitro under different carbon dioxide environment

323

N Joshi, A Dave, S Vyas & S D Purohit

 

 

 

An efficient in vitro protocol for clonal multiplication of Ginger – var. Varada

328

R Kavyashree

 

 

 

Short Communications

 

Desiccation of callus enhances somatic embryogenesis and subsequent shoot regeneration in sugarcane

332

Ajinder Kaur & S S Gosal

 

 

 

Biodegradation of leaf litter of tree species in presence of cow dung and earthworms

335

A K Sannigrahi

 

 

 

Instructions to Contributors

339

 

 

 

 

 

AUTHOR INDEX

 


Ajungla L

317

Ali M

253

Amena S

280

Annamalai N

291

Anwar A

286

 

 

Balasubramanian T

291

Barmukh R B

317

Boobathy S

272

 

 

Dave A

323

Dayanand A

280

Dubey A

276

 

 

Garcia V V

304

Ghorui S K

276

Ghosh M

298

Gosal S S

332

 

 

Hariharan A G

253

Hynniewta M

311

 

 

Iqbal S

286

Jain S

253

Jayalakshmi S

291

Joshi N

323

 

 

Kashyap S K

276

Kathiresan K

272

Kaur A

332

Kavyashree R

328

Kumar D S

266

Kumar T T A

272

 

 

Laskar M A

311

Lingappa K

280

 

 

Mishra N

253

 

 

Nazeer R A

266

Nikam T D

317

 

 

Panda S

259

Panda Su

259

Pathak S

298

Patil P P

317

Prabhakar M

280

Purohit S D

323

 

 

Qadar S A U

286

 

 

Rao C S

311

 

 

Sambasivam S

266

Sannigrahi A K

335

Saravanan R

266

Shanmugam A

266

Shireen E

286

Sinha J

259

 

 

Thavasi R

291

 

 

Vanan T T

266

Vishalakshi N

280

Vyas S

323

 

 

Yuliana N

304

 

 

 

 

 

Indian Journal of Biotechnology

Vol 8, July 2009, pp 253-258

 

 

Catalytic antibodies as potential therapeutics

Murtuza Ali*, A G Hariharan, Neeraj Mishra1and Sanjay Jain

Smriti College of Pharmaceutical Education, Dewas Naka, Nipania, Indore 452 010, India

1Department of Pharmaceutical Sciences, Dr H S Gour Vishwavidyalaya, Sagar 470 003, India

Received 25 April 2008; revised 3 October 2008; accepted 20 December 2008

The idea that enzymes can be generated by employing antibodies complimentary to haptenic groups, resembling the transition state of a given reaction, laid down the foundation of catalytic antibodies. New possibilities arise for their therapeutic applications, because of  high degree of reaction specificity, greater affinity towards transition state analog and the latent ability to block unwanted protein-protein interactions. Antibody directed abzyme prodrug therapy (ADAPT) will largely replace antibody directed enzyme prodrug therapy (ADEPT) for selectively delivering chemotherapeutic agents to the affected tissues. They can enzymatically cleave specific surface proteins and sugars on viruses or tumour cells, thereby disrupting the invaders. They had also been successfully used to detoxify drugs like cocaine and methamphetamine. Desired reaction selectivity can be induced in antibodies to exhibit a wide range of chemical reactions, so that they would prove to be a milestone in human endeavour to treat genetic diseases, which show brighter prospects for their therapeutic applications in near future. This review aims at analyzing the updated information on biochemical and mechanistic implications of catalytic antibodies with special reference to their therapeutic application and the advances made in these areas.

Keywords: Abzyme; antibodies; hapten; transition state; therapeutics

 

 

Indian Journal of Biotechnology

Vol 8, July 2009, pp 259-265

 

 

The saga of cytotoxin evolution–Switching of destructive role to a constructive role

Subhamay Panda1*, Santamay Panda2 and Jayanta Sinha3

1*Department of Biological Sciences, Gupta College of Technological Sciences, Asansol 713 301, India

2Department of Physics, B C College, Asansol 713 304, India

3Department of Zoology, B B College, Asansol 713 303, India

Received 30 May 2008; revised 27 November 2008; accepted 20 February 2009

Snake venom contains the toxin proteins, cytotoxins. Cytotoxins exert their effect upon the target cells by interacting with membrane lipids and proteins. Ultimate objective of a cytotoxin is to destroy the target cells. These cytotoxins contain cysteine residues responsible for disulphide linkage between them. Similar variety of peptides enriched with cysteine is also found in many other organisms. But interestingly, in those cases they never have a cell destructive function, in turn, they act to be cell-friendly. In this work, we analysed the cytotoxins and related peptides in terms of amino acid percentage profile, multiple sequence alignment, codon usage, isoelectric point determination, protein secondary structure prediction and phylogenetic tree construction through different softwares. Among all the interesting results, lysine profile was very much informative. High amounts of lysine are conserved in all the cytotoxins whereas in other related peptides it is in less numbers. Phylogenetic tree showed a stepwise dynamic evolution of these interesting molecules. This paper, therefore, showed that there is a great possibility to turn harmful natural peptides into a beneficial engineered molecule for the betterment of lives of mankind.

Keywords: Cytotoxin, cysteine, lysine, molecular evolution, phylogenetic analysis, snake

 

 

Indian Journal of Biotechnology

Vol 8, July 2009, pp 266-271

 

 

Isolation, purification and biochemical characterization of conotoxin from
Conus figulinus Linnaeus (1758)

R Saravanan1*, S Sambasivam1, A Shanmugam1, D Sathish Kumar2, T Tamil Vanan2 and R A Nazeer3

1Centre of Advanced Study in Marine Biology, Annamalai University, Paraingipettai 608 502, India

2Department of Biotechnology & 3School of Biotechnology, SRM University, Kattankulathur 603 203, India

Received 24 July 2008; revised 15 January 2009; accepted 22 March 2009

Cone snails are remarkable for the extent and diversity of gene-encoded peptide neurotoxins that are expressed in their venom apparatus. The protein content of the crude toxin extract of Conus figulinus Linneaus was found to be 1900 μg/mL. The crude extract (dilution up to 10-5) expressed hemolytic activity. The crude extract subjected to gel filtration chromatography yielded 60 fractions; the fractions 7, 12 and 55 showed significant peaks at 280 nm. The fractionated toxin was then characterized by performing SDS-PAGE having the lower peptides ranging from 10 to 43 kDa; two lower peptides below 14 kDa have been identified. The total RNA and purified mRNA were characterized by Agarose gel electrophoresis and for total RNA two prominent bands of 18s and 28s were obtained of which 28s showed double intensity than the other. For mRNA a single band of 6000 base pairs was obtained.

Keywords: Conus figulinus,cone snail, chymotrypsin, toxin, trypsin, total RNA, mRNA

 

 

Indian Journal of Biotechnology

Vol 8 July 2009, pp 272-275

 

Isolation of symbiotic bacteria and bioactive proteins from the marine sponge, Callyspongia diffusa

 

S Boobathy*, T T Ajith Kumar and K Kathiresan

Centre of Advanced Study in Marine Biology, Annamalai University, Parangipettai 608 502, India

Received 3 April 2008; revised 3 November 2008; accepted 28 January 2009

The marine sponge, Callyspongia diffusa, collected near Mumbai coast, was studied for symbiotic bacterial association and biologically active proteins. Four bacterial species (Pseudomonas aeruginosa, Escherichia coli, Vibrio parahaemolyticus and V. cholera) were found associated with the sponge. Crude protein from the sponge was extracted at concentration of 1.43 mg/mL in methanol and 1.62 mg/mL in aqueous extract. Antibacterial activity of the extracts against the four symbiotic bacteria inhibited the growth of only V. cholera. Both extracts exhibited hemolytic activity on chicken erythrocytes at 6.99 HT/mg for methanolic and 8.64 HT/mg for aqueous extracts. On SDS-PAGE, the crude protein yielded eight bands in methanolic extract and five bands in aqueous extract, with molecular weight ranging from 14.4 to 116 kDa with three well defined bands of 19.5, 39.0, 66.2 kDa in both the extracts.

Keywords: Marine sponge, symbiotic bacteria, bioactive proteins, hemolytic assay, Callyspongia diffusa

 

 

Indian Journal of Biotechnology

Vol 8, July 2009, pp 276-279

 

 

Differentiation of Staphylococcus aureus strains based on 16S-23S ribosomal RNA intergenic space polymorphism

Ashok Dubey, S K Ghorui1 and S K Kashyap*

College of Veterinary and Animal Science, Bikaner 334 001, India

1National Research Centre on Camel, Jorbeer, Bikaner 334 001, India

Received 29 January 2008; revised 22 December 2008; accepted 10 March 2009

Discrimination of Staphylococcus aureus strains, isolated from camel abscesses and mastitic milk of camel, cattle and goats, on the basis of 16S-23S ribosomal RNA intergenic space polymorphism was carried out. Two sets of primers were used for amplification of DNA of intergenic space; the one having a highly conserved sequence in eubacterial 23S rRNA transcript, while the other having less conserved sequence of 16S rRNA, reported earlier by other workers. Of the two sets of primers used, amplification could be achieved with one set of primers. Of 60 strains of S. aureus tested, amplification could be achieved in only 18 strains. In these strains the most frequent bands of DNA were of 350, 500, 750 and 1500 base pairs. Polymorphism was noted in the number of the rRNA transcripts and size of the 16S-23S rRNA intergenic space, as evident by variable band pattern in different strains of S. aureus.

Keywords: Ribosomal RNA, 16S-23S rRNA intergenic space, Staphylococcus aureus

 

 

Indian Journal of Biotechnology

Vol 8, July 2009, pp 280-285

 

 

Production of alkaline protease from Streptomyces gulbargensis and its application in removal of blood stains

N Vishalakshi1, K Lingappa*, S Amena, M Prabhakar and A Dayanand

Department of Microbiology, Gulbarga University Gulbarga 585 106, India

Received 1 August 2008; revised 12 January 2009; accepted 15 March 2009

The alkaline protease obtained from a newly isolated strain of Streptomyces gulbargensis was used for the washing of surgical instruments. The isolate showed b-haemolysis. Therefore, the isolate was employed for the production of thermo stable alkaline protease enzyme using wheat bran as the substrate under solid state fermentation. The characterization studies of the enzyme showed that it is active at 45°C and pH 9.0 with casein as the substrate. The wash performance analysis of blood stains on cotton fabrics and on surgical instruments showed an increase in the reflectance as the time increased with the enzyme treatment. The removal of blood stains completely was observed at 20 min incubation of cotton cloths and surgical instruments.

Keywords: Alkaline protease, blood stains, surgical instruments, S. gulbargensis, wheat bran

 

 

Indian Journal Biotechnology

Vol 8, July 2009, pp 286-290

 

 

Optimization of protease production from newly isolated strain of Bacillus sp. PCSIR EA-3

Shah Ali Ul Qadar1*, Erum Shireen2, Samina Iqbal3 and Abida Anwar3

1Institute of Sustainable Halophyte Utilization and 2Department of Biochemistry
University of Karachi, Karachi, Pakistan3

Pharmaceutical Research Center, PCSIR Laboratories Complex, Karachi, Pakistan

Received 10 July 2008; revised 12 January 2009; accepted 16 March 2009

A newly isolated strain of Bacillus sp. PCSIR EA-3 showed maximum protease production in 48 h. Cell free filtrate (CFF) from the strain was investigated for its proteolytic activity. The isolated strain exhibited highest proteolytic activity when grown on skim milk agar; average area of zone of inhibition was 480 mm2. Enzyme showed maximum activity at 35°C and pH 7.0 with glucose as an important medium component. It was strongly activated by metal ions, such as, Ca+2 ion. Protease from PCSIR EA-3 can easily be used in food industry, especially in cheese manufacturing due to its thermostability at neutral pH.

Keywords: Bacillus sp., calcium ion, culture optimization, protease

 

 

Indian Journal of Biotechnology

Vol 8, July 2009, pp 291-297

 

 

Thermostable and alkaline tolerant xylanase production by Bacillus subtilis isolated form marine environment

N Annamalai, R Thavasi1*, S Jayalakshmi and T Balasubramanian

CAS in Marine Biology, Annamalai University, Parangipettai 608502, India

1Department of Chemical and Biological Sciences, Polytechnic Institute of New York University
Six Metrotech Center, Brooklyn 11201, New York, USA

Received 5 September 2008; revised 30 December 2008; accepted 12 March 2009

The aim of the present study was to isolate a thermostable and alkaline tolerant xylanase producing strain from an estuarine environment, and to produce, purify and characterize the enzyme. The bacterium, Bacillus subtilis isolated from the estuarine environment was grown in shake flasks to derive the optimum culture conditions and also cultured in lab scale fermentor to obtain more xylanase. Maximum enzyme production (128 U/mL) was recorded in stationary phase (36 h) of the culture. The enzyme was purified using ammonium sulphate precipitation (60% saturation), followed by DEAE-cellulose column chromatography. The enzyme was optimally active at 55oC and pH 9.0. Influence of metal ions on enzyme activity revealed that, Fe2+, Ca2+, and Mg2+ greatly enhanced the enzyme activity to 238, 148, 208 U/mL, respectively; whereas Hg2+ (0 U/mL) and EDTA (18 U/mL) strongly inhibited the enzyme activity. Based on the results obtained, xylanase isolated in this study may be useful in pulp pre-bleaching process to remove the hemicelluloses. The use of thermostable alkaline tolerant xylanase for enzyme assisted pulp bleaching could greatly reduce the need for pH and temperature readjustment, thus offering enormous technical and economic advantages.

Keywords: Alkaline tolerant, Bacillus, marine, thermostable, xylan, xylanase

 

 

Indian Journal of Biotechnology

Vol 8, July 2009, pp 298-303

 

 

Biopolymer based intervention for Salmonella in water

Moushumi Ghosh1* and Santosh Pathak

Department of Biotechnology and Environmental Sciences, Thapar University, Patiala 147 004, India

Received 10 July 2008; revised 27 November 2008; accepted 20 February 2009

This study evaluated the potential of a bacterial polymer for binding and removal of Salmonella spp. currently deemed as a biowarfare agent in water. The biopolymer previously characterized as a polysaccharide with flocculating activity is produced extracellularly by the bacterium, Klebsiella terrigena. The biopolymer could effectively remove 3 log cfu/mL of Salmonella typhimurium ATCC 23564, within a period of 30 min from amongst other indicator (Escherichia coli DH5α and Enterococcus faecalis ATCC 35550) and enteropathogens of moderate concern (Staphylococcus aureus ATCC 9144) spiked in water samples at ambient temperature. The optimum dose of biopolymeric flocculant for removal of Salmonella was 2 mg/L. FISH and CLSM demonstrated that the removed S. typhimurium was selectively bound to the biopolymer matrix. The results indicate a significant possibility of using this biopolymer for rapid detection and removal of key biowarfare agents (for instance Salmonella) transmitted through water.

Keywords: Biopolymer, biowarfare, flocculation, polysaccharide, Klebsiella terrigena, Salmonella Typhimurium

 

 

Indian Journal of Biotechnology

Vol 8, July 2009, pp 304-310

 

 

Influence of Pediococcus acidilactici as a starter on the flavour of tempoyak (fermented durian)

Neti Yuliana1* and Virgilio V Garcia2

1Jurusan Teknologi Hasil Pertanian (THP), Fakultas Pertanian,

Universitas Lampung (UNILA). Jalan Sumantri Brojonegoro #1 Bandar Lampung, Indonesia

2 Institute of Food Science and Technology, University of the Philippine at Los Banos, Philippines

Received 24 July 2008; revised 28 November 2008; accepted 10 February 2009

Pediococcus acidilactici UP02 was used as a starter for the fermentation of tempoyak (Durio zibethinus Murr.), a traditional Indonesian fermented fruit. The flavour of tempoyak was compared to those in spontaneous fermentation (no starter added). The results showed that addition of P. acidilactici UP02 in tempoyak fermentation decreased the quantity of sulphur components, reduced the sugar content, and increased various non-sulphur compounds as well as, non-volatile acidity and contents of organic acids (lactic, malic, and acetic). Sensory evaluation showed that there was no significant difference among the samples for sourness and aroma. However, the P. acidilactici UP02-inoculated sample was more acceptable than the sample with no-starter treatment. On the basis of the results of GC-MS, the flavour components of tempoyak with P. acidilactici UP02 treatment were observed to have 48 compounds, mainly composed of dimethyl disulfide and diethyl disulfide followed by ethane,1-1-bis ethylthio.

Keywords: Pediococcus acidilactici, tempoyak, malic acid, sulphur components

 

 

Indian Journal of Biotechnology

Vol 8, July 2009, pp 311-316

 

 

In vitro propagation of Citrus indica Tanaka—An endangered progenitor species

M A Laskar*, M Hynniewta and C S Rao1

Department of Biotechnology and Department of Botany1, St Anthony’s College, Shillong 793 001, India

Received 17 April 2008; revised 26 December 2008; accepted 27 February 2009

A method for in vitro propagation of Citrus indica Tanaka by shoot organogenesis from leaf-derived callus was developed. Regenerative calli were induced on MS medium supplemented with 0.01 mg L‑1 TDZ and 0.1 mg L‑1 NAA. Shoots were regenerated on WPM medium supplemented with 0.5 mg L‑1 BAP, 0.25 mg L‑1 TDZ and 0.25 mg L‑1 NAA. Regenerated shoots were rooted on MS medium supplemented with 1.0 mg L‑1 NAA. Sixty per cent of the rooted plantlets were acclimatized successfully under ex situ conditions.

Keywords: Citrus indica endangered, ex situ adaptibility, leaf-derived callus, progenitor, shoot organogenesis

 

 

Indian Journal of Biotechnology

Vol 8, July 2009, pp 317-322

 

 

Influence of biotic and abiotic elicitors on accumulation of hyoscyamine and scopolamine in root cultures of Datura metel L.

L Ajungla, P P Patil, R B Barmukh1 and T D Nikam

Department of Botany, University of Pune, Pune 411007, India

1Modern College of Arts, Science and Commerence, Shivajinagar, Pune 411 005, India

Received 18 August 2008; revised 30 December2008; accepted 17 March 2009

Leaf-derived root cultures of Datura metel L., established on B5 medium containing 1.2 µM IAA, were employed to study the influence of biotic (Aspergillus niger, Alternaria sp., Fusarium monoliforme and yeast extract) and abiotic (salicylic acid, AlCl3, CaCl2, NaCl and Na2SO4) elicitors on the growth and production of hyoscyamine and scopolamine, the medicinally important tropane alkaloids. The hyoscyamine and scopolamine contents in control root cultures were 1.39 mg/g dw and 0.069 mg/g dw, respectively. The highest hyoscyamine (4.35 mg/g dw) and scopolamine (0.28 mg/g dw) accumulation was obtained in cultures treated with 500 μM salicylic acid, followed by treatment with 0.75 g L-1 yeast extract (3.17 mg/g dw hyoscyamine & 0.16 mg/g dw scopolamine) and 250 μM AlCl3 (2.49 mg/g dw hyoscyamine and 0.11 mg/g dw scopolamine).

Keywords: Datura metel, elicitors, hyoscyamine, scopolamine, tropane alkaloids

 

 

Indian Journal Biotechnology

Vol 8, July 2009, pp 323-327

 

 

Growth and shoot proliferation in Chlorophytum borivilianum Sant. et Fernand. in vitro under different carbon dioxide environment

N Joshi, A Dave, S Vyas and S D Purohit*

Plant Biotechnology Laboratory, Department of Botany, Mohanlal Sukhadia University, Udaipur 313 001, India

Received 2 September 2008; revised 25 November 2008; accepted 1 February 2009

In vitro growth and shoot proliferation in Chlorophytum borivilianum Sant. et Fernand. were studied in a controlled CO2 environment. The cultures were grown on BA supplemented MS medium with or without 3% sucrose. A range of CO2 concentrations (0.0, 0.6, 10.0 and 40.0 g m-3) was controlled in small chambers by using solutions of NaHCO3, Na2CO3, KHCO3 and K2CO3. In order to maintain a CO2-free environment, a saturated solution of KOH was kept in the chambers. It was observed that the growing shoot cultures required either sucrose in the medium as a carbon source or an environment with controlled CO2 concentration. Complete absence of a carbon source caused severe yellowing of shoots and death within 15 d. The growth of shoot cultures at 40.0 g m-3 CO2 was comparable to the growth obtained with 3.0% sucrose in the medium. With both CO2 and sucrose being available, the best response was obtained at 40.0 g m-3 CO2 in the chamber. At this concentration the increase in number of shoots was nearly double the standard rate obtained when exposed to the natural CO2 level and sucrose supplemented medium. Total fresh and dry wt and shoot number were the maximum under this condition.

Keywords: Carbon dioxide enrichment, Chlorophytum borivilianum, in vitro growth, micropropagation, photoautotrophy, shoot culture

 

 

Indian Journal Biotechnology

Vol 8, July 2009, pp 328-331

 

 

An efficient in vitro protocol for clonal multiplication of Ginger – var. Varada

R Kavyashree*

Plant Biotechnology Unit, Annexe-Department of Botany, Bangalore University, Bangalore 560 056, India

Received 22 August 2008; revised 28 November 2008; accepted 5 February 2009

 

An efficient, in vitro multiplication protocol was developed for Zingiber officinale Rosc., var. Varada through direct regeneration of vegetative buds. Multiple shoots were induced from vegetative buds on LSBM fortified with BAP (17.76 μM) with 96% initiation response. The repeated subculture resulted in rapid shoot multiplication at the average rate of 4-fold per culture. The well-developed multiple shoots formed roots on the same medium after 2-3 passages of subculture, thus eliminating the step of in vitro rooting. The statistical data pertaining to multiple shoot and root formation revealed highest mean number of 19.1 and 12.3, respectively. The regenerated plantlets were successfully established in the field with 86% survival frequency after few days of indoor acclimatization.

Keywords: Vegetative bud, in vitro multiplication, survival frequency, Zingiber officinale

 

 

Indian Journal of Biotechnology

Vol 8, July 2009, pp 332-334

 

 

Desiccation of callus enhances somatic embryogenesis and subsequent shoot regeneration in sugarcane

Ajinder Kaur and S S Gosal*

School of Agricultural Biotechnology, Punjab Agricultural University, Ludhiana 141 004, India

Received 6 September 2007; revised 11 September 2008;
 accepted 15 February 2009

Callus cultures were established in three commercial sugarcane varieties, viz., CoJ 64, CoJ 83 and CoJ 86, from spindle leaf segments on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D; 4 mg/L) and 6-benzylaminopurine (BAP; 0.5 mg/L). The calli were sub-cultured on MS+2,4-D (2 mg/L)+BAP (0.5 mg/L)+agar (0.8% w/v) medium (control) and MS+2,4-D (2 mg/L)+BAP (0.5 mg/L)+agar (1.6% w/v) medium (treatment) to study the effect of desiccation caused by double agar on somatic embryogenesis. Per cent somatic embryogenesis observed in treatment-calli of sugarcane varieties CoJ 64, CoJ 83 and CoJ 86 was 90, 90.63 and 89.66, respectively; while in control-calli the corresponding figures were 66.67, 64.52 and 63.33. Likewise, shoot regeneration from desiccated calli on MS+BAP (0.5 mg/L) medium was also higher over non-desiccated control, i.e., 84.27, 86.52 and 83.13% as compared to 54.35, 56.25 and 50.38%, respectively in CoJ 64, CoJ 83 and CoJ 86. Thus, this fairly simple double agar medium provided an alternative method for improving somatic embryogenesis and, hence, regeneration frequency of sugarcane callus.

Keywords: Callus, desiccation, regeneration, Saccharum officinarum L., somatic embryogenesis, sugarcane

 

 

Indian Journal of Biotechnology

Vol 8, July 2009, pp 335-338

 

 

Biodegradation of leaf litter of tree species in presence of cow dung and earthworms

A K Sannigrahi*

Proof and Experimental Establishment, DRDO,
Ministry of Defence, Chandipur, Balasore 756 025, India

Received 17 April 2008; revised 27 November 2008;
accepted 25 February 2009

The dry leaves of Psidium guajava Linn., Ficus benghalensis Linn., Codiaeum variegatum Linn., Polyalthia longifolia Sonner., Eucalyptus citriodora Hook., Caesalpinia pulcherrima Linn., Syzygium cumini Linn., Artocarpus heterophyllus Lamk., Musa paradisiaca Linn., Plumeria rubra Linn., Litchi chinensis Gaertn., Pinus insularis Endl., Elaeocarpus sphaericus Gaertn., Bombax ceiba Linn., Streblus asper Lour. and Dalbergia sissoo Roxb. were converted to vermicomposts in about 5 to 8 months while those of Mangifera indica Linn., Terminalia chebula Retz., Lagerstroemia speciosa Linn. and Tectona grandis Linn. took about 10 months. However, Leucaena leucocephala Lamk. leaves took only 2 months to become vermicompost. These vermicomposts contained: N, 0.9 to 1.9; P, 0.5 to 1.2; K, 0.9 to 2.5; Na, 0.8 to 3.5; Ca, 0.9 to 4.9 and S, 0.9 to 2.2%. The vermicompost of Bombax ceiba flowers produced in 3.5 months contained: N, 1.2; P, 0.6; K, 1.8; Na, 1.4; Ca, 0.4 and S, 0.9%. Nutritional status of these vermicomposts varied with the variation of tree species. This information would motivate farmers for vermicomposting of dry leaves instead of burning.

Keywords: Avenue trees, biodegradation, earthworm, fruit trees, leaf litter, Perionyx excavatus, vermicompost