Indian J Exp Biol

JULY 2002

CODEN: IJEB (A6) 40(7) 749-860 (2002)

ISSN: 0019-5189

 

 

 

Indian Journal of Experimental Biology

http : // www.niscom.res.in; http : // www.bioline.org.br/

 

 

 

 

VOLUME 40

CODEN : IJEB (A6) 40(7) 749-860 (2002)

NUMBER 7

JULY 2002

ISSN : 0019-5189

 

CONTENTS

View Point

 

 

Novel parasite (nematode) transglutaminase  Potential drug target

R Chandrashekar

753

 

 

 

 

Review Articles

 

 

Role of rhizobial biosynthetic pathways of amino acids, nucleotide bases and vitamins in symbiosis

Gursharn S Randhawa & Raad Hassani

 

755

 

 

 

 

Validation of traditional claim of Tulsi, Ocimum sanctum Linn. as a medicinal plant

S K Gupta, Jai Prakash & Sushma Srivastava

765

 

 

 

 

Papers

 

 

Localisation of identical organophosphorus pesticide degrading (opd) genes on genetically dissimilar indigenous plasmids of soil bacteria: PCR amplification, cloning and sequencing of opd gene from Flavobacterium balustinum

Sita Somara, Bramanandam Manavathi, Christoph C Tebbe &

Dayananda Siddavatam

 

 

774

 

 

 

 

Short-term androgen deprivation does not alter CaR and VDR mRNA expression in duodenal mucosa in male rats

S Tiwari, S K Gupta, M Mehrotra, G Agarwal , P K Awasthi & M M Godbole

 

780

 

 

 

 

Effect of phosphocreatine on H+ extrusion, pHi and dimorphism in Candida albicans

Nikhat Manzoor, M Amin & Luqman A Khan

785

 

 

 

 

Control of hyperglycemia and retardation of cataract by mulberry (Morus indica L.) leaves in streptozotocin diabetic rats

B Andallu & N Ch Varadacharyulu

 

791

 

 

 

 

Growth behaviour and bioproduction of indole acetic acid by a Rhizobium sp. isolated from root nodules of a leguminous tree Dalbergia lanceolaria

A C Ghosh & P S Basu

 

796

 

 

 

 

Secretion of ligninperoxidase by Penicillium citrinum, Fusarium oxysporum and

Aspergillus terreus

Meera Kumari, R S S Yadav & K D S Yadav

 

802

 

 

(Contd)

 

Effect of low level exposure of lead and cadmium on hepatic estradiol metabolism in female rats

Anil Pillai, Laxmipriya, Ami Rawal & Sarita Gupta

 

807

 

 

 

 

Screening of Indian plants for biological activity: Part XVI

A K Goel, D K Kulshreshtha, M P Dubey & S M Rajendran

812

 

 

 

 

Notes

 

 

Antimicrobial potentiality of a phenothiazine group of antipsychotic drug-prochlorperazine

Rupa Mazumder, Sharmila Ray Chaudhuri & Avijit Mazumder

828

 

 

 

 

Synergistic effect of ayurvedic pearl preparation on enhancing effectiveness of

antibiotics

Manisha Kulkarni, J Y Deopujari & H J Purohit

 

831

 

 

 

 

SEM study on cytotoxic effect of monocrotophos (MCP) on lungs of rat

Sangeeta, S M Handa & P K Mittal

835

 

 

 

 

Use of RAPD in assessing the genetic variability in Spodoptera litura

S Janarthanan, S Seshadri, K Kathiravan & S Ignacimuthu

839

 

 

 

 

Entomopoxvirus of cotton bollworm, Helicoverpa armigera (Hbn.)

K Narayanan

842

 

 

 

 

Mass production of polyhedral occlusion bodies of NPV of Helicoverpa armigera in relation to dose, age and larval weight

K Narayanan & S Jayaraj

 

846

 

 

 

 

In vitro regeneration of Acacia catechu Willd. from callus and mature nodal

explants  An improved method

Manisha Thakur, D R Sharma, Kamlesh Kanwar & Anil Kant

 

850

 

 

 

 

Cyanobacterial N2 fixation in presence of nitrogen fertilizers

A E Mekonnen, Radha Prasanna & B D Kaushik

854

 

 

 

 

Effect of lead on Na+, K+-ATPase activity in Penaeus indicus postlarvae

858

 

C Satyavathi & Y Prabhakara Rao

 

 

 

 

 

 

 

 

 

Indian Journal of Experimental Biology

Vol. 40, July 2002, pp. 753-754

 

 

 

View Point

 

Novel parasite (nematode) transglutaminase  Potential drug target

R Chandrashekar

 

 

Indian Journal of Experimental Biology

Vol. 40, July 2002, pp. 755-764

 

 

 

Review Articles

 

Role of rhizobial biosynthetic pathways of amino acids, nucleotide bases
and vitamins in symbiosis

Gursharn S Randhawa* & Raad Hassani

 

 

Rhizobia require the availability of 20 amino acids for the establishment of effective symbiosis with legumes. Some of these amino acids are synthesized by rhizobium, whereas the remaining are supplied by the host plant. The supply from plant appears to be plant-type specific. Alfalfa provides arginine, cysteine, isoleucine, valine and tryptophan, and cowpea and soybean provide histidine. The production of ornithine and anthranilic acid, the intermediates in the biosynthetic pathways of arginine and tryptophan, respectively, seems to be essential for effective symbiosis of Sinorhizobium meliloti with alfalfa. The expression of ilvC gene of S. meliloti is required for induction of nodules on the roots of alfalfa plants. An undiminished metabolic flow through the rhizobial pathways for the synthesis of purines and pyrimidines and the synthesis of biotin, nicotinic acid, riboflavin and thiamine by rhizobium appear to be requirements for normal symbiosis. To the best of our knowledge, this is the first review article on the role of rhizobial biosynthetic pathways of amino acids, nucleotide bases and vitamins in rhizobium-legume symbiosis. The scientific developments of about 35 years in this field have been reviewed.

 

Indian Journal of Experimental Biology

Vol. 40, July 2002, pp. 765-773

 

Validation of traditional claim of Tulsi, Ocimum sanctum Linn.
as a medicinal plant

S K Gupta*, Jai Prakash & Sushma Srivastava

 

In several ancient systems of medicine including Ayurveda, Greek, Roman, Siddha and Unani, Ocimum sanctum has vast number of therapeutic applications such as in cardiopathy, haemopathy, leucoderma, asthma, bronchitis, catarrhal fever, otalgia, hepatopathy, vomiting, lumbago, hiccups, ophthalmia, gastropathy, genitourinary disorders, ringworm, verminosis and skin diseases etc. The present review incorporates the description of O. sanctum plant, its chemical constituents, and various pharmacological activities.

 

 

Papers

Indian Journal of Experimental Biology

Vol. 40, July 2002, pp. 774-779

 

Localisation of identical organophosphorus pesticide degrading (opd) genes on genetically dissimilar indigenous plasmids of soil bacteria: PCR amplification, cloning and sequencing of opd gene from Flavobacterium balustinum

Sita Somara, Bramanandam Manavathi, Christoph C Tebbe & Dayananda Siddavatam

 

Received 19 November 2001; revised 28 February 2002

Plasmid borne organophosphorus pesticide degrading (opd) gene of Flavobacterium balustinum has been amplified using polymerase chain reaction (PCR) and the resulting PCR product (1.25 Kb) was cloned in pUC18. Further, a detailed restriction map was determined to PCR product and subcloned as overlapping restriction fragments. The nucleotide sequence was determined for all subclones to obtain complete sequence of PCR amplified fragment. The sequence showed 98% similarity to opd genes cloned from other soil bacteria isolated from diversified geographical regions. The protein sequence predicted from the nucleotide sequence was almost identical to parathion hydrolase, a triesterase involved in hydrolysis of triester bond found in variety of op-pesticides. The signal sequence of parathion hydrolase contained recently discovered twin arginine transport (tat) motif. It appears that tat motif plays a critical role in membrane targeting of parathion hydrolase.

 

 

Indian Journal of Experimental Biology

Vol. 40, July 2002, pp. 780-784

 

Short-term androgen deprivation does not alter CaR and VDR mRNA expression in duodenal mucosa in male rats

S Tiwari, S K Gupta, M Mehrotra, G Agarwal , P K Awasthi & M M Godbole

 

Received 4 December 2001; revised 26 February 2002

Androgen deprivation is associated with decline in intestinal calcium absorption. The effect of androgen on CaR and VDR intestinal mucosa has not yet been studied. Calcium homeostasis, areal bone mineral density (aBMD, dual energy X-ray absorptiometry) and expression of CaR and VDR mRNA in duodenal mucosa of orchidectomized (ORX) and sham operated (Sham) adult Sprague Dawley rats at 4 week have been studied. There was no significant difference in serum calcium, alkaline phosphatase, calcidiol and calcitriol levels between both the groups. Serum testosterone (T) (ng/dl) and inorganic phosphorous (iP) (mg/dl) levels were significantly lower in ORX rats. As compared to sham rats, ORX rats had significant decline in
in-vitro aBMD at proximal, middle and distal tibia, proximal, mid and distal femur and femoral neck (P < 0.05). Northern blot analysis revealed no significant alteration in the CaR and VDR mRNA expression in duodenal mucosa in ORX rats. CaR and VDR mRNA expression in duodenal mucosa is therefore, not affected by physiological concentrations of testosterone in rats.

 

Indian Journal of Experimental Biology

Vol. 40, July 2002, pp. 785-790

 

Effect of phosphocreatine on H+ extrusion, pHi and dimorphism
in Candida albicans

Nikhat Manzoor, M Amin & Luqman A Khan*

 

Received 13 July 2001; revised 26 December 2001

Candida albicans is an opportunistic pathogen. Its proliferation in human hosts is believed to be controlled by immunologic mechanisms. The plasma membrane of the fungus posseses an H+-ATPase (PM-ATPase) which actively extrudes protons to generate an electrochemical gradient which is used in co-transport of nutrients. This ATPase is associated with the growth, dimorphism and pathogenicity of the fungus. The physiological concentration of phosphocreatine (PCr) is 20-35 mM in skeletal muscles. H+-extrusion in Candida cells was strongly inhibited by PCr; 44% at 20mM and 69% at 40mM. H+-extrusion was stimulated 6.2-fold in the presence of 10mM glucose. This glucose stimulated extrusion was inhibited significantly by PCr; 36% at 20mM and 53% at 40mM. The intracellular pH pattern of cells destined to differentiate was greatly altered in the presence of PCr. Evagination time for control cells was between 90-120 min. PCr, delayed dimorphism, reduced the population of cells differentiating to hyphae and also reduced the length of hyphae after each time interval. Only 60% differentiation was observed with 10mM PCr and 40% for higher PCr concentration even after 210 min. Direct interaction of PM-ATPase and PCr has been demonstrated by difference spectrum measurement employing stopped flow spectrophotometer. It can be concluded that PCr may be playing a significant role in checking growth and pathogenesis of C. albicans.

 

Indian Journal of Experimental Biology

Vol. 40, July 2002, pp. 791-795

 

Control of hyperglycemia and retardation of cataract by mulberry
(Morus indica L.) leaves in streptozotocin diabetic rats

B Andallu

 

and

N Ch Varadacharyulu

 

Received 18 June 2001; revised 21 December 2001

Dried leaf powder of mulberry (M. indica L.) when given along with the diet at 25% level to streptozotocin induced diabetic male Wistar albino rats for 8 weeks, controlled hyperglycemia, glycosuria, albuminuria and retarded onset of retinopathy. Untreated diabetic rats showed hyperglycemia, glycosuria, albuminuria and developed lenticular opacity after 8 weeks of experimental period.

 

 

Indian Journal of Experimental Biology

Vol. 40, July 2002, pp. 796-801

 

 

Growth behaviour and bioproduction of indole acetic acid by a Rhizobium sp.
isolated from root nodules of a leguminous tree Dalbergia lanceolaria

A C Ghosh & P S Basu

 

Received 28 February 2001; revised 5 February 2002

The Rhizobium sp. isolated from healthy and mature root nodules of a leguminous tree, Dalbergia lanceolaria Linn.f., preferred mannitol and KNO3 for growth as carbon and nitrogen sources, respectively. The bacterium produced a high amount (22.3 mg/ml) of indole acetic acid (IAA) from L-tryptophan supplemented basal medium. Growth and IAA production started simultaneously. IAA production was maximum at 20 hr when the bacteria reached the stationary phase of growth. Cultural requirements were optimized for maximum growth and IAA production. The IAA production by the Rhizobium sp. was increased by 270.8% over control when the medium was supplemented with mannitol (1%,w/v), SDS (1mg/ml), L-asparagine (0.02%,w/v) and biotin (1mg/ml) in addition to L-tryptophan (2.5 mg/ml). The possible role of IAA production in the symbiosis is discussed.

 

 

Indian Journal of Experimental Biology

Vol. 40, July 2002, pp. 802-806

 

 

Secretion of ligninperoxidase by Penicillium citrinum, Fusarium oxysporum
and Aspergillus terreus

Meera Kumari, R S S Yadav & K D S Yadav

 

Received 8 May 2001; revised 12 February 2002

Secretion of ligninperoxidase [E.C.1.11.1.7] by Penicillium citrinum, Fusarium oxysporum and Aspergillus terreus in liquid culture growth medium has been demonstrated. Enzymatic characteristics like Km , pH and temperature optima using veratryl alcohol as the organic substrate of ligninperoxidases from above sources have been determined. Km values using veratryl alcohol as substrate for enzymes from P. citrinum, F. oxysporum and A. terreus were 69, 64 and 60M respectively. Km values using H2O2 as the variable substrate were 64, 72 and 80M.The pH optima were 4.0, 2.3 and 2.0 respectively. The values of temperature optima were 30, 25 and 22C for the enzymes from P. citrinum, F. oxysporum and A. terreus respectively.

 

 

Indian Journal of Experimental Biology

Vol. 40, July 2002, pp. 807-811

 

 

Effect of low level exposure of lead and cadmium on hepatic estradiol metabolism in female rats

Anil Pillai, Laxmipriya, Ami Rawal & Sarita Gupta

 

Received 9 April 2001; revised 8 February 2002

Toxic effect of metal cations on female reproduction and gonadal functions was studied. Adult synchronized female rats were treated intraperitoneally with lead acetate and cadmium acetate separately and in combination (0.025, 0.05 and 0.1 mg/kg body wt) for 15 days. The metabolizing enzymes (17b-hydroxy steroid oxidoreductase and UDP glucoronyl transferase) activities decreased with increasing dose showing significant change compared to control. Also, significant decrease in cytochrome P450 (CYP450) content was found after the treatment. Displacement of zinc bound to metallothionein was more in cadmium treated rats compared to other groups. In all these parameters, treatment in combination of lead and cadmium showed intermediate results indicating some kind of competition between the two metals. But the histological studies showed that combined treatment caused more cytotoxic effect than cadmium and lead alone. These results indicated that metal cations tested did have a direct inhibitory effect on metabolizing enzyme activities.

 

 

Indian Journal of Experimental Biology

Vol. 40, July 2002, pp. 812-827

 

Screening of Indian plants for biological activity*: Part XVI

A K Goel, D K Kulshreshtha, M P Dubey & S M Rajendran

 

Received 15 October 2001; revised 16 January 2002

Alcoholic extracts of 288 of plant materials from 199 plant species have been tested for various biological activities including chemotherapeutic and pharmacological screening. Biological activities, ranging from moderate to good degree, have been observed in 61 plants extracts. Follow up studies have been carried out in these extracts and some of them have shown moderate degree of activities at this Institute. However, none of the extracts was found to be good enough for further development. Results of the present studies, alongwith chemical investigations on different species of similar genera which were screened earlier, are also discussed.

 

 

Notes

Indian Journal of Experimental Biology

Vol. 40, July 2002, pp. 828-830

 

 

 

 

Antimicrobial potentiality of a phenothiazine group of antipsychotic
drug-prochlorperazine

Rupa Mazumder, Sharmila Ray Chaudhuri & Avijit Mazumder

 

Received 26 April 2001; revised 11 March 2002

The antipsychotic drug, prochlorperazine (Pcp), was tested for its antimicrobial efficacy against 103 strains belonging to both gram positive and gram negative bacteria. The drug was found to possess maximum activity against Staphylococcus aureus, Vibrio cholerae and Shigella spp. Pcp was moderately active against E.coli but most of the strains belonging to Bacillus spp, Klebsiella spp, Salmonella spp and Lactobacillus spp were found to be resistant to this drug. The drug was tested for its mode of antibacterial activity against Shigella dysenteriae 1 and it was found to be bacteriostatic in action. In in vivo studies, Pcp offered significant protection to Swiss albino mice at concentrations of 0.75 mg/g (P<0.01) and 1.5 mg/g (P<0.001) body weight when challenged with 50 median lethal dose of Salmonella typhimurium NCTC 74. Thus the result depicts that prochlorperazine may emerge as a strong antimicrobial drug to replace the conventional antibiotics and to overcome the problem of drug resistance.

 

 

Indian Journal of Experimental Biology

Vol. 40, July 2002, pp. 831-834

 

Synergistic effect of ayurvedic pearl preparation on enhancing effectiveness
of antibiotics

Manisha Kulkarni, J Y Deopujari & H J Purohit

 

Received 19 November 2001; revised 12 February 2002

Studies were carried out with ayurvedic preparations derived from pearl, which include preparations bhasma and pishti. The synergistic effect to reduce the dose of antibiotic was tested against E .coli the test bacterium with ampicillin antibiotic by bore well and disks diffusion methods. It was observed that pearl preparations do not show any antibacterial activity but when used at 200 mg/ml concentration with antibiotics, then even at sub-lethal dose, the antibiotic has effectively shown the results with reduced contact time. The protocol was also tested with the other bacteria like, Pseudomonas aeruginosa, Vibrio cholarae, Salmonella typhi, and Staphylococcus aureus and has shown similar results. The pearl bhasma synergistic effect was also tested with other antibiotics such as erythromycin, kanamycin, and ampicillin.

 

 

Indian Journal of Experimental Biology

Vol. 40, July 2002, pp. 835-838

 

SEM study on cytotoxic effect of monocrotophos (MCP) on lungs of rat

Sangeeta, S M Handa & P K Mittal

 

Received 23 November 2001; revised 19 February 2002

Monocrotophos (MCP) on oral administration (0.28 mg/100 g of body wt. i.e. 1/5th of LD50) to female rats for 15 and 30 days damaged alveolar walls lined by type II cells (great alveolar cells); clara cells (non-ciliated cells) lining bronchiolar epithelium; and emphysematous lesions due to loss of inter-alveolar walls. This led to increase in surface tension in lung due to decrease in secretion of surfactant as a result of necrosis of great alveolar cells and clara cells resulting in hypoxia. This effect was time dependent. In R group (15 days without pesticide after 30 days daily oral treatment), the toxic effects mentioned above still persisted which revealed non-repair of necrosis caused by MCP.

 

Indian Journal of Experimental Biology

Vol. 40, July 2002, pp. 839-841

 

 

Use of RAPD in assessing the genetic variability in Spodoptera litura

S Janarthanan, S Seshadri, K Kathiravan & S Ignacimuthu

 

Received 20 July 2001; revised 19 February 2002

A polymerase chain reaction based assay to distinguish six different ecotypes of the Spodoptera litura, a sporadic insect pest and a predominant defoliator of various crops was carried out. A total of 40 random primers were screened to reveal the existence of polymorphism between the populations. Among them eight showed scorable banding patterns and three primers (OPA-01, OPA-05, OPM-01) exhibited distinguishable banding patterns. However, Chengalpattu and Chennai populations revealed their closed relatedness and Coimbatore population stood distantly from others.

 

Indian Journal of Experimental Biology

Vol. 40, July 2002, pp. 842-845

 

 

Entomopoxvirus of cotton bollworm, Helicoverpa armigera (Hbn.)

K Narayanan

 

Received 10 September 2001; revised 16 November 2001

Occurrence of an Entomopoxvirus (EPV) from a lepidopteran insect viz;. cotton bollworm, H. armigera (HaEPV) along with gross pathological symptoms is reported for the first time in India. Histopathological study revealed that the fat body being the most favoured site of infection followed by haemocytes and gut epithelium. HaEPV was found to be not cross infective to six of the agricultural lepidopteran insect pests except for the potato black cutworm, Agrotis segetum registering 100% mortality showing typical symptom. Further, safety of HaEPV was shown against beneficial insect like mulberry silkworm, Bombyx mori and an useful insect general predator, Chrysoperla carnea.

 

Indian Journal of Experimental Biology

Vol. 40, July 2002, pp. 846-849

 

 

Mass production of polyhedral occlusion bodies of NPV of Helicoverpa armigera in relation to dose, age and larval weight

K Narayanan

 

and

S Jayaraj

 

Received 8 August 2001; revised 16 November 2001

A significant difference was noticed in the yield of polyhedral occlusion bodies (POBs) in various larval instars of H. armigera when three different doses of the nuclear polyhedrosis virus (NPV) were administered. The yield of POBs from a single larva ranged from 0.35  106 to 25033.33  106 with a mean of 18422.33  106 for fourth instar inoculated. Positive correlation existed between larval weight and number of POBs recovered. The regression analysis indicated POBs recovered responded with predictable manner to the weight of different larval instars and the various concentration of virus administered. The medium lethal time increased in the instars of the larva advanced with a minimum of 3.5 and maximum of 8 days in the first and fifth instars respectively.

 

 

Indian Journal of Experimental Biology

Vol. 40, July 2002, pp. 850-853

 

In vitro regeneration of Acacia catechu Willd. from callus and mature
nodal explants 
 An improved method

Manisha Thakur, D R Sharma, Kamlesh Kanwar & Anil Kant

 

Received 6 September 2001; revised 5 February 2002

Callus was derived from cultured cotyledons on MS medium supplemented with 2,4-D (0.25 mg/l) and NAA (0.25 mg/l). Plantlets were regenerated from the callus and nodal explants on MS medium containing BAP (2.0 mg/l) and Kn (2.0 mg/l), and further multiplied on the same medium. Addition of adenine sulphate (25.0 mg/l), ascorbic acid (20.0 mg/l) and glutamine (150.0 mg/l) in the medium resulted in enhanced axillary branching. Multiple shoots formed after 6 weeks were separated and subcultured in the fresh medium of same composition. For rhizogenesis, microshoots of 2.0-2.5 cm length were dipped in sterilized IAA solution (10 mg/l) for 24 hr followed by transfer to half strength MS medium containing activated charcoal (0.02%) resulting in rooting (75%) within 8 weeks. The rooted plants were transferred to pots containing sterilized soil and sand mixture for hardening and 71% survival was recorded. Fifty true to type plantlets of A. catechu could be obtained within seven months of culture establishment.

 

 

Indian Journal of Experimental Biology

Vol. 40, July 2002, pp. 854-857

 

Cyanobacterial N2 fixation in presence of nitrogen fertilizers

A E Mekonnen, Radha Prasanna & B D Kaushik

 

Received 8 August 2001; revised 21 March 2002

Anabaena oryzae ARM 570 was examined for its growth (chlorophyll and protein), heterocyst frequency, nitrogenase (acetylene reduction) activity, ammonia excretion, and glutamine synthetase and nitrate reductase in response to two levels of urea-N vis--vis N2-N. Growth of cyanobacterium increased with duration of incubation. Reduction in heterocyst frequency (40%) was observed at 30 ppm of urea-N, whereas at 60 ppm of urea-N, filaments were completely devoid of heterocysts and no nitrogenase activity was observed. Maximum excretion of ammonia occurred at 30 ppm of urea-N, which was concomitant with minimum glutamine synthetase activity. These results suggested that A. oryzae could be effectively utilized in cyanobacterial biofertilizer programme even in the presence of combined nitrogen, for improving N-budget in rice cultivation.

 

 

Indian Journal of Experimental Biology

Vol. 40, July 2002, pp. 858-860

 

Effect of lead on Na+, K+-ATPase activity in Penaeus indicus postlarvae

C Satyavathi & Y Prabhakara Rao

 

Received 22 June 2000; revised 9 January 2002

In vivo effect of lead on Na+, K+-ATPase was studied in plasma membrane/mitochondrial fraction of P. indicus postlarvae (PL), exposed to 30 days to a sublethal concentration (1.44 ppm) of lead. A significant (P < 0.05) decrease in the enzyme activity was observed for exposed PL with respect to their controls at different intervals except 24hr. Further the substrate (ATP) and ion (Na+ and K+)-dependent kinetics of Na+, K+-ATPase was studied with the plasma membrane/mitochondrial fractions of control and 30 days exposed PL. The apparent KM and Vmax values were calculated to determine the nature of inhibition. Both the control and exposed PL showed almost the same apparent KM values in the presence of different substrate or ion concentrations indicating that lead interacts with the enzyme at a different binding site.