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1. Journal of Cell and Tissue Research 14(3): 4471- 4475 (2014)
Male factor infertility, oxidative stress and phytochemicals: A critical review
Patil, R.B., Patil S.R. and Pillai, M.M.
Department of Zoology, Veer Wajekar Arts, Science and Commerce College, Phunde (Uran,
Navi Mumbai) 400 702. E. mail: sachintendulkar.patil@gmail.com
Abstract: Excessive production of Reactive Oxygen Species (ROS) in the testes has been a major contributory factor to the infertility. Oxidative stress has attracted enormous interest in researchers in recent past. Reactive oxygen species are continuously formed in the cells by various metabolic & physiological processes. Oxidative stress is result of imbalance between ROS level & label of antioxidants in the body which can lead to sperm DNA damage, in viability, deformity and eventually infertility, however many evidences demonstrate sperm capacitation and acrosome reaction are associated with extracellular production of superoxide anions. Apart from self-defense antioxidant system against ROS, recent research provided some strong evidence of consuming dietary supplement containing polyphenols, flavonoids etc.This article of ROS, impact of oxidative stress on sperm physiology & antioxidant strategies to reduce oxidative stress.
Key words: Male infertility factor, Oxidative stress
2. Journal of Cell and Tissue Research 14(3): 4477- 4483 (2014)
Cytological and histopathological studies of mesenchymal and melonotic neoplasms in dogs
Kotrappa, Y.M. Suguna, R., Girish, B.C., Byregowda, S.M. and Manjunatha, K.
Liveon Biolabs Private Limited, Plot No. 46 & 47, II Phase, Water Tank Road, KIADB, Industrial Area, Kasaba Hobli, Antharasanahalli, Tumkur 572106. E. mail: girishbekkare@gmail.com,
Abstract: The incidence of neoplasia has assumed an increasing trend in domestic animals especially in canines due to their longevity of life. Neoplasms of mesenchymal tissue origin are found to occur with high incidence in dogs. The current work was undertaken with the objectives of studying the cytological and histopathological features of non round cell and non epithelial neoplasms and comparing them as diagnostic tools. The mesenchymal and melanotic tumors diagnosed in the study included fibrosarcoma (#5), malignant melanoma (#5), lipoma (#3), chondrosarcoma (#2), osteosarcoma (#1) and malignant schwannoma (#1). Cytologically varying degrees of cellular anaplasia, pleomorphism and mitotic activity were recorded. Microscopically the interlacing bundles of spindle to stellate shaped cells with elongated plumpy nuclei (fibrosarcoma), varying number of brownish to blackish colored granules (malignant melanoma) presence of mature uniform sized adipocytes with eccentrically placed nuclei (Lipoma), hyperchromatic nuclei, prominent nucleoli and occasional binucleated cells (chondrosarcoma), presence of bony spicules with osteoid material (Osteosarcoma) and bands of densely packed spindle shaped cells with oval or round hyperchromatic nuclei (malignant Schwannoma) were observed. Cytology in complementary with histological characterization resulted in better diagnosis of mesenchymal neoplasms.
Key words: Dogs, Malignant melanoma, Mesenchymal neoplasms
3.Journal of Cell and Tissue Research 14(3): 4485- 4489 (2014)
Polymerase chain reaction assay for diagnosis of Escherichia coli mastitis in Murrah buffaloes
Charaya, G., Kumar, A., Sharma, A., Singh, M. and Goel, P.
Department of Veterinary Medicine, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar 125004, Haryana. E. mail: anshusharma_dr@yahoo.com,
Abstract: Mastitis is a multietiological disease of lactating animals responsible for huge economic losses throughout the world. E. coli is the most common cause of severe clinical form of mastitis. Early and correct detection of E. coli infection by suitable test(s) is imperative for timely initiation of treatment. In the present study, PCR assay was standardized and applied on total of 202 quarters milk samples of 52 Murrah buffaloes for detection of E. coli. Results of PCR assay were compared with cultural examination. Sixty four samples showed presence of E. coli by using 16S to 23S rRNA specific primers whereas on culture examination only 57 samples were found positive for E. coli. This might be due treatment being going on or presence of very small number of organisms being present. The PCR assay has been found to be rapid, sensitive, reliable and less labour intensive. This method may be used as an alternative to culture examinations assay or can also be used in conjunction with cultural examination so that antimicrobial sensitivity can be carried out for initiation of treatment with suitable antibiotics.
Key words: Murrah buffaloes, Mastitis, E. coli
4.Journal of Cell and Tissue Research 14(3): 4491- 4494 (2014)
Association of selected humoral and cell mediated immuno mediators with postpartum reproductive disorders in buffaloes
Mili, B. and Pandita, S.
Dairy Cattle Physiology Division, National Dairy Research Institute, Karnal 132001 (Haryana).
E. mail: dr.bhabesh87@gmail.com,
Abstract: The late pregnancy and early lactation certainly is the most interesting task for management of uterine disorders. The gradual decline of humoral immunity (IgG and IgM) to cell mediated immune response associated with oxidative stress around parturition may contribute to uterine infections in buffalo. For this, 25 Murrah buffaloes postpartum were selected from National Dairy Research Institute (NDRI) cattle herd. Through clinical examination 5 buffaloes were found to be infected with metritis, 6 with endometritis and 8 exhibited Mastitis. Remaining 6 were apparently healthy. Blood samples were drawn by jugular venipuncture in sterile heparinised vacutainer tubes from each healthy buffalo at 6.00 A.M. in the morning on days +7, +14, +21, +28, +35, +42, +49, +56 postpartum. The blood samples were also collected from unhealthy buffaloes twice in sterile heparinised vacutainers on alternate day as and when the symptoms of reproductive abnormalities were noticed. The IgG and total antioxidant activity (TAA) concentrations (p<0.05) were significantly low in buffaloes infected with metritis, endometritis and mastitis cases. However, plasma nitric oxide (NO) was significantly high (p<0.05) in buffaloes suffered from endometritis and mastitis cases as against the control groups but not in case of metritis cases. This study revealed that plasma IgG, TAA and NO in postpartum infected buffaloes differed significantly from control/ healthy group. Thus, these blood parameters may be used for early assessment of postpartum infection in buffaloes.
Key words: IgG, IgM, Postpartum, Buffalo
5. Journal of Cell and Tissue Research 14(3): 4495- 4499 (2014)
Polymorphism of CD14 gene and its association with mastitis in murrah buffalo
Baro, D., Sangwan, M.L., Khepar, A., Wani, S.A., Kumari, A. and Batra, K.
Department of Animal Biotechnology, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, 125004. E. mail: dao.haru@yahoo.com,
Abstract: The present investigation was undertaken to study the genetic polymorphism in CD14 gene and its association with mastitis by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique in 50 Murrah buffaloes (including 25 animals suffering/suffered from mastitis) maintained at CIRB, Hisar, and TVCC, LUVAS, Hisar, Haryana. Genomic DNA was isolated by Phenol Chloform Isoamyl alcohol (PCI) method. DNA quality was checked by 0.7% agarose gel electrophoresis. PCR was standardized to amplify partial CD14 gene with designed primer. Digestion of PCR products of partial CD14 gene with FauI, TaqáI and MspI restriction endonucleases, detected xx, yy and zz genotypes respectively. Monomorphism was observed with all restriction fragments and hence association of CD14 gene with mastitis could not be established. Cloned product of CD14 gene was sequenced which has been found to be of 539 bp long and sequence data generated was compared with the sequences available in NCBI database to study homology with other species. The query sequence of CD14 gene showed maximum identity to Bubalus bubalis [99%], followed by Bos taurus [98%], Ovis canadensis [98%], Ovis aries [97%], Capra hircus [93%] and least with Sus scrofa [84%].
Key words: CD14 gene, Murrah buffaloes
6. Journal of Cell and Tissue Research 14(3): 4501- 4504 (2014)
Effect of uterine immunomodulation on serum Amyloid-A concentration and conception rate in cyclic non breeding cows
Sahoo, S. and Mohanty, D.N.
Department of Animal Reproduction Gynaecology and Obstetrics, College of Veterinary Science and Animal Husbandry, OUAT, Bhubaneswar 751003, Odisha. E. mail: saraswat.vet06@gmail.com,
Abstract: Twenty one repeat breeding cows were screened by white side test to eliminate endometritis and they were assigned to three treatment protocols with equal number of seven animals in each group. Cows of control group were administered with 50 ml of normal saline, cows in the group second received 20 ml of fresh colostrum and in group third 10 ml of non pathogenic E. coli. All treatments were given as intra uterine medication. Blood samples from each experimental animal were collected for evaluation of serum amyloid A concentration during pre- and post- treatment periods, consequent to various therapies as per standard procedures. In addition to this, the conception rate for each treatment regimen was evaluated for efficacy.The serum amyloid A values showed a receding trend in all the experimental groups with significant variation (p<0.01) was marked in the colostrum and non pathogenic E. coli treated group.
Key words: Serum amyloid, Cyclic non breeder
7. Journal of Cell and Tissue Research 14(3): 4505- 4508 (2014)
Multi-trait selection indices for genetic improvement in productivity of egg type chicken
Tomar, A.K., Poonia, J.S., Dubey, T., Chaudhari, M. and Kumar, P.
Department of Animal Genetics and Breeding, College of Veterinary Science, Lala Lajpat Rai
University of Veterinary and Animal Sciences, Hisar 125004 (Haryana).
E. mail: tripurari.dube@gmail.com,
Abstract: The data, on 2416 pullets, progenies of 252 sires of White Leghorn over five generations (2008-09 to 2012-13), maintained at the poultry breeding farm of Department of Animal Genetics and Breeding, LUVAS, Hisar, were utilized for the present study BW20, AFE, EW40, EN40, BW40 and EM40 were used in various combinations for construction of selection indices. The effect of hatch within a gene-ration and generation was significant (P d” 0.01) on all the traits. A total of 35 multi-trait selection indices incorporating BW20, AFE, EW40, EN40, BW40 and EM40 were constructed. The index I1 which incorporated all six traits was found to be most efficient and considered as a standard selection index. Index I1 with selection intensity 1.4 was expected to produce aggregate genetic gain ( Δ H) of 33.44 with RIH value of 0.6565. The expected genetic change in individual traits (ΔGi) were 2.20 g, -0.80 days, -0.54 g, 2.70 eggs, -15.13 g, 38.47 g, respectively for BW20, AFE, EW40, EN40, BW40 and EM40. Highest contribution to aggregate genetic gain ( Δ H) was observed from EN40 (23.15%) followed by BW40 (7.51%), BW20 (5.92%), EW40 (3.84%), AFE (0.32%) and least was from EM40 (0.08%) indicating the relative importance of the traits. The index I2, I8, I18 and I30 incorporating five, four, three and two traits with relative efficiency of 99.91%, 94.04%, 93.92% and 91.93%, respectively may be consid-ered as indices of choice for overall genetic improvement of egg type chicken.
Key words: Egg type chicken, Multi-trait selection index
8. Journal of Cell and Tissue Research 14(3): 4509- 4513 (2014)
Low levels of aflatoxin induced hepatic lesions and their amelioration with N-Acetylcysteine in
broiler chicken
Yalagod, S. G.,Narayanaswamy, H.D.,Satyanarayana, M.L., Rao, S., Yathiraj, S. and Rathnamma, D.
Department of Pathology, Veterinary College, Hebbal Bangalore 560024, Karnataka;
E. mail: drsgyalagod@rediffmail.com,
Abstract: The present trial was conducted to study the effect of low dietary levels of AFB1 and ameliorative effect of N-Acetylcysteine (NAC) supplementation on gross and histological alterations in the liver. One hundred and forty four commercial broiler chicks were randomly divided in to six groups. The first Four groups had AFB1 levels of 0.0 (control), 0.25 ppm, 0.5 ppm and 1 ppm. The other two groups were supplemented with NAC (800 mg/kg) of which one group had dietary aflatoxin B1 (AFB1) inclusion at 1 ppm. The diets were fed for 28 days from the day of hatch. Grossoly the liver was enlarged, icteric, mottled, soft and friable. Histopathologically the liver showed Vacuolar degeneration, necrosis, bile duct epithelial hyperplasia, fatty change and infiltration of mononuclear cells. The results suggested that NAC provides protection against negative effects of aflatoxins.
Key words: Aflatoxin, Hepatic lesion, N-acetylcysteine
9.Journal of Cell and Tissue Research 14(3): 4515- 4518 (2014)
Cytogenetic studies of chromosomes of Yak
Saikhom, R.,Sahoo, A.K., Shekhar, S.and Dalai, N.
Department of Animal Genetics and Breeding, W.B.U.A.F.S., 37 K. B. Sarani , Kolkata- 700037.
E. mail: reshmasaikhom@gmail.com
Abstract: The present investigation chromosomes of male and female yaks were analysis. 40 yaks (20 males and 20 females) were selected for this purpose. The blood samples were collected incubated and cultured using appropriate media and specific methods. The samples were analyzed for chromosome number and morphology, relative length of the chromosome, arm ratio and centromere index of X and Y chromosomes and chromosomal abnormalities in male and female yaks, if any. The diploid number of metaphase chromosomes in male as well as in female yak to be 2n= 60, out of which 58 are autosomes and 2 being sex chromosomes. From the centromeric position, karyotyping studies revealed that all the 29 pairs of autosomes were acrocentric and the sex chromosomes, X and Y were submetacentric and metacentric respectively. The autosomal length for 1st to 29th pairs of chromosomes were 6.12 ± 0.09 to 1.24 ± 0.05 and 6.28 ± 0.08 to 1.42 ± 0.06 in male and female yak respectively and did not reveal any significant difference. The mean absolute length of the sex chromosome XX were 5.18 ± 0.08 in female and that of sex chromosome X and sex chromosome Y in case of male were 4.96 ± 0.07 and 2.14 ± 0.07 respectively. The arm ratio of X-chromosome was 2.84 ± 0.05 in male and 2.69 ± 0.003 in female. The centromere index of X-chromosome was found to be 28.25 ± 0.6 in male and 25.46 ± 0.5 in female yak. The arm ratio of Y-chromosome was 1.48 ± 0.001 and the centromeric index was 32.25 ± 0.05. Chromosomal abnormalities were not found in male and female yak in this study.
Key words: Chromosomes, Yaks
10. Journal of Cell and Tissue Research 14(3): 4519- 4524 (2014)
Assessment of different template preparation methods for nested pcr based detection of marek’s disease virus of poultry
Kumar, A., Chandraprakash, M., Maan, N. S., Mahajan, N. K., Sunayna., Batra, K., Ghosh, A., Nanda, T. and Maan, S.
Department of Animal Biotechnology, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana. E. mail: ak.hau.abt@gmail.com,
Abstract: High performance DNA preparation from liver tissue is of major interest for identification of the Gallid herpes virus 2 (GaHV-2), genome by the polymerase chain reaction (PCR). In this study we evaluated the impact of four widely used DNA extraction protocols on the performance of molecular diagnosis of GaHV-2. Four DNA extraction protocols using guanidine isothiocyanate (GIT), TRIzol, phenol: chloroform: isoamyl alcohol (PCI) and chelexR100 resin were compared for use in nested polymerase chain reaction (nPCR) based detection of GaHV-2, responsible for causing Marek’s disease in poultry. A total of 20 GaHV-2 positive liver samples were processed and viral DNA was extracted using above mention methods. PCI method was found to be the most sensitive method for detection of GaHV-2 DNA in liver samples using nPCR as 20 out of 28 samples (20/28) were found positive, followed by modified GIT (18/28), TRIzol (15/28) and Chelex R100 resin (4/28). Although, average yield of nucleic acid was highest in case of PCI method, average quality was highest in case of modified GIT. We have found that the DNA extraction method plays an important role in success of downstream molecular applications like PCR. Therefore, we recommend the use of PCI treatment of samples for routine diagnosis of GaHV-2 DNA in clinical samples using nPCR.
Key words: DNA extraction, Marek’s disease,
11. Journal of Cell and Tissue Research 14(3): 4525- 4529 (2014)
Phylogeny of macrobrachium species using mitochondrial 16S ribosomal DNA
Sharma, C.,Krishna, G., Pavan Kumar, A.and Nayak, S.K.
Mangalore University Mangalagangotri Karnataka 574199. E. mail: charufgb@gmail.com
Abstract: Mitochondrial partial 16s ribosomal DNA was used to infer the evolutionary relationship between selected Macrobrachium species. From 517 characters, 361 characters are variable and 331 characters are parsimony informative. The parameters specified under optimum model were nucleotide frequencies of A = 0.31, C = 0.21, G= 0.16, T = 0.30, reversal rates of change for A < > C = 0.3509, A < > G = 5.1208, A < > T = 1.0000, C < > G = 0.3509, C < > T = 3.2990, gamma shape distribution α = 0.617 and a significant proportion of invariable sites (pinv = 0.20). Across all species, no substitution saturation was found as the level of transitional base substitutions was higher than the level of transversions over all genetic distances. The phylogenetic trees reconstructed by four methods i.e., distance, maximum parsimony, maximum likelihood and bayesian inference, yielded similar topologies with a slight variation in bootstrap values. The maximum parsimony, bayesian inference method resulted in soft polytomies in tree topology and these might be due to weakness of marker or less number of species. Tree topologies resulted by NJ, MP, ML and BI analysis supported the monophyly of the genus Macrobrachium with high bootstrap values. The out group Exopalaemon formed as paraphyletic to Macrobrachium with significant bootstrap value. Principally all species clustered into 5 clades as per their geographic location and evolutionary relationship. M. rosenbergii, M. gangeticum, M. idella, M. villosimanus, M. lamarrei, M .malcomsonii and M. sankollii (Species endemic to India; India group) formed one cluster. Species from East Asia viz., M. nipponense, M. inflatum, M. maculatum, M. shokitai, M. asperulum, M. pinguis, M. anhuiense formed another cluster.
Key words: Molecular phylogeny, Macrobrachium, 16s rRNA
12. Journal of Cell and Tissue Research 14(3): 4531- 4534 (2014)
Genetic diversity analysis of lentil (Lens culinaris Medik.) germplasm using molecular marker
Singh, A.K., Kumar, P.,Singh, J., Rani, R., Rani, A., Shukla, P.K. and Misra, P.
Sam Higginbottom Institute of Agriculture, Technology and Sciences, Allahabad 211007 (UP).
E. mail: pragatimisra3@rediffmail.com,
Abstract: Genetic relationship of thirty eight lentil (Lens culinaris Medik.) germplasm studied using RAPD markers. Thirty RAPD primers were selected for molecular characterization of lentil germplasm. Out of thirty primers only 18 primers were amplified with thirty eight germplasm of lentil. Genetic diversity was studied on the basis of different band pattern and based on band pattern analysis dendogram was formed. The Unweighted pair group for arithmetic average analysis (UPGMA) was used for cluster analysis. The range of similarity was found between 0.421 to 0.971. Our results suggested that genetic relationships in lentil germplasm using RAPD banding data may be useful for plant characterization.
Key words: Genetic diversity, Germplasm, Lentil, RAPD marker.
13. Journal of Cell and Tissue Research 14(3): 4535- 4538 (2014)
Bacopa monniera improves the proteins of lysosomal and non-lysosomal fractions of prostate gland reduced in aged mice
Kalamade, V. I.
Yashwantarao Chavan College of Science, Karad 415124 (M.P.).E. mail: vahidakalamade@yahoo.co.in,
Abstract: Aging is unavoidable process in every living organism in which slow and gradual deterioration of tissues takes place. Free radicals are the major causative agents of aging. Prostate gland is the most affected organ during the process of aging. Various proteins produced by prostate are the markers and can be used to diagnose the prostatic disorders. They are reduced during aging and in pathological disorders of prostate. Disturbance in oxidant- antioxidant balance may be one of the causes of reduction in proteins level. Many plants contain antioxidants which can be used to compensate the reduced level of antioxidants in aging. Bacopa monniera is antioxidant rich, antiaging herb. Hence present work is undertaken to investigate whether Bacopa leaf extract can maintain normal level of proteins in prostate and protect it from pathological abnormality. In the present work biochemical study of proteins was carried out in prostate of aging mice and in Bacopa treated mice. It was found that Bacopa can maintain proteins at almost normal level.
Key words: Bacopa monniera, Prostate gland
14. Journal of Cell and Tissue Research 14(3): 4539- 4545 (2014)
ISSR-based molecular characterization of Triticum eastivum L. cultivars
Sharma, D., Seth, P., Saharan, V., Joshi, A., Maloo, S.R. and Khandelwal, S.K.
Department of Molecular Biology and Biotechnology, Maharana Pratap University of Agriculture and Technology, Udaipur 313 001 (Rajasthan). E. mail: deepakbagda77@gmail.com,
Abstract: Molecular genetic markers are widely used tools in genotype and species identification. Genetic relationship study was performed with 20 ISSR primers among 12 Indian wheat accessions (6 salt tolerant and 6 salt susceptible). Genomic DNA from wheat genotypes were analyzed using inter simple sequence repeats (ISSR) markers. Thirteen ISSR primers produced 78 amplified DNA fragments ranging in size from 100–2200 base pairs, 6 fragments were monomorphic (7.7%) and 72 fragments were polymorphic (92.3%) with an average of 6 fragments per primer and 5.53 polymorphic fragments per primer. Six primers were found to be highly informative as they gave unique bands to differentiate between salt susceptible and salt tolerant genotypes. From ISSR profiles similarity matrix was obtained and Jaccard’s similarity coefficient was observed in between 0.40 to 0.77 and on this basis a dendogram was constructed with UPGMA method. Easy handling, high information levels and reliability are the features that justify the utility of ISSR markers in DNA fingerprinting and genetic variability analysis which is highly useful for finding genes controlling agronomically important traits in wheat
Key words: Molecular markers, Triticum eastivum L
15. Journal of Cell and Tissue Research 14(3): 4547- 4550 (2014)
Inter simple sequence repeats (ISSR) based polymorphism for powdery mildew resistance in green gram (Vigna radiata (L.) wilczek)
Bainade, P.S., Kale, A.A., Kumbhar, S.D. and Deshmukh, S.G.
Department of Agriculture Botany, M.P.K.V., Rahuri, (M.S.) 413722. E. mail: pradeep.s.bainade@gmail.com,
Abstract: The present study was undertaken to identify ISSR markers associated with powdery mildew resistance in green gram (Vigna radiata (L.) Wilczek) cross Kopargaon (susceptible) and BPMR-48 (resistant). Bulk segregant analysis was employed to identify ISSR marker linked to powdery mildew resistant gene of BPMR-48. A total of 75 ISSR markers were used for identification of polymorphic markers between the DNA bulks of resistant and susceptible F3 individuals and their parents. Among the 75 ISSR markers studied, only 54 markers were found to amplify in all green gram DNA extracted. Four markers were found polymorphic between the two parents (Kopargaon and BPMR-48) whereas, only one marker ISSR 834 was found polymorphic between the parents as well as resistant and susceptible bulk, indicating its possible linkage in BSA.
Key words: ISSR marker, Green gram, powdery mildew resistance
16. PAPER
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17. Journal of Cell and Tissue Research 14(3): 4559- 4564 (2014)
Molecular studies on the transmission of Sri Lankan Cassava Mosaic Virus (SLCMV) in cassava by Bemisia tabaci collected from cassava and cassava based crops
Kumar, J., Rabindran, R., Singh, H. and Kumar, P.
Department of Plant Pathology, Centerfor Plant Protection Studies, Tamil Nadu Agricultural University, Coimbatore 641003, Tamil Nadu. E. mail: jitpatho@gmail.com,
Abstract: Cassava mosaic disease, caused by cassava mosaic geminiviruses are transmitted by Bemisia tabaci. Field surveys were conducted in cassava growing areas of Tamil Nadu during 2008-2009. Results showed that a there was high infection (89.09%) due to use of infected planting material (stem cuttings), while the whitefly borne infection was found to be very low (10.01%). Disease symptoms were generally mild. There was no change in disease incidence over the survey period. The B. tabaci were collected from cassava, tomato and bhendi separately and colonies reared on the same host. The meristem derived virus free cassava plant produced from apical meristem culture was studied to determine their ability to transmit Sri Lankan cassava mosaic virus (SLCMV) from cassava to cassava. Virus free plants were confirmed by polymerase chain reaction (PCR) using geminivirus degenerate primers. For the transmission studies virus acquisition access period (AAP) of 24 h on virus infected cassava leaves and 48 h virus inoculation access periods (IAP) on virus free healthy leaves were given to the all whiteflies that were reared on different host. SLCMV was absolutely transmitted by whiteflies reared on cassava but tomato and bhendi whiteflies did not transmitted SLCMV. The results were confirmed by using multiplex PCR. The transmission ability of this Hemipteran insect with SLCMV revealed that they were able to transmit the viruses from cassava to cassava effectively. Aim of this study was to observe that the tomato and bhendi whiteflies are able to transfer cassava mosaic virus (CMV) in comparison with the white flies of cassava.
Key words: Bemisia tabaci, Cassava mosaic virus
18. Journal of Cell and Tissue Research 14(3): 4565- 4569 (2014)
Effect of growth factors BAP, IBA and NAA on initiation, proliferation and formation of shoots and roots of Dendrobium cv. Banyat pink
Priyadarshini, S.,Salam, J.S.and Mohanty, Ch.R.
Department of Horticulture, Orissa University of Agriculture and Technology, Bhubaneswar 751003, Orissa. E. mail: jekendrasalam@rediffmail.com,
Abstract: The effect of growth regulators, benzylamino purine (BAP), Indole butyric acid (IBA) and naphthalene acetic acid (NAA) on initiation and proliferation of shoots and roots of Dendrobium cv. Banyat Pink were studied. Number of protocorms proliferated into shoots (9.37) and leaves per plantlet (15.15) were significantly higher in MS medium fortified with 2.0 mg-l BAP followed by 1.5 mg-l BAP (5.36). Maximum number of shoots (2.24 - 2.62) per tube were formed in MS medium fortified with BAP in the range of 0.5-2.5mg-l whereas, the lowest number (1.66) of shoots were observed with higher concentration of 3.0 mg-l BAP. 1mg-l IBA showed maximum values in number of leaves per tube (29.67), number of leaves per plantlet (5.90) and leave length (1.05cm). NAA alone or in combination with IBA could not have any significant effect on the leave development. Maximum number of roots were recorded in MS mediums supplemented with 1-1.5mg-l IBA alone (7.42-7.78) or in combination with 1.0 mg-l IBA and 1.0mg-l NAA (8.00). Number of roots/plantlet was highest in MS medium fortified with 1.0 mg-l IBA (1.68) followed by 1.5mg-l IBA(1.37), while it was minimum with the MS medium supplemented with 2.0mg-l IBA(0.27). In Dendrobium cv. Banayat Pink, BAP could not improve the leaf length as compared to control (0.71 cm) instead it was observed to have inhibitory effect as the length of leaves were decreasing with the increased in concentration of BAP. IBA alone is sufficiently effective in the development of roots than NAA alone or in combinatrion with IBA.
Key words: Dendrobium, Banyat Pink, Growth hormones
19. Journal of Cell and Tissue Research 14(3): 4571- 4576 (2014)
Application of RAPD markers in genetic diversity analysis of groundnut
Vadodariya Gopal, D., Sharma, H., Chahar, S., Maloo, S.R. and Tandel, D.H.
Department of Plant Pathology, N.M. College of Agriculture, Navsari Agricultural University, Navsari
396 450, Gujarat. E. mail: gplvadodariya@gmail.com,
Abstract: Estimation of genetic diversity and identification of groundnut cultivars and breeding lines has been done using 15 RAPD primers, among which 13 were polymorphic. A total of 78 bands were obtained and 71 (91.02%) showed polymorphism between cultivars. On an average 6 bands per primer were obtained. Cluster analysis based on Jaccard’s similarity coefficient using UPGMA grouped all the cultivars into seven major clusters. Further, the groups/ clusters obtained by dendrogram could also be distinguished by similarity for the morphological characteristics within each group. Such an association may be used for improvement of yield and other traits in groundnut.
Key words: RAPD markers, Groundnut, Molecular diversity
20. Journal of Cell and Tissue Research 14(3): 4577- 4580 (2014)
The role of peroxidase and polyphenoloxidase isoenzymes in resistance to bacterial leaf blight (Xanthomonas campestris pv. Cyamopsidis) in clusterbean (Cyamopsis tetragonoloba (L.) Taub.)
Kalaskar, S.R., Shinde, A.S., Dhembre, V.M., Sheikh, W.A., Patil, V.S., Rathod, A.H. and Acharya, S.
Department of Plant Molecular Biology and Biotechnology, Centre of Excellence for Research on Pulses, Sardarkrushinagar Dantiwada Agricultural University, Sardarkrushinagar, Gujarat.
Abstract: Clusterbean, also known as guar (Cyamopsis tetragonoloba (L.) Taub), is a major arid legume and among the major diseases bacterial leaf blight (Xanthomonas campestris pv. cyamopsidis) is the most dreaded and limits clusterbean productivity. In the Present investigation activity of the isoenzyme peroxidase (PO) and polyphenoloxidase (PPO) were estimated in healthy and bacterial leaf blight affected samples of the individual 12 genotypes of clusterbean. The range of 0.42 to 1.07 and 0.08 to 0.36 changes in OD/min/g of fresh weight tissues in healthy plants and 0.19 to 1.20 and 0.09 to 0.52 changes in OD/min/g of fresh weight tissues in bacterial leaf blight affected plants was observed for peroxidase and polyphenoloxidase activity respectively. Decrease in the peroxidase and polyphenoloxidase activity in bacterial leaf blight infected susceptible plant as compared to bacterial leaf blight affected resistant plants and contrast in resistant plants were observed. The decrease was maximum in most susceptible genotype viz; PNB i.e 0.75 to 0.19 (Peroxidase) and 0.22 to 0.09 (Polyphenoloxidase). Thus increase or no change in peroxidase activity after infection can be taken as a marker for resistance to bacterial leaf blight.
Key words: C. tetragonoloba, Xanthomonas, Clusterbean
21. Journal of Cell and Tissue Research 14(3): 4581- 4586 (2014)
Proximate composition, isozyme and glutenin protein variation among Indian wheat cultivars
Patil, V.R.,Singh, C. and Talati, J.G.
Department of Biochemistry, B.A. College of Agriculture, Anand, Agricultural University, Anand 388 110, Gujarat.. E. mail: mrvishalpatil@gmail.com,
Abstract: Biochemical variation among 12 Indian aestivum and durum wheat cultivars were revealed by proximate composition and electrophoretic separation of isozyme and glutenin protein. Flour qualities of wheat cultivars for different end use products were studied using different qualitative parameters. Among them protein, â-carotene and lysine-tryptophan content showed high polymorphism in cultivars and found efficient for quality testing of flour. The study of esterase, peroxidase, and polyphenol oxidase isozymes provides interesting markers for species differentiation, but it is less useful for studies of within species genetic variation. Glutenin protein fractionation on SDS-PAGE found efficient technique to detect polymorphism among wheat cultivars according to their dough quality. HMW-GS allele also showed wide genetic variation between cultivars of species T. aestivum and T. durum, this can be a useful marker for species identification.
Key words: Glutenin protein, Wheat cultivars
22. Journal of Cell and Tissue Research 14(3): 4587- 4590 (2014)
In vitro regeneration of Curcuma amada (Roxb.) through shoot tip culture
Bimal, R., Rani, S. and Priyadarshini, M.
Biotechnology Lab, University Department of Botany, B.R.A. Bihar University, Muzaffarpur
842001 (Bihar). E. mail: raageeva@gmail.com,
Abstract: The present paper reports in vitro micro-propagation of Curcuma amada (Roxb.) commonly known as mango ginger because of its typical mango like flavour and ginger like appearance. The rhizomatous part of the plant is a rich source of many pharmaceutically important chemicals. The plant is propagated only vegetatively. Since there is a great demand of rhizome the novel technique of tissue culture holds promise for in vitro micro-propagation. In current contribution in vitro regeneration potentials of shoot tips of perennial rhizomatous herb is investigated. The excised buds cultured on MS medium supplemented with BAP (11.12µm) and NAA (5.37µm) proliferated 7-9 shoot buds in 1-2 weeks of culture. The regenerated shoots sub-cultured on MS medium containing IBA (4.90µm) produced well developed roots in 10-12 days of culture. The healthy plantlets with proper roots were transferred to pots. The survival frequency of the regenerated plants transplanted in the soil was 75-80%.
Key words : Curcuma amada, shoot tip culture
23. Journal of Cell and Tissue Research 14(3): 4591- 4594 (2014)
In vitro culture of floral heads and induction of vivipary in Eclipta alba Hassk
Bimal, R., Priyadarshini, M. and Rani, S.
Biotechnology Lab, University Department of Botany, B.R.A. Bihar University, Muzaffarpur
Abstract: The members of sunflower family (Asteraceae) are characterized by a highly specialized type of flower arrangement or inflorescence called head or capitulum and its ultimate advantage seems to be the mass pollination, setting of seeds in most heads for multiplication in large numbers and continuity of species. The fertilized flower heads or capitula of different ages of Eclipta alba were cultured in liquid Murashige and Skoog’s medium for morphogenetic studies. The heads collected after fertilization and cultured on MS plain nutrient medium induced vivipary i.e. the seeds geminated directly on floral heads. The early stage and immature embryos complete their full development viviparously in seeds embedded in heads leading to synchronized germination into plantlets. The induction of vivipary in E. alba by flower head culture may prove to be an important potential system to study molecular basis of seed germination and vivipary. This is the first report of vivipary in attached or detached heads in E. alba.
Key words : Eclipta alba, Capitula culture, Vivipary
24. Journal of Cell and Tissue Research 14(3): 4595- 4600 (2014)
Influence of genotype, explant and growth regulators onin vitro regeneration of brinjal (Solanum melongena L.)
Jadhav, M.A., Jadhav, A.S., Pawar, B.D., Kale, A.A. and Kute, N.S.
State Level Biotechnology Centre, Mahatma Phule Krishi Vidyapeeth, Rahuri, 413722:
E. mail: jadhavmandar1@gmail.com.
Abstract: A reproducible and highly efficient plant regeneration protocol using shoot tip and hypocotyl explant in four brinjal cultivars, viz. ‘Manjarigota’, ‘Ruchira’, ‘Poona selection’ and ‘Krishna kathi’ was developed. Considerable genotypic variation in response to plant growth regulators used was observed in both explants. The MS medium with 2.0 mg/l BAP, 1.0 mg/l Kn was found to be the best regeneration medium for Manjarigota, whereas MS medium with 2.0 mg/l BAP, 0.1 mg/l IAA was found to be the best regeneration medium for Ruchira, Poona selection and Krishna kathi with regards to regeneration efficiency, days to callus initiation, percent callus induction, days to shoot initiation and number of shoots per explant. Earliest callus induction (9.3 days) with shoot tip explant was observed in Manjarigota whereas with hypocotyl explant it was earliest (9.6 days) in Krishna kathi. Poona selection showed highest callus formation (90 and 81.3%) with shoot tip and hypocotyl explants respectively. Direct shoot initiation without formation of callus was observed on media supplemented with BAP and NAA. Earliest shoot initiation (14.6 days) with shoot tip explant was observed in Manjarigota and Ruchira whereas with hypocotyls, it was observed in Ruchira (15.3 days). Among explants used shoot tip showed highest shoot multiplication (73.4%) followed by hypocotyls (70.1%) in Manjarigota. Highest numbers of shoots per explant (3.8) were obtained with shoot tip explants in Manjarigota. Earliest in vitro root initiation was observed within 14.6 days on MS medium with 1.0 mg/l IBA in 14.6 days. The in vitro rooted plantlets were successfully established in polycarbonated polyhouse.
Key words: Solanum melongena, Growth regulators
25. Journal of Cell and Tissue Research 14(3): 4601- 4608 (2014)
RNA interference for insect pest management-recent developments: A review
Shah, M.A., Khan, A.A. and Mir, G.M.
Division of Entomology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar, Srinagar, 190 025. E. mail: khubaib20@gmail.com,
Abstract: RNA interference (RNAi) is a post-transcriptional gene regulatory mechanism that controls gene expression (gene silencing) at the mRNA level within living cell. RNAi has become the first choice in functional genomic research on insects, but it also has considerable potential for the control of pest insects. It was postulated that in order for an insecticidal RNAi-based strategy to be effective it must have as its target a gene required for a process vital to the host. For this purpose, efficient methods of dsRNA delivery and choice of the gene to be knocked down are key operations. Currently, delivery of dsRNA via transformed host plants, virus host system, transformed bacterial feeding and spraying has been found to be effective in principle. High throughput target gene screening employing techniques such as RNA-seq, DGE-tag and RIT-seq have enabled the large scale screening of potential target genes from the insect genome and by each day new targets are being evaluated for insect pest management
Key words: RNAi, pest management, dsRNA
26. Journal of Cell and Tissue Research 14(3): 4609- 4612 (2014)
Incidence and biochemical characterization of Aeromonas species isolated from retail fish and chicken in north Kolkata region
Praveen, P.K.,Debnath,C., Pramanik, A.K., Shekhar, S.and Dalai, N.
Department of Veterinary Public Health and Epidemiology, Faculty of Veterinary and Animal Sciences,
West Bengal University of Animal and Fishery Sciences, Kolkata - 37 (West Bengal).
E. mail: drpraveenvet2005@gmail.com,
Abstract: In the present study 179 samples from fish (Gills) and chicken (Raw meat) were processed for bacteriological examination. Ampicillin Dextrin Agar (ADA) media was used for isolation of Aeromonads from fish and chicken. A total of 31 (17.32%) isolates were obtained from 179 samples of fish and chicken in North Kolkata Region. All isolates were positive for oxidase, catalase and arginine decarboxylase, negative for ornithine decarboxylase, grew on 0% NaCl solution and produced acid from mannitol indicating the presence of Aeromonas organism. In 31 isolates highest isolation were achieved from fish 24 (18.89%), followed by chicken 7 (13.46%) out of which 29 (93.54%), 1 (3.22%), 1 (3.22%) were recognized as three species of Aeromonas namely A. hydrophila, A. sobria and A. caviae respectively through biochemical characterization including aesculin hydrolysis, Voges-Proskauer (VP) and gas from glucose (TSI) tests. Strains which showed positive for all the three tests were identified as A. hydrophila, whereas A. sobria were positive for TSI and VP but negative for aesculin hydrolysis. In contrast A. caviae were negative for TSI and VP but positive for aesculin hydrolysis.
Key words: Aeromonas, Chicken, Fish, Isolation
27. Journal of Cell and Tissue Research 14(3): 4613- 4616 (2014)
Antibiotic sensitivity and virulence potential study of Aeromonas species isolated from retail fish and chicken in and around Kolkata
Praveen, P.K., Debnath, C., Pramanik, A.K., Shekhar, S., Dalai, N. and Rai, R.
Department of Veterinary Public Health and Epidemiology, Faculty of Veterinary and Animal Sciences,
West Bengal University of Animal and Fishery Sciences, Kolkata - 37 (West Bengal).
E. mail: drpraveenvet2005@gmail.com,
Abstract: In present study, 31 Aeromonas isolates were tested for antibiotic sensitivity and virulence potential. The antibiogram of the isolates revealed that uniform resistance among isolates against vancomycin, augmentin and nalidixic acid (100%). On the contrary, rifampicin, ceftrixone, chloramphenicol, streptomycin, ampicillin were found as a group of antibiotics for which more than 80% sensitivity could be recorded. Likewise, tetracycline, levofloxacin, clindamycin, amikacin, ciprofloxacin and cefpodoxime were moderately sensitive antimicrobials against isolated Aeromonads. More than 40% of the isolates were found resistant to co-trimoxazole and gentamicin. In case of virulence study haemolysin test indicated that 18 (58.06%) isolates out of 31 tested were positive in this test. Majority of the haemolysin positive isolates were obtained from fish (83.33%) samples as compared to chicken (16.67%). With the growing importance of Aeromonas as an emerging pathogen, it is important to combat this organism. It is indisputable that Aeromonas strains may produce many different putative virulence factors, such as enterotoxins, haemolysins or cytotoxins. It has been established that haemolysin is a virulence factor contributing to the pathogenesis of A. hydrophila infection.
Key words: Aeromonas, Antibiotic
28. Journal of Cell and Tissue Research 14(3): 4617- 4620 (2014)
Molecular traceability of non-descript sheep meat using polymerase chain reaction
Narendra Babu, R.,Thulasi, G., Robinson, J.J.A.and Ruban, S.W.
Department of Meat Science and Technology, Madras Veterinary College, Chennai - 7
E. mail: nbabumst2@gmail.com,
Abstraxt: A study was undertaken to assess the use of PCR technique as analytical tool for meat species identification. Mutton samples were taken as fresh, frozen, cooked and processed from indigenous non-descript local sheep breed in Tamil Nadu. The extraction of DNA was carried out using simple lysis based extraction procedure and a commercial DNA extraction kit. Samples with OD ratio between 1.7 to 1.9 were considered good in terms of concentration and purity and were used for PCR amplification. Electrophoresis of amplified PCR products in 3% agarose gel and then gel documentation revealed species specific bands with 335 bp when common forward primers and species specific reverse primers were used. Cooking and further processing did not affect the efficacy of PCR amplification. Sequencing of purified PCR product was done by automated sequencing procedure using BigDye terminator V 3.1 cycle and AB1 3700 DNA analyzer. The results of sequence analysis and comparison with the sequences for mitochondrial cytochrome b gene showed 98-99% identity with that published in National Centre for Biotechnology Information (D 84205.1).
29. Journal of Cell and Tissue Research 14(3): 4621- 4626 (2014)
Plant regeneration from immature embryo-derived callus of barley (hordeum vulgare L.): Effect of auxins and cytokinins
Tiwari, V.K.,Cocking, E.C. and Davey, M.R.
Plant Genetic Manipulation Group, Department of Life Science, University of Nottingham, University Park, Nottingham NG7 2RD, England.E. mail: vkt786@rediffmail.com,
Abstract: Embryogenic callus was induced from culturing immature embryos of three barley cvs. Dissa, Golden Promise and Igri. Maximum embryogenic callus formation was achieved by culturing immature embryos on CC and LS medium containing 2 mg/litre and 2.5 mg/litre 2,4-dichlorophenoxyacetic acid (2,4-D) respectively. Embryogenic calli were frequently produced distinguished somatic embryos on these medium. Highest frequency of somatic embryo’s germination was occurred on MS medium supplemented with 0.5 mg/litre kinetin and 3% glucose in cv. Dissa. This medium also induced regeneration in the calli of cv. Golden Promise and Igri. The age of callus also influenced regeneration with high frequencies occurring after 6 wk in culture. Subsequently, plant regeneration was achieved by transferring germinated somatic embryos on hormone-free MS medium. It indicated that plant regeneration was achieved via somatic embryogenesis and organogenesis. Plants regenerated from immature embryos were grown to maturity in soil.
Key words: Hordeum vulgare L., Somatic embryos, Callus culture
30. Journal of Cell and Tissue Research 00(0): 0000- 0000 (0000)
This paper has been withdrawn
31. Journal of Cell and Tissue Research 14(3): 4633- 4640 (2014)
Isolation, characterization and identification of methyl parathion degrading bacteria from
industrial waste
Vadukia, M.R., Tomar, R.S., Parakhia, M.V., Malviya. B.J., Rathod, V.M., Thakkar, J.R., Bhalara, R.L. and Golakiya, B.A.
Department of Biotechnology, Junagadh Agricultural University, Junagadh 362001, Gujarat.
E. mail: mvparakhia@gmail.com,
Abstract: The present study focus on isolation of bacteria which have capacity to bioremediation of methyl parathion from pesticide contaminated soil. This was attempted by isolating 45 bacteria from industrial waste and methyl parathion (MP) enriched soil. Out of the screened 45 bacterial isolates, 21 grew on mineral salts medium containing 500 ppm methyl parathion as the sole carbon source. The most promising methyl parathion degrading bacterium was MPD-21 which was completely degraded up to 500 ppm methyl parathion within 24 hrs. MPD-14, MPD-1, MPD-17 and MPD-2 were the other bacteria which degraded the pesticide by 189 mg/l, 185.1 mg/l, 182.5mg/l and 175 mg/l respectively in 24 hrs. All isolates were characterized by morphological, biochemical and phenotypical assays. Based on HPLC result five highly efficient isolates were characterized by 16S rRNA gene sequencing. According to the results MPD-21 was identified as Pseudomonas putida, the similarity of MPD-17 with Enterobacter cloacae sub spp.
Key words: Methyl parathion, Degrading bacteria
32. Journal of Cell and Tissue Research 14(3): 4641- 4646 (2014)
Isolation and bio-analytical characterization of chloropyrifos degrading bacteria
Parmar, K.J., Tomar, R.S., Parakhia, M.V., Malviya. B.J., Rathod, V.M., Thakkar, J.R., Kothari, V.V., Bhatt, A.J., Bhalara, R L. and Golakiya, B.A.
Department of Biotechnology, Junagadh Agricultural University, Junagadh 362001, Gujarat.
E. mail: mvparakhia@gmail.com, Cell: 09979164149
Abstract: Pesticide residue in food is burning problem in present era. To identify remedy for this nineteen chlorpyrifos degrading bacteria were isolated from an organophosphosphate pesticides manufacturing area of Gujarat. All isolates were compared by biochemical, morphological, phenotypical and analytical methods. The isolate utilized chlorpyrifos as the sole source of carbon and phosphorus for its growth and hydrolyzed chlorpyrifos to 3,5,6-trichloro-2-pyridinol. These isolates grow at concentrations of chlorpyrifos up to 500 ppm. Biodegradation study of chlorpyrifos was performed using high performance liquid chromatography (HPLC) using 300ppm chlorpyrifos concentration and the sample was collected at different time interval (at 6 hr, 12 hr, 18 hr and 24 hr). The most promising chlorpyrifos degrading bacteria was CPD-1, CPD-4, CPD-9, CPD-15 and CPD-18 which degraded 70%, 71%, 66%, 80% and 83% respectively with in 24 hr. Five highly efficient isolates were identified based on 16s rRNA sequencing and identified CPD-1, CPD-4, CPD-9 and CPD-15 as Enterobacter aerogenes, Citrobacter gillenii, Citrobacter braakii and Citrobacter murliniae respectively.
Key words: Chlorpyrifos, Chlorpyrifos degrading bacteria, 16s rRNA
33. Journal of Cell and Tissue Research 14(3): 4647- 4652 (2014)
Effect of plant growth regulators on in vitro regeneration from different explants in clusterbean
Cyamopsis tetragonoloba (L.) TAUB
Meghwal, M.K., Kalaskar, S.R., Rathod, A.H., Tikka, S.B.S. and Acharya, S.
Department of Genetics and Plant Breeding, Chimanbhai Patel College of Agriculture and Centre of
Excellence for Research on Pulses, Sardarkrushinagar Dantiwada Agricultural University,
Sardarkrushinagar, Gujarat, E. mail: drsampatkalaskar@gmail.com, Cell: 09923724612
Abstract: Cotyledonary node, hypocotyl and epicotyls of clusterbean were used as explant which were cultured on Murashige and Skoog’s basal medium supplemented with Auxin (2, 4-D, NAA and IBA), cytokinin (BA and Kn) and Gibberellin (GA3). Lower level of 2, 4-D and BA were found good in terms of production of friable and morphogenic callus induction from explant like cotyledonary nodes and hypocotyls of clusterbean. Among the different media concentration tried, MS + 1.2 mg/l 2, 4-D + 0.6 mg/l BA recorded minimum number of days for callus initiation. The shoots were directly regenerated from cultured epicotyle explants. The hormonal combination of 2 mg/l BA + 2 mg/l Kn + 1 mg/l GA3 was most promising for enhanced branching and regeneration of multiple shoot. The regenerated shoot of 3 to 4 cm in length was found ideal for the induction of roots. The results revealed that the MS + 1.2 mg/l NAA was the most effective for optimum root induction in regenerated shoots
Key words:Cyamopsis tetragonoloba, Callus culture, Growth regulators
34. Journal of Cell and Tissue Research 14(3): 4653- 4658 (2014)
Callus culture and organogenesis in Fir (Abies pindrow Royle)
Bhat, S.J.A., Gangoo, S.A., Geelani, S.M., Qasba, S.S. and Parray, A.A.
Faculty of Forestry, Sher-e-Kashmir University of Agricultural Science and Technology, Kashmir 191121. E. mail: javaidahmad44@yahoo.com
Abstract: A protocol for callus induction and regeneration of complete plantlets from nodal and internal stem segment explants of Fir (Abies pindrow Royle) was developed. For callus formation explants (nodal and internodal stem segments) were inoculated on MS medium supplemented with different concentration of 2, 4-D, NAA and BAP alone and in combination were kept in dark for 3 weeks. MS medium supplemented with NAA (3.00 mg l-1) resulted in highest percentage (10.33%) of callus induction on internodal stem explant with bark removed. Callus cultures were kept in growth chamber for 4±1 weeks on same medium for multiplication. Multiplied callus cultures were kept on regeneration medium containing different concentrations of TDZ, BAP alone and in combination with IBA. Maximum shoot regeneration percentage (24.00 %), average number of shoots (5.00) was obtained in MS medium supplemented with BAP (5.00 mg l-1) plus IBA (0.50 mg l-1) and length of longest shoot (15.00 mm) was recorded on MS medium supplemented with BAP (1.50 mg l-1). Rooting in regenerated microshoots was achieved in the MS half medium containing IBA (4.00 mg l-1) and activated charcoal (200 mg l-1).
Key words: Callus, Abies pindrow Royle
35. Journal of Cell and Tissue Research 14(3): 4659- 4664 (2014)
In vitro propagation of Stevia rebaudiana (Bert.) A natural, non caloric sweetener herb
Autade, R.H., Fargade, S.A., Borhade, P.G., Udmale, S.K. and Choudhary, R.S.
Department of Plant Biotechnology, College of Agricultural Biotechnology, Loni, ITI Campus, Chandrapur
Road, Tal. Rahata, Dist. Ahmednagar, 413736 (Maharashtra) E. mail: rishiz.autade@gmail.com,
Abstract: Stevia rebaudiana is a medicinally important, zero-caloric value, sweet tasted and an antidiabetic herb. In vitro micropropagation of Stevia has been done by using nodal segments as an explant that were cultured on Murashige and Skoog media supplemented with five different concentrations of benzylaminopurines (BAP) (0.0-2.0mg/l) in combination with kinetin (KN) (0.0-2.0 mg/l). After, five weeks of initiation, maximum numbers of shoots (5.16±0.76) were obtained on media supplemented with BAP (1.0 mg/l) + KN (1.0 mg/l). Best shoot length (7.00±0.64 cm) was observed on media containing BAP (0.5 mg/l) + KN (0.5 mg/l) with the highest 95% survival rate. Shootlets were regenerated through axillary shoot proliferation on MS media with same hormone concentrations those were used for establishment of explants. Maximum number of shoots (5.28±0.75) and shoots length (7.45 ± 0.24cm)were recorded on MS media containing BAP (1.5mg/l) + KN (1.5 mg/l) and BAP (2.0mg/l) + KN (2.0mg/l)with a survival rate 85% and 95% respectively. The single shoots were transferred for rooting to half MS media containing indole 3-butyricacid (IBA) (0.0-2.5mg/l). Most efficient rooting response with average length 1.68 ± 0.3cm and number of roots (3.60 ± 0.29) were observed onhalf MS media supplemented with 1.0mg/l IBA, where survival rate was 90%. The well-rooted Stevia plantlets transferred to green house condition for primary hardening that showed promising survival rate.
Key words: Stevia rebaudiana, Micropropagation
36. Journal of Cell and Tissue Research 14(3): 4665- 4669 (2014)
In vitro studies on effect of corm weight and availability of moisture on plant development in saffron (Crocus sativus L.)
Salwee, Y. and Nehvi, F.A.
Saffron Research Station, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Kashmir 191121 (J. and K.). E. mail: f.nehvi@rediffmail.com,
Abstract: Significant impact of corm weight was observed on enhancing number of sprouts located in apicular, sub apicular and auxiliary regions. Increased number of sprouts particularly in sub-apicular and auxiliary region was observed to be detrimental for induction of more number of productive flowering sprouts and exhibited significant relationship with increased number of vegetative nonproductive sprouts. 1-3 flowers/sprout were recorded from apicular regions in corms weighing above 8 g whereas, sub-apicular regions showed maximum 1-2 flowers/spathe. Corms weighing below 7 g exhibited only vegetative sprouts, thereby, confirming importance of corm weight on flower induction. Number of leaves/corm ranged from, 2.0 (1-2g) - 21.9(>15 g) with maximum contribution of sprouts in apicular region (2-10.6) followed by sub-apicular (2-7.3) and auxiliary sprouts (1.6-4). Similar positive effects of corm weight and moisture availability was observed on root development and cataphyll length. Maximum number of lateral/terminal contractile roots (5.9) were observed in corms weighing above 15 g with adequate moisture followed by 4.3 in corms weighing 8-14 g, whereas, in other category of corms weighing less than 7 g only 2 contractile roots were observed, thereby, confirming that heavier corms confirm to better potential of daughter corm production. Available moisture was found to be a critical factor for cataphyll length and root development irrespective of corm weight but proportionate increase was more in bigger corms rather than smaller corms.
Key words: Saffron (Crocus sativus L), Corm weight
37. Journal of Cell and Tissue Research 14(3): 4671- 4676 (2014)
Standardization of selective lethal dose of antibiotics (Hygromycin and Paromomycin) for selection of transgenics in durum wheat
Verma, L.,Bains, N.S.and Gosal, S.S.
School of Agricultural Biotechnology, Punjab Agricultural University, Ludhiana 141004,
Punjab. E.mail: vermaleela@yahoo.co.in,
Abstract: In transformation process antibiotic resistance genes are introduced along with the gene of interest and used for selecting the transformed cells from the non-transformed ones. Selective lethal dose of two antibiotics i.e. hygromycin and paromomycin was standardized for three durum wheat varieties viz., PDW314, PDW291 and PDW233. Antibiotic sensitivity tests were carried out on callus and seedlings from immature embryos of durum wheat. To determine the selective lethal dose of antibiotic for selective killing of non-transformed cells or tissues of durum wheat after Particle gun and Agrobacterium mediated transformation, different concentration of hygromycin and paromomycinwas added into the callusing medium (0, 15, 20, 25, 30, 35 and 40 mg/l) and the basal medium for seedlings (0, 5, 10, 15, 20, 25 and 30 mg/l).The calli cultured on the concentration of 25 mg/l hygromycin and 20 mg/l of paromomycin was found to be suitable. While in case of seedling culture, the basal medium supplemented with 15 mg/l of hygromycin and paromomycin was found to be most effective and these concentrations were used afterwards for the selection of explant transformed with hygromycin phosphotra-nsferase (hpt) and neomycin phosphotransferase II (npt II) selectable genes.
Key words: Durum wheat, Hygromycin, Paromomycin, Callus culture
38. Journal of Cell and Tissue Research 14(3): 4677- 4680 (2014)
Rosette formation: A tool for T-lymphocyte enumeration and assessment of innate immunity
Dewangan, G., Hirpurkar, S.D., Giri, D.K. and Kashyap, D.K.
Department of Veterinary Microbiology, College of Veterinary Science and Animal Husbandry, Anjora, Durg, (C.G.) 491001. E. mail: govinadewangan@gmail.com,
Abstract: Rosettes formation technique (SRBC) was employed for the assessment of lymphocyte per cent. Development of accurate methods for enumeration of T and B lymphocytes in human and animal peripheral blood during the past decades has led to a plethora of papers on T and B lymphocytes in various pathologic states. Lymphocytes can be divided into T and B lymphocytes on the basis of selective binding capacity of T-lymphocytes with Sheep RBC (SRBC). On the basis of enumeration of T-lymphocyte innate immunity status of Non-discript (ND) and Sahiwal (S) cattle was compared. Rosette forming T-lymphocytes were enumerated in all the four groups of animals. The mean (±SE) values of T-lymphocytes (per cent) in ND-I, ND-II, S-I and S-II was 41.04±0.47, 56.83±0.99, 35.79±0.93 and 56.62±0.76, respectively. A comparative study on T-lymphocytes showed that there was significant increase (P<0.01) in T-lymphocytes count with respect to age in both breeds. The T-lymphocytes count of ND calves was significantly (P<0.01) high when compared with Sahiwal calves whereas breed difference was not observed in adult animals.
Key words: T-lymphocytes, Innate immunity
39. Journal of Cell and Tissue Research 14(3): 4681- 4684 (2014)
A microbial, antibiogram and haematological study of mastitic cows
Padhy, A., Dalai, N., Shekhar, S., Sahu, A.R.,Sahoo, S. and Kashyap, D.K.
Department of Veterinary Microbiology, Arawali Veterinary College, Sikar, Rajasthan.
E. mail: dearlita123@gmail.com,
Abstract: In the present study 32 milk samples from mastitic cows were processed for microbial and antibiogram test and blood samples were analysed for haemotological study. Out of 32 samples, Staphylococcus aureus accounted as a causative agent of 68.75% followed by E. coli (18.75%) and Streptococcus spp. (12.5%). From antibiogram test highest resistance was noticed in Ceftizidime, Penicillin and Tetracycline for Staphylococcus aureus, E. coli and Streptococcus spp. respectively. Gentamicin was found to be highest susceptible antibiotic for all the three types of isolates. Haematological study revealed the significant increase in ESR and WBC and subsequently significant decrease in PCV, Hb and RBC (p<0.05).
Key words: Mastitic cows,Haemotology
40. Journal of Cell and Tissue Research 14(3): 4685- 4690 (2014)
Molecular diversity in chickpea (Cicer arietinum L.) genotypes by ISSR markers
Deshmukh, S.G., Harer, P.N. and Sawant, P.V.
Department of Agricultural Botany, Post Graduate Institute, Mahatma Phule Krishi Vidyapeeth, Rahuri
413722, Ahmednagar (Maharashtra). E. mail: deshmukhsonalkumar@yahoo.co.in
Abstract: Forty two genotypes were selected for molecular diversity analysis. Out of 25 ISSR primers, 13 were polymorphic which produced a total of 78 reproducible loci with an average of 6.00 loci per primer which gives 95.50% polymorphism. The number of loci generated by each primer varied from 3 (ISSR 866) to 10 (ISSR 809 and ISSR 880). The similarity coefficient between the genotypes varied from 0.45 to 0.90 indicating moderate to high diversity among the genotypes. The UPGMA based clustering analysis using Dice similarity coefficient grouped forty two Chickpea genotypes into three major clusters. Genotypes inside the each cluster represent the similarity in genetic constitution while their grouping in different clusters represents diversity.
Key words: Molecular Diversity, ISSR Marker, Chickpea
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