Indian Journal of Biotechnology

 

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VOLUME8

NUMBER4

OCTOBER2009

CODEN: IJBNAR 8(4) (2009) 343-476

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

 

  CONTENTS

 

 

Review

 

Retroviral vectors and gene therapy: An update

349

S K Maurya, Sushant Srivastava & R K Joshi

 

Papers

 

Promoter region polymorphism of CYP11B2 (344 C>T) gene in healthy volunteers of South Indian Tamilian population

358

S Rajan, P Ramu, D G Shewade & C Adithan

 

Transformation of tomato using biolistic gun for transient expression of the b-glucuronidase gene

363

D Ruma, M S Dhaliwal, Ajinder Kaur & S S Gosal

 

Genetic diversity and relationships among tea (Camellia sinensis) cultivars as revealed by RAPD and ISSR based fingerprinting

370

S C Roy & B N Chakraborty

 

Molecular characterization of Pestalotiopsis spp. associated with tea (Camellia sinensis) in southern India using RAPD and ISSR markers

377

Sarvottam D Joshi, R Sanjay, U I Baby & A K A Mandal

 

Development and molecular characterization of interspecific hybrids of Jatropha curcas ´
J. intergerrima

384

R S Dhillon, M S Hooda, M Jattan, V Chawla, M Bhardwaj & S C Goyal

 

Assessment of genetic diversity among Podophyllum hexandrum genotypes of the North-western Himalayan region for podophyllotoxin production

391

Md Afroz Alam, Pallavi Gulati, Aswini K Gulati, Gyan P Mishra & Pradeep K Naik

 

Amyloglucosidase from Rhizopus mold and b-glucosidase from sweet almond catalysed syntheses of riboflavinyl glycosides

400

Ramaiah Sivakumar &  Soundar Divakar

 

A comparative study on citric acid production kinetics of two Yarrowia lipolytica strains in two different media

408

S Karasu Yalcin, M T Bozdemir & Z Y Ozbas

 

Cost-effective fermentative production of calcium lactate using BISS (below Indian standard sugar) and Spirulina hydrolysate

418

A K Yadav, A B Chaudhari & R M Kothari

 

Studies on microbial transformation of albendazole by soil fungi

425

G Shyam Prasad, S Girisham & S M Reddy

 

Agrobacterium rhizogenes-mediated hairy root production in tea leaves [Camellia sinensis (L.) O. Kuntze]

430

K M Mariya John, Sarvottam D Joshi, A K A Mandal, S Ram Kumar & R Raj Kumar

 

Isolation of morphovariants through plant regeneration in Agrobacterium rhizogenes induced hairy root cultures of Plumbago rosea L.

435

K Satheeshkumar, Binoy Jose, E V Soniya & S Seeni

 

Induction of morphogenic callus and multiple shoot regeneration in Momordica cymbalaria Fenzl.

442

T D Nikam, S G Ghane, J N Nehul & R B Barmukh

 

Micropropagation of Dendrobium transparens L. from axenic pseudobulb segments

448

H Sunitibala & Rajkumar Kishor

 

Short Communications

 

In silico docking of ligand 3-hydroxy methyl xylitol with target protein ZnT-8 involved in type II diabetes

453

P Praveena & S Ignacimuthu

 

Clinical and pathological status of haemoglobinopathies among pregnant women in southern Orissa

456

Amrita Panda, B Praveen & Satpal Singh Bisht

 

List of Referees

459

Annual Author Index

465

Annual Subject Index

468

Instructions to Contributors

473

 

AUTHOR INDEX

 

 


Adithan C

358

Alam M A

391

 

 

Baby U I

377

Barmukh R B

442

Bisht S S

456

Bozdemir M T

408

 

 

Chakraborty B N

370

Chaudhari A B

418

 

 

Dhaliwal M S

363

Divakar S

400

 

 

Ghane S G

442

Girisham S

425

Gosal S S

363

Gulati A K

391

Gulati P

391

 

 

Ignacimuthu S

453

 

 

John K M M

430

Jose B

435

Joshi R K

349

Joshi S D

377

Joshi S D

430

 

 

Kaur A

363

Kishor R

448

Kothari R M

418

Kumar R R

430

Kumar S R

430

 

 

Mandal A K A

377

Mandal A K A

430

Maurya S K

349

Mishra G P

391

 

 

Naik P K

391

Nehul J N

442

Nikam T D

442

 

 

Ozbas Z Y

408

 

 

Panda A

456

Prasad G S

425

Praveen B

456

Praveena P

453

 

 

Rajan S

358

Ramu P

358

Reddy S M

425

Roy S C

370

Ruma D

363

 

 

Sanjay R

377

Satheeshkumar K

435

Seeni S

435

Shewade D G

358

Sivakumar R

400

Soniya E V

435

Srivastava S

349

Sunitibala H

448

 

 

Yadav A K

418

Yalcin S K

408


 

Review

 

Indian Journal of Biotechnology

Vol 8, October 2009, pp 349-357

Retroviral vectors and gene therapy: An update

S K Maurya*, Sushant Srivastava and R K Joshi

Department of Veterinary Biochemistry, College of Veterinary Science and Animal Husbandry
Narendra Deva University of Agriculture and Technology, Kumarganj, Faizabad 224 229, India

Received 20 February 2008 ; revised 3 February 2009 ; accepted 17April 2009

Gene therapy aims at treatment of diseases by transfer of genetic material into specific cells of a patient. So the transduction of appropriate target cell is critical. Retroviruses infect nearly every cell in the target population and become integrated in host cell genome for a stable expression. All retroviruses have only three structural genes gag, pol and env, except for lentiviruses where two other genes tat, rev and four accessory genes are required (vif, vpu, nif, vpr). There are two components of retroviral vector system—i) Packaging cell lines, which provide the products of gag, pol & env genes but are unable to package itself as they lack the y sequence, and ii) retroviral vectors where gag, pol and env are deleted but y sequence along with LTR is present. Several other types of vectors are also described.

Keywords: Gene therapy, retroviruses, vector

 

 

Papers

 

Indian Journal of Biotechnology

Vol 8, September 2009, pp 358-362

Promoter region polymorphism of CYP11B2 (344 C>T) gene in healthy volunteers of South Indian Tamilian population

S Rajan, P Ramu, D G Shewade and C Adithan *

Pharmacogenomics Laboratory, Department of Pharmacology, JIPMER, Pondicherry 605 006, India

Received 7 October; revised 18 March; accepted 25 May 2009

CYP11B2 gene encodes aldosterone synthase enzyme, which produces aldosterone. The objective of study was to establish the allele and genotype frequency of CYP11B2 (344 C>T) gene polymorphism in healthy volunteers of South Indian Tamilian population. Authors carried out the study in 424 unrelated healthy volunteers including both male and female subjects. Genotyping was done by PCR-RFLP method. The observed genotype and allele frequencies were compared with other major ethnic groups. The influence of blood pressure was also analyzed with respect to the genotypes. The allele frequency was 39.9 and 601% for wild type C allele and variant T allele, respectively. The diastolic blood pressure was found to be significantly higher in the variant TT homozygous genotypes in the total study subjects and also in the male subjects (p<0.05). Interethnic difference in the genotype and allele frequencies of CYP11B2 gene polymorphism was observed in the South Indian Tamilian population.

Keywords: Aldosterone, blood pressure, CYP11B2, genotype, polymorphism

Indian Journal of Biotechnology

Vol 8, October 2009, pp 363-369

Transformation of tomato using biolistic gun for transient expression of the
β-glucuronidase gene

D Ruma1, M S Dhaliwal1, Ajinder Kaur2 and S S Gosal2*

1Department of Vegetable Crops and 2Department of Plant Breeding, Genetics and Biotechnology,
 Punjab Agricultural University, Ludhiana 141 004, India

Received 24 March 2008; revised 19 March 2009; accepted 25 May 2009

We report for the first time, conditions for the biolistic transformation of tomato for the introduction of β-glucuronidase gene (gusA) into different explants viz., shoot tips, hypocotyls and cotyledons of genotype IPA-3. The respective per cent plant regeneration in different explants was 95.16, 79.30 and 90.14% on MS media supplemented with BAP (2.0 mg L-1) and Kn (1.0 mg L-1). The influence of physical parameters of particle gun on rates of transient GUS expression have been investigated viz., quantity of DNA, distance between the microcarrier launch assembly and target tissues; and the biological factors associated with the target tissues i.e., effect of osmoticum (mannitol), pre-bombardment culture period and post-bombardment culture period. Maximum GUS expression of 25.00, 33.00 and 22.20% was respectively recorded when 10-d-old shoot tips, hypocotyls and cotyledons were bombarded with 18 mL of DNA suspension. A firing distance of 7.5 cm was found to be most suitable recording 34.12, 36.56 and 22.69% transient GUS expression in shoot tips, hypocotyls and cotyledons, respectively. Addition of osmoticum into the culture medium reduced per cent GUS expression significantly in all the explants even at 1 molar concentration. Pre-culture of explants prior to bombardment also deduced the transformation efficiency, however post bombardment culture period of one day resulted in maximum GUS expression in all the explants.

Keywords: Lycopersicon esculentum, biolistic transformation, GUS assay, cotyledons, shoot tips, hypocotyls,
β-glucuronidase gene

Indian Journal of Biotechnology

Vol 8, October 2009, pp 370-376

Genetic diversity and relationships among tea (Camellia sinensis) cultivars as revealed by RAPD and ISSR based fingerprinting

S C Roy* and B N Chakraborty

DRS Department of Botany, University of North Bengal, PO-NBU, Siliguri 734 013, India

Received 29 September 2008; revised 18 March 2009; accepted 15 May 2009

Genomic fingerprinting in 21 tea genotypes was carried out using 7 ISSR and 12 RAPD primers. Polymorphism was 88.54% in ISSR fingerprinting, but was 77.77% in case of RAPD based fingerprinting. Dendrogram was constructed on the basis of genetic similarity matrix using the UPGMA algorithm, which showed three clusters (China type, Assam type and Cambod type). The genetic diversity over all groups (HT) on an average was 0.38, diversity within populations (HS) was 0.27, and genetic differentiation (GST) between populations over all loci was 0.25. The China variety had shown the largest within-group diversity (HS = 0.285-0.291), while the Cambod tea had the least diversity (HS = 0.193-0.207) and moderate diversity existed in Assam tea (HS = 0.223-0.241). Interpopulation gene flow [Nm = 0.5(1 – GST)/GST] was 0.76; Nm < 1.0 shows the limited genetic exchange among populations.

Keywords: Dendrogram, genetic diversity, gene flow, RAPD-ISSR fingerprinting, tea

Indian Journal of Biotechnology

Vol 8, October 2009, pp 377-383

Molecular characterization of Pestalotiopsis spp. associated with tea
(Camellia sinensis) in southern India using RAPD and ISSR markers

Sarvottam D Joshi, R Sanjay, U I Baby and A K A Mandal*

UPASI-TRF, Tea Research Institute, Nirar Dam BPO, Valparai 642 127, India

Received 24 September 2008; revised 17 February 2009; accepted 23 April 2009

The genus Pestalotiopsis is second most important fungal pathogen causing grey blight disease in tea plant
(Camellia sinensis). Due to grey blight disease, total tea crop loss is estimated to be 17% in southern India. In the present study, 42 Pestalotiopsis isolates were collected from five different regions in southern India. Among them, 22 isolates showed diverse morphological characters like colour, size and length of conidia and virulence. Genetic diversity was studied for these 22 isolates using two molecular marker systems (RAPD and ISSR). In RAPD, a total of 255 loci were generated and all were polymorphic in nature and the band size ranged from 0.2 to 3.0 kb. In ISSR, 194 amplified loci were observed and all were polymorphic as like RAPD and the band size ranged from 0.25 to 3.2 kb. Using Jaccard’s similarity coefficient matrix, highest similarity of 95.9% and 92.6% was observed between AP-8 and AP-9 isolates in both RAPD and ISSR markers, respectively. Lowest similarity was observed between AP-14 and NP-5 (8.8%) in RAPD matrix but in ISSR matrix lowest similarity (18.5%) was between AP-14 and EN-5. The UPGMA clustering of both methods was comparable. The results indicate that, within southern India, the diversity of Pestalotiopsis was high both morphologically and genetically.

Keywords: Camellia sinensis; Pestalotiopsis, RAPD, ISSR, phylogenetic, PCR, tea

 

Indian Journal of Biotechnology

Vol 8, October 2009, pp. 384-390

Development and molecular characterization of interspecific hybrids
of Jatropha curcas × J. integerrima

R S Dhillon1*, M S Hooda1, M Jattan2, V Chawla2, M Bhardwaj2 and S C Goyal3

1Department of Forestry, 2Department of Genetics and 3Department of Botany, CCS Haryana Agricultural University
Hisar 125 004, India

Received 23 September 2008; revised 5 March 2009; accepted 6 May 2009

Reciprocal crosses between J. curcas and J. integerrima were attempted. The interspecific hybrids were successful only when the former was used as seed parent. The F1 hybrids exhibited a wide range of variation for vegetative and reproductive traits suggesting considerable heterogeneity between the parental plants. Some of the characters such as stem type, branching habit, leaf size and shape of the hybrids resembled the female parent while others like leaf pigmentation, fruit and seed size, flower colour, resembled the male parent. The parents and their 23 hybrid plants were subjected to random amplified polymorphic DNA (RAPD) analysis using 61 random decamer primers. Out of 61 primers, the amplification was obtained in 51 primers producing 384 bands. The similarity value between the parents (41.4%) was less than those between parents and hybrids. Cluster analysis based on Jaccard’s similarity coefficient outgrouped the female parent from rest of the genotypes at a similarity coefficient of 0.55. The 2-Dimensional and 3-Dimensional scaling by principal component analysis (PCA) grouped some hybrids with male parent, some with female parent while others in between both the parents.

Keywords: Jatropha, interspecific hybrids, RAPD markers

Indian Journal of Biotechnology

Vol 8, October 2009, pp 391-399

Assessment of genetic diversity among Podophyllum hexandrum genotypes of the North-western Himalayan region for podophyllotoxin production

Md Afroz Alam1, Pallavi Gulati1, Aswini K Gulati2, Gyan P Mishra1 and Pradeep K Naik1*

1Department of Bioinformatics and Biotechnology, Jaypee University of Information Technology, Waknaghat, Solan 173 215, India 2Himachal Pradesh Forest Department, Talland, Shimla 171 001, India

Received 22 August 2008; revised 15 January 2009; accepted 20 March 2009

Podophyllum hexandrum (Indian Mayapple) is an important medicinal plant valued all over the world. Genetic diversity among the 28 genotypes of P. hexandrum distributed in 11 geographical regions from Himachal Pradesh (a part of the North-western Himalaya) was analyzed using RAPD markers. The genetic diversity was high among the genotypes as measured by percentage of polymorphic bands (PPB = 92.37%) and Shanon information index (I = 0.50). The mean coefficient of gene differentiation (Gst) was 0.69, indicating that 33.77% of the genetic diversity resided within the genotypes. Analysis of molecular variance indicated that the source of variation among the groups was 53% and among the genotypes of groups was 47%. An overall value of mean estimated number of gene flow (Nm = 0.22) indicated that there was limited gene flow among the genotypes. The existence of variation among the 28 genotypes as observed through podophyllotoxin content proved to be coupled with geographical altitude (r = 0.92) and local ecological conditions (temperature, rainfall, humidity, soil pH, etc.) but not on genetic basis (r = -0.55). Based on the observed genetic variations among the genotypes of Podophyllum, we recommend for their in situ conservation and germplasm collection expeditions in future conservation plans.

Keywords: Podophyllum hexandrum, podophyllotoxin, genetic diversity, RAPD

Indian Journal of Biotechnology

Vol 8, October 2009, pp. 400-407

Amyloglucosidase from Rhizopus mold and b-glucosidase from sweet almond catalysed syntheses of riboflavinyl glycosides

Ramaiah Sivakumar and Soundar Divakar*

Fermentation Technology and Bioengineering Department, Central Food Technological Research Institute, Mysore 570 020, India

Received 1 May 2008; revised 16 January 2009; accepted 20 March 2009

Riboflavinyl glycosides were synthesized enzymatically using amyloglucosidase isolated from Rhizopus mold and b-glucosidase from sweet almond to produce more water soluble riboflavinyl derivatives. The reaction conditions were optimized in terms of incubation period, pH, buffer, enzyme and substrate concentrations. With D-glucose, D-galactose, D-mannose, D-ribose, sucrose, maltose and lactose, both the enzymes gave yields in the range of 5-40%. Amyloglucosidase catalysis gave C1a and C1b glycosides of D-glucose, D-galactose, D-mannose, D-ribose, maltose, C6-O-arylated products of D-glucose and maltose and C1-O-arylated product of sucrose. However, b-glucosidase gave C1b-glucoside, C1b-lactoside and C1a and C1b glycosides of D-galactose and D-mannose. Riboflavinyl glycosides showed angiotensin converting enzyme inhibitory activities, with IC50 values in the 0.8±0.04-2.08±0.1 mM range.

Keywords: Amyloglucosidase, carbohydrates, b-glucosidase, regioselectivity, riboflavin, riboflavinyl glycosides, Rhizopus, sweet almond

Indian Journal of Biotechnology

Vol 8, October 2009, pp 408-417

A comparative study on citric acid production kinetics of two
Yarrowia lipolytica strains in two different media

S Karasu Yalcin1, M T Bozdemir2 and Z Y Ozbas3*

1Food Engineering Department, Faculty of Engineering and Architecture, Abant Izzet Baysal University, Golkoy 14280, Bolu, Turkey

2Chemical Engineering Department and 3Food Engineering Department, Faculty of Engineering, Hacettepe University
Beytepe 06532, Ankara, Turkey

Received 4 November 2008; revised 25 February 2009; accepted 24 April 2009

Microbial production of citric acid was performed by Yarrowia lipolytica NBRC 1658 and a domestic strain
in the media containing glucose or fructose as substrates. The study was carried out in a batch system. The non-competitive substrate inhibition model was proposed for growth of the yeasts in both substrate media. Higher dry mass and lower Ks values were obtained in glucose media when compared to fructose media. The highest citric acid concentration (65.1 g/L) was obtained with the domestic strain in the medium containing 200 g/L fructose. In the media containing fructose, maximum citric acid concentration and productivity values determined with the domestic strain were aproximately two-fold greater than those obtained with NBRC 1658 strain. The required initial concentration of glucose or fructose at which best citric acid production properties were observed changed between 100-200 g/L for both of the strains. The ratio of citric acid to isocitric acid was found to change between 11.70-16.62.

Keywords: Batch fermentation, citric acid, growth kinetics, production kinetics, Yarrowia lipolytica, yeast

 

Indian Journal of Biotechnology

Vol 8, October 2009, pp 418-424

Cost-effective fermentative production of calcium lactate using BISS
(below Indian standard sugar) and Spirulina hydrolysate

A K Yadav, A B Chaudhari and R M Kothari1*

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

Received 6 October 2008; revised 23 February 2009; accepted 4 May 2009

Batch fermentation of BISS (below Indian standard sugar) in combination with Spirulina hydrolysate (SH) has been successfully explored at 50 L scale to replace costly glucose and yeast extract (YE) medium as a source of complex nutrients to produce calcium lactate economically by using Lactobacillus delbruckii. This has led to optimization of a protocol for cost-effective production of calcium lactate. Reduction in duration of fermentation has potential to provide additional benefits of (i) more batches per unit capital investment and time, (ii) less consumption of utilities, and (iii) higher output per unit labour cost.

Keywords:   Lactobacillus delbruckii, BISS, calcium lactate, Spirulina hydrolysate

Indian Journal of Biotechnology

Vol 8, October 2009, pp 425-429

Studies on microbial transformation of albendazole by soil fungi

G Shyam Prasad1, S Girisham* and S M Reddy

Department of Microbiology, Kakatiya University, Warangal 506 009, India

Department of Pharmaceutics, Kakatiya Institute of Pharmaceutical Sciences, Mdl-Hasanparthy (Dist Warangal), India

 

Received 8 September 2008; revised 5 March 2009; accepted 3 May 2009

Soil enrichment technique was followed to isolate the fungi capable of performing biotransformation of albendazole. Among the 5 fungi isolated, Aspergillus fumigatus, A. niger and Penicillium chrysogenum could transform albendazole to one metabolite and Fusarium moniliforme could transform albendazole to two metabolites. The transformation was confirmed by HPLC. Based on LC-MS-MS analysis, the metabolites formed were predicted to be albendazole sulfoxide and albendazole sulfone. The results support that the soil enrichment is a promising technique for isolation of fungi with industrial applicability, viz. production of active metabolites from drugs.

Keywords: Albendazole sulfoxide, albendazole sulfone, biotransformation, fungi

Indian Journal of Biotechnology

Vol 8, October 2009, pp 430-434

Agrobacterium rhizogenes-mediated hairy root production in
tea leaves [Camellia sinensis (L.) O. Kuntze]

K M Mariya John, Sarvottam D Joshi, A K A Mandal*, S Ram Kumar and R Raj Kumar

Plant Physiology Division, UPASI Tea Research Foundation, UPASI Tea Research Institute
Nirar Dam BPO, Valparai 642 127, India

Received 24 September 2008; revised 17 February 2009; accepted 1 May 2009

Leaves of tea [Camellia sinensis (L.) O. Kuntze] plants were transformed with Agrobacterium rhizogenes strain (MTCC 532) and hairy roots were induced. Among the different concentrations of acetosyringeone tested, 300 µM/L was found to enhance the transformation frequency upto 70%. Murashige and Skoog (MS) medium supplemented with 30 g/L maltose and indole-3-acetic acid (IAA) at 5 mg/L was found suitable for hairy-root culture and accumulation of phenolic compounds. Confirmatory studies were carried out by PCR analysis using rol C gene primer for transformation. Amplification of the specific gene was noted in the transformant at 540 bp. High performance liquid chromatography (HPLC) analysis confirmed the higher levels of catechins and their fractions in roots arising from the transformed tissues. Catechin fractions, viz., EC, ECG, EGC and EGCG were detected both in untransformed leaves and hairy roots produced by Agrobacterium infected cells.

Keywords: Agrobacterium rhizogenes, Camellia sinensis, catechin, hairy root, rol C gene, secondary metabolites

Indian Journal of Biotechnology

Vol 8, October 2009, pp 435-441

Isolation of morphovariants through plant regeneration in Agrobacterium rhizogenes induced hairy root cultures of Plumbago rosea L.

K Satheeshkumar1*, Binoy Jose1, E V Soniya2 and S Seeni1

1Plant Biotechnology Division, Tropical Botanic Garden and Research Institute, Palode, Thiruvananthapuram 695 562, India

2Rajiv Gandhi Center for Biotechnology, Poojappura, Thiruvananthapuram 695 014, India

 

Received 21 August 2008.; revised 20 January 2009; accepted 25 March 2009

In vitro raised shoots of Plumbago rosea L. were infected with A4 strain of Agrobacterium rhizogenes to initiate hairy root formation, which produced 3.0±0.33 hairy roots per incision on explants in 20 d incubation. Southern blot analysis confirmed the integration of T-DNA into the genome of the roots. The hairy roots were cultured on MS agar medium supplemented with 2.0 mg/L BAP to induce the formation of shoots (3.2±0.24) of 0.2-0.4 cm length in 7-8 wks. Isolated shoots were multiplied through sub culturing in the presence of 0.5 mg/L BAP and the resultant shoots were subjected to combined elongation (3.29±0.16 cm) and rooting (12.6±0.57) in a medium supplemented with 0.1 mg/L IBA. The rooted plants were invariably abnormal with short internodes and wrinkled leaves showing 34.5% establishment in 2 months after transplantation in polybags and rearing under 75% sunlight in a shade house. Out of 38 plants transferred to the field, 20 (52.6%) survived and grew over 10-month period to reveal variations in morphological and growth characters. The growth of all the hairy root-derived plants was slow to varied extent, 13 among them retaining actively growing hairy roots and shoots having short internodes and expanded leaves, one with emerging normal shoot and a tuberous root in the midst of the abnormal shoot and hairy roots and the rest with hairy roots as well as 1-2 normal roots and abnormal shoots having wrinkled leaves. The biomass production ability of the transformed plants contributed by foliage and root characters was poor compared to plants raised from the nodal explants of normal shoot cultures. Southern blot analysis of DNA further confirmed the presence of bacterial T-DNA in these established plants in the field after 10 months. The demonstrated hairy root regeneration system including field establishment of the plants may be useful for scoring new variations especially in non-seed setting P. rosea.

Keywords: Agrobacterium rhizogenes, Plumbago rosea, hairy roots, morhovariants, regeneration

 

Indian Journal of Biotechnology

Vol 8, October 2009, pp 442-447

Induction of morphogenic callus and multiple shoot regeneration
in Momordica cymbalaria Fenzl.

T D Nikam1*, S G Ghane1, J N Nehul2 and R B Barmukh3

1Department of Botany, University of Pune, Pune 411 007, India
2Dada Patil Rajale College, Adinathnagar (Dist Ahmednagar) 414 505, India
3Modern College of Arts, Science and Commerce, Shivajinagar, Pune 411 005, India

Received 9 July 2008; revised 2 February 2009; accepted 5 April 2009

An in vitro propagation protocol for a wild vegetable and an ethnomedicinal plant, Momordica cymbalaria Fenzl., has been developed. The influences of 0.0-5.0 mM 6-BA, 0.0-20.0 mM Kn alone and in combination with 2.5- 5.0 mM IAA,
2.5-5.0
mM NAA and 2.5-5.0 mM 2,4-D on in vitro multiple shoot production from node, internode and leaf explants was studied. The maximum number of indirect (callus interspersed) regeneration of multiple shoots (9.0±0.5 shoots per explant) was achieved from leaf explants on MS medium enriched with 2.5 mM BA alone. Further, large-scale shoot
formation (35±3.4 shoots per culture) was achieved by repeated subculturing of leaf-callus on shoot regeneration medium (MS+2.5
mM BA). The capacity of large-scale shoot regeneration remained constant in the callus over a period of 2 years. The best root induction (100%) and survival (88%) was achieved on hormone free half strength MS medium. Addition of IAA or NAA in rooting medium induced callus formation in the shoots. Field established plants showed uniform growth and were morphologically identical to the parental stock.

Keywords: Callus, ethnomedicinal, in vitro propagation, Momordica cymbalaria, vegetable

Indian Journal of Biotechnology

Vol 8, October 2009, pp 448-452

Micropropagation of Dendrobium transparens L. from
axenic pseudobulb segments

H Sunitibala* and Rajkumar Kishor

Medicinal Plants and Horticultural Resources Division, Institute of Bioresources and Sustainable Development
Takyelpat, Imphal 795 001, India

Received 1 September 2008; revised 14 January 2009; accepted 2 April 2009

An efficient protocol for micropropagation of Dendrobium transparens L. using the axenic pseudobulb segments, derived from in vitro germinated seedlings, was developed. The immature embryos from 120-d-old capsules after pollination were first germinated on half-strength Murashige and Skoog (MS) basal medium supplemented with 1 mg L-1 of naphthalene acetic acid (NAA) + 2 mg L-1 of benzyl aminopurine (BAP). For multiple shoot induction, the axenic nodal segments excised from 120-d-old seedlings were cultured on the medium containing different concentrations and combinations of NAA and BAP. The multiplied shoots were then transferred to half-strength MS medium supplemented with 1 mg L-1 of indole-3 acetic acid (IAA) or indole-3 butyric acid (IBA) or NAA. Best response for shoot multiplication was achieved when the medium was supplemented with 1 mg L-1 NAA and 2 mg L-1 BAP, while that for rooting with 1 mg L-1 IAA. The plantlets were hardened off and more than 90% of survived plants were released to the shade house. The plantlets flowered 1 yr after transplantation.

Keywords: Axenic pseudobulb segment, benzyl aminopurine, Dendrobium transparens, in vitro, microshoot

 

Short Communications

 

ndian Journal of Biotechnology

Vol 8, October 2009, pp 453-455

 

In silico docking of ligand 3-hydroxy methyl xylitol with target protein ZnT-8 involved in type II diabetes

P Praveena and S Ignacimuthu*

Entomology Research Institute, Loyola College
Chennai 600 034, India

Received 2 April 2008; revised 23 February 2009;
 accepted 25 March 2009

ZnT-8 protein has been localized in insulin secretory granules. An automated molecule model for the target protein
ZnT-8 was generated at Swiss 3D modeling. Ramachandran plot was used to determine the angles. ADMET properties were calculated for the ligand, 3-hydroxy methyl xylitol. Pre ADMET hyperchem tool was used to find the QSAR properties of the ligand. In silico docking of ligand 3-hydroxy methyl xylitol with target protein ZnT-8 involved in type II diabetes was carried out to assess the efficacy of the ligand in binding to the target. The study indicated high affinity between the ligand and the target protein, suggesting that 3-hydroxy methyl xylitol is a good drug to control blood glucose levels.

Keywords: In silico docking, ligand, target protein, type II diabetes Indian Journal of Biotechnology

 

Indian Journal of Biotechnology

Vol 8 October 2009, pp 456-457

Clinical and pathological status of haemoglobinopathies among pregnant women in southern Orissa

Amrita Panda, B Praveen and Satpal Singh Bisht*

Department of Biotechnology, Roland Institute of Pharmaceutical Sciences, Berhampur 760 010, India

Received 24 April 2008; revised 6 March 2009;
accepted 5 May 2009

Sixty-two pregnant women were categorized into four age groups and investigated to know the prevalence of haemoglobinopathies among them in and around Berhampur using sickling test, naked eye single tube red cell osmotic fragility test (NESTROFT), and haemoglobin electrophoresis. Out of the
62 pregnant women four cases of sickle cell trait and one of
β-thalassaemia trait was found. There was no significant difference recorded in the blood cell indices between normal and sickle cell trait in pregnant women. Sickle cell haemoglobinopathy is prevalent among the general, scheduled caste followed by other backward class groups of southern Orissa and less HbS% was observed among the pregnant women which indicate the probable interaction of sickle hemoglobin with
α- thalassemia.

Keywords: Electrophoresis haemoglobinopathies, sickle cell, NESTROFT