Indian Journal of Biotechnology      
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VOLUME 9

CODEN: IJBNAR 9(3) (2010) 229-344

NUMBER 3

JULY 2010

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

 

CONTENTS

 

 

Review

 

Recent advances in molecular breeding of drought tolerance in rice (Oryza sativa L.)

233

Khela Ram Soren, Kishwar Ali, Vandana Tyagi & Aruna Tyagi

 

Papers

 

Evolutionary interrelationships among polyomaviruses based on nucleotide and amino acid variations

252

Soumen Bhattacharjee

 

Anti-HIV, anti-tubercular and mutagenic activities of borrelidin

265

D V R N Bhikshapathi, D R Krishna & V Kishan

 

Efficiency of Bemisia tabaci (Gennadius) populations from different plant-hosts for acquisition and transmission of cotton leaf curl virus

271

V K Gupta, Rakesh Sharma, Satnam Singh, Jawala Jindal & V K Dilawari

 

RAPD analysis in mungbean [Vigna radiata (L.) Wilczek.] II: A comparison of efficiency parameters of RAPD primers

276

Madhu Saini, Subhadra Singh, Z Hussain & V K Sikka

 

Evaluation of genetic diversity among Jatropha curcas (L.) by RAPD analysis

283

K Subramanyam, Dowlathabad Muralidhara Rao, N Devanna, A Aravinda & V Pandurangadu

 

Biotization with Piriformospora indica and Pseudomonas fluorescens improves survival rate, nutrient acquisition, field performance and saponin content of micropropagated Chlorophytum sp.

289

S K Gosal, A Karlupia, S S Gosal, I M Chhibba & A Varma

 

Genotyping by ERIC-PCR of Escherichia coli isolated from bovine mastitis cases

298

Vivek Prabhu, S Isloor, M Balu, V V S Suryanarayana & D Rathnamma

 

Siderophore production by Alcaligenes feacalis and its application for growth promotion
in Arachis hypogaea

302

R Z Sayyed, N S Gangurde, P R Patel, S A Joshi & S B Chincholkar

 

Studies on biphasic biomethanation of spoiled mango puree

308

R D Kirtane, P C Suryawanshi, A B Chaudhari & R M Kothari

 

Polishing of biomethanated spent wash (primary treated) in constructed wetland: A bench scale study

313

P K Singh, K P Sharma, Shweta Sharma, R C Swami & Subhasini Sharma

 

Plant regeneration from alginate-encapsulated somatic embryos of Dalbergia sissoo Roxb.

319

Ajay Kumar Singh & Suresh Chand

 

Short Communications

 

Intraspecific variation in the internal transcribed spacer (ITS) regions of rDNA in Withania somnifera (Linn.) Dunal

325

Bilal Ahmad Mir, Sushma Koul, Arun Kumar, Maharaj K Kaul, Amarjit Singh Soodan &
Soom Nath Raina

 

Direct organogenesis from hypocotyl explants of Psoralea corylifolia L. — An endangered medicinal plant

329

P Baskaran & N Jayabalan

 

Micropropagation of Verbesina encelioides — An invasive weed

333

Satish C Jain, Renuka Jain & Renu Singh

 

Somatic embryogenesis from leaf explants of soapnut (Sapindus mukorossi Gaertn.)

336

N S Philomina

 

Instructions to Contributors

341

 

AUTHOR INDEX

 

 


Ali K

233

Aravinda A

283

 

 

Balu M

298

Baskaran P

329

Bhattacharjee S

252

Bhikshapathi D V R N

265

 

 

Chand S

319

Chaudhari A B

308

Chhibba I M

289

Chincholkar S B

302

 

 

Devanna N

283

Dilawari V K

271

 

 

Gangurde N S

302

Gosal S K

289

Gosal S S

289

Gupta V K

271

 

 

Hussain Z

276

 

 

Isloor S

298

Jain R

333

Jain S C

333

Jayabalan N

329

Jindal J

271

Joshi S A

302

 

 

Karlupia A

289

Kaul M K

325

Kirtane R D

308

Kishan V

265

Kothari R M

308

Koul S

325

Krishna D R

265

Kumar A

325

 

 

Mir B A

325

 

 

Pandurangadu V

283

Patel P R

302

Philomina N S

336

Prabhu V

298

 

 

Raina S N

325

Rao D M

283

Rathnamma D

298

Saini M

276

Sayyed R Z

302

Sharma K P

313

Sharma R

271

Sharma S

313

Sharma Sw

313

Sikka V K

276

Singh A K

319

Singh P K

313

Singh R

333

Singh S

271

Singh Su

276

Soodan A S

325

Soren K R

233

Subramanyam K

283

Suryanarayana V V S

298

Suryawanshi P C

308

Swami R C

313

 

 

Tyagi A

233

Tyagi V

233

 

 

Varma A

289


Indian Journal of Biotechnology

Vol 9, July 2010, pp 233-251

 

 

Recent advances in molecular breeding of drought tolerance
in rice ( Oryza sativa L.)

Khela Ram Soren, Kishwar Ali, Vandana Tyagi 1 and Aruna Tyagi*

Division of Biochemistry, Indian Agricultural Research Institute, New Delhi 110 012, India

1 Germplasm Exchange Unit, National Bureau of Plant Genetic Resources, New Delhi 110 012, India

Received 28 March 2008; revised 1 December 2009 ; accepted 20 February 2010

Rice is an ideal plant species for genomic studies for its relative small genome size (~430 Mb), diploid origin (2x=24) and close relationship with other important crops. Rice has been grown under diverse ecological conditions and gets exposed to different environmental stresses like drought, salinity, cold, etc. Drought is generally avoided in irrigated rice production system but it is more prone to 63.5 mha of rainfed rice grown annually in different parts of world. Severe osmotic stress causes detrimental changes in cellular components. Yet in response to various environmental stresses, plants have developed different physiological and biochemical strategies to adapt stress conditions, such as, stress associated changes in metabolites and amino acids (proline), amines (glycin-betaine and polyamines), and variety of sugar and sugar alcohols (manitol and trehalose). There is also activation of cascade of molecular networks involved in stress perception, signal transduction and the expression of specific stress related genes. To understand these genetically complex mechanisms of abiotic stress tolerance, an integrated approach of molecular breeding, classical physiology and conventional breeding is necessary, and the present review is an effort to deal these issues.

Keywords: Drought stress, osmoprotectent, rice, transcription factor

 

Indian Journal of Biotechnology

Vol 9, July 2010, pp 252-264

 

Evolutionary interrelationships among polyomaviruses based on nucleotide and amino acid variations

Soumen Bhattacharjee

Department of Zoology, University of North Bengal, Raja Rammohunpur, Darjeeling 734 013, India

Received 20 November 2009; revised 23 December 2009; accepted 26 February 2010

Polyomaviruses (PyVs), members of the family Polyomaviridae , are strictly vertebrate resident viruses implicated in various pathogenic conditions including neoplasias, especially in human. This study aims to review both nucleotide and amino acid sequences of large T-antigen (LTAg), virion protein 1 (VP1) gene and also whole genomes of all known PyVs till date, in ascertaining phylogenetic relationships among themselves. The study detected genomic recombinations within the oncoprotein LTAg coding sequences of PyV group, and identified unique nucleotide and amino acid positions in the conserved regions of PyV-LTAg coding sequence, which can facilitate strain differentiations in clinical samples.

Keywords: Large T-antigen (LTAg), polyomavirus, viral protein 1 (VP1

 

Indian Journal of Biotechnology

Vol 9, July 2010 , pp 265-270

 

Anti-HIV, anti-tubercular and mutagenic activities of borrelidin

D V R N Bhikshapathi, D R Krishna and V Kishan *

University College of Pharmaceutical Sciences, Kakatiya University, Warangal 506 009 India

Received 2 January 2009; revised 5 October 2009; accepted 20 December 2009

Borrelidin, an antibiotic found to be an angiogenesis inhibitor, is a potential drug candidate due to its antiangiogenic activity and other biological activities. Keeping in view the scope of the antibiotic, we have performed some of the biological activities like effect of borrelidin on Mycobacterium tuberculosis H 37 Rv ( ATCC 27294 ) by BACTEC 460 TB and multidrug resistant M. tuberculosis . In these studies, the minimum inhibitory concentration (MIC) of borrelidin against
M. tuberculosis H 37 Rv was found to be 3.12 µg/mL. When compared with known antituberculous drugs like rifampicin, isoniazid, ethambutol and streptomycin, borrelidin showed higher MIC on M. tuberculosis H 37 Rv (ATCC 27294) . The
MIC was, however, lower than that of pyrazinamide. The inhibitory activity on multidrug resistant
M. tuberculosis was, observed at 256 µg/mL for borrelidin, rifampicin and isoniazid. The anti-HIV reverse transcriptase activity of borrelidin was tested by ELISA test, and mutagenic effect of borrelidin was performed by Ames test. Borrelidin showed moderate activity against anti-HIV reverse transcriptase enzyme. Results of Ames test indicated that borrelidin was non-mutagenic up to 500 µg/mL concentration.

Keywords: Borrelidin, Mycobacterium tuberculosis, mutagenicity, anti-HIV reverse transcriptase enzyme

 

 

Indian Journal of Biotechnology

Vol 9, July 2010, pp 271-275

 

Efficiency of Bemisia tabaci (Gennadius) populations from different plant-hosts for acquisition and transmission of cotton leaf curl virus

V K Gupta*, Rakesh Sharma, Satnam Singh, Jawala Jindal and V K Dilawari

Insect Molecular Biology Unit, Department of Entomology, Punjab Agricultural University, Ludhiana 141 004, India

Received 23 June 2009; revised 1 December 2009; accepted 5 February 2010

Whitefly ( Bemisia tabaci ) populations collected from five crop plants, viz brinjal, cotton, potato, tomato, soybean and a weed, Sida sp., and maintained on respective host plants were studied for virus acquisition from diseased cotton plant and its subsequent transmission to healthy cotton plants. The presence of virus in whitefly and diseased plants was established by PCR amplification of CLCuV-specific DNA primers (P 1800-500 ). The virus acquisition from the diseased cotton plants as well as its transmission to healthy cotton plant to cause disease was 100 per cent in the case of cotton-specific whitefly. Compared to this, using whitefly from other host-plants, both the values for acquisition of virus as well as its subsequent transmission decreased, being higher with soybean-specific whitefly (80 and 70%, respectively) and minimum with tomato whitefly (20% each). However, the actual virus transmission efficiency (ATE), when expressed after taking into account the fraction of whitefly samples that did not acquire virus, though was found to be 100 per cent in the case of cotton-, tomato- and brinjal-whitefly, it remained more than 67% with whitefly from other crops/weed. The study revealed that whitefly populations that are specific to different host-plants, differ primarily in their efficiency for acquiring CLCuV and once the virus is acquired it is efficiently transmitted to healthy cotton plants to cause cotton leaf curl disease (CLCuD). The results hold significance in control of CLCuD through appropriate management of whitefly on alternate host plants/weeds, both in and around the cotton growing areas.

Keywords: Bemisia tabaci, plant hosts, CLCuV transmission, whitefly

 

Indian Journal of Biotechnology

Vol 9, July 2010, pp 276-282

 

RAPD analysis in mungbean [ Vigna radiata (L.) Wilczek.]
II: A comparison of efficiency parameters of RAPD primers

Madhu Saini 1 , Subhadra Singh 1 *, Z Hussain 2 and V K Sikka 1

1 Department of Genetics, CCS Haryana Agricultural University, Hisar 125 004, India

2 National Bureau of Plant Genetic Resources, Pusa Campus, New Delhi 110 012, India

Received 24 March 2009; revised 10 November 2009; accepted 24 January 2010

The study was undertaken to identify the efficient RAPD primers that could differentiate a set of 39 mungbean [ Vigna radiata (L.) Wilczek] genotypes. Various efficiency parameters, namely, discriminatory power (D), discriminatory power with infinite sample size (D L ), diversity index (DI), resolving power (R p ) and marker index (MI) were studied for 30 primers. The relationship between the parameters was studied using Spearman rank correlation coefficient. The parameters D and D L were the most efficient, followed by R p , MI and DI. DI was not a good parameter in those cases where a set of primers with intermediate range of variation in their efficiency were used. None of the 30 primers alone could differentiate all 39 mungbean genotypes. However, a methodology could be designed to find the primer/primers combination to differentiate all 39 genotypes. The primers S-1 and S-2 were more efficient with iso-frequency distribution of most of their banding patterns and a combination of any one of the primers with S-1 or S-2 could identify all the genotypes. The study showed how to select the minimum number of efficient primers aimed to distinguish all genotypes cost-effectively.

Keywords : Discriminatory power, diversity index, marker index, mungbean, rank correlation, RAPD pattern, resolving power

 

Indian Journal of Biotechnology

Vol 9, July 2010, pp 283-288

 

Evaluation of genetic diversity among Jatropha curcas (L.) by RAPD analysis

K Subramanyam 1 , Dowlathabad Muralidhara Rao 2 *, N Devanna 3 , A Aravinda 1 and V Pandurangadu 1

1 Department of Biochemistry, Oil Technological Research Institute
Jawaharlal Nehru Technological University, Anantapur 515 001, India

2 Department of Biotechnology, Sri Krishnadevaraya University, Anantapur 515 055, India

3 Department of Chemistry, Jawaharlal Nehru Technological University, Anantapur 515 001, India

Received 14 October 2008; revised 18 September 2009; accepted 25 November 2009

The genetic diversity and pedigree analysis of Jatropha curcas (L.) was evaluated through RAPD markers. A total number of 10 accessions, collected from different ecoclimatic zones of India, were screened with 43 random decamer primers to evaluate polymorphism. Selected 10 primers generated 125 bands, 76 of which were found to be polymorphic. The amplification products ranged from 9-14 bands for different primers. Each primer produced on an average 12.5 bands per primer, of which 7.6 were polymorphic. UPGMA method was used to construct dendrogram and genetic distance matrix was determined using squared Euclidean distances. Cluster analysis of data using UPGMA algorithm placed the ten accessions into two main clusters, the genetic dissimilarity matrix between genotypes ranged from 0-8 and the principal component analysis placed the 10 accessions into three groups.

Keywords:    Cluster analysis, DNA fingerprinting, genetic diversity, Jatropha curcas , PCR, principal component analysis, RAPD

 

Indian Journal of Biotechnology

Vol 9 July 2010, pp 289-297

 

Biotization with Piriformospora indica and Pseudomonas fluorescens improves survival rate, nutrient acquisition, field performance and saponin content of micropropagated Chlorophytum sp .

 

S K Gosal 1 , A Karlupia 1 , S S Gosal 2 *, I M Chhibba 3 and A Varma 4

1 Department of Microbiology, 2 School of Agricultural Biotechnology & 3 Department of Soils

Punjab Agricultural University, Ludhiana, 141 004, India

4 Amity Institute of Microbial Technology, Amity University, Noida, 201 303, India

Received 20 April 2009; revised 4 November 2009; accepted 20 January 2010

Micropropagated plantlets usually exhibit high mortality rate upon their transfer to soil as a result of transplantation shock caused by abiotic and biotic stresses and weak root system in the absence of beneficial microflora. Biotization of micropropagated Chlorophytum sp . with the fungus, Piriformospora indica and the bacterium, Pseudomonas fluorescens, improved plantlet survival rate, growth parameters, field performance, P content and the micronutrient acquisition. Biotized plants showed root colonization of P. indica in cortical cells of roots and exhibited the presence of pear shaped spores and hyphae as well as rhizospheric colonization of P. fluorescens. Microbial biotization enhanced plant survival up to 91.2% by dual inoculation over uninoculated control (78.8%), on transfer from laboratory to green house. Biotized field grown plants exhibited increase in root length, number of lateral roots, shoot dry weight, leaf length, number and dry weight of fleshy roots in dual inoculation at P F which were significantly better over single as well as uninoculated control. Plants inoculated with P. indica exhibited maximum chlorophyll content (8.76 mg g - 1 ) while maximum P content (0.26%) was observed in dual inoculated plants, which was at par with P. indica alone even at low phosphorus . Higher saponin content was observed with both, P. indica alone as well as dual inoculations. Maximum acquisition of Cu (40 µg g -1 ) was observed in P. indica inoculated plants at P F level, which was at par with dual inoculation. In dual inoculated plants, the highest contents of Fe (4405 µg g -1 ), Zn (135 µg g -1 ) and Mn (160 µg g -1 ) were observed. Thus, microbial biotization improved survival and nutrient uptake by micropropagated Chlorophytum plants.

Keywords:    Chlorophytum , micropropagation, micronutrient, Piriformospora indica , Pseudomonas fluorescens , saponin

 

Indian Journal of Biotechnology

Vol 9, July 2010, pp 298-301

 

Genotyping by ERIC-PCR of Escherichia coli isolated from bovine mastitis cases

Vivek Prabhu 1 , S Isloor 1 *, M Balu 2 , V V S Suryanarayana 3 and D Rathnamma 1

1 Department of Microbiology, Veterinary College, KVAFSU, Hebbal, Bangalore 560 024, India

2 Project Directorate on Animal Disease Monitoring and Surveillance, Hebbal, Bangalore 560 024, India

3 Molecular Virology Laboratory, Indian Veterinary Research Institute, Hebbal, Bangalore 560 024, India

Received 1 September 2009; revised 24 December 2009; accepted 25 February 2010

Mastitis is an important problem in dairy farms and pathogen Escherichia coli has a world-wide importance. In the present study, authors have shown that E. coli strains isolated from bovine mastitis cases could be differentiated using a PCR with enterobacterial repetitive intergenic consensus sequences (ERIC) primers. In all, 40 strains of E. coli from bovine mastitis cases were subjected for ERIC-PCR. Of these, 37 showed amplicons ranging from 350 to >3000 bp. The PCR profile generated showed polymorphism in 37 strains. An intense amplicon of 1300 bp was seen in all the strains, except E. coli O27 (code M10) and O69 (M33). Based on ERIC-PCR profiles, of 37 E. coli strains, 22 were found to be distributed among 4 genotypes, whereas each of the remaining 15 strains showed unique genotypic pattern. The study emphasizes the utility of ERIC-PCR in intraserotype differentiation of strains based on their genotype and, thus, it is complimentary to serotyping. Furthermore, it was possible to differentiate strains of the same serotype into different genotypes. PCR amplification with ERIC primers was a fast and reliable method for differentiation and identification of E. coli strains. The advantage of this method compared to serotyping is the fact that different genotypes could be found even in strains within the same serotype or in untypable strains

Keywords: Bovine mastitis, Escherichia coli , ERIC-PCR, strain differentiation

 

Indian Journal of Biotechnology

Vol 9, July 2010, pp 302-307

 

Siderophore production by Alcaligenes faecalis and its application for growth promotion in Arachis hypogaea

R Z Sayyed 1 * N S Gangurde, P R Patel, S A Joshi and S B Chincholkar 2

1 Department of Microbiology, PSGVP Mandal's S I Patil Arts, G B Patel Science and STSKVS Commerce College, Shahada, Dist Nandurbar, 425 409, India

2 Department of Microbiology, School of Life Sciences, North Maharashtra University, Jalgaon 425 001, India

Received 16 June 2009; revised 19 November 2009; accepted 29 January 2010

A rhizobacterium isolated from groundnut rhizosphere and identified as Alcaligenes faecalis BCCM 2374 produced siderophore during 24 h submerged growth in modified succinic acid medium (SM) prepared in tap water without deferration. For the optimized production of siderophore in the presence of starch and sucrose, the optimum parameters found were as follows: slight acidic p H (5.5), 35 o C temperature and 1% inoculum. While alkaline p H (8.5), 37 o C temperature, 5% inoculum in the presence of fructose, xylose and glucose gave optimum growth, succinic and citric acids, NH 4 SO 4 , NH 4 NO 3 and NH 4 Cl supported growth as well as siderophoregenesis. Amino acids also showed varying effects on growth and siderophore production. Inoculation of A. faecalis enhanced seed germination (8.75%), root length (9.35%), shoot length (16%) and chlorophyll content (8.0%) in Arachis hypogaea over control treatment under pot culture conditions.

Keywords : Alcaligenes faecalis , Arachis hypogaea , siderophore, optimization, groundnut

 

Indian Journal of Biotechnology

Vol 9, July 2010, pp 308-312

 

Studies on biphasic biomethanation of spoiled mango puree

R D Kirtane*, P C Suryawanshi, A B Chaudhari 1 and R M Kothari #

Jain Hi-Tech Agri Institute, Jain Irrigation Systems Ltd., Jalgaon 425 001, India
1 School of Life Sciences, North Maharashtra University, Jalgaon 425 001, India

Received 23 September 2008; revised 14 September 2009; accepted 15 November 2009

In a pilot-scale (175 L) study, spoiled mango puree (8.0-12.0% total solids) was subjected to biphasic biomethanation. For this purpose, an efficient phase separation between puree hydrolysis/acidification and its methanogenesis was achieved by using three acidogenic reactors (ARs) in series, operated aerobically at ambient temperature. The hydrolysate coming out of the AR series contained high percentage of volatile fatty acids (VFAs, 15,000-20,000 mg L -1 ), indicating efficient conversion of sugars. Subsequently, this hydrolysate was fed to two methanogenic reactors (MRs), operated anaerobically at 35±1.0°C. About 30% digestate from MRs was further recycled to ARs for enhancing the efficiency of hydrolytic activity. This arrangement permitted the maintenance of optimum p H in ARs at 4.5-5.5 range and an efficient anaerobic digestion in MRs as indicated by p H of the digestate in 7.2-7.8 range.

The optimized conditions permitted, (i) an increase in feedstock initially from 2.0 to 10.0 L/d, (ii) methanogenic transformation of 15,000-20,000 mg L -1 VFAs, (iii) generation of 100±20 L d -1 biogas containing 65% methane initially and 550±25 L d -1 biogas with 78% methane upon stabilization over 180 d, (iv) digestate leaving MRs with 600-1200 mg L -1 VFAs, and (v) around 90% efficiency in terms of VFAs utilized.

Keywords: Acidification, biphasic methanogenesis, methane-rich biogas, spoiled mango puree

 

 

Indian Journal of Biotechnology

Vol 9, July 2010, pp 313-318

 

Polishing of biomethanated spent wash (primary treated)
in constructed wetland: A bench scale study

P K Singh 1 , K P Sharma 1 *, Shweta Sharma 2 , R C Swami 1 and Subhasini Sharma 2

1 Departments of Botany and 2 Zoology, University of Rajasthan, Jaipur 302 004, India

Received 16 June 2009; revised 24 November 2009; accepted 28 January 2010

This paper describes polishing of primary treated diluted spent wash (COD = 4000 - 10,000 mg/L) in the batch fed downflow constructed wetlands ( surface area = 615 cm 2 , depth = 27.5 cm ) filled 3/4 th with a mixture (1:3) of coarse river sand and stone grit (length=2.0-3.0 cm, width=1.0-1.5 cm) and planted with Phragmites karka . The concentrated spent wash (COD > 5000 mg/L) was found toxic to shoots resulting in their etiolation in dose dependent manner. The reduction in COD (54-63%) and BOD (58-70%) values and their loads (COD = 78-98%, BOD = 81-95%) in the effluents were significant and independent of influent concentrations whereas colour removal (34-82%) was concentration dependent. Despite of the marked reduction in pollutant loads, effluents failed to meet discharge standards that limits wetland scope merely for reducing spent wash volume (COD = < 5000 mg/L) after which effluents may further be treated conventionally.

Keywords: Constructed wetland, COD, BOD, duckweed assay, fish assay, spent wash

 

Indian Journal of Biotechnology

Vol 9, July 2010, pp 319-324

 

Plant regeneration from alginate-encapsulated somatic embryos of
Dalbergia sissoo Roxb.

Ajay Kumar Singh 2 and Suresh Chand 1 *

Plant Tissue Culture and Genetics Research Group, School of Life Sciences, Devi Ahilya University,
Khandwa Road, Indore 452 017, India

Received 16 April 2009; revised 6 October 2009; accepted 26 December 2009

A method has been developed for plant regeneration by encapsulation of somatic embryos obtained from callus cultures derived from semi-mature cotyledon explants of Dalbergia sissoo Roxb. (family Fabaceae). Embryogenic callus was developed from cotyledon pieces on Murashige and Skoog (1962) medium supplemented with 9.04 m M 2,4-dichlorophenoxyacetic acid and 0.46 m M kinetin. The somatic embryos were induced from embryogenic callus on hormone free ½-MS medium with 2% sucrose. Cotyledonary stage somatic embryos were encapsulated using sodium alginate (2.5%) and calcium chloride (75 m M ) as gelling matrix. The highest frequency (43.3%) for conversion of encapsulated somatic embryos into plantlets was achieved on ½-MS medium with 2% sucrose. Plantlets with well developed shoots and roots were established in pots containing autoclaved mixture of peat moss and soil. Under ex vitro conditions, the conversion of encapsulated somatic embryos into plantlets was also achieved when these were directly sown in autoclaved peat moss moistened with ½-MS0 medium.

Keywords: Dalbergia sissoo , encapsulation, legume tree, somatic embryogenesis, synthetic seeds

 

Indian Journal of Biotechnology

Vol 9, July 2010, pp 325-328

 

Intraspecific variation in the internal
transcribed spacer (ITS) regions of rDNA in Withania somnifera (Linn.) Dunal

 

Bilal Ahmad Mir 1 , Sushma Koul 1 *, Arun Kumar 1 ,
Maharaj K Kaul 1 , Amarjit Singh Soodan 2 and
Soom Nath Raina 3

1 Biodiversity and Applied Botany Division, Indian Institute of Integrative Medicine, Jammu 180 001, India

2 Department of Botanical and Environmental Sciences, Guru Nanak Dev University, Amritsar, India

3 Cellular and Molecular Cytogenetics Laboratory, University of Delhi, Delhi 110 007, India

Received 2 June 2009; revised 1 December 2009;
accepted 5 February 2010

Intraspecific variation in ITS regions of the rDNA among the five wild and five cultivated genotypes of Withania somnifera , were evaluated at nucleotide sequence level using restriction fragment length polymorphism (RFLP). The entire internal transcribed spacer (ITS1-5.8S-ITS2) region was first amplified by PCR and then cleaved with four different restriction enzymes (EcoRV, Hinf I, Afa I & Hae III). R estriction endonuclease digests, types, and sequence length composition of ITS 1 and ITS 2 of nuclear ribosomal DNA provided discrete differences between the cultivated and wild genotypes. A 710 bp single amplified product was obtained in all the five wild genotypes whereas, two ITS bands named as ITS type A and B of 709 bp and 552 bp, respectively were obtained in the five cultivated genotypes. A single deletion at 672 position was noted in ITS type A of cultivated genotypes. There was no restriction site in 552 bp ITS band for all the four restriction enzymes used. The variation of ITS at amplification as well as digestion level is in conformity with morphological and phytochemical differences in W. somnifera genotypes .

Keywords: Withania somnifera , RFLP, internal transcribed spacer (ITS), intraspecific variation

 

Indian Journal of Biotechnology

Vol 9, July 2010, pp 329-332

 

Direct organogenesis from hypocotyl explants of Psoralea corylifolia L.—An endangered medicinal plant

 

P Baskaran* and N Jayabalan

Department of Plant Science, School of Life Sciences, Bharathidasan University, Tiruchirappalli 620 024, India

Received 31 July 2009; revised 1 December 2009;
accepted 2 February 2010

A protocol has been developed for in vitro regeneration of Psoralea corylifolia L. through adventitious shoot regeneration from hypocotyl explants cultured on Murashige and Skoog (MS) medium supplemented with various combinations and concentrations of plant growth regulators. The highest response was obtained on MS medium containing 3.0 µ M benzyladenine (BA), 1.0 µ M naphthalene acetic acid (NAA), 5.0 µ M ascorbic acid (AA) and 100 mg L -1 casein hydrolysate (CH). The regenerated shoots were rooted on half-strength MS basal medium supplemented with 3.0 µ M indole-3-butyric acid (IBA). Rooted shoots were effectively transferred to garden soil, farmyard soil and sand (2:1:1 ratio) mixture. Subsequently, the plants were successfully acclimatized. During acclimatization, the plants were irrigated with 50 mL one-eighth strength MS basal salt solution devoid of sucrose and inositol, but supplemented with 3.0 µ M IBA and 100 mg L –1 bavistin (BVN). The plants were subsequently established in the field.

Keywords: Hypocotyl, plant regeneration, Psoralea corylifolia

 

Indian Journal of Biotechnology

Vol 9, July 2010, pp 333-335

 

Micropropagation of Verbesina encelioides—An invasive weed

Satish C Jain 1 *, Renuka Jain 2 & Renu Singh 1

1 Medicinal Plants and Biotechnology Laboratory,
Department of Botany and 2 Department of Chemistry,
University of Rajasthan, Jaipur 302 004, India

Received 10 July 2008; revised 29 March 2010;
accepted 3 May 2010

An efficient protocol for in vitro micropropagation of V. encelioides (Cav.) Benth. & Hook. fil. ex Gray , a medicinal weed, through hypocotyl segments of seedlings was developed on Murashige and Skoog's (MS) medium supplemented with various concentrations of a-naphthalene acetic acid (NAA, 0.50-15.00 mg/L), 6-benzylaminopurine ( BAP, 1.00-3.00 mg/L) and kinetin(Kn, 0.20-0.60 mg/L). Highly organized callus induction and proliferation were achieved on MS medium+NAA (1.00 mg/L)+BAP (3.00 mg/L). Optimum shoot multiplication (13.00±0.28) was achieved on MS medium+NAA (0.50 mg/L)+BAP (1.00 mg/L) with the shoot height of 1.80±0.15 cm. Similarly, efficient root induction (8.30±0.40) was achieved on MS medium+indole 3-acetic acid (IAA, 1.00 mg/L) with the root length of 2.60± 0.34 cm as 83% of the microshoots produced roots. These plantlets were hardened in a controlled plant growth chamber, prior to ex-vitro transfer. No phenotypic differences were observed among the regenerates.

Keywords : Callusing, micropropagation, plant regeneration, Verbesina encelioides

 

Indian Journal of Biotechnology

Vol 9, July 2010, pp 336-337

 

 

Somatic embryogenesis from leaf explants of soapnut (Sapindus mukorossi Gaertn.)

 

N S Philomina*

Department of Botany, Yogi Vemana University, Kadapa 516 003, India

Received 5 October 2009; revised 27 November 2009; accepted 25 January 2010

Plant regeneration through somatic embryogenesis has been developed in an economically important forest tree, Sapindus mukorossi Gaetin. Calli obtained by culturing young leaf explants on MS medium containing growth regulators, 2,4-D (6.7 m M )
and Kn (9.0 m M ), when subjected to reduced levels of 2,4-D (2.2 m M )+Kn (4.6 m M ), produced numerous somatic embryos. Somatic embryos developed into complete plantlets on MS medium devoid of growth regulators. The regenerated plantlets were successfully established in the soil with 90% survival frequency after a few days of acclimatization.

Keywords : Soapnut, somatic embryogenesis, callus, regeneration, Sapindus mukorossi