Indian Journal of Biotechnology

 

 

 

Total visitors:5,241 since 24-01-07

VOLUME 6

CODEN:IJBNAR 6(1) 1-132 (2007)

NUMBER 1

JANUARY 2007

ISSN:0972-5849

 

CONTENTS

 

Papers

 

 

Phylogenetic analysis of divergent structural organization of nucleotide binding domain encoded by resistance genes and gene homologs in the family Fabaceae

9

 

        IPC Code: Int. Cl.8 C12N15/09, 15/29

 

 

        J Basak, S Kundu & A Pal

 

 

 

 

 

Assessment of genetic diversity among some Indian wheat cultivars using random amplified polymorphic DNA (RAPD) markers

18

 

        IPC Code: Int. Cl.8 C12N15/10

 

 

        Sapna Grewal, Pushpa Kharb, Rekha Malik, Sunita Jain, & R K Jain

 

 

 

 

 

Multiple sequence analysis of polygalacturonases and invertases and phase shift in conserved motifs

24

 

        IPC Code: Int. Cl.8 C12N9/26, 9/38, 9/40

 

 

        P Palanivelu

 

 

 

 

 

Standardization of different parameters for ‘particle gun’ mediated genetic transformation of sugarcane (Saccharum officinarum L.)

31

 

        IPC Code: Int. Cl.8 A01H15/00; C12N15/9

 

 

        Ajinder Kaur, M S Gill, Raman Gill & S S Gosal

 

 

 

 

 

Assessment of canine distemper virus infection in vaccinated and unvaccinated dogs

35

 

        IPC Code: Int. Cl.8 C12N15/11, 15/12, 15/45

 

 

        D Latha, S R Srinivasan, P S Thirunavukkarasu, L Gunaselan, P Ramadass &
R B Narayanan

 

 

 

 

 

Genotypic characterization of infectious bronchitis viruses from India

41

 

        IPC Code: Int. Cl.8 C12N15/11, 15/13, 15/50

 

 

        K Suresh Kumar, G Dhinakar Raj, A Raja & P Ramadass

 

 

 

 

 

Molecular characterization of Begomovirus infecting sweet pepper in Oman

45

 

        IPC Code: Int. Cl8 C12N15/11, 15/34

 

 

        Akhtar J Khan, Nadiya A Al-Saady, Madleen S Al-Mahruki, Muna Al-Oufi &
Ali M Al-Subhi

 

 

 

 

 

Nitrogen and phosphorous scavenging potential in microalgae

52

 

        IPC Code: Int. Cl.8 C02F3/32

 

 

        Nirbhay Kumar Singh & Dolly Wattal Dhar

 

 

 

 

 

Induction of sporulation by sulphate limitation in Nostoc ANTH, a symbiotic strain capable of colonizing roots of rice plants

57

 

        IPC Code: Int. Cl.8 C12N3/00

 

 

        Omarlin Kyndiah & Amar Nath Rai

 

 

 

 

 

Expression of an antimicrobial peptide (MSI-99) confers enhanced resistance to Aspergillus niger in transgenic potato

63

 

        IPC Code: Int. Cl.8  C12N15/09, 15/63

 

 

        T R Ganapathi, S B Ghosh, N H S Laxmi & V A Bapat

 

 

 

 

 

Synthesis of ethyl oleate employing synthetic hydrogel-immobilized lipase of Bacillus coagulans MTCC-6375

68

 

        IPC Code: Int. Cl.8 C12N9/20, C12N11/04

 

 

        S S Kanwar, R K Kaushal, M L Verma, Y Kumar, W Azmi, R Gupta,S S Chimni & G S Chauhan

 

 

 

 

 

Comparison of ethanol and temperature tolerance of Zymomonas mobilis strain in glucose and molasses medium

74

 

        IPC Code: Int. Cl.8 C12P7/06

 

 

        Parmjit S Panesar, Satwinder S Marwaha & John F Kennedy

 

 

 

 

 

Studies on bioemulsifier production from marine Streptomyces sp. S1

78

 

        IPC Code: Int. Cl.8 C12N9/20

 

 

        C R Kokare, S S Kadam, K R Mahadik & B A Chopade

 

 

 

 

 

Polyamines stimulate shoot bud proliferation and multiplication in Achras sapota grown in culture

85

 

        IPC Code: Int. Cl.8 A01H5/12

 

 

        S D Purohit, A Singhvi, R Nagori & S Vyas

 

 

 

 

 

Studies on varietal difference, tissue specificity and developmental variation of esterase and peroxidase isozymes in pearl millet [Pennisetum glaucum  (L.) R. Br.]

91

 

        IPC Code: Int. Cl.8 C12N9/08, 9/16

 

 

        K H Smila, M Johnson & M Rajasekarapandian

 

  

 

 

 

Initiation of embryogenic suspension culture and plant regeneration in onion (Allium cepa L.)

100

 

        IPC Code: Int. Cl.8 A01H4/00, 5/10

 

 

        S Tiwari, M K Tripathi, U K Khare & R Rana

 

 

 

         

 

Degradation of polyaromatic hydrocarbons by mixed culture isolated from oil contaminated soil—A bioprocess engineering study

107

 

        IPC Code: Int. Cl.8 B09C1/10

 

 

        Ruma Roy, Raja Ray, Ranjana Chowdhury & Pinaki Bhattacharya

 

 

Short Communications

 

 

Optimization of culture media and cultural conditions for the production of extracellular lipase by Bacillus coagulans

114

 

        IPC Code: Int. Cl.8 C12N9/20

 

 

        M P Prasanth Kumar & A K Valsa

 

 

 

 

 

Detection of a gene for iron containing superoxide dismutase in the terrestrial cyanobacterium Tolypothrix byssoidea

118  

 

        IPC Code: Int. Cl.8 C12N15/9, 15/11

 

 

        Siba P Adhikary & Gabriele Klug

 

 

 

 

 

Standardization of in vitro protocol in Chrysanthemum cv. Madam E Roger for development of quality planting material and to induce genetic variability using g-radiation

121 

 

        IPC Code: Int. Cl.8 A01H4/00, 5/02; C12N15/01

 

 

        Pratibha Misra & S K Datta

 

 

 

 

 

Direct regeneration from apical bud explants of Withania somnifera Dunal

125

 

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

 

 

        I Sivanesan

    

 

 

 

 

Instructions to Contributors

129

 

 

 

 


 

AUTHOR INDEX

 

 


Adhikary S P

118

 

Al-Mahruki M S

45

 

Al-Oufi M

45

 

Al-Saady N A

45

 

Al-Subhi A M

45

 

Azmi W

68

 

Bapat V A

63

 

Basak J

9

 

Bhattacharya P

107

 

Chauhan G S

68

 

Chimni S S

68

 

Chopade B A

78

 

Chowdhury R

107

 

Datta S K

121

 

Dhinakar Raj G

41

 

Ganapathi T R

63

 

Ghosh S B

63

 

Gill M S

31

 

Gill R

31

 

Gosal S S

31

 

Grewal S

18

 

Gunaselan L

35

 

Gupta R

68

 

Jain R K

18

 

Jain S

18

 

Johnson M

91

 

Kadam S S

78

 

Kanwar S S

68

 

Kaushal R K

68

 

Kaur A

31

 

Kennedy J F

74

 

Khan Akhtar J

45

 

Kharb P

18

 

Khare U K

100

 

Klug G

118

 

Kokare C R

78

 

Kundu S

9

 

Kyndiah O

57

 

Kumar Y

68

 

Latha D

35

 

Laxmi N H S

63

 

Mahadik K R

78

 

Malik R

18

 

Marwaha S S

74

 

Misra P

121

 

Nagori R

85

 

Narayanan R B

35

 

Pal A

9

 

Palanivelu P

24

 

Panesar P S

74

 

Prasanth Kumar M P

114

 

Purohit S D

85

 

Rai A N

57

 

Raja A

41

 

Rajasekarapandian M

91

 

Ramadass P

35, 41

 

Rana R

100

 

Ray R

107

 

Roy R

107

 

Singh N K

52

Singhvi A

85

Sivanesan I

125

Smila K H

91

Srinivasan S R

35

Suresh Kumar K

41

Thirunavukkarasu P S

35

Tiwari S

100

Tripathi M K

100

Valsa A K

114

Verma M L

68

Vyas S

85

Wattal Dhar D

52


 

 

Indian Journal of Biotechnology

Vol 6, January 2007, pp 9-17

 

 

Phylogenetic analysis of divergent structural organization of nucleotide binding domain encoded by resistance genes and gene homologs in the family Fabaceae

J Basak, S Kundu and A Pal*

In higher plants, disease resistance (R) genes are present in abundance and majority of these encode proteins with a nucleotide-binding site (NBS). N-terminal end of the NBS is either endowed with TIR or by Non-TIR sequences. We have cloned and sequenced one yellow mosaic virus-resistance linked R gene homolog (RGH) from Vigna mungo, line VM-1, GenBank accession number AY297425. Later, two other RGHs from YMV resistant lines, V. mungo WBU 108 and V. radiata Pusa 9072 were selectively amplified using R gene targeted degenerate primers and were cloned subsequently (AY301991 and trIQ7XZT9, respectively). Characterization of these three RGHs and analysis of a total of 221 R genes and RGHs lead to the question of the evolution and distribution of the R genes/RGHs in the family Fabaceae, to which the primary hosts of the YMV belong. These 14 species mainly comprised grain legumes and the R gene phylogeny were reconstructed. The phylogenetic analyses indicate that two-third of the sequences are of the TIR-NBS type, while about one third represent the Non-TIR subfamily. Simultaneous presence of the TIR and the Non-TIR domains within the Fabaceae indicates divergent evolution and heterogeneity within the NBS domain. The plausible mechanism of continued diversification of the NBS sequences within the family Fabaceae has been substantiated by the supportive published evidences. It is presumed that such cases of divergent architectures of NB-domains of R genes within a single species or in the cultivars of the species has been influenced considerably through human interference during domestication, as against, evolution and random selection in nature. This finding reflects that the successful introgression of the functional R gene could be possible to the disease susceptible cultivars within the tribes Phaseoleae and Trifoleae.

Keywords: disease resistance gene, Fabaceae, grain legume, nucleotide binding domain, gene phylogeny

IPC Code: Int. Cl.8 C12N15/09, 15/29

Indian Journal of Biotechnology

Vol 6, January 2007, pp 18-23

 

 

Assessment of genetic diversity among some Indian wheat cultivars using random amplified polymorphic DNA (RAPD) markers

Sapna Grewal, Pushpa Kharb*, Rekha Malik, Sunita Jain and R K Jain

Genetic relationship study was performed with RAPD primers among 20 Indian wheat accessions (17 hexaploids, 2 tetraploids and 1 diploid). A total of 372 bands were observed using 25 RAPD primers, of which 323 (86.8%) were polymorphic. The total number of amplification products per primer ranged from 5 (OPO-02) to 30 (OPH-01) with an average of 14.8±1.15 bands per primer. The size of PCR products ranged from 162 bp to 2529 bp. A significant corelation (0.933, p<0.01) was observed between the total number of bands and the number of polymorphic bands. Sixteen primers amplified 30 unique bands for specific genotypes that would prove to be highly useful in identification of genotype and designing future breeding strategy. The similarity coefficient values ranged from 0.52 to 0.82, indicating high genetic variability among the wheat genotypes. The 20 wheat genotypes were grouped into two major clusters. Cluster I had the diploid genotype T. urartu, tetraploid variety WH 896 and hexaploid variety PBW 175. Rest of the 16 hexaploid varieties and a tetraploid variety (Khapli) were placed in the other major cluster.

Keywords: wheat, RAPD, genetic diversity, PCR, varietal identification

IPC Code Int, Cl.8: C12 N15/10

Indian Journal of Biotechnology

Vol 6, January 2007, pp 24-30

 

 

Multiple sequence analysis of polygalacturonases and invertases and
phase shift in conserved motifs

P Palanivelu*

Multiple sequence analysis by programs, such as ClustalW and T-COFFEE, is one of the important tools in bioinformatics. These programs are used to predict conserved motifs among related or unrelated proteins and nucleic acids that are of functional importance. However, it is observed that the conserved motifs in protein sequence analysis from some organisms especially from archae bacteria, extremophilic organisms and fungi do not align with other conserved sequences all the time and the conserved motif(s) in these organisms may be slightly shifted, either side of the conserved motif aligned for all the other organisms. Even one protein sequence is not aligned with others; the ClustalW or T-COFFEE does not mark the whole block as the conserved motif even though the particular motif is conserved among others. This is a serious problem as the investigator may miss important and functional motif(s) because of such misalignments. But a careful manual analysis of the aligned sequences will solve such anomaly. This is substantiated with two examples; one from polygalacturonase sequences analysis and the other from invertase sequences analysis using ClustalW, T-COFFEE, DIALIGN and MAFFT algorithms.

Keywords: ClustalW, conserved motifs, DIALIGN, invertases, MAFFT, phase shift, polygalacturonases, T-COFFEE

IPC Code: Int. Cl.8 C12N9/26, 9/38, 9/40

Indian Journal of Bioteechnology

Vol 6, January 2007, pp 31-34

 

 

Standardization of different parameters for ‘particle gun’ mediated genetic transformation of sugarcane (Saccharum officinarum L.)

Ajinder Kaur, M S Gill, Raman Gill and S S Gosal*

Three different parameters, viz. quantity of DNA, size of target tissue and distance between microcarrier launch assembly and target tissue (firing distance), for ‘particle gun’ (Bio-Rad) mediated transformation were optimized using embryogenic calli of sugarcane (target tissue), gusA (reporter gene) and hpt (hygromycin resistance gene). Of 12, 18 and
24 µL of DNA (gusA and hpt) suspension used per two bombardments, highest (33.33%) GUS expression was observed with 18 µL of suspension carrying 1.88 µg DNA. Bombardments made into 2-5 mm calli exhibited 55.55% GUS expression, while no expression was obtained in 7.5-12 mm calli after 7 h of their incubation in X-gluc. Out of different firing distances, viz. 2.5, 5.0, 7.5 and 10.0 cm, maximum intensity of GUS expression was recorded when calli were bombarded at a distance of 5.0 cm from the microcarrier launch assembly in the ‘particle gun’.

Keywords: embryogenic calli, GUS assay, particle gun’ bombardment, sugarcane, transformation

IPC Code: Int. Cl.8 A01H5/00; C12N15/9

 

Indian Journal of Biotechnology

Vol 6, January 2007, pp 35-40

 

 

Assessment of canine distemper virus infection in vaccinated and unvaccinated dogs

D Latha, S R Srinivasan, P S Thirunavukkarasu, L Gunaselan, P Ramadass and R B Narayanan*

Canine distemper is the most prevalent viral disease of dogs, which has high mortality rate. Vaccination of dogs against canine distemper is the only measure to control the disease. The aim of the present study was to find out the effect of commercial live attenuated vaccines in the control of distemper and the importance of the annual vaccination against distemper based on the occurrence of infection. One hundred and sixty conjunctival samples, collected from dogs with clinical symptoms suggestive of canine distemper, were screened by Dot-Enzyme Linked Immunosorbent Assay (Dot-ELISA) for the presence of canine distemper virus.  The application of Dot-ELISA in screening the samples was validated using Indirect Immunofluorescence assay as the standard technique. Out of the 160 samples tested by Dot-ELISA, 112 (70%) were positive for canine distemper. The results of Dot-ELISA used for epidemiological study based on sex, age and vaccination status showed that dogs of both the sexes and 1-5 years of age were more susceptible to Canine distemper virus infection and there is a lack of regular annual vaccination of dogs which results in the high incidence of distemper in dogs of 1-5 years of age. The present study suggests that vaccination followed by regular annual boosters against distemper would effectively protect the dogs from the disease.

Keywords:  Dot-ELISA, canine distemper, canine distemper virus, vaccination

IPC Code: Int. Cl.8 C12N15/11, 15/12, 15/45

Indian Journal of Biotechnology

Vol 6, January 2007, pp 41-44

 

 

Genotypic characterization of infectious bronchitis viruses from India

K Suresh Kumar, G Dhinakar Raj*, A Raja and P Ramadass

Infectious bronchitis (IB) is an acute, highly contagious and economically important disease of chickens. In India, IB vaccinations are carried out using vaccines belonging only to the Massachusetts 41 (Ma41) serotype. However, the vaccine strain used may not always be protective and it becomes necessary to continuously monitor the viruses causing disease. There has been anecdotal evidence of IB outbreaks even in vaccinated flocks in India. Tissue suspensions were inoculated intra-allantoically into 11-day-old embryonated chicken eggs. The allanotic fluids collected at 48 h post-inoculation (PI) were used in reverse transcription-polymerase chain reaction (RT-PCR) to amplify a part of S1 gene. The partial S1 gene products obtained were sequenced. All the seven isolates had sequences with 94.8-98.8% homology with the vaccine strain, H120. The cross-neutralization tests were used to identify the serotype of the field isolates. All the four isolates showed greater than 50% antigenic relatedness value indicating their classification in Massachusetts’s serogroup. The commercial vaccines conferred protection to the viruses isolated.

Keywords: infectious bronchitis virus, chickens, genotyping, antigenic relatedness, protectotyping

IPC Code: Int. Cl.8 C12N15/11, 15/13, 15/50

Indian Journal of Biotechnology

Vol 6, January 2007, pp 45-51

 

 

Molecular characterization of Begomovirus infecting sweet pepper in Oman

Akhtar J Khan*, Nadiya A Al-Saady, Madleen S Al-Mahruki, Muna Al-Oufi and Ali M Al-Subhi

Whitefly transmitted tomato yellow leaf curl is one of the most devastating viral disease of cultivated sweet pepper (Capsicum frutescens grossum) and other vegetables in Oman. Infected sweet pepper plants showed typical begomovirus symptoms as upward leaf curling, interveinal and leaf chlorosis, and growth stunting. Begomovirus infecting sweet pepper in Oman was detected by polymerase chain reaction (PCR) using begomovirus specific degenerate primers (PAL1v1978/PAR1c496 and AV494/AC1048). Core region (74-604 bp) of coat protein gene of the begomovirus was amplified by PCR with tomato yellow leaf curl virus (TYLCV) specific degenerate primers (TycpV369/TycpC1023). Core region of coat protein gene contains highly conserved regions and is used to identify the begomovirus infecting sweet peppers. Virus identification was performed by percent sequence identity and parsimony analysis using core coat protein gene sequences of sweet pepper virus with complete genome, core region of coat protein and coat protein gene sequences from reference begomoviruses. The core region sequence identity of coat protein gene of sweet pepper virus from Oman was 92.2, 96.5, 94.0, 93.8, and 96.5% with TomGV-Lebanon, TYLCV-Guadeloupe, TYLCV-Israel, TYLCV-Kuwait, and TYLCV-Mexico, respectively. Phylogenetic trees and percent sequence identity with reference to begomoviruses permitted the identification of sweet pepper virus as TYLCV based on tree position and extent of sequence identity. Phylogenetic analysis revealed that sweet pepper tomato yellow leaf curl virus clustered with its closest relatives from Middle East regions but formed a separate strain.

Keywords: sweet pepper, begomovirus, core region of coat protein gene

IPC Code: Int. Cl.8 C12N15/11, 15/34

Indian Journal of Biotechnology

Vol 6, January 2007, pp 52-56

 

 

Nitrogen and phosphorous scavenging potential in microalgae

Nirbhay Kumar Singh* and Dolly Wattal Dhar

Growth potential and N and P scavenging ability was examined in microalgal strains grown under secondary treated sewage effluent. The mean dry weight and the pigments were highest in microalgae under standard BG-11 medium in comparison to those grown in sewage effluent. There was a marked reduction in available nitrogen and phosphorous with the growth of microalgae in the sewage water. A significant correlation coefficient between N and P removal and dry weight and pigments has indicated the usefulness of sewage effluent for cultivation of microalgae with the efficient N and P scavenging ability. Chlorella vulgaris was most efficient in scavenging ammonical nitrogen while nitrate-scavenging ability was highest in Oscillatoria. All the microalgal genera also removed available phosphorous efficiently.

Keywords: nutrients, scavenging, potential, microalgae, nitrogen, phosphorus

IPC code: Int. Cl. 8 C02F3/32

 

Indian Journal of Biotechnology

Vol 6, January 2007, pp 57-62

 

 

 

Induction of sporulation by sulphate limitation in Nostoc ANTH, a symbiotic strain capable of colonizing roots of rice plants

Omarlin Kyndiah and Amar Nath Rai*

Nostoc ANTH is a symbiotically compatible cyanobacterium that associates with rice plants and carries out associative N2-fixation. Investigations were carried out to induce profuse sporulation in the cyanobacterium for use as inocula in rice paddies. Impacts of pH and temperature changes, addition of various carbon sources and limitation of phosphate and sulphate on akinete formation were studied. Among these, only phosphate and sulphate limitation induced akinete formation in Nostoc ANTH. Under both the conditions all vegetative cells eventually became akinete. However, induction of akinete differentiation was quicker and resulted in more profuse akinetes differentiation in response to sulphate limitation than to phosphate limitation. These akinetes showed long-term viability (upto 5 years) and excellent germination frequency
(90-95 %). This is the first report on induction of akinete formation by sulphate limitation.

Keywords: Nostoc ANTH, cyanobacteria, akinetes, akinete differentiation, sulphate limitation

 

Indian Journal of Biotechnology

Vol 6, January 2007, pp 63-67

 

 

Expression of an antimicrobial peptide (MSI-99) confers enhanced resistance to Aspergillus niger in transgenic potato

T R Ganapathi, S B Ghosh, N H S Laxmi and V A Bapat*

MSI-99, a synthetic substitution analogue of antimicrobial peptide magainin, was used to impart enhanced resistance against fungal pathogen, Aspergillus niger in transgenic potato cultivars, Kufri Jyothi and Kufri Bahar. Internodal stem segments were transformed with Agrobacterium tumefaciens strain EHA105. Integration of the transgene, MSI-99 in the transgenic plants of both the cultivars was confirmed by PCR and PCR-Southern analysis, exhibiting 180 bp fragment and hybridization with labelled probe pSAN168. Bioassay studies using detached leaves of transgenics showed the enhanced resistance against A. niger.

Keywords: antimicrobial peptides, Aspergillus niger, MSI-99, transgenic potato

IPC Code: Int. Cl.8 C12N15/09, 15/63

Indian Journal of Biotechnology

Vol 6, January 2007, pp 68-73

 

 

Synthesis of ethyl oleate employing synthetic hydrogel-immobilized lipase of Bacillus coagulans MTCC-6375

S S Kanwar*, R K Kaushal, M L Verma, Y Kumar, W Azmi, R Gupta, S S Chimni and G S Chauhan

Ten polymeric hydrogels were chemically synthesized by varying the concentrations of copolymer (DMA) and cross-linker (MBAm) molecules. An alkaline lipase of Bacillus coagulans MTCC-6375 was immobilized onto a poly
(MAc-co-DMA-cl-MBAm)-hydrogel support at pH 8.5 and temperature 55oC in 16 h. The bound lipase possessed 7.6 U.g-1 (matrix) lipase activity with a specific activity of 18 U.mg-1 protein. Hydrogel bound-lipase catalyzed esterification of oleic acid and ethanol to synthesize ethyl oleate in n-nonane. Various kinetic parameters were optimized to produce ethyl oleate using immobilized lipase. The optimal parameters were bound enzyme/substrate (E/S) ratio 0.62 mg/mM, ethanol/oleic acid 100 mM:75 mM or 100 mM:100 mM, incubation time 18 h and reaction temperature 55
°C that resulted in approximately 53% conversion of reactants into ethyl oleate in n-nonane. However, addition of a molecular sieve to the reaction mixture promoted the conversion to 58% in 18 h in n-nonane, which was equivalent to 55 mM of ethyl oleate produced.

Keywords:   Poly (MAc-co-DMA-cl-MBAm)-immobilized lipase, synthetic hydrogel lipase, Bacillus coagulans MTCC-6375, ethyl oleate synthesis

IPC Code: Int. Cl.8 C12N9/20, C12N11/04

 

Indian Journal of Biotechnology

Vol 6, January 2007, pp 74-77

 

 

Comparison of ethanol and temperature tolerance of Zymomonas mobilis
strain in glucose and molasses medium

Parmjit S Panesar, Satwinder S Marwaha and John F Kennedy*

The effect of temperature and exogenous ethanol was evaluated on the ethanol production potential of Zymomonas mobilis MTCC 2428 as an alternative to the classic and widespread use of Saccharomyces cerevisiae. At 30°C, this strain produced the maximum amounts of ethanol, i.e. 4.0 and 3.3% (v/v) after utilizing 98 and 83% (w/v) sugar on glucose and molasses medium, respectively. While at higher temperatures (35 & 40°C), the ethanol production by the strain decreased. Moreover, the strain was able to operate in the presence of 8% (v/v) ethanol in glucose medium at 30°C; whereas it tolerated only 6 and 2% (v/v) ethanol at 35 and 40°C, respectively. At the respective temperatures, further higher concentrations of the supplemented ethanol were found to be inhibitory to the isolate.

Keywords: Zymomonas mobilis, glucose, molasses, ethanol

IPC Code: Int. Cl.8 C12P7/06

IPC Code: Int. Cl.8 C12N3/00

 

Indian Journal of Biotechnology

Vol 6, January 2007, pp 78-84

 

 

Studies on bioemulsifier production from marine Streptomyces sp. S1

C R Kokare*, S S Kadam, K R Mahadik and B A Chopade

Out of the 80 strains of actinomycetes isolated and screened from Alibag, Janjira and Goa coastal regions of India, 56 showed lipase activity. Six potential strains were studied for bioemulsifier production by using oils and hydrocarbons as substrates. Streptomyces sp. S1 isolated from Goa showed maximum bioemulsifier production of 200 EU/mL. It also showed significant growth on maltose yeast extract medium. The fermentation conditions were optimized. Maximum bioemulsifier production was obtained at an initial pH 7, temperature 28°C, 120 rpm, sodium chloride concentration 3% (w/v) and time 14 d. Bioemulsification activity was significant against growth media containing 1% (v/v) toluene (361.2 EU/mL). A medium containing 1% (v/v) toluene showed appreciable reduction in surface tension (42.6 dynes/cm). Critical micelle concentration (CMC) of purified composite bioemulsifier was 0.3 mg/mL. The bioemulsifier having 82% protein, 17% polysaccharide and 1% reducing sugar was unstable at 10 and 50°C; but was found to be stable at room temperature (28°C). The optimized fermentation process produced a bioemulsifier yield of 3.8 g/L.

Keywords: Streptomyces sp., bioemulsifier, surface tension, critical micelle concentration, toluene

IPC Code: Int. Cl.8 C12N9/20

Indian Journal of Biotechnology

Vol 6, January 2007, pp 85-90

 

 

Polyamines stimulate shoot bud proliferation and multiplication in Achras sapota grown in culture

S D Purohit*, A Singhvi, R Nagori and S Vyas

In vitro shoot bud proliferation and multiplication was stimulated in shoot clusters of Achras sapota when polyamines were incorporated in the Schenk and Hildebrandt’s (SH) medium in combination with 6-benzylaminopurine (BA). Of the three polyamines–putrescine (Put), spermidine (Spd) and spermine (Spm), Put proved to be highly stimulatory. With Put at 0.1 mM concentration in combination with 8.87 µM BA, a little less than three-fold rate of shoot multiplication could be achieved while BA alone could not evoke such response. The stimulatory effect of Put was followed by Spm. The involvement of polyamines in the above response was confirmed by suppression of process by use of their biosynthesis specific inhibitors–α-difluoromethylornithine (DFMO) and α-difluoromethylarginine (DFMA). DFMO at 1.0 mM concentration caused more effective suppression of shoot proliferation as compared to DFMA at the same concentration. The inhibitory effect could be reversed by incorporation of additional polyamines in the medium. Stimulated shoot bud proliferation (more than that obtained with Put) was also observed when GA3 was incorporated in place of polyamines in the medium. However, its consistent use was not found beneficial in the long run.

Keywords:Achras sapota var. Cricket Ball, axillary bud break, cotyledonary node, in vitro propagation, polyamines

IPC Code: Int. Cl.8 A01H5/12

 

 

Indian Journal of Biotechnology

Vol 6, January 2007, pp 91-99

 

 

Studies on varietal difference, tissue specificity and developmental variation of esterase and peroxidase isozymes in pearl millet [Pennisetum glaucum (L.) R. Br.]

K H Smila, M Johnson* and M Rajasekarapandian

Varietal difference, tissue specificity and developmental variation of four varieties of pearl millet [Pennisetum glaucum (L.) R. Br.], viz. ICMV 221, ICMV 155, IP 4021 and BK 560, were studied using isoenzymes analyses (esterases and peroxidases) at 3 different developmental stages (on 20th, 40th and 60th d for esterase and 25th, 50th and 75th d for peroxidases). A number of tissue specific isoforms (present in the root and leaf tissues of each varieties) of both esterases and peroxidases were observed. At each stage of development, different isozyme banding patterns were obtained, which implies the role of specific isoforms in differentiation and plant development. RM values of the isozymic banding pattern ranged from 0.04 to 0.99.

Keywords: esterase, isoenzymes, pearl millet, peroxidase

IPC Code: Int. Cl.8 C12N9/08, 9/16

Indian Journal of Biotechnology

Vol 6, January 2007, pp 100-106

 

 

 

Initiation of embryogenic suspension culture and plant regeneration in onion (Allium cepa L.)

S Tiwari*, M K Tripathi, U K Khare and R Rana

A rapidly growing and maintainable embryogenic suspension culture of Allium cepa L. was accomplished with two different approaches, either by culturing mature embryos directly or by transferring embryogenic callus cultures in liquid media. The cultures obtained were inundated with clumps of proliferating globular embryos with very little non-embryogenic tissues. The number and size of somatic embryo clumps was measured to quantify growth of embryogenic tissues under various conditions. The suspensions were subcultured every 15 d by replacing the old medium with an equal volume of fresh medium. Initiation and proliferation of such embryogenic suspension culture depended upon the genotypes, method of suspension culture initiation and various exogenous growth regulators supplemented to the culture medium at variable quantity. For the establishment of suspension cultures, the liquid MS media fortified with 4.0 mg L1 2,4-D in combination with 0.5 mg L-1 BAP was found to be the most effective. For subsequent subculturing, the reduced level of 2.0 mg L-1 2,4-D in combination with 0.5 mg L-1 BAP promoted faster development of embryos. Frequent and efficient plantlet regeneration occurred on MS medium with 0.5 mg.L-1 each of NAA, BAP and kinetin. Regenerated plants were found to be phenotypically normal and true to the type.

Keywords: Allium cepa, embryo culture, cell suspension culture, somatic embryo

IPC Code: Int. Cl.8 A01H4/00, 5/10

 

Indian Journal of Biotechnology

Vol 6, January 2007, pp 107-113

 

 

Degradation of polyaromatic hydrocarbons by mixed culture isolated from oil contaminated soil—A bioprocess engineering study

Ruma Roy, Raja Ray, Ranjana Chowdhury, Pinaki Bhattacharya*

In the present investigation, decomposition of polyaromatic hydrocarbons was studied using mixed culture. Mixed strains were isolated from soil of petrol stations of different Indian cities and the best performing strains from these sources were used for subsequent bioprocess study using simulated mixture of anthracene and naphthalene as carbon source in methanol solution. The cell growth curve and substrate depletion time history curve obtained from batch fermentative process show that the reaction engineering behaviour of the systems under study can well be represented by classical substrate uninhibited Monod’s model. In a separate attempt the intrinsic kinetic parameters mmax and KS were evaluated following differential analysis of experimental data.

Keywords: biodegradation, bioprocess engineering, isolation of bacterial strains; oil contaminated soil, polyaromatic hydrocarbons

IPC Code:     Int. Cl.8 1309 C1/10

Short Communications

 

Indian Journal of Biotechnology

Vol 6, January 2007, pp 114-117

 

 

Optimization of culture media and cultural conditions for the production of extracellular lipase by Bacillus coagulans

M P Prasanth Kumar and A K Valsa*

An extracellular lipase producing bacterium, identified as Bacillus coagulans based on the morphological and biochemical characters, was isolated from the soil. The maximum growth and lipase production was obtained after 72 h of bacterial culture. While the optimum pH and temperature for the production of lipases was 7 and 30oC, respectively. Peptone and olive oil were found to be the best nitrogen and carbon sources, respectively. Na+ ions stimulated lipase production than K+ and Mg++.

Keywords: Bacillus coagulans, carbon, cultural conditions, extracellular lipase, nitrogen.

IPC Code: Int. Cl.8 C12N9/20

Indian Journal of Biotechnology

Vol 6, January 2007, pp 118-120

 

 

Detection of a gene for iron containingsuperoxide dismutase in the terrestrial cyanobacterium
Tolypothrix byssoidea

Siba P Adhikary* and Gabriele Klug

Using oligonucleotide probes deduced from a conserved region of amino acids of SOD (superoxide dismutase) proteins, one SOD gene was detected in the cyanobacterium, Tolypothrix byssoidea, isolated from the exposed rock surface of Sun temple, Konark. Amplification of specific genes of the genomic DNA and sequencing of the PCR product yielded a sodB gene of 292 bp size. Southern hybridization confirmed the presence of a single sodB gene in T. byssoidea.

Keywords: FeSOD, sodB gene, southern analysis, cyanobacteria, Tolypothrix byssoidea, terrestrial habitat

IPC Code: Int. Cl.8 C12N9/20

 

Indian Journal of Biotechnology

Vol 6, January 2007, pp 121-124

 

 

Standardization of in vitro protocol in Chrysanthemum cv. Madam E Roger for development of quality planting material and to induce genetic variability using g-radiation

Pratibha Misra and S K Datta*

‘Madam E Roger’, a greenish white large flowered chrysanthemum cultivar- was selected for in vitro propagation and mutagenesis to induce further genetic variability. A protocol has been standardized to develop large-scale quality planting material for commercial exploitation. Shoot bud differentiation could be achieved in ray florets in the presence of 0.5 mg L-1
NAA+ 2.0 mg L-1 BA. The differentiated shoots were further proliferated in the same medium, rooted in the presence of
0.5 mg L-1 NAA and hardened to grow in the field. Genetic variability in the form of a solid mutant of yellow colour has been obtained from this cultivar, when the freshly inoculated ray florets were treated with 1.0 Gy
g-radiation dose.

Keywords: Chrysanthemum, in vitro mutagenesis, quality planting material, shoot differentiation, g-radiation, ray florets

IPC Code: Int. Cl.8 A01H4/00, 5/02; C12N15/01

 

Indian Journal of Biotechnology

Vol 6, January 2007, pp 125-127

 

 

Direct regeneration from apical bud explants of Withania somnifera Dunal

I Sivanesan*

Effecient multiplication of Withania somnifera was achieved through culture of shoots tips of mature plants on Murashige and Skoog (MS), Schenk and Hildebrandt (SH) and Gamborg (B5) media. However, MS medium was found superior to SH and B5. MS medium supplemented with BAP+IAA (each at 2.0 mg/L) was optimal for induction of shoot buds; whereas, MS supplemented with 0.3 mg/L GA3 was the most suitable for shoot elongation. Elongated shoots were rooted on half strength MS containing with 2.0 mg/L IBA. The rooted plantlets were successfully established in field.

Keywords: apical bud explants, direct regeneration, endangered plants, micropropagation, Withania somnifera

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