Indian Journal of Experimental Biology

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VOLUME 52

NUMBER 7

JULY 2014

CODEN: IJEB (A6) 52 (7) 669-758 (2014)

ISSN: 0019-5189 (Print); 0975-1009 (Online)

CONTENTS

 

Papers

 

 

 

Abnormal lipid metabolism in collagen-induced arthritis rat model: In vitro, high resolution NMR spectroscopy based analysis

673

 

 

Niraj Kumar Srivastava, Shikha Sharma, Rudra N. Purusottam, Neeraj Sinha, Rameshwar Singh & Deepak Sharma

 

 

 

Effects of ursolic acid on glucose metabolism, the polyol pathway and dyslipidemia in non-obese type 2 diabetic mice

683

 

 

Jin Lee, Hae-In Lee, Kown-Il Seo, Hyun Wook Cho, Myung-Joo Kim, Eun-Mi Park & Mi-Kyung Lee

 

 

 

Anti-bacterial activity of Achatina CRP and its mechanism of action

692

 

 

Sandip Mukherjee, Soma Barman, Shuvasree Sarkar, Narayan Chandra Mandal & Shelley Bhattacharya

 

 

 

Evaluation of hepatoprotective activity of vasicinone in mice

705

 

 

Chaitali Sarkar, Sankhadip Bose & Sugato Banerjee

 

 

 

Characterization of oleic acid-induced acute respiratory distress syndrome model in rat

712

 

 

Aparna Akella, Parul Sharma, Ratna Pandey & Shripad B Deshpande

 

 

 

Anti-hyperglycemic activity of Rutin in streptozotocin-induced diabetic rats: An effect mediated through cytokines, antioxidants and lipid biomarkers

720

 

 

Netaji T Niture, Ansar A Ansari & Suresh R Naik

 

 

Influence of environmental hypertonicity on the induction of ureogenesis and amino acid metabolism in air-breathing walking catfish (Clarias batrachus, Bloch)

728

 

 

Bodhisattwa Banerjee, Gitalee Bhuyan and Nirmalendu Saha

 

 

 

Isolation of mosquitocidal bacteria (Bacillus thuringiensis, B.sphaericus and B.cereus) from excreta of arid birds

739

 

 

Subbiah Poopathi, K Thirugnanasambantham, C Mani, K Ragul & SM Sundarapandian

 

 

 

A simple and cost effective liquid culture system for the micropropagation of two commercially important apple rootstocks

748

 

 

Mohina Mehta, Raja Ram & Amita Bhattacharya

 

 

 

Large scale propagation of an exotic edible bamboo, Phyllostachys pubescens Mazel ex H. De Lehale (Moso Bamboo) using seeds

755

 

 

Anil Sood, Harleen Kaur Nadha, Sangita Sood, Shivani Walia & Om Parkash

 

 

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Editor痴 Note

 

Indian Journal of Experimental Biology in Open Access Mode

 

The Indian Journal of Experimental Biology (IJEB) is now an open access journal in the repository, NISCAIR Online Periodicals Repository (NOPR) [http://nopr.niscair.res.in].

Full text of all articles published in IJEB from 1999 onwards can now be accessed at NOPR in the open access mode. Papers in the current issue shall be uploaded immediately. Papers published in earlier years shall be added soon.

NOPR is based on DSpace, a digital repository software, and allows document browsing, document searching and various search options like title, author name, keywords, year,issue, etc.

 

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Erratum

Aromatase activity in brain and ovary: Seasonal variations correlated with circannual gonadal cycle in the catfish, Heteropneustes fossilis, by Neerja Aggarwal, Shashi Vadan Goswami, Preeti Khandelwal & Neeta Sehgalク Indian J Exp Biol, Vol. 52, May 2014, pp. 527-537.

The figure 4 appearing on p. 532 may be replaced with the following figure:

 

 

 

 

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Author Index

Akella Aparna

712

Ansari Ansar A

720

 

 

Banerjee Bodhisattwa

728

Banerjee Sugato

705

Barman Soma

692

Bhattacharya Amita

748

Bhattacharya Shelley

692

Bhuyan Gitalee

728

Bose Sankhadip

705

 

 

Cho Hyun Wook

683

 

 

Deshpande Shripad B

712

 

 

Kim Myung-Joo

683

 

 

Lee Hae-In

683

Lee Jin

683

Lee Mi-Kyung

683

Mandal Narayan Chandra

692

Mani C

739

Mehta Mohina

748

Mukherjee Sandip

692

 

 

Nadha Harleen Kaur

755

Naik Suresh R

720

Niture Netaji T

720

 

 

Om Parkash

755

 

 

Pandey Ratna

712

Park Eun-Mi

683

Poopathi Subbiah

739

Purusottam Rudra N.

673

 

 

Ragu K

739

Raja Ram

748

 

 

Saha Nirmalendu

728

Sarkar Chaitali

705

Sarkar Shuvasree

692

Seo Kown-Il

683

Sharma Deepak

673

Sharma Parul

712

Sharma Shikha

673

Singh Rameshwar

673

Sinha Neeraj

673

Sood Anil

755

Sood Sangita

755

Srivastava Niraj Kumar

673

Sundarapandian SM

739

 

 

Thirugnanasambantham K

739

 

 

Walia Shivani

755

 

 

Keyword Index

Achatina fulica

692

Acute lung injury

712

Amino acid metabolism

728

Ammonia

728

Antidiabetic activity

720

Antioxidants

720

Apoptosis-like-death

692

Apple rootstocks

748

Arid-Birds

739

 

 

B9 and MM106

748

Bacillus cereus

739

Bacillus sphaericus

739

Bacillus thuringiensis

739

Bamboo

755

Bioassays

739

 

 

Cholesterol

673

Clarias batrachus

728

Collagen-induced arthritis

673

Cost effective

748

C-reactive protein

692

 

 

Dyslipidemia

683

 

 

Edible shoots

755

Environmental hypertonicity

728

Ex vitro propagation

755

Excreta

739

 

 

Gene Sequence

739

 

 

Healthy plants

748

Hepatoprotective

705

 

 

In vitro propagation

755

 

 

Justicia adhatoda

705

 

 

Kidney

683

 

 

Lipid components

673

Lipid extraction

673

Lipid profile

720

Liquid culture

748

Liver

683

 

 

Mannitol

728

Metabolic enzymes

692

Metabolism

673

Mice

705

Micropropagation

748

Minute ventilation

712

Mosquitoes

739

NMR spectroscopy

673

 

 

Ornithine-urea cycle

728

Oxidative stress

673

Oxidative stress

692

 

 

P/F ratio

712

Phospholipids

673

Phyllostachys pubescens

755

Phylogenetic analysis

739

Polypol pathway

683

Pro-inflammatory cytokines

720

Pulmonary edema

712

 

 

Rutin

720

 

 

Streptozotocin

720

Surfactant

712

 

 

Urea

728

Ursolic acid

683

 

 

Vasicinone

705

 

 

Walking catfish

728

 

 

Correspondent author is marked by *

 

 

Indian Journal of Experimental Biology

Vol. 52, July 2014, pp. 673-682

 

 

 

Abnormal lipid metabolism in collagen-induced arthritis rat model: In vitro, high resolution NMR spectroscopy based analysis

Niraj Kumar Srivastavaa, Shikha Sharmaa, Rudra N. Purusottamb, Neeraj Sinhab,
Rameshwar Singha & Deepak Sharmaa*

aNeurobiology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi 110 067, India

bCenter of Biomedical Magnetic Resonance,

Sanjay Gandhi Postgraduate Institute of Medical Sciences Campus, Lucknow 226 014, India

Received 1 November 2013; revised 16 April 2014

Collagen-induced arthritis (CIA) was induced in female Wistar rats by intradermal injection of porcine immunization grade native collagen type II (Chondrex). Development and progression of CIA was monitored by studying histopathological, radiographical and biochemical features of arthritic manifestations in the knee joints, hind limb and blood plasma. In addition, oxidative stress status of arthritic animals was determined by measuring lipid peroxidation and the antioxidant enzymes: catalase, superoxide dismutase and glutathione peroxidase. High resolution proton NMR spectroscopy was employed for the analysis of lipid components in the lipid extracts of the joint tissue and plasma of collagen-induced arthritic and control rats. Triglyceride levels showed significant decreases in plasma (1.7 times) but were unchanged in the joint tissue of CIA rats as compared to control. One-dimensional proton NMR spectra showed a 6.2 times reduction in the quantity of choline-containing phospholipids in the plasma of CIA as compared to control rats. There was a 1.6 times elevation of choline-containing phospholipids in the joint tissue of CIA rats as compared to controls. Induction of arthritis showed a 4.0 times reduction in the level of total cholesterol in the plasma and 1.6 times elevation in the joint tissue of CIA rats as compared to controls. The ratio of saturated fatty acids to unsaturated fatty acids was 1.5 times significantly higher in joint tissue and 2.1 times significantly higher in plasma of CIA rats as compared to controls. The results demonstrated significantly altered lipid patterns in the joint tissue and plasma of collagen-induced arthritic rats as detected by one- and two-dimensional NMR spectroscopy compared with controls.

 

 

Indian Journal of Experimental Biology

Vol. 52, July 2014, pp. 683-691

 

 

Effects of ursolic acid on glucose metabolism, the polyol pathway and dyslipidemia in non-obese type 2 diabetic mice

Jin Lee1,, Hae-In Lee1,, Kown-Il Seo1, Hyun Wook Cho2, Myung-Joo Kim3, Eun-Mi Park3 & Mi-Kyung Lee1,*

Department of 1Food and Nutrition, and 2Biology, Sunchon National University,
255 Jungang-ro, Suncheon, Jeonnam, 540-950, Korea

3Department of Hotel Cusine, Suseong College, Daegu, 706-022, Korea

Received 1 November 2013, Revised 6 March 2014

Ursolic acid (UA) is a pentacyclic triterpenoid compound that naturally occurs in fruits, leaves and flowers of medicinal herbs. This study investigated the dose-response efficacy of UA (0.01 and 0.05%) on glucose metabolism, the polyol pathway and dyslipidemia in streptozotocin/nicotinamide-induced diabetic mice. Supplement with both UA doses reduced fasting blood glucose and plasma triglyceride levels in non-obese type 2 diabetic mice. High-dose UA significantly lowered plasma free fatty acid, total cholesterol and VLDL-cholesterol levels compared with the diabetic control mice, while LDL-cholesterol levels were reduced with both doses. UA supplement effectively decreased hepatic glucose-6-phosphatase activity and increased glucokinase activity, the glucokinase/glucose-6-phosphatase ratio, GLUT2 mRNA levels and glycogen content compared with the diabetic control mice. UA supplement attenuated hyperglycemia-induced renal hypertrophy and histological changes. Renal aldose reductase activity was higher, whereas sorbitol dehydrogenase activity was lower in the diabetic control group than in the non-diabetic group. However, UA supplement reversed the biochemical changes in polyol pathway to normal values. These results demonstrated that low-dose UA had preventive potency for diabetic renal complications, which could be mediated by changes in hepatic glucose metabolism and the renal polyol pathway. High-dose UA was more effective anti-dyslipidemia therapy in non-obese type 2 diabetic mice.

 

 

Indian Journal of Experimental Biology

Vol. 52, July 2014, pp. 692-704

 

 

Anti-bacterial activity of Achatina CRP and its mechanism of action

Sandip Mukherjee1, Soma Barman2, Shuvasree Sarkar1, Narayan Chandra Mandal2 & Shelley Bhattacharya1*

1Environmental Toxicology Laboratory, Department of Zoology (Centre for Advanced Studies),
2Microbiology, Mycology & Plant Pathology Laboratory,
Department of Botany, Visva-Bharati University, Santiniketan 731 235, India

Received 28 June 2013; revised 4 April 2014

The physiological role of C-reactive protein (CRP), the classical acute-phase protein, is not well documented, despite many reports on biological effects of CRP in vitro and in model systems in vivo. It has been suggested that CRP protects mice against lethal toxicity of bacterial infections by implementing immunological responses. In Achatina fulica CRP is a constitutive multifunctional protein in haemolymph and considered responsible for their survival in the environment for millions of years. The efficacy of Achatina CRP (ACRP) was tested against both Salmonella typhimurium and Bacillus subtilis infections in mice where endogenous CRP level is negligible even after inflammatory stimulus. Further, growth curves of the bacteria revealed that ACRP (50 オg/mL) is bacteriostatic against gram negative salmonellae and bactericidal against gram positive bacilli. ACRP induced energy crises in bacterial cells, inhibited key carbohydrate metabolic enzymes such as phosphofructokinase in glycolysis, isocitrate dehydrogenase in TCA cycle, isocitrate lyase in glyoxylate cycle and fructose-1,6-bisphosphatase in gluconeogenesis. ACRP disturbed the homeostasis of cellular redox potential as well as reduced glutathione status, which is accompanied by an enhanced rate of lipid peroxidation. Annexin V-Cy3/CFDA dual staining clearly showed ACRP induced apoptosis-like death in bacterial cell population. Moreover, immunoblot analyses also indicated apoptosis-like death in ACRP treated bacterial cells, where activation of poly (ADP-ribose) polymerase-1 (PARP) and caspase-3 was noteworthy. It is concluded that metabolic impairment by ACRP in bacterial cells is primarily due to generation of reactive oxygen species and ACRP induced anti-bacterial effect is mediated by metabolic impairment leading to apoptosis-like death in bacterial cells.

 

 

 

Indian Journal of Experimental Biology

Vol. 52, July 2014, pp. 705-711

 

 

Evaluation of hepatoprotective activity of vasicinone in mice

Chaitali Sarkara, Sankhadip Bosea & Sugato Banerjeeb*

aGupta College of Technological Sciences, College of Pharmacy, Asansol 713 301, India

bDepartment of Pharmaceutical Sciences, Birla Institute of Technology, Mesra, Ranchi 835 215, India

Received 8 January 2013; revised 4 April 2014

Justicia adhatoda (vasaka) leaves have long been used in Indian Ayurvedic system of medicine as antitussive. Its crude extract has been previously reported to have hepatoprotective activity. Vasicinone was isolated from leaves of J. adhatoda, column purified and characterized using, TLC UV, FT-IR and 1H NMR. The isolated vasicinone was evaluated for hepatoprotective activity using (CCl4)-induced acute hepatotoxicity model in mice. CCl4 treatments lead to significant increase in SGOT, SGPT, ALP levels. Pre-treatment with vasicinone and silymarin (25 mg/kg/day for 7 days) significantly decreased these enzyme levels. Histopathology of the livers from vasicinone and silymarin pre-treated animals showed normal hepatic cords and absence of necrotic changes suggesting pronounced recovery from CCl4 induced liver damage. Both vasicinone and silymarin significantly decrease the CCl4 mediated increase in pentobarbital indiced sleeping time in experimental animals, thus indicating recovery of liver function. Based on the above results it can be concluded that vasicinone may act as hepatoprotective in mice and warrants further investigation on human volunteers.

 

 

Indian Journal of Experimental Biology

Vol. 52, July 2014, pp. 712-719

 

 

Characterization of oleic acid-induced acute respiratory distress
syndrome model in rat

Aparna Akella1, Parul Sharma1, Ratna Pandey & Shripad B Deshpande*

Department of Physiology, Institute of Medical Sciences, Banaras Hindu University, Varanasi 221 005, India

Received 9 December 2013; revised 21 April 2014

Animal studies using oleic acid (OA) model to produce acute respiratory distress syndrome (ARDS) have been inconsistent. Therefore, the present study was undertaken to establish an acute model of ARDS in rats using OA and to characterize its effect on cardio-respiratory parameters and lethality. The trachea, jugular vein and femoral artery of anesthetized adult rats were cannulated. A dose of OA (30-90 オL; iv) was injected in each animal and changes in respiratory frequency (RF), heart rate (HR) and mean arterial pressure (MAP) were recorded. Minute ventilation and PaO2/FiO2 (P/F) ratio were also determined. At the end, lungs were excised for determination of pulmonary water content and histological examination. At all doses of OA, there was immediate decrease followed by increase in RF, however at 75 and 90 オL of OA, RF decreased abruptly and the animals died by 63 ア 8.2 min and 19 ア 6.3 min; respectively. In all the groups, HR and MAP changes followed the respiratory changes. The minute ventilation increased in a dose-dependent manner while the values of P/F ratio decreased correspondingly. Pulmonary edema was induced at all doses. Histological examination of the lung showed alveolar damage, microvascular congestion, microvascular injury, infiltration of inflammatory cells, pulmonary edema and necrosis in a dose-dependent manner. With these results, OA can be used to induce different grades of ARDS in rats and OA doses of 50, 60 and 75 オL resemble mild, moderate and severe forms of ARDS respectively. Hence, OA model serves as a useful tool to study the pathophysiology of ARDS.

 

Indian Journal of Experimental Biology

Vol. 52, July 2014, pp. 720-727

 

 

Anti-hyperglycemic activity of Rutin in streptozotocin-induced diabetic rats:
An effect mediated through cytokines, antioxidants and lipid biomarkers

 

Netaji T Niture1, Ansar A Ansari1 & Suresh R Naik2*

1Sinhgad College of Pharmacy, Vadgaon (Bk), Pune 410 041, India

2Sinhgad Institute of Pharmaceutical Sciences, Kusgaon (Bk), Lonavala, Pune 411 401, India

Received 29 August 2012, revised 2 May 2014

Administration of rutin (50 and 100 mg/kg) and pioglitazone (10 mg/kg) orally for 3 weeks treatment significantly improved body weight, reduced plasma glucose and glycosylated hemoglobin, pro-inflammatory cytokines (IL-6 and
TNF-α), restored the depleted liver antioxidant status and serum lipid profile in high fat diet + streptozotocin induced
type 2 diabetic rats. Rutin treatment also improved histo-architecture of ゚ islets and reversed hypertrophy of hepatocytes. Rutin exhibited significant antidiabetic activity, presumably by inhibiting inflammatory cytokines, improving antioxidant and plasma lipid profiles in High fat diet + streptozotocin induced type 2 diabetic model and may be useful as a diabetic modulator along with standard antidiabetic drugs. However, such effects need to be confirmed on human subjects in
clinical condition.

 

 

Indian Journal of Experimental Biology

Vol. 52, July 2014, pp. 728-738

 

 

Influence of environmental hypertonicity on the induction of ureogenesis
and amino acid metabolism in air-breathing walking catfish
(Clarias batrachus, Bloch)

Bodhisattwa Banerjee, Gitalee Bhuyan & Nirmalendu Saha*

Biochemical Adaptation Lab., Department of Zoology, North-Eastern Hill University, Shillong 793 022, India

Received 11 November 2013; revised 28 April 2014

Effect of environmental hypertonicity, due to exposure to 300 mM mannitol solution for 7 days, on the induction of ureogenesis and also on amino acid metabolism was studied in the air-breathing walking catfish, C. batrachus, which is already known to have the capacity to face the problem of osmolarity stress in addition to other environmental stresses in its natural habitats. Exposure to hypertonic mannitol solution led to reduction of ammonia excretion rate by about 2-fold with a concomitant increase of urea-N excretion rate by about 2-fold. This was accompanied by significant increase in the levels of both ammonia and urea in different tissues and also in plasma. Further, the environmental hypertonicity also led to significant accumulation of different non-essential free amino acids (FAAs) and to some extent the essential FAAs, thereby causing a total increase of non-essential FAA pool by 2-3-fold and essential FAA pool by 1.5-2.0-fold in most of the tissues studied including the plasma. The activities of three ornithine-urea cycle (OUC) enzymes such as carbamoyl phosphate synthetase, argininosuccinate synthetase and argininosuccinate lyase in liver and kidney tissues, and four key amino acid metabolism-related enzymes such as glutamine synthetase, glutamate dehydrogenase (reductive amination), alanine aminotransaminase and aspartate aminotransaminase were also significantly up-regulated in different tissues of the fish while exposing to hypertonic environment. Thus, more accumulation and excretion of urea-N observed during hypertonic exposure were probably associated with the induction of ureogenesis through the induced OUC, and the increase of amino acid pool was probably mainly associated with the up-regulation of amino acid synthesizing machineries in this catfish in hypertonic environment. These might have helped the walking catfish in defending the osmotic stress and to acclimatize better under hypertonic environment, which is very much uncommon among freshwater teleosts.

 

 

Indian Journal of Experimental Biology

Vol. 52, July 2014, pp. 739-747

 

 

 

Isolation of mosquitocidal bacteria (Bacillus thuringiensis, B.sphaericus and B.cereus) from excreta of arid birds

Subbiah Poopathi*, K Thirugnanasambantham, C Mani, K Ragul & SM Sundarapandian#

Unit of Microbiology and Immunology, Vector Control Research Centre (Indian Council of Medical Research),
Indira Nagar, Puducherry 605 006, India

#Department of Ecology and Environmental Sciences, School of Life-Sciences, Puducherry University, Puducherry 605014, India

Received 11 November 2013; revised 2 April 2014

Mosquitocidal bacteria are environmentally friendly alternatives to chemical insecticides for controlling mosquitoes and therefore, there have been tremendous world-wide efforts to identify novel mosquitocidal bacteria from natural environment. In the present study, excreta from arid-birds were analyzed for identifying mosquitocidal bacteria. The selection of sample for bacterial screening is significant, because, arid-birds are the unique living species and gathering the foods from variety of sources from environment. Out of 1000 samples examined, twelve bacterial strains were identified as mosquitocidal and the 16S rRNA gene sequence alignment depicted that these isolates belonged to Bacillus species (Bacillus thuringiensis, B.sphaericus and B.cereus). Toxicity assay against mosquito vectors have shown that these isolates are potential. The B. sphaericus VCRC-B547 (NCBI: JN377789) has shown a higher toxicity against Cx. quinquefasciatus, An. stephensi, and Aed. aegypti. Result from SDS-PAGE has shown that there was considerable difference in the protein profiles among the new bacterial isolates. Phylogenetic tree with branch length 0.05 revealed three distinct groups with homology among the closely related Bacillus strains. This study therefore throws considerable interest on the diversity of microbial organisms from arid birds and its application in mosquito control.

 

 

Indian Journal of Experimental Biology

Vol. 52, July 2014, pp. 748-754

 

 

A simple and cost effective liquid culture system for the micropropagation of two commercially important apple rootstocks

Mohina Mehta, Raja Ram & Amita Bhattacharya*

Division of Biotechnology, CSIR-Institute of Himalayan Bioresource Technology (IHBT), Palampur, 176 061, India

Received 10 July 2013; revised 5 March 2014

The two commercially important apple rootstocks i.e., MM106 and B9 were micropropagated using a liquid culture system. Three different strengths of 0.8% agar solidified PGR free basal MS medium were first tested to optimize the culture media for both the rootstocks. Full strength medium (MS0) supported maximum in vitro growth, multiplication, rooting and survival under field conditions as opposed to quarter and half strength media. When three different volumes of liquid MS0 were tested, highest in vitro growth, multiplication, rooting and also survival under field conditions were achieved in 20 mL liquid MS0. The cost of one litre of liquid medium was also reduced by 8 times to Rs. 6.29 as compared to solid medium. The cost of 20 mL medium was further reduced to Rs. 0.125.

 

 

Indian Journal of Experimental Biology

Vol. 52, July 2014, pp. 755-758

 

 

Large scale propagation of an exotic edible bamboo, Phyllostachys pubescens Mazel ex H. De Lehale (Moso Bamboo) using seeds

Anil Sood*, Harleen Kaur Nadha, Sangita Sood1, Shivani Walia1 & Om Parkash

Division of Biotechnology, CSIR-Institute of Himalayan Bioresource Technology (IHBT) Palampur 176 061, India

Received 14 March 2013; revised 2 May 2014

For ex vitro propagation, seeds of P.pubescens were treated with different concentrations of gibberellic acid (GA3) and germination of seeds was tested both in plastic pots as well as by direct sowing in the nursery beds. Maximum seed germination was achieved when treated with 200 mgL1 (w/v) GA3. For in vitro propagation, an exposure of nodal explants from in vitro raised seedlings to 0.2 mgL1 1朴henyl3(1,2,3釦hiadiazol5幌l) urea and 1 mgL1 kinetin supplemented medium for 30 days and thereafter to hormone free Murashige and Skoog basal medium resulted in axillary shoot proliferation. For rooting, in vitro raised shoots were exposed to MS medium containing 2 mgL1 indole-3-butyric acid for 15 days and then shifted to hormone free medium. On an average, 2.8 shoots were obtained in 75% of the cultures within 4 weeks. Such in vitro raised plants were successfully hardened and shifted to field conditions.