CTR Publication :: Past Abstracts

2. Journal of Cell and Tissue Research 14(1): 4009- 4017 (2014)

In vivo antioxidative effect of bamboo (Bambusa arundinacea), leaves extract on arsenic induced hepatic oxidative stress

Kalia, K., Chiragini, H.M. and Sood, P.P.

BRD School of Biosciences, Sardar Patel University, Vallabh Vidyanagar 388 120, Gujarat
E. Mail: kirankalia_in@yahoo.com

Abstract: The present study has been carried out to investigate the protective role of the bamboo leaves methanolic extract (BLME) against arsenic induced hepatic oxidative insult. Sodium arsenite at a dose of 5.55 mg/ kg body weight (b.w.) per day was preferred as the source of arsenic in our study. We have included various liver cell injury markers like serum glutamate pyruvate transaminase (SGPT), serum glutamate oxaloacetate transaminase (SGOT) and oxidative stress markers as a parameter to measure degree of oxidative damage viz. advanced oxidation end product (AOPP), reduced glutathione (GSH), thiobarbituric acid reactive substances (TBARS), and healing due to bamboo leaves extract. Activities of enzymes, like superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), responsible for antioxidant defense system were also estimated. Bamboo leaves extract at three different doses i.e. 50 mg, 250 mg and 500 mg per kg b.w. for last fifteen days after arsenic treatment prior to sacrifice altered most of the above parameters of oxidative stress caused by arsenic. Bamboo leaves extract eliminate the effect of arsenic induced oxidative stress by being a potent free radical scavenger.

Key words: Bamboo leaves, Arsenic, Hepatic protection

3. Journal of Cell and Tissue Research 14(1): 4019- 4026 (2014)

Molecular characterization of ampicillin resistant poultry isolates of Salmonella typhimurium

Verma, J. K., Pilkhwal, D., Tamuly, S., Rajesh Kumar, Avdesh Kumar and Saxena, M. K.

Animal Biotechnology Center, Department of Veterinary Physiology and Biochemistry, College of
Veterinary and Animal Sciences, G.B Pant University of Agriculture and Technology Pantnagar 263145.
E. mail: g2biotech@rediffmail.com

Abstract : Multiple drug resistance and high genetic variation among the field isolates are one of the two major constrains in eradication and control of Salmonella Typhimurium from poultry. In the present study 51 isolates of Salmonella typhimurium which were isolated from eggs were analyzed for their responses against commonly used sixteen antibiotics. Genetic variation among these isolates was studied by using ERIC-PCR, REP-PCR and AP-PCR. Most of the isolates were possessing resistance for more than three antibiotics. Maximum resistance was exhibited for Ampicillin and Augmentin. Among three (ERIC-PCR, REP-PCR and AP-PCR), AP-PCR shown highest discrimination ability D value 0.89 . On combined molecular-typing discriminative index (D value 0.984) was achieved. Present findings indicates that these isolates having high genetic variation and multiple drug resistance which may cause serious health problems in countries like India where hygienic conditions are not very good.

Key words: Salmonella Typhimurium, Poultry isolates

5. Journal of Cell and Tissue Research 14(1): 4033- 4036 (2014)

Effect of marinating chicken meat with garlic paste at room temperature on common food borne pathogens

Sudarshan, S., Fairoze, M. N., Badhe, S. R. and Prabha, R.

Department of Livestock Products Technology, Veterinary College, Bidar 585 401;
E. mail: drsudi.lpt@gmail.com,

Abstract: The enhancement of flavour in food by addition of spices has been practiced from time immemorial. The role of spices as a biopreservative is one of the important aspect noticed in recent years. The incidence of pathogens in chicken meat especially of salmonella and Escherichia coli has been reported in India and elsewhere. Among the commonly used spices, the role of garlic in different forms as an antibacterial agent containing food stuff has been well documented. The present study was under taken with an objective of determining the antibacterial efficacy of garlic paste on common food borne pathogens in chicken meat by marinating under ambient temperature. Chicken meat was marinated with 1.5 and 2.5 percent of garlic paste (w/w) wrapped in aluminum foil for different durations. Microbial analysis was carried out at 0, 2, 4, and 6 hr interval of marination at room temperature (26 ± 20 C). Among the different groups of garlic paste concentrations and time combinations, 2.5 percent garlic paste and 4 hr duration of marination showed significant (P< 0.01) reduction in viable count of total bacteria, staphylococci, E. coli and salmonella count.

Key words: Garlic paste, Chicken meat, Food borne pathogens

6. Journal of Cell and Tissue Research 14(1): 4037- 4042 (2014)

Relative abundance of cell organelles in steroidogenic cells of corpus luteum of goat in different reproductive phases

Batra, S. and Sharma, R.K.

Department of Zoology, Kurukshetra, University, Kurukshetra 136119. Haryana,
E mail: soniabatra2dec@rediffmail.com1

Abstract: Variations in relative abundance of different cell organelles of steroidogenic cells from seven categories of goat corpus luteum viz. small (1-5 days), medium (6-10 days), large (11-15 days), regressing (16-21 days), previous cycle, penultimate cycle and pregnancy (<30 days), have been analysed. In granulosa luteal cells, smooth endoplasmic reticulum variations were not prominent. Rough endoplasmic reticulum increased from small to large category and was most abundant in corpus luteum of pregnancy. The number of mitochondria, Golgi complex and secretory granules increased from small to large category. The maximum frequency was observed during the pregnancy. Vesicles were prominent in the previous and penultimate categories. The numbers of lipid droplets were maximum in corpus luteum of the previous cycle followed by the regressing category while lysosomes out numbered in corpus luteum of the penultimate cycle. In theca luteal cells, smooth endoplasmic reticulum and number of mitochondria increased from the small to large category and maximum numbers were observed during the pregnancy. Golgi complexes were abundant in corpora lutea of the large category and that of the pregnancy while vesicles showed their preponderance in corpus luteum of the penultimate category. Numbers of lipid droplets were more in corpus luteum of the previous cycle. Lysosomes were abundant in number in theca luteal cells of corpus luteum of the regressing category. Secretory granules were higher in number in corpora lutea of the large and pregnancy categories. These variations in relative abundance of cell organelles in steroidogenic cells shall be discussed in relation to their physiological obligations and endocrine profile at specific stage of reproductive cycle.

Key words: Corpus luteum, Cell organelles, Goat ovary
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7. Journal of Cell and Tissue Research 14(1): 4043- 4048 (2014)

Polymorphism, dynamics and ultrastructure of steroidogenic cells of caprine corpus luteum

Batra, S. and Sharma, R.K.

Reproductive Biology laboratory, Department of Zoology, Kurukshetra University,Kurukshetra 136119 (Haryana). E. mail: soniabatra2dec@rediffmail.com1

Abstract: Seven categories of corpora lutea i.e. small ( 1-5 days), medium (6-10 days), large (11-15 days), regressing (16-21days) and of previous cycle, penultimate cycle and pregnancy (<30 days) of goat (Capra hircus) were analyzed for cellular polymorphism, dynamics and ultrastructural variations. Histologically, corpus luteum was comprised of two types of steroidogenic cells. The diameter of granulosa luteal cells increased from 33.00 m in small to 42.00 m in large category. The theca luteal cells dimensions increased from 19.25 to 28.00 m. The cell diameters of both these categories of cells showed maxima during pregnancy whereas corpora lutea of previous and penultimate cycles revealed the minimum values. The relative abundance of different types of steroidogenic and non-steroidogenic cells were studied in different reproductive phases of the goat. Blood capillaries and the endothelial cells showed statistically significant variations in different phases. Ultrastructurally, both theca and granulosa luteal cells revealed variations in the relative abundance of different of cell organelles. Mitochondria, smooth endoplasmic reticulum, Golgi complex and secretory granules increased with the increase in granulosa cell diameter during pregnancy. The lysosomes, lipid droplets and vesicles were maximum in the theca luteal cells showing minimum values of diameters in penultimate cycle.

Key words: Goat ovary, Corpus luteum

8. Journal of Cell and Tissue Research 14(1): 4049- 4053 (2014)

Molecular studies on meat quality gene in Bandur sheep

Sunilkumar, M.A., Nagaraja, C.S., Jayashankar, M.R., Fairoze, N. and Veeregowda, B.M.

Department of Animal Genetics and Breeding, Veterinary College, Hebbal, Bangalore 560024,
Kvafsu, Karnataka. E. mail: sunilvetagb@gmail.com, Cell: 9632641299

Abstract: Calpastatin (CAST) has been known as candidate gene in muscle growth efficiency and meat quality. A study was conducted with the objective of analyzing the genetic variation at the CAST region of Bandur breed of sheep by using PCR-RFLP technique. Genomic DNA was isolated from blood samples of 79 Bandur animals. The genetic diversity of ovine CAST locus was investigated by PCR-RFLP. A 622 bp fragment of exon 1C/1D from domain 1 region including the intron was amplified. The size of the amplified product was same in all the animals studied indicating conservation of DNA at this locus. Two restriction endonucleases MspI and NcoI were used to detect the genetic variation of the experimental unit in the CAST gene. CAST/MspI polymorphism was observed as three patterns which allowed the identification of two alleles viz., A and B. The allelic frequencies for A and B alleles were 0.74 and 0.26 respectively. The population studied was in Hardy- Weinberg equilibrium. CAST/NcoI polymorphism also showed three genotypes with two alleles, viz., A and B, the gene frequencies were 0.697 and 0.303 respectively. The NcoI digested the allele B amplicon, but not allele A. The population studied was in Hardy- Weinberg equilibrium. The sequence analysis indicated that there is high homology between the present result and the published ovine CAST sequences. There was query coverage to the extent of 95%. The present study revealed the genetic variability at CAST gene locus in Bandur sheep. Further study is suggested to understand the association of this gene with meat quality traits.

Key words: Bandur, Calpastatin gene, Polymorphism

10. Journal of Cell and Tissue Research 14(1): 4059- 4064 (2014)

CXCR2 gene polymorphism and its association with mastitis in murrah buffalo

Wani, S.A., Sangwan, M.L., Kumar, A., Dar, M.A., Dhubay, T. and Kumari, A.

Division of Animal Biotechnology, Lala Lajpat Rai University of Veterinary and Animal Science, Hisar 125,004 (Hariyana). E. mail: wanisajad759@gmail.com, Cell 07599172038

Abstract: Present study was carried out on interleukin-8B receptor gene of Murrah buffalo with an objective to find polymorphism of interleukin-8B receptor gene and its association with mastitis. Genomic DNA was isolated from randomly selected lactating buffalo by phenol-chloroform isoamyl alcohol method. Quality and quantity of DNA was checked and it was found to be of good quality as determined by agarose gel electrophoresis. A self designed set of primers covering 459 bp nucleotides of IL-8B receptor gene which was used and PCR products of 459 bp were obtained. PCR-RFLP was done for all the samples from healthy and mastitis animals using Msp I, AvaII, BlpI restriction enzymes. Pattern of resolution of digested fragments by all the restriction enzymes was found same i.e xx, yy, and zz respectively in both two groups of animals. All monomorphic bands were found. Target sequence IL-8B receptor gene of Murrah buffalo was sequenced which has been found to span over 459 bp long. The multiple sequence alignment between four healthy and four mastitic animals choosen randomly revealed variation at one places. M2 (mastitic) animal showed substitution of C by G at position no. 6. Basic local alignment search tool (BLAST) analysis revealed sequence identity of target region (IL-8B receptor gene) of Murrah buffalo was, 99% with Bos taurus× Bos indicus (F927834.1), 98% with Bos taurus (DQ328664.1), 97% with Ovis aries (XM004005420.1) and 93% with Sus scrofa (AK230995.1). No significant association of Interleukin-8B receptor gene with mastitis was found. In phylogenetic tree, the target sequence of interleukin-8B receptor gene of Bubalus bubalis are found more closely related to Bos taurus× Bos indicus and Bos taurus than to Ovis aries and Sus scrofa

Key words: Murrah buffalo, Interleukin-8B receptor,

11. Journal of Cell and Tissue Research 14(1): 4065- 4068 (2014)

Study of prolactin gene polymorphism in Murrah buffalo

Biradar, S.M., Unaune, K.P., Dodamani, S., Mhatre, P.S., Londhe, S.P., Pawar, V.D., Sawane, M.P. and Umrikar, U.D.

Department of Animal Genetics and Breeding, Bombay Veterinary College, Parel, Mumbai 400012.
Maharashtra. E. mail: drsuryakantbiradar.com, Cell: 089514 02881

Abstract: Prolactin hormone plays an important role in the initiation of lactation. This is a protein molecule translated from prolactin gene located on chromosome 23 and consists of 5 exons and 4 introns encoding the 199 amino acid mature protein. Prolactin-Rsa I in exon 3 was found to be associated with milk yield in exotic dairy cattle. Therefore, the present study was undertaken to detect such polymorphism for prolactin locus (exon 3) using PCR-RFLP in Murrah buffaloes. The genomic DNA was isolated from 50 whole blood samples of Murrah buffaloes of Chitale Dairy Farms, Maharashtra. The PCR amplification of 156 bp prolactin gene (exon 3) fragment was carried out using bovine specific primers, Forward -5’ CGA GTC CTT ATG AGC TTG ATT CTT 3’ and Reverse -5’ GCC TTC CAG AAG TCG TTT GTT TTC 3’. Bovine specific primers successfully amplified 156 bp prolactin gene (exon 3) in Murrah buffalo indicating the sequence homology of that region of gene between cattle and buffalo. Amplified PCR products of prolactin gene (exon 3) after digestion with restriction enzyme Rsa I revealed intact product of 156 bp indicating that there was no restriction site for Rsa I in amplified prolactin gene (exon 3). As observation of only one allele i.e. ‘A’ in studied population in the present study indicating monomorphic nature of the locus showing only AA genotype which may be a characteristic of Murrah buffalo.

Key words: Prolactin gene, Murrah buffalo

12. Journal of Cell and Tissue Research 14(1): 4069- 4072 (2014)

Effect of paclobutrazol and sucrose on in vitro corm formation in saffron (Crocus sativus)

Zaffar, G., Ahmad, M., Shahida, I., Razvi, S. M., Habib, M., and Ahmad, A

Division of Genetics and Plant Breeding Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir Shalimar, Srinagar 191 121 (J &K) E. mail: sahilmushtaqdar@rediffmail.com

Abstract: Saffron being triploid ( 2n = 3x=24) is sterile and vegetative propagation ( through corms) is the exclusive mode of propagation, this method is slow and the corms can be infected by corm rot disease which gets multiplied during replanting process, affecting corm growth and development which consequently leads to reduction in flower and saffron yields. In vitro propagation of saffron through cormogenesis can be an efficient alternative method for large scale propagation of pathogen free corms if a reproducible and efficient in vitro micro-propagation protocol is available. To develop a protocol for in vitro micro corm formation in saffron, effects of different concentrations of sucrose in combination with paclobutrazol were investigated. Embryogenic callus derived by culturing leaf segments and corm sections on Murashige and Skoog (MS) medium supplemented with 1.0 mg l-1 2,4-D and 1 mgl -1 BA. Shoots were regenerated from callus by sub culturing on MS medium fortified with 1.0 mgl-1 BA and 1.0 mgl-1 NAA. In vitro regenerated shoots from callus were cultured in Murashige and Skoog (MS) medium supplemented with Paclobutrazol (PAC) 0-10 mg1-1 and sucrose (60-120 g1-1). Higher concentrations of PAC (5mgl -1 ) along with 90 g1-1 sucrose resulted in formation of relatively larger micro corms. There was a significant interaction between paclobutrozol and sucrose for micro-corm weight. The protocol for micro-corm formation can be used as tool for mass multiplication of disease free planting material for up scaling cultivation of saffron.

Key words: Saffron (Crocus sativus), Micro corm

13. Journal of Cell and Tissue Research 14(1): 4073- 4076 (2014)

Micropropagation of saffron (Crocus sativus l.): A review

Ahmad, M., Razvi, S.M., Zaffar, G., Nengroo, Z.I., Mir, S.D., Wani, B.A., Ameeque, A. and Habib, M.,

Division of Genetics and Pant Breeding, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir Shalimar campus, Srinagar 191 121. E. mail: sahilmushtaqdar@rediffmail.com

Abstract: Tissue culture is useful method for large scale production of healthy corms and also avenues for creating variability through in vitro use of mutagens and polyploidizing agents. Induction of callus and subsequent regeneration of plants is suggested as possible means of introducing new variation. In vitro micropopagation for product formation its utility is increasing the amount of crocin, picrocrocin and safranal. Selection at cellular level is likely to help in isolating cell lines rich in these three chemical compounds which account for popularity of saffron stigma. Like many other cormous and bulbous plants, meristem culture and regeneration through tissue culture is the most effective method for mass production of healthy and pathogen-free saffron plant materials. In vitro corm production through somatic embryogenesis as an effective method for plant regeneration could be one of the important aspects of tissue culture application in saffron.

Key words: Crocus sativus L., In vitro Micropopagation

14. Journal of Cell and Tissue Research 14(1): 4077- 4082 (2014)

Efficient regeneration system for a very low proliferating diploid (AB) banana CV Elakki bale (Syn neypoovan)

Meenakshi, S., Sowmya, H.D., Usharani, T.R., Vijayalakshmi, Pavithra, S. and Sukhada, M.

Indian institute of horticultural research, Hessarghatta lake post, Bangalore-560098, Karnataka.
E. mail: sukhada.mohandas@gmail.com, Cell: 094480 01836

Abstract: Banana cv Elakki Bale (syn NeyPoovan AB) is the most favoured cultivar of Southern India. The cultivar is known for its special taste, aroma and the long keeping quality of its fruits. The demand for planting material of this cultivar is huge and tissue cultured plantlets are not available as the multiplication rate is very low. As the cultivar includes a B genome the multiplication rate is slower when compared to those with A genome. Regeneration through meristem culture offers disease free planting materials against banana bunchy top virus, leaf spot disease and panama wilt. Therefore, the present study was taken up to develop regeneration protocol which gives high rate of shoot proliferation. MS medium containing 5 mgl-1 BAP, kinetin 1 mgl-1 (B17) was found to be the best in shoot proliferation with a multiplication rate of 4.1 among the different concentrations tested. Multiplication rate increased after 3rd subculture and did not decline even after 7th subcultures as it is a common problem in other varieties of banana. Plants were regenerated and successfully hardened. Somaclonal variation studied in few field grown plants was within the admissible limit.

Key words: Micro propagation; Diploid banana

15. Journal of Cell and Tissue Research 14(1): 4083- 4088 (2014)

Molecular characterization of date palm (Phoenix dactylifera) using combined marker analysis grown in Kutch region of India

Srivashtav, V.S., Solanki, V.H., Patel, H.K. and Kansara, R.

Department of Plant Molecular Biology and Biotechnology, Navsari Agricultural University, Navsari 396450, Gujarat. E. mail: vishal_bt85@yahoo.com

Abstract: In Date palm (Phoenix dactylifera L.) molecular characterization to find the best genotype and male/female identification at an early stage before flowering is of great economic significance to the farmers. Genetic finger printing using molecular markers is an important tool. In present investigation four types of markers RAPD, ISSR, SSR, ITS gene were applied in 5 unknown genotypes of Date palm (female 1, female 2, female 3, Male 2 small and Male1 tall). Four type of markers (highly polymorphic) with two best primers for each marker (RAPD, ISSR, SSR and ITS), were used. Amplification of genomic DNA of 5 Date palm genotypes yielded 32 scorable fragments out of which 17 were polymorphic, with an average of 4.25 polymorphic fragments per primer. Number of amplified fragments ranged from 3 (BNL-448) to 10 (OPG-2 and ITS-3) and varied from ~200bp to ~4kb. Percentage of polymorphism ranged from 20% (ITS-3) to a maximum of 100% (BNL-448). Cluster analysis using UPGMA contains two clusters A and B with Jaccard’s similarity matrix with low value of 0.660 in that female 2 genotype was distinct. On the other hand, cluster A contains female 3 and Male 1 tall with highest value of 0.810 and Male 2 small genotype having similarity value of 0.790. In cluster A1 female 1 genotype with similarity value of 0.750 was found diverse in cluster A.

Key words: Date Palm, Markers (RAPD, ISSR, SSR, ITS)

16. Journal of Cell and Tissue Research 14(1): 4089- 4098 (2014)

Sequence homology based classification and characterization of microrna families from selected plant species

Mahale, B.M., Gund, S.V., Daspute, A. and Borphukan, B.

Department of Biotechnology, University of Agricultural Sciences, Dharwad 580005, Karnataka;
E-mail: barkumahale@gmail.com, Cell:08971608525

Abstract: MicroRNAs (miRNAs) regulate many important aspects of plant development. It suggests their key roles in the evolution of plants. Evolutionary divergence studies of miRNAs finds its importance for the same reason. Although, miRNAs are 20-22 nt RNA molecules, their mature sequences play pivotal role in target identification. Studies exploring miRNA sequence characteristics are expected to open new vistas in understanding complex nature of small RNA biology and evolution. Using MEGA4 software, we identified phylogenic relationship between miRNAs, collected from three species independently, based on their sequence similarities. We attempted to group 243 Arabidopsis miRNAs into five distinct groups based on their phylogenic key positions and sequence characteristics. Following same analysis, the 345 rice miRNAs were classified into four groups, whereas 148 of sorghum in six well distinct miRNA groups. Further, the most generalized nucleotide sequence was identified for each miRNA group from each species using Position Weight Matrix analysis.

Key words: Arabidopsis thaliana, MicroRNAs

17. Journal of Cell and Tissue Research 14(1): 4099- 4103 (2014)

In vitro regeneration of pigeon pea {Cajanus cajan (l.) millsp.} genotype GT-102 using apical meristem

Parekh, M.J., Mahatma, M. K. And Kapadia, C.V.

Department of Plant Molecular Biology and Biotechnology, Navsari Agricultural University, Navsari-396450, Gujarat. Gujarat. E. mail: maheshmahatma@gmail.com, Cell: 097250 18784

Abstract: A protocol for plant regeneration via apical meristem was developed in high-yielding local pigeon pea genotype GT-102. Multiple shoots were induced directly from apical meristem explants on Murashige and Skoog’s (MS) medium supplemented with different concentrations of BAP, Kinetin and TDZ either singly or in combination. Type of cytokinin and its concentration influenced the frequency and number of multiple shoots. BAP was better for multiple shoots induction compared to other cytokinin. The well-multiplied shoots were elongated on MS medium supplemented with different concentrations of cytokinins and with GA3. Further well elongated plantlets were transferred in MS medium supplemented with different concentrations of IBA. Regenerated plants were transferred to soil and grown to maturity with 80% survival and these tissue culture-raised plants produced viable seeds. This protocol to produce multiple numbers of shoots with a high frequency and their subsequent conversion to whole plants offers potential for use in gene transfer and development of transgenic in this important grain legume.

Key words: Pigeon pea, Genotype gt-102

18. Journal of Cell and Tissue Research 14(1): 4105- 4111 (2014)

In vitro protocol for induction of gametic embryos formation in maize (Zea mays l.)

Choudhary, P., Jhajharia, A. and Singh, D.

Department of Plant Breeding & Genetics, College of Agriculture, Jawaharlal Nehru Krishi Vishwa
Vidyalaya, Jabalpur 482 004 (M.P.); E. mail: arunstatistics@gmail.com, Cell: 087651 20512

Abstract: The aim of the present study was to establish a protocol for the transformation of microspores and gametic embryos in maize. The response to isolated microspore culture of the three androgenic genotypes HKI-1011, HKI-1128 and HKI-1344 was studied. The response of the three genotypes to anther culture and the ability of their gametic embryos to undergo somatic embryogenesis and further regeneration of double haploid plants were also studied. The results revealed that the microspore culture approach results in a very poor androgenic response, despite the improvement achieved by adding activated charcoal (a max. of 43 embryos per 80000 microspores). The anther culture protocol, results in a high number of gametic embryos (a max. of 810 embryos per 100 anthers) that undergo somatic embryogenesis and regenerate into diploid plants at a high rate. For these reasons, preferentially gametic embryos from anther culture, rather than microspores, should be considered as targets for genetic transformation.

Key words: Gametic embryos, Maize

19. Journal of Cell and Tissue Research 14(1): 4113- 4116 (2014)

In vitro regeneration of lathyrus (Lathyrus sativus L.) as influenced by mutagen treatment

Bainade, P.S., Patil, S.R., Deshmukh, S.G. and Sawant, P.V.

Department of Botany, College of Agriculture, Nagpur 440005, India
E. mail: pradeeps.bainade@gmail.com, Cell: 94041 47345

Abstract: In vitro plant regeneration has been obtained from leaf segments of 10-15 days old seedling, which were used as explant and B5 basal media + 2.0 mg/l NAA + 0.5 mg/l BAP, was used for callus induction and differentiation. The leaf explant of Lathyrus cv. NLK- 40 exhibited 92% response to callusing and it took 30-32 days to produce full grown green compact callus. The fully grown callus then treated with different combinations of EMS as 0, 0.1, 0.2, 0.4, 0.6, 0.8 and 1.0 % for 30 and 60 minutes. Out of these treatments the callus growth was observed only in treatments control, 0.1% EMS for 30 and 60 minutes and 0.2% EMS for 30 and 60 minutes. Shoot differentiation ranged from 33.33 to 62.22 % and it was found only in treatments control, 0.1% EMS for 30 and 60 minutes within 3-4 weeks. The shoot buds that developed 2-3 cm long were excised and cultured on ½ MS basal + 0.01 mg/l IBA which developed roots within 3 weeks.

Key words : Lathyrus, In vitro regeneration, Mutagenesis

20. Journal of Cell and Tissue Research 14(1): 4117- 4122 (2014)

Therapeutic potentials of a herbal formulation in paracetamol induced hepatotoxicity in albino rats

Radhika, J., Sangeetha, D., Yuvarani, S. and Jothi, G.

Department of Biochemistry, Srimad Andavan Arts and Science College, Tiruchirappalli 620005.
E. mail: radiarun2005@yahoo.co.in. Cell: 077080 07402

Abstract: Liver disease is a major threat to public health and is a worldwide problem. Disease are mainly caused by pollutants, viral infection, auto immune disorders, alcohol and drugs. The present study was designed to evaluate the therapeutic potentials of a polyherbal formulation in drug induced liver damage in Albino rats. Wistar albino rats were divided into six groups comprising of six rats each. Group I served as a normal control. Group II was induced with a single dose of paracetamol (2g/kg body weight), Group III & IV animals were induced with paracetamol and treated with the herbal formulation under study (100mg and 200mg/kg body weight respectively) for 15 days, Group V treated with formulation (200mg/kg body weight) alone and Group VI induced with paracetamol and treated with Liv-52 (3ml/kg body weight). The following biochemical parameters viz., hepatic marker enzymes (AST, ALT, ALP, and GGT), Na+ /K+ ATPase, serum bilirubin, total protein liver glycogen and lipid profile were analyzed. The results depict an elevated level of enzyme markers, serum bilirubin, serum and hepatic cholesterol, triglycerides, serum phospholipids and lowered levels of total protein, liver glycogen, hepatic phospholipids and Na+/K+ ATPase in the paracetamol induced rats when compared to normal control. The administration of the formulation led to the normal functioning of the liver with the restoration of the parameters to near normal in the Group III and IV animals. The present study suggest that the treatment with the polyherbal formulation comprising of Coriandrum sativum L., Tephrosia purpurea Linn. and Tinospora cordifolia (Willd.) in the ratio 2:1:1, enhances the recovery of the liver from the paracetamol induced damage.

Key words: Paracetamol, Hepatotoxicity, Herbal formulation

21. Journal of Cell and Tissue Research 14(1): 4123- 4130 (2014)

Effect of growth regulators on in vitro propagation of Aloe barbadensis mill. and assessment of genetic fidelity through RAPD markers

Sahoo, S. and Rout, G.R.

Department of Agricultural Biotechnology, College of Agriculture, Orissa University of Agriculture & Technology, Bhubaneswar-751003, Odisha. E. mail: grrout@rediffmail.com, 094373 08014

Abstract: A successful protocol was developed for mass propagation of Aloe barbadensis Mill., an important medicinal plant. Multiple shoots were induced in apical and axillary meristems on Murashige and Skoog (1962) medium supplemented with 1.0 – 2.0 mg/l 6-benzylaminopurine (BAP), 0.25 -0.5 mg/l Kinetin and 3% sucrose. Inclusion of 0.5 mg/l 1-napthaleneacetic acid (NAA) enhances the rate of shoot elongation and multiplication. The rate of multiplication was higher when the cultures were incubated under continuous light rather than the 16h photoperiod. Rooting was readily achieved upon transferring the microshoots onto ½ strength MS basal semi-solid medium supplemented with 0.5 mg/l NAA or IBA after 8 - 10 days of culture. Micropropagated plantlets were acclimatized and successfully grown in soil mixture. About 90% of micropropagated plantlets were hardened in the polyhouse and successfully established in the soil. Random ampliûed polymorphic DNA marker was used to detect the variability among the micropropagated plants developed through in vitro. The results showed that there was no polymorphism among the micropropagated plants. This study will help for propagation of quality planting material of Aloe barbadensis for commercialization.

Key words: Growth regulators, Aloe barbadensis

22. Journal of Cell and Tissue Research 14(1): 4131- 4136 (2014)

Histological studies on antler of spotted deer (Axis axis) and horns of black buck (Anteplope cervicapra) with that of horns of domestic cattle

Sridevu, P.C., Prasad, R.V., Narayana Bhat, M., Jayashankar, M.R. and Ramkrishna, V.

Veterinary College, Hebbal, Bangalore560024, , Karnataka. E. mail: rvprashwin@gmail.com, Cell: 09449855893

Abstract: The present study was conducted on antlers of spotted deer and on horns of black buck and cattle with the objective of differentiating them from each other. In the horizontal ground section of antler four distinct zones were observed namely subvelvet zone, zone of osteonic bone, transition zone and spongiosa zone from periphery to the center of the antler. Similar type of zonation was also observed at the tip of the tine of antler. Such zonations were not observed either in the transverse ground section of burr and pedicle of the antler, as well as in Cattle and Black buck horn core. The transverse section of the soft tissue located between the horn sheath and horn core of cattle showed stratified squamous epithelium towards horn sheath. Underlying this epithelial layer was a thin layer of highly vascularised loosely arranged dermis composed of irregularly oriented collagen fibres.

Key words: Antlers; Horns, Axis axis, Antelope cervicapra

23. Journal of Cell and Tissue Research 14(1): 4137- 4141 (2014)

Antioxidant activity of Asteracantha longifolia in DMBA induced mammary cancer in Sprague dawley rats

Santhosh Kumar, C.N., Shridhar, N.B., Jagadeesh, S.S., Sanganal, J.S., Suguna, R. and Ansar Kamran, C.

Department of Veterinary Pharmacology and Toxicology, Hebbal, Bangalore 560 024, Karnataka Veterinary, Animal and Fisheries Sciences University, Bidar-585401, Karnataka.
E. mail: sridhar_vet@rediffmail.com, Cell: 09448059777

Abstract: Antioxidants play an important role in inhibiting and scavenging free radicals, thus providing protection to human against infections and degenerative diseases. Current research is directed towards natural antioxidants originated from plants due to safe therapeutics. Asteracantha longifolia (Family-Acanthaceae), commonly known as ‘Hygrophila’ being used as folklore medicine. Plant has been reported to posses in vitro antioxidant activity and anticancer in nature. Hence the present study was evaluated for antioxidant activity of petroleum ether extract of A. longifolia by chemically inducing mammary tumor using 7, 12-Dimethylbenz (a) anthracene (DMBA) in Sprague Dawley rats. Test groups were treated with A. longifolia extract @ 600, 900 and 1200 mg/kg and the antioxidant activity is compared with the group of rats administered with Vitamin C (ascorbic acid) @ 200mg/kg body weight for 45 days. Mammary tumor, liver and kidney tissue were processed for antioxidant enzyme assay. The antioxidant activity were correlating with increased dose of plant extract, the data obtained in present study suggests that the extract of A. longifolia have potent antioxidant activity and prevent oxidative damage to major biomolecules and afford significant protection against oxidative damage seen in cancer.

Key words: Asteracantha longifolia, Mammary tumor, antioxidant.

24. Journal of Cell and Tissue Research 14(1): 4143- 4148 (2014)

An improved plant regeneration system of Ocimum sanctum l. - An important indian holy basil plant

Sharma, N. K., Choudhary, R.C. and Kumar, M.

Department of Agricultural Biotechnology and Molecular Biology, FBS & H, Rajendra Agricultural University, Bihar, Pusa, Samastipur 848 125. E. mail: nkbiotech@yahoo.co.in, Cell: 07387616018

Abstract: An efficient, rapid and large scale propagation of a multipurpose medicinal herb, Ocimum sanctum through in vitro culture of nodal segments from mature plants has been developed. Shoot proliferation was initiated from nodal explants cultured on MS medium supplemented with various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzyladenine (BA) (0.5-2.0 mgl-1), kinetin (KN) (0.5-2.0 mgl-1) and with combinations such as {BA (0.5 mgl-1) + IAA (0.10-1.00 mgl-1)}. The maximum number of shoots (11.33 ± 0.27), with average length 5.46 ± 0.10 cm was recorded with MS medium containing 1.5 BA mgl-1. Shoot culture was established by repeated subculturing of the original nodal segment explants on shoot multiplication medium after each harvest of newly formed shoots. In this way, 30-35 shoots were obtained from a single explant after 3 months. The single shoots were transferred for rooting to ½ MS medium containing 1.5 mgl-1 IBA. The in vitro raised plantlets with well-developed shoots and roots were successfully established in plastic cups containing garden soil, farm yard manure and sand (1:1:1) and were grown in green house with 90% survival rate. Thus, this micropropagation protocol can be used for its rapid commercial propagation of O. sanctum L. and for future genetic improvement studies.

Key words: Ocimum sanctum L., In vitro regeneration

25. Journal of Cell and Tissue Research 14(1): 4149- 4152 (2014)

Effect of diverse levels of cell culture confluences on development of hand made cloned
buffalo embryos

Shah, F., Sadeesh, E.M., Gupta, M., Ghosh, K. And Yadav, P.S

Department of Veterinary Physiology and Biochemistry, LLRUVAS, Hisar 125 004;
E. mail: rsherwal@gmail.com, Cell: 092533 50342

Abstract: The main purpose of the experiment was to compare the efficiency of three different somatic cell culture confluences (60-70%; 70-80% and 80-90%) on developmental rates of zona free HMC buffalo embryos. Cultured ear pinna fibroblast from new born Murrah buffalo calf were harvested in above mentioned three different confluences and were used as donor cells in this experiment. From present study, 60-70% of cell confluence gave overall cleavage rate of 64.24±7.1 and blastocyst formation rate 9.7±0.3, while 70-80% and 80-90% confluence gave overall cleavage rate of 56.11 ± 3.1, 69.84 ± 7.79 and blastocyst rate 16.1 ±3.9, 24.65 ± 4.1 respectively. The results showed that there was insignificant difference (P>0.05) in cleavage rate but there was significant difference (P< 0.05) in blastocyst formation between cloned embryo derived from three different cell culture confluences. This suggests that cell culture confluence of donor cell may affect cloning efficiency. However, the pregnancy rate and the development to the term of cloned embryos derived from three different cell culture confluences need further investigation.

Key words: Buffalo embryo, Cloned embryos

26. Journal of Cell and Tissue Research 14(1): 4153- 4157 (2014)

Efficacy of growth hormone for callus induction in Calendula officinalis, L

Nikam, S.L. and Khan S.J.

The Institute of Science, Mumbai. 15, Madam Cama Road, Mumbai 40032 .
E. mail: sush_nik@rediffmail.com

Abstract: The high demand for plant material from Calendula officinalis ,L in the production of herbal medicines and cosmetics, turns the technique of plant tissue culture into one of the alternatives for improvement of crop over short period of time. The present work is based on developing a protocol for the callus induction in Calendula officinalis from leaf and ray floret explants. The sterilized explants were inoculated in MS media containing various combinations of auxins such as naphthalene acetic acid (NAA), indole 3-acetic acid (IAA), indole 3-butyric acid (IBA), 2,4 dichlorophenoxyacetic acid (2,4 D) and cytokinins such as kinetin, benzylaminopurines (BAP) and TDZ (tidiazuron). The highest rate of callus induction for Calendula leaf explants was achieved on media supplemented with 2,4D (2mg/L) and KIN (5mg/L) with compact, fast growing, fluorescent green coloured callus. Whereas ray floret explants showed development of best callus growth in dark condition with hormonal combination IAA (0.5mg/L) and TDZ (5mg/L) with compact, brown colored callus. However, Leaf is the most potential explants for callus induction.

Key words: Calendula officinalis, Callus induction

27. Journal of Cell and Tissue Research 14(1): 4159- 4164 (2014)

Ascorbic and citric acids in combination resolve the problems encountered in micro-propagation of litchi from shoot tips

Pankaj, K., Pandey, S.K. and Roy, K.A.

P. G. Department of Biotechnology, T. M. Bhagalpur University, Bhagalpur 812007.
E. mail: pankajeworld@gmail.com, Cell: 093867 95532

Abstract: A major problem in the micro propagation of litchi is the secretion of polyphenols into the medium by the tissues once excised. This secretion undergoes oxidation to produce substances that kill the tissues. The first visual symptom of deterioration is skin browning which has been reported to appear due to polyphenol oxidase activity on anthocyanins resulting the production of brown coloured by-products. Litchi plant is rich in phenolics compounds which are catalysed by enzyme polyphenol oxidase and make the medium brown. Ascorbic and citric acids, potent antioxidants, have been frequently used for browning control. It reduces the polyphenol oxidase enzyme and control browning. The combination of ascorbic acid with citric acid acted as powerful antioxidants inhibiting the polyphenol oxidase activity. It is also acted as vitamins which are able to check the browning and also initiate the proliferation of cells of the explants. In the present experiment, it is noticed that excision and cutting of explants under anoxygenic condition followed by soaking in 160mg/l ascorbic acid and 160mg/l citric acid solution subsequently culturing on basal medium amended with combination of ascorbic acid (250mg/l) and citric acid (250mg/l) is suitable protocol to eliminate the browning by reducing the polyphenol oxidase enzyme activity of Litchi. Thus the amendent of MS medium with ascorbic and citric acid for browning control in vitro culture of litchi explant is considered as standard one for micropropagation of shoot tips.

Key words: Litchi micro propagation, Ascorbic acid, Citric acid

28. Journal of Cell and Tissue Research 14(1): 4165- 4171 (2014)

Effect of hormones, sera, follicular fluid and oocyte quality on in vitro nuclear maturation and fertilization rate of indian water buffalo (Bubalus bubalis ) oocytes

Sadeesh, E. M., Shah, F., Balhara, A. K., Thirumaran., S. M. K., Balhara, S., Kumar, D., Yadav, S., Singh, I. and Yadav, P. S.

Division of Animal Physiology and Reproduction, Central Institute for Research on Buffaloes, Hisar 125 001.
E. mail: sadeeshcirb@gmail.com,

Abstract: The object of this study was to identify effect of follicular fluid (FF), gonadotropin hormones (PMSG + hCG), sera and quality of oocytes on cumulus expansion, nuclear maturation and fertilization of Indian buffalo (Bubalus bubalis) oocytes. The cumulus oocyte complexes (COCs), denuded cumulus oocytes (DCOs) were collected from slaughterhouse ovaries, matured in three different groups of maturation media. Media used for IVF and subsequent development of embryo was same in all groups. The results revealed that significant improved (P<0.05) cumulus expansion and polar body formation rates of COCs in maturation media with hormones, FF, and FBS as compared to maturation media with hormones plus FF and maturation media without hormones plus FBS (91.9 ± 1.3, 81.6 ± 4.4, 31.4 ± 5.5; 79.0 ± 0.9, 70.4 ± 7.1, 14.6 ± 8.6 cumulus expansion and polar body formation rates respectively), significant improvement (P<0.05) in cleavage and blastocyst rates were also observed when matured oocytes from maturation media with hormones, FF and FBS subjected to IVF as compared to maturation media with hormones plus FF and maturation media without hormones plus FBS (49.2 ± 9.2, 41.7 ± 2.9, 7.4 ± 3.7, 24.2 ± 4.7, 9.2 ± 1.6, 0 respectively). The IVM rates of DCOs in the maturation media with hormones, FF and FBS as compared to maturation media with hormones plus FF and maturation media without hormones plus FBS were 16.7 ± 3.3, 6.7 ± 3.3, 3.3 ± 3.3, respectively. In conclusion, addition of PMSG + hCG+ FF in MM supplemented with FBS can successfully improve in vitro maturation and fertilization of Indian buffalo (Bubalus bubalis) oocytes; for nuclear maturation, the use of COCs is crucial

Key words: Bubalus bubalis, Oocytes, Follicular fluid, Hormones, Sera
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29. Journal of Cell and Tissue Research 14(1): 4173- 4179 (2014)

Wound healing, antimicrobial and anti fungal activities of ethanolic leaf extract of Aegle marmelos in rats

Fahmeeda, B., Jayaprakashreddy, G., Shivakrishna and Rajitha, K.

Department of Pharmacology, St.Peter’s College of Pharmacy, Madikonda, Warangal
506 142 (A.P). E. mail: shiva_krishnapabba@yahoo.com
.
Abstract: Aegle marmelos is a medicinal plant commonly known as bale. This plant contains different components of therapeutic value. The ethanolic extract of Aegle marmelos showed the presence of phytoconstituents like flavonoids, triterpenoids, coumarins, sterols, triterpenoids, alkaloids with other constituents. In present investigation, antibacterial, antifungal and wound healing effects of ethanolic extract of leaves of Aegle marmelos has been studied. This extract was formulated 4% Aegle marmelos ethanolic leaf extract ointment and was studied for its wound healing property against an excision wound on the skin of Wistar albino rats. The activity was compared with 2% framycetin ointment. Study shows faster wound healing with 4% Aegle marmelos ethanolic leaf extract ointment. On 12th day biochemical parameters like hydroxyproline, collagen and hexosamine were determined from excised tissue and they were significantly increased in extract treated groups. Healed skin was also subjected to histopathological studies to examine the microscopic changes and it also supported the wound healing on application of extract ointment in treated group. Histopathological observations showed increase in granulation and rapid collagen turnover. It was concluded that wound healing actibity of ethanolic leaf extract of a Aegle marmelos was better and faster than framycetin ointment.

Key words: Aegle marmelos,Wound healing