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1. Journal of Cell and Tissue Research 12(3) 3265-3271 (2012)
Unlike norepinephrine and serotonin, dopamine disor-ganizes direct cell-cell communication in hepatocyte cultures
Brodsky, V.Y., Konchenko, D, S., Zvezdina, N.D., Malchenko, L.A. and Dubovaja, T.K
Institute of Developmental Biology, Russian Academy of Sciences and Russian State Medical University,
26 Vavilov st., 117334 Moscow, Russia. E. mail: brodsky.idb@bk.ru
Abstract: Primary cultures of rat hepatocytes on slides were studied in serum-free supplemented medium. Ultradian protein synthesis rhythm was used as a marker of direct cell-cell communication resulting in synchronization of individual oscillations and formation of a common rhythm of the cell population. Dense synchronous and sparse non-synchronous cultures were used to determine the effect of dopamine on the kinetics of protein synthesis. Treatment (30-40 min) of synchronous dense cultures with 1-10 µM dopamine suppresses the protein synthesis rhythm relative to control non-treated cultures. The dopamine effect is maintained at least 5 hours but in a day the rhythm of protein synthesis is completely restored. Medium conditioned by normal dense cultures initiated synchronization of sparse cultures. The effect was not observed with medium conditioned by dense cultures pretreated with dopamine. After addition of gangliosides, melatonin or norepinephrine to the medium in cultures pretreated with dopamine the rhythm was restored. The data are discussed with respect to mechanisms of organization and disorganization of protein synthesis oscillations and accordingly of the direct cell-cell communication especially regarding in vivo conditions and ageing.
Key words: Cell-cell communication, Protein synthesis rhythm, Neurotransmitters, Dopamine, Hepatocytes
2. Journal of Cell and Tissue Research 12(3) 3273-3278 (2012)
Detection of apoptosis during follicular atresia in Caprine ovary by tunel and fluorescence microscopy
Bhardwaj, J.K. and Sharma, R.K.
Reproductive Physiology Laboratory, Department of Zoology, Kurukshetra University,
Kurukshetra 136119. E. mail: jkbkuk@gmail.com
Abstract: Apoptosis is an intrinsic form of cell death whereby the animal cells can self dest-ruct. Many hormones and growth factors are capable of inducing or facilitating programmed cell death under physiological or pathological conditions. Therefore, the present study was carried out to understand the mechanistic machinery of apoptosis and to investigate is it the changed estrogen (E2) over progesterone (P4) ratio exclusively responsible for the induction of apoptosis in granulosa cells. Granulosa cells smears from different categories of follicles were subjected to TUNEL assay and fluorescent assay for specific detection of apoptotic granulosa cells. The ladder pattern of fragmented apoptotic DNA was also observed. The estrogen and progesterone titre was carried out by direct radio immunoassay. The relative frequency of apoptotic granulosa cells in healthy, slightly atretic and atretic follicles was 12 + 1%, 34 + 1% and 52 + 1% respectively. E2/P4 : > 1 in healthy follicles while E2/P4 < 1 in atretic follicles. In situ 3' end labeling and ladder assay of fragmented DNA in apoptotic cells confirmed that the phenomenon of cell death was apoptosis. Imbalance between E2 and P4 in follicular fluid is involved in the initiation of granulosa cells apoptosis and hence follicular atresia.
Key words: Apoptosis; Granulosa cells; Ovary; Atresia
3. Journal of Cell and Tissue Research 12(3) 3279-3284 (2012)
Association of protein profiling and agronomic traits in chickpea as revealed using SDS-PAGE
Singh, P.K., Kumar, A., Srivastava, N., Agarwal, R.M. and Bhagyawant, S.S.
School of Studies in Biotechnology, Jiwaji University, Gwalior. E. mail: sbhagyawant@yahoo.com,
Abstract: SDS-PAGE was employed for elaborating genetic diversity in 20 cultivars of chickpea including one wild accession. Electrophoretic patterns of the protein are correlated to agronomic characteristics thus used to fingerprint genetic makeup. A total of three hundred and three polypeptides bands were reliably scored following such proteomic assay. The polypeptide profile for each Cicer accession was observed to be strain specific and genotype differed in grouping was estimated by constructing a dendogram. The analysis assorted all accessions in 4 clusters. In the present study, a biochemical protein marker for pod-borer is differentiated. The relationships between seed storage banding profile and agronomic traits can thus forecast the selection of parents in chickpea crop improvement program.
Key words: Germplasm, SDS- PAGE, Chickpea, Seed storage protein
4. Journal of Cell and Tissue Research 12(3) 3285-3289 (2012)
Screening of BMP15 (FecX) fecundity genein prolific Raighar goats of Odisha
Palai, T.K.,Maity, A., Bisoi, P.C.,Polley, S., Mukharjee, A. and De, S.
Department of Biochemistry, C.V.Sc & A.H., OUAT, Bhubaneswar 751 003, Odisha.
E. mail: tapan.palai@gmail.com
Abstract: BMP15 (FecX) is an established fecundity gene in sheep. The present study was undertaken to find the contribution of BMP15 towards prolificacy of Raighar goats. Blood of 101 does with history of high prolificacy was collected and subjected to DNA isolation. Further T-ARMS-PCR (tetra-primer amplification refractory mutation system) and agarose Gel electrophoresis used to analyse the samples. In Raighar goat, among the five BMP15 SNPs studied none were found to be polymorphic. Therefore, prolificacy in case of Raighar goat is not due to the mutation at any of the BMP 15 loci. It is thought to search for other genes or loci in goat fecundity.
Key words: Raigarh Goat, Prolificacy, BMP 15, Mutation
5. Journal of Cell and Tissue Research 12(3) 3291-3295 (2012)
Decellularization of buffalo bone to prepare bone scaffolds for effective bone tissue engineering
Pathak, R., Amarpal, Tiwari, A.K., Kurade, N.P. and Amar Nath
Division of Surgery, Indian Veterinary Research Institute, Izatnagar 243122, UP E. mail: rekhasurgery@rediffmail.com
Abstract: To prepare the porous buffalo bone scaffolds for effective bone tissue engineering, in this study we attempted to decellularize bone for rendering the bone acellular. For this work buffalo femur bone was collected from local abattoir to render the bone acellular, three decellularization methods A): Acetone: Ethanol, B): SDS treatment C): Freeze and thawing methods were adopted and their effectivity was evaluated through histopathology and scanning electron microscopy (SEM). Cancellous bone appeared as network of homogenous red stained osteoid material of trabeculae with no cells in the intercommunicating spaces in histological sections for all the treatment methods adopted. SEM results indicate that not much significant disturbances in extracellular matrix morphology and having an average pore size of 83.402 μm to 297.437μm and total porosity was found to be around 85%. Study demonstrates and established decellularization protocols to make the buffalo bone acelluler, that might be useful to study the biomaterial and cell interactions.
Key words: Decellularization; Composite scaffold; Buffalo bone
6. Journal of Cell and Tissue Research 12(3) 3297-3305 (2012)
Identification of a yellowing inducing strain of Fusarium oxysporum F. sp. Ciceris from India
Joshi, N.S., Subramanian, R.B. and Rao, K.S.
B. R. Doshi School of Biosciences, Sardar Patel Maidan, Vadtal Road, Post Box No. 39, Sardar Patel
University, Vallabh Vidyanagar-388120, Gujarat. E. mail: neharajiv1215@gmail.com
Abstract: Identification of physiological race of Fusarium oxysporum f. sp. ciceris (Foc) Em. J-15 was carried out by comparing its effect on an Indian set of differential cultivars with a reference isolate (Foc race 2). Chickpea cultivars JG-62, GG-1, GG-4, JCP-27 and WR-315 were used. The results of the pathogenicity tests indicated that Foc J-15 induced yellowing symptoms in the susceptible cultivar (JG-62). Protein profile on SDS-PAGE revealed that the protein synthesis in Foc J-15 treated plants was induced to a normal level whereas a suppressed induction of proteins was observed in the Foc race 2 treated plants. Molecular analysis revealed the presence of race 0 specific bands of 0.9 kb, OPI-09; 0.39 kb, OPF-12 and 0.65 kb, OPF-16 with Foc-J-15. In this study, a 450 bp product was sequenced and identified as a sugar transporter in Foc J-15 by modifying the existing primers coding for Fol Avr1 gene. The combined analysis of the results obtained from the pathogenicity tests and RAPD based molecular trials indicated the field isolate to be a strain inducing yellowing.
Key words: Fusarium oxysporum F. sp
7. Journal of Cell and Tissue Research 12(3) 3307-3309 (2012)
Studies on pathology of chlorpyriphos toxicity in chick embryos
Roopadevi, Y.S. Narayanaswamy, H.D. Chandrashekhara, N. and Sathyanarayana, M.L.
LRIC, Nagamangala, Department of Animal Husbandry, Bangalore, Karnataka 560 024. E. mail: ncsrvet@gmail.com
Abstract: The Chlorpyriphos was inoculated to seven day-old chick embryos of Girirani strain at the rate of 750, 1000 and 1250 μg per embryo. The embryo pathology was studied on 12th, 15th and 18th day of incubation. Chlorpyriphos produced degeneration and necrosis of hepatocytes in the liver, degeneration of tubular epithelium in the kidney, swelling of cardiac myocytes, degeneration and desquamation of mucosal epithelial cells of proventriculus indicating the potential cause of multiorgan toxicity.
Key words: Chlorpyriphos, Chick embryo
8. Journal of Cell and Tissue Research 12(3) 3311-3316 (2012)
Hypolipidemic effects of bees wax on serum lipid parameters in induced hyperlipidemic male albino rabbits
Purohit, A., Joshi, K., Kotru, B. and Kotru, S.
Department of Zoology, JNVU University, Jodhpur -342 005, E-mail: drdiafoot@gmail.com
Abstract: Policosanol is a natural mixture of higher aliphatic primary alcohols, extracted from natural sources like bees wax. The present study was designed to investigate the possible hypolipidemic effects of crude bees wax on cholesterol fed rabbits and to compare with standard drug available in the market statin. The rabbits were first made exogeneously hyperlipidemic by giving them high fat diet and cholesterol powder (500mg/kg body weight) in 5 ml of coconut oil orally for 15 days and then were administered with drugs like bees wax and statin. In the hyperlipidemic control rabbits there were an apparent reduction in the animal body weight and a significant increase in serum total cholesterol (78.67%),triglyceride (27.24 %), LDL-c(78.56 %) and VLDL-c(38.04 %) with a concomitant non significant variation in serum HDL-c (25.45 %). Whereas oral treatment of animals with bees wax for about 45 days in a dose of 50mg/Kg body weight lead to non significant variation in the final animal body weight and decrease in serum total cholesterol (93.26 %), triglyceride (85.33%), LDL-c (95.10%) and VLDL-c (85.30%) with a slightly non significant increase in HDL-c. In conclusion, bees wax was found nearly equal in efficacy with that of statin standard drug when compared with hyperlipidemic control animals on all serum lipid parameters. Thus bees wax offers promising hypolipidemic effects with no drug related disturbances in safety medications that may be mainly attributed to its potent antiatherosclerotic potential. Further studies will be needed in future in order to determine which one or more of its active constituents has the main hypolipidemic effects.
Key words: Beeswax, Lipids, Cholesterol
9. Journal of Cell and Tissue Research 12(3) 3317-3322 (2012)
Evaluation of hypoglycemic effect of cow urine distillate in streptozotocin induced diabetic rat model
Kadagi. M., Jayakumar, K., Shridhar, N.B., Narayana Swamy, H.D., Narayana Swamy,M. and Manjunatha, K.P.
Department of Pharmacology and Toxicology, Veterinary College, Hebbal, KVAFSU, Bangalore 560024.
E. mail: mahesh.sk35@gmail.com
Abstract: The present study was designed to determine the hypoglycemic activity of cow urine distillate in streptozotocin induced diabetic rats. The Wister albino rats were divided into five groups (n=10). Group I served as normal control. Diabetes was induced in group II to V by injecting streptozotocin. Group II served as Diabetic control. Group III as a diabetic positive control. Group IV and V were administered with cow urine distillate @ 0.5 and 1 ml/kg respectively. Body weight was measured and blood samples were collected on day 0, 7, 14, 21 and 28 and analysed for various biochemical and hematological parameters. Necropsied rats were subjected for histopathology. There was significant (P< 0.001) increase in the blood glucose in streptozotocin treated rats and significant decrease in triglycerides, total cholesterol and HDL level in cow urine distillate treatment groups when compared with the diabetic control group. Histopathological examination of pancreas revealed regeneration and hyperplasia of the islets cells in treatment groups.
Key words: Strptozotocin, Hypoglycemic activity, Cow urine distillate
10 Journal of Cell and Tissue Research 12(3) 3323-3326 (2012)
Muscarinic vs histaminic activity in smooth muscle contraction of caprine ascending colon
Dash, J.R., Datta, B.K., Patra, P.H., Khan, M., Sar, T.K. and Mandal, T.K.
Department of Pharmacology and Toxicology, West Bengal University of Animal and Fishery Sciences, Mohanpur Campus, Nadia, West Bengal 741252. E. mail: pharma.tkm@gmail.com
Abstract: Muscarinic cholinergic system is abundantly expressed and mediates important roles in gastrointestinal tract and different muscarinic receptors have been identified in different parts along the gastrointestinal tract in different species. The present work was aimed to investigate the primary receptors actively involved in smooth muscle contraction of colon in goat. It was observed that a) a single dose of atropine (10 µM) effectively reduced the normal colon motility by >70%. b) Acetylcholine (10nM-100 µM) induced sustained and increased longitudinal contraction of the smooth muscle fibres of the colon in a dose dependent manner with a maximum response at 100µM (EC50 = 1.76±2.82 µM, n=6) compared to a poor effect by histamine (10nM-100 µM), EC50 = 4.18±1.05 µM, n=6. c) IC50 value for atropine in this tissue was 0.5nM (n=6) and atropine (10 µM) preincubation shifted Ach induced contractile response curve towards right which led us to confirm that longitudinal contraction of colon of goat may be primarily due to muscarinic receptors.
Key words: Ascending colon, Muscarinic, Histaminic, Smooth muscle
11 Journal of Cell and Tissue Research 12(3) 3327-3331 (2012)
Sequential pathology of bluetongue virus infection in chicken embryos
Manjunatha, K. P., Suguna Rao., Byregowda, S. M., Satyanarayana, M. L., Purushotham, K. M., Narayanaswamy, H. D. and Ansar Kamran, C.
Department of Pathology, Veterinary College, Hebbal, KVAFSU, Bangalore-560024. E. mail: manjukpvet@gmail.co.in
Abstract: The study was conducted by experimentally infecting chicken embryos with bluetongue virus by intravenous route and sequential pathology of infection was studied by histopathology, immunohistochemistry, antigen capture ELISA and PCR from 24 to 120 hours post infection. The BTV infected chicken embryos revealed mortality only after 48 hours PI. The organs subjected for pathological studies revealed grossly congestion, hemorrhages and edema in various organs. Among the microscopic lesions, congestion and haemorrhages were more prominently noticed during 24 to 48 hours PI, and edema in the later hours of study. Among various organs subjected for Immunohistochemistry the positive reaction was observed in kidney and liver with highest concentration in kidney. The antigen capture ELISA and PCR detected the virus in brain during 48 hours PI and in liver and kidney from 72 hours PI.
Key words: Chicken embryos, Bluetongue virus
12 Journal of Cell and Tissue Research 12(3) 3333-3336 (2012)
Antioxidants status and the effect of amla juice supplementation in smokers and non-smokers
Goswami, K., Patel, B.G., Dave, P.H., Goswami, G. and Patel, N.V.
Department of Food Science and Nutrition, ASPEE College of Home Science and Nutriton, Sardar Krushinagar Dantiwada Agricultural University, S. K. Nagar, Gujarat. E. mail: bgpatel62@gmail.com
Abstract: In order to evaluate the effect of smoking habit on various body’s antioxidant (SOD, lipid peroxidation, vitamin-C), twenty four healthy male smokers and same number of non-smokers in the age group of 30 to 35 years were selected and antioxidant parameters were estimated and statistically compared. Smokers were consuming low quantity of antioxidant rich food than non-smokers. Due to higher oxidative stress in smokers, the level of lipid peroxidation (LPO) was increased and level of Vitamin-C was found to be decreased. Level of Superoxide dismutase (SOD) was found to be significantly (P < 0.001) higher in smoker as compared to non-smokers. This may be due to induction of SOD in smokers. The level of Vitamin C in smokers was increased significantly (17.52 %; P < 0.001) as compared to non-smokers (1.6 %; P < 0.001) and level of LPO was decreased significantly in smokers (17.07 %; P < 0.001) as compared to non-smokers after the administration of amla juice (1 glass daily) to all the subjects for 15 days. The improved antioxidant status in smokers after Amla juice supplementation may be due to high antioxidant content in Amla juice. Thus, the results of the present study can be concluded that a regular supplementation of 200 ml of amla juice would be beneficial for the improvement of antioxidant status which in turn reduces the risk of smoking related health hazards.
Key words: Smokers, Non-smokers,Amla juice
13 Journal of Cell and Tissue Research 12(3) 3337-3342 (2012)
An efficient protocol for in vitro shoot induction andsomatic embryogenesis through callus using root explants in Daucus carota l, subsp. Halophilus
Ojha, A., Kumar, S., Singh, M.K.and Bind, D.M.
Plant Tissue Culture R & D, Department of Biotechnology, Meerut Institute of Engineering, and Technology, Meerut 250005. E. mail: ashishbt26@gmail.com
Abstract: A protocol for induction of callus and somatic embryogenesis from root explants of Daccus carota L, sub sp. halophilus was evaluated. Root explants were cultured on MS medium supplemented with different concentration and combinations of hormones. The influence of plant growth regulators (PGRs) including 2,4-dichlorophenoxyacetic acid (0.25-2.50 mg/L-1 2,4-D) on callus induction from root explants was studied. 2,4-dichlorophenoxyacetic acid (0.5-2.5 mg/L-1 and (0.25-0.75 mg/L-1 kinetin were used for somatic embryogenesis. The highest (100%) callus induction was found on Murashige and Skoog (MS) medium supplemented with 2, 4-D (0.75 mg/L-1). The highest rate (100%) of callus multiplication in MS + 2, 4-D (1.5 mg/L-1) + (0.25 mg/L-1) kinetin was observed. The highest frequency of embryogenic callus emerged on MS medium supplemented with MS + 2, 4-D (0.5 mg/L-1) + Kn (0.75 mg/L-1). Mean of 13 somatic embryos per embryogenic callus were obtained. Multiple shoot were induced from the callus of D. carota by culturing them in MS medium supplemented with 1.5 mg/L-1 of 6-benzylaminopurine (BAP). Averages of 12 shoots were developed from per callus. The present study is focused on shoot induction and embryogenesis through callus using root explants and facilitates mass propagation, applied to culture of the roots of wild Daucus carota.
Key words: Daucus carota L., Root explants callus, Somatic embryogenesis
14 Journal of Cell and Tissue Research 12(3) 3343-3348 (2012)
Evaluation of antifungal, antibacterial and antioxidant activities of Amphineuron extensus, Hedyotis diffusa and Vitex negundo
Bhuyan, D.J. and Barooah, M.S.
Department of Food and Nutrition, College of Home Science, Assam Agricultural University, Jorhat 785013, Assam.
E. mail: deeprkn@gmail.com Mobile: 09435050682
Abstract: In the present study, the antifungal and antibacterial properties of three selected plants namely Amphineuron extensus, Hedyotis diffusa and Vitex negundo used in the preparation of rice beer starter culture cakes were evaluated against Aspergillus niger, Aspergillus flavus and E. coli. A. extensus showed the highest inhibition zone against A. niger at 20% concentration whereas H. diffusa showed the lowest MIC value of 1.25%. In case of A. flavus, V. negundo showed the highest Inhibition zone at 20% concentration and all the plant extracts exhibited MIC value of 5%. A. extensus showed the highest inhibition zone against E. coli followed by V. negundo and H. diffusa at 20% concentration. A. extensus also exhibited the lowest MIC value of 1.25% against E. coli. The antioxidant capacity of the methanol extracts was also estimated using DPPH method. V. negundo has the highest antioxidant capacity in comparison to other two plant extracts. These findings suggest the potential antimicrobial and antioxidant properties of the plant extracts and also their possible contribution to the overall medicinal and therapeutic benefits of rice beer.
Key words: Amphineuron extensus, Hedyotis diffusa, Vitex negundo, Antimicrobial, Antioxidant
15 Journal of Cell and Tissue Research 12(3) 3349-3355 (2012)
Assessment of isozyme diversity in the medicinal plant isabgol (Plantago ovata forsk.)
Kaswan, V., Joshi, A. and Maloo, S.R.
Department of Molecular Biology and Biotechnology, Rajasthan College of Agriculture, Maharana Pratap University of Agriculture and Technology, Udaipur-313001(Raj.) E. mail: vineetkaswan@gmail.com
Abstract: Isabgol is considered to be one of the important medicinal plants in India. In the present study 24 genotypes were collected from different geographical locations of India and three enzymes viz. esterase, peroxidase and superoxide dismutase were screened by native polyacrylamide gel electrophoresis (PAGE) technique to study the variation in isozymatic patterns. Only nine bands were resolved for young (28 days after sowing) leaf samples. The isozyme profile for esterase showed 4 bands with one polymorphic band whereas, peroxidase and superoxide dismutase profiles showed 2 and 3 monomorphic bands, respectively. The pooled data for Jaccard’s similarity coefficient values revealed that the genetic distance derived from isozyme analysis was very low (0-11%) due to the small number of polymorphic bands. The above study inferred that all genotypes might have some ancestral/phylogenic relationship due to migration/inflow of genes etc. and esterase banding pattern has offered a rapid and reliable method for estimation of variability/similarity between different genotypes that could be utilized for the gene pool conservation and also by the breeders for further improvement of this unique medicinal plant.
Key words: Isabgol, Plantago ovata, Isozyme, Genetic diversity
16 Journal of Cell and Tissue Research 12(3) 3357-3360 (2012)
Effectiveness of auxins in inducing in vitro adventitious root formation in Tylophora indica (burm. f.) Merrill
Rashmi, M. P., Vinaya, M., Vedamurthy, A. B. and Nayeem, A.
P. G. Department of Biotechnology, The Oxford College of Science, HSR Layout, Bangalore 560102.
E. mail: nayeemqaadri@gmail.com
Abstract: Tylophora indica is commonly known as Antamul or Indian ipecac, traditionally used as a folk remedy in treatment of bronchial asthma, bronchitis, rheumatism allergies, inflammation and as an Antidote. In this study, the effectiveness of type and concentrations of auxins such as indole acetic acid (IAA), indole butyric acid (IBA) and naphthaleneacetic acid (NAA), on the induction of adventitious roots from leaf explants of T. indica at different concentrations were tested on full strength Murashige and Skoog (MS) medium. The results indicated that, the best auxin was IAA at an optimal concentration a 1 mg/L as measured in terms of percentage of explants forming roots and the number of roots formed per explant. This study reports the first successful adventitious roots induction in T. indica, which promise a high potential of large scale commercial production in bioreactor for the pharmaceutical industries.
Key words: Adventitious root, T. indica, Medicinal plant, Tissue culture
17 Journal of Cell and Tissue Research 12(3) 3361-3366 (2012)
In vitro evaluation of tomato cultivars for salt tolerance
Baviskar, S.B., Patil, S.R. and Bainade, P.S.
M. P. K. V. Rahuri 440001, (M.S.). E. mail: sapana_bawiskar@yahoo.co.in
Abstract: Shoot tip of tomato cultivars Pusa Ruby, PKM-1 and Selection-22 was used for culturing in the MS basal media + 10 per cent CW fortified with BAP, kinetin and IAA in different concentration and combination to evaluate tomato cultivars for its suitability to saline conditions. NaCl at 50, 100, 150 and 200 mM concentration was used in the media. Shoot differentiation influenced by the different treatments of growth regulators indicated that treatment T11 (MS + 3.0 mgl-1 Kinetin + 0.4mgl-1 + IAA + 10% CW) was significantly superior. Germination of tomato seed occurred in all the three cultivar in MS + 0 and 50mM NaCl and was totally inhibited in MS + 100, 150, 200mM NaCl. Differentiation studies indicated that the tomato shoot differentiated in 0, 50 and 100mM NaCl and was inhibited from 150mM NaCl onwards. The shoot differentiation traits like days required for shoot initiation, number of shoot buds culture-1 and number of shoots elongated was decreased as salt concentration increased from 0 to 100mM NaCl. The shoots obtained from media with salt took more number of days to initiate roots as compared to the shoots obtained from media without salt in all the cultivars. Cultivar Pusa Ruby and Selection-22 was found to show better response as compared to PKM-1 for different aspects of shoot differentiation in 0, 50 and 100mM NaCl and hence identified as the cultivars suitable for growing in saline area.
Key words: In vitro, Tomato cultivars, Salt tolerance
18 Journal of Cell and Tissue Research 12(3) 3367-3372 (2012)
Cultivation of strawberry (Fragaria x ananassa Duch.) cv. chandler in specic climatic conditions of Jammu (J&K). 1: Mass propagation
Sharma, S., Wali, V. K., Sharma, M., Gupta, V.and Khajuria, S.
Division of Fruit Science, Faculty of Agriculture, Chatha Campus, Jammu- 180009. E. Mail: munishsangra@gmail.com
Abstract: A micro propagation method is described for strawberry (Fragaria x ananassa Duch.) Cv. Chandler. Vegetative buds were surface sterilized with HgCl2 (0.1 %) for 3 minutes which gave minimum contamination with maximum culture establishment. Of various treatment combinations, Murashige and Skoog (MS) medium + BAP 1.00 mg/l with GA3 2.00 mg/l resulted in maximum establishment of cultures in lesser time. MS medium + BAP 2.00 mg/l + GA3 2.00 mg/l gave maximum multiple shoots. Maximum rooting was obtained on MS medium (half strength) supplemented with IBA 1.00 mg/l and activated charcoal 200 mg/l.
Key words:, Strawberry, Micropropagation
19 Journal of Cell and Tissue Research 12(3) 3373-3376 (2012)
Cultivation of micro propagated plants of strawberry (Fragaria x ananassa Duch.) cv. Chandler in specic climatic conditions of Jammu (J&K). 2: Hardening and acclimatization
Sharma, S., Wali, V.K., Sharma, M., Gupta, V. and Khajuria, S.
Division of Fruit Science, Faculty of Agriculture, Chatha Campus, Jammu- 180009. E. mail: munishsangra@gmail.com
Abstract: The experimental plant material of Cv. Chandler was raised through tissue culture on Murashige and Skoog medium using vegetative buds as explants. The MS medium supplemented with BAP 2.00 mg/l + GA3 2.00 mg/l was used for shoot multiplication. The rooting was obtained on MS (half strength) medium fortified with IBA 1.00 mg/l and activated charcoal 200 mg/l. The in vitro rooted plantlets were hardened and acclimatized by using different treatments. Plants transplanted at the age of 4 weeks after root initiation gave maximum survival (100.00 %) during transplanting. These plants were hardened in glass beaker and polythene bags singly or in cluster. The maximum survival during hardening (100.00 %) was observed by covering the plantlets with glass beaker individually and kept in culture room. Out of various potting mixture tried, the potting mixture containing soil: sand and FYM (1:1:1 v/v/v) gave maximum height and survival of plantlets.
Key words: Fragaria x ananassa, Hardening
20 Journal of Cell and Tissue Research 12(3) 3377-3382 (2012)
Factors affecting germination and seedling growth of an endangered forest tree Wrightia tomentosa (Roxb.) Roem. & Schult. through in vitro zygotic embryo culture
Srinivas, P., Samatha, T., Shyamsundara Chary, R., Rajinikanth, M. and Rama Swamy, N.
Plant Biotechnology Research Lab, Department of Biotechnology, Kakatiya University, Warangal 506 009 (A P).
E. mail: swamynr.dr@gmail.com
Abstract: The species Wrightia tomentosa has medicinal value and importance in toy making industry. Due to its over exploitation the species has become an endangered. The present investigation reports on a protocol for conservation of W. tomentosa through zygotic embryo culture. The zygotic embryos were cultured on MS (Murashige & Skoog), WPM (Woody Plant Medium) and B5 (media. The germination percentage was found more on MS medium compared to WPM and B5 media. When the zygotic embryos cultured on full and half strength MS media containing 15-30 g/L sucrose it showed less percentage of germination and more days for germination. Maximum percentage (96%) of germination was obtained by culturing zygotic embryos on ¼ strength MS medium containing 7.5gm/L sucrose in comparison to the other media used. Germination was also affected by the orientation of zygotic embryos in culture. The zygotic embryos placed vertically on the medium were germinated early with more percentage and with healthy seedlings. The in vitro germinated seedlings were acclimatized in the culture room, transferred to research field and maintained under shady conditions. The survival percentage of plantlets was found to be 90% and the plants were morphologically normal, healthy and similar to the mother plant. This protocol can be useful for overcoming seed dormancy and also for rapid multiplication and conservation of an endangered forest tree W. tomentosa using zygotic embryo culture.
Key words: Wrightia tomentosa, Zygotic embryo culture
21 Journal of Cell and Tissue Research 12(3) 3383-3386 (2012)
Effect of kanamycin on growth of hypocotyl tissue of brinjal (Solanum melongena l. cv. Pusa purple long)
Bardhan, S.K., Sharma, C. and Srivastava, D.K.
Department of Biotechnology, Dr Y. S. Parmar University of Horticulture and Forestry, Nauni, Solan
173 230 (HP). E. mail: chhaya_uhf@yahoo.co.in
Abstract: Aminoglycoside antibiotic kanamycin is widely used as a selective agent in the genetic transformation with the neomycin phosphotransferase-II gene as the selective marker gene was evaluated. Successful plant transformation requires effective regeneration and selection systems. Increasing concentration of kanamycin generally resulted in full or partial inhibition of growth of plant tissue. A gradual decline in fresh weight in hypocotyl explants was observed with the increase in kanamycin concentration. Kanamycin strongly inhibited regeneration even at low doses. 50 mg/l kanamycin did not completely inhibit the growth at the initial stage, but later on callus formation and shoot regeneration was affected. In this paper, effects of kanamycin were studied in order to reveal effect mechanism of kanamycin on growth and development of hypocotyl tissue of brinjal
Key words: Kanamycin, Hypocotyl, Brinjal
22 Journal of Cell and Tissue Research 12(3) 3387-3393 (2012)
Morphological and histopathological changes in papaya due to virus isolate from middle gangetic plains of India
Singh, V. and Shukla, K.
D. D. U. Gorakhpur University, Gorakhpur 273 009 (U.P.). E. mail: vimlatalan@gmail.com
Abstract: The aim of this investigation was to study the morphological and histopathological changes induced by virus isolate infecting Carica papaya. L. in the middle gangetic plains of India. The samples of infected host were collected from 15 districts of eastern Uttar Pradesh representing this belt. Based on the visible symptoms, particle morphology (by Electron microscopy and Immunosorbent electron microscopy) and serology (DAC-ELISA and DIBA), the virus isolate was characterized to be Papaya ringspot virus (PRSV). The virus induced ringspot, distortion and mottling symptoms on foliage, stunting in stem, fragile roots and deteriorated fruit quality and quantity. Histopathological studies of 90 days old symptomatic leaf, stem and root sample revealed extensive deformation of parenchyma tissues in leaf, poor differentiation of vascular tissue without disintegration, increased number of calcium oxalate crystals in stem, reduced size of xylem cells in roots. The root tissues also showed increased lignifications and reduced protoxylem. The PRSV isolate under study did not produce any necrotic symptom as reported in some studies. Hyperplasia in cells externally visible as ringspots was observed. Hypertrophy in cells due to poor differentiation of secondary tissues caused weak and fragile stem and roots in host. The disease was very widespread in the study region causing 71-86% yield losses. The devastating impacts of the disease have restricted this crop to kitchen gardens only.
Key words:Papaya ringspot virus, Carica papaya L
23 Journal of Cell and Tissue Research 12(3) 3395-3400 (2012)
In vitro callus induction and plantlet regeneration in Fig (Ficus carica l.)
Dhage, S.S., Pawar, B.D., Chimote, V.P., Jadhav, A.S. and Kale, A.A.
State Level Biotechnology Centre, Mahatma Phule Krishi Vidyapeeth, Rahuri 413 722, Maharashtra.
E. mail: bhau.raje@gmail.com
Abstract: In vitro plant regeneration was tried in four fig cultivars namely Brown Turkey, Conadria, Deanna and Poona Fig using leaf explants obtained from in vitro established shoots. Optimum callusing was observed on MS medium supplemented with 2.0 mg/l thidiazuron (TDZ) and 4.0 mg/l 2-iso-pentenyl adenine (2 iP). Among the cultivar studied, Brown Turkey showed maximum response to callusing (85.8 %) and showed shooting on same medium. Earliest callus induction was observed on MS medium supplemented with 2.0 mg/l TDZ and 2.0 mg/l IBA (Indole-3-butyric acid). Shooting was induced from callus on transfer to MS medium supplemented with 0.5 mg/l á-naphthalene acetic acid (NAA) and 7.0 mg/l TDZ. Shoots developed roots following transfer to half strength MS medium supplemented with 1.0 mg/l IBA and 2 g/l activated charcoal.
Key words: In vitro, Callus culture, Ficus carica
24 Journal of Cell and Tissue Research 12(3) 3401-3408 (2012)
Somatic embryogenesis and plant regeneration in cotton cv. Coker-312(Gossypium hirsutum L.)
Sangannavar, P.A., Katageri, I.S.,Vamadevaiah, H.M. and Khadi, B.M
Department of Genetics and Plant Breeding, University of Agricultural Sciences, Dharwad 580005, Karnataka.
E. mail: ikatageri@yahoo.com
Abstract: Between hypocotyl and cotyledon as a explants, Callus induction rate (days taken for callus induction), callus induction response (callus induction on number explants and amount of callus produced) and nature of callus in combination with different plant growth regulators in MS media, use of hypocotyls was found more useful by culturing on MS medium supplemented with 0.1 mg/l 2, 4-D plus 0.5 or 1.0 mg/l kinetin. Callus induction was early, high in response and callus was friable in nature, this friable callus is useful in formation of somatic embryogenesis. Emergence of torpedo or cotyledonary stage embryos was observed in 0.01 mg/l 2, 4-D plus 0.1 mg/l kinetin added to MS medium, 8-9 weeks after cultures of friable calli. Maturation of cotyledonary stage embryos to produce normal plantlets with both shoots and roots was observed in MS salts without any plant growth regulators. Incubation of plantlets cultured on MS salts for 4 weeks found beneficial in establishment of plants in ex vitro condition. Additionally after transplanting plantlets to soil and peat mixture (1:1), placing them in plant growth chamber for a week before shifting them to ex vitro condition was found most useful in establishment of more number of plants under ex vitro condition.
Key words: Somatic embryogenesis, Plant regeneration, Cotton
25 Journal of Cell and Tissue Research 12(3) 3409-3416 (2012)
In vitro response of five high reserpine yielding genotypes of Rauvolfia serpentina roxb. (L.) Benth through axillary bud culture
Mishra, Y. and Bouddha, S.
Tropical Forest Research Institute, P.O- R.F.R.C, Mandla Road, Jabalpur 482021 (M.P). E. mail: m_yogiraj@yahoo.com
Abstract: The biotechnological tools are important to select, multiply, and conserve the critical genotypes of medicinal plants. The present study was undertaken to compare in vitro response of five superior genotypes of Rauvolfia serpentina. These genotyopes were namely, GO-MN, GO-SG, OR-AG, KL-PB and KL-AJ, collected from Maina (Goa, GO-MN), Sigone (Goa GO-SG), Angul (Orissa, OR-AG), Perumboover (Kerala, KL-PB) and Anjanakund (Kerala, KL-AJ) and reported to possesss reserpine ranging from 0.07% to 0.09%. The in vitro responses of these genotypes were evaluated in terms of sprouting of axillary buds, shoot multiplication, rooting and hardening. GO-MN genotype was recorded with maximum sprouting (84%) and number of shoot/explant (3.10). The MS medium supplemented with 1µM NAA + 10 µM BA was recorded with highest shoot multiplication rate of 9.66 fold with maximum shoot length (9.33) and node number (5.78) in GO-MN genotype. GO-MN invariably emerged as the best genotype to produce 81% rooting on ½ B5 medium whereas GO-SG and KL-AJ produced at par root number (7) on ½ MS medium. The regenerated plantlets were successfully acclimatized in pre-sterilized soilrite without any pathogenecity and transferred to the field with 100% survival. The developed protocol can be used for en masse propagation and conservation of selected high reserpine yielding genotypes of Rauvolfia serpentina.
Key words: Genotype, Rauvolfia serpentina, Reserpine
26 Journal of Cell and Tissue Research 12(3) 3417-3422 (2012)
Effect of Azadirachta indica leaf extract on callus induction and its enhancement for the plantlet regeneration in Carica papaya L. (var. Pusa dwarf)
Srivastava, N., Tiwari, N. and Sharma, V.
Department of Bioscience and Biotechnology, Banasthali University, P.O. Banasthali Vidyapith, 304022, Dist. Tonk, Rajasthan. E. mail: vinaysharma30@yahoo.co.uk
Abstract: Effect of various concentrations of Azadirachta indica L. leaf extract (ALE) on the frequency of callus induction and regeneration of green plantlets in Carica papaya L. (var. Pusa dwarf) was investigated using modified MS media (MMS media) supplemented with IAA alone and in combinations with BAP. Different explants (leaf, petiole and stem cuttings) from 10th day old seedlings were used for maximum callus induction, growth of protocorm like bodies (PLBs) and its regeneration capacity. Out of three concentrations (50,100 and 150 mg l-1) of A. indica leaf extract , 100 mg l-1 inoculated with stem explants was found to be optimal for the callus induction and its regeneration capacity.
Key words: Azadirachta indica L., Plantlet regeneration, Carica papaya
27 Journal of Cell and Tissue Research 12(3) 3423-3429 (2012)
Production of andrographolide from callus and cell suspension culture of Andrographis paniculata
Sharma, S. N.and Jha, Z.
Department of Plant Molecular Biology & Biotechnology, IGKV, Raipur 492 006, India
E, mail : shivsharma.bt@gmail.com
Abstract: Andrographolide is the principal component of the medicinal plant Andrographis paniculata which is attributed for its diverse pharmacological properties. Traditionally andrographolide was extracted from the leaves, stems and other parts of the plant. Leaf contains maximum andrographolide content (2-3%) in compare to other parts of plant. In the present study callus and cell suspension culture was established and explored for andrographolide content on them. Callus was induced by culturing leaf discs on Murashige and Skoog (MS) medium with different concentration of 2,4-D and combination of 2,4 D + Kinitin, 2,4D +NAA and BAP + NAA. Best callus induction were obtained at lower concentration of 2,4-D (0.5 and 1.0 mg/l), combination of 2,4D+NAA (1+1mg/l) 2,4 D + Kin (1.0+0.5mg/l) and BAP + NAA (1.0+1.0mg/l). Initiation of cell suspension culture of A. paniculata was established by inoculating fresh friable fragments of callus on MS medium supplemented with respective callus grown medium. Callus from the 2,4D +NAA (1+1mg/l) was friable and best to release cells in suspension culture. Cells were successfully subcultured every 20-21 days on liquid MS media supplemented with respective callus grown medium. The highest amount of andrographolide (32.40 ±2.22) is found in 2, 4-D + NAA (1mg+1mg) cell suspension culture followed by 2,4 D+Kin (1.0mg+0.5 mg) (31.95 ±2.21). These studies provided an efficient way to produce andrographolide from A. paniculata callus as well as cell suspension culture. The developed method was used for the large quantity production of andrographolide.
Key words: Andrographis paniculata, Andrographolide, Callus culture
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