Botany - Mphil Thesis

Permanent URI for this collectionhttps://kr.cup.edu.in/handle/32116/30

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End date: 2016Has files: Yes

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    Selection for Salt Tolerance in Petunia Grandiflora
    (Central University of Punjab, 2014) Najar, Raoof Ahmad; Saini, R.G.
    Petunia grandiflora a native of South America is a popular and high value ornamental plant in the world. Soil salinity is the major abiotic stress in semi-arid Malwa region of Punjab, adversely affecting its productivity, survival and quality. In order to develop salt resistant varieties of P. grandiflora, an attempt was made to select salt tolerant seedlings from varieties Violet Blue, Giant California and Nana compecta by using ex-vitro and in-vitro methods. Seedlings were subjected to salt treatments of 100 mM, 200 mM, 300 mM, 400 mM, 500 mM and 600 mM for durations of 4 hr, 6 hr, 12 hr and 24 hours. Salt treatment reduced shoot length, leaf number and survival percentages and delayed days to 50% flowering. Variety Nana Compecta was identified as tolerant to salt and variety Giant California was most sensitive to soil salinity. In in-vitro method of gradual increase in NaCl concentration (0, 25, 50, 75 and 150 mM) from low to high level was found to be a better approach for selecting salt tolerant calli as compared to direct method in which direct transfer of calli to high salt concentrations (50, 100, 150, 200, 250 and 300 mM) was found to be detrimental to callus survival and growth. Treatment of calli upto 50 mM salt concentrations was good for shoot regeneration in all the three varieties. Protocol was standardized for callus induction, direct shoot induction from leaves, shoot regeneration from callus and root induction from shoots. For callus induction, 2 mg/l 2, 4-D and 0.5 mg/l kinetin was optimum for variety Violet Blue and 1 mg/l both BAP and NAA was optimum for varieties Giant California and Nana Compecta. For direct shoot induction from leaves 2 mg/l BAP and 0.1 mg/l NAA were good for all the three varieties. For shoot regeneration from calli, 2 mg/l BAP in combination with 0.1 IAA for variety Violet Blue and 1 mg/l both BAP and NAA for varieties Giant California and Nana Compecta were ideal. For root induction, 0.5 mg/l NAA and 0.1 mg/l IBA were ideal for all the three varieties. Seeds from plants grown ex-vitro showing tolerance to salt were harvested and stored for further testing.
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    Development and characterization of genomic microsatellite markers in Melia azedarach
    (Central University of Punjab, 2014) Thakur, Sapna; Bhardwaj, Pankaj
    Melia azedarach is ecologically imperative species known for its innumerable biological benefits such as antiviral, anthelminthic, antibacterial, etc. In this study, we developed 43 genomic microsatellite markers from (AG)n enriched library and subsequently employed 23 of them for genetic diversity and population structure analysis of Melia azedarach growing in Indian Thar desert. Fourteen populations encompassing 95 genotypes were selected for analysis and we found a moderate level of diversity (Na = 3.211, Ho = 0.558, He = 0.549, P = 94.41%) in them. Gene diversity (h) among population pairs varied from 0.566 to 0.714 with very low overall genetic differentiation (F = 0.021). The highest value of ΔK estimated using STRUCTURE indicated 2 subpopulations (K=2) and admixed cluster occupied maximum area (75.79%) under Bar plot. Genetic distance based UPGMA dendrogram also identified 2 major clusters among 14 Melia azedarach populations. UNJ tree based on genetic dissimilarity clustered genotypes from different population together. No significant correlation between geographical and genetic distance was found in present study (Rxy = 0.261, P = 0.18). Allele frequency distribution under “mode-shift” indicator was normal L-shaped, suggesting populations under study are not experiencing any recent bottleneck. This study laid the foundation for more precise inference about the biogeography and management of M. azedarach in the Indian Thar Desert
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    To study the effects of insulin and metformin on pc-3 cell line
    (Central University of Punjab, 2016) Singh, Pushpendra; Bast, Felix
    Dietary habits, genetic factors, hormonal factors and environmental factors are the independent risk factors for prostate cancer as well as diabetes. Androgen is the primary growth factor for the prostate cancer initiation and progression, however, non androgen peptide growth factor like insulin and insulin growth factor also involved in the prostate cancer as well as diabetes. Insulin and insulin growth factor are peptide that regulates metabolism, growth, cellular proliferation and apoptosis. The anti-diabetic drug metformin is rapidly emerging as a potential anti-cancer agent that improves insulin homeostasis and decreased growth and cellular proliferation of the prostate cancer cell line. Thus it is necessary to understand the growth promoting role of insulin on prostate cancer cell line and the possible influences of metformin on the proliferation of prostate cancer cell line in the presence and absence of insulin has been studied.
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    Development and chacterization of genomic microsatellite markers in rhdodendron arboreum
    (Central University of Punjab, 2014) Choudhary, Shruti; Bhardwaj, Pankaj
    Rhododendron arboreum, a key species inhabiting Indian Himalayas, has ecological and economic importance. Global climate change and anthropogenic activities can pose a threat to the biodiversity richness. Population genetics characteristics are the fundamentals of conservation and management practices. However, no genetic background for the species is presently available. To provide an insight into the population structure of the species, an effective marker resource was developed. A genomic library enriched for microsatellites was constructed using biotin- streptavidin hybridization technique, followed by sequencing of positive clones. With 54% enrichment rate, 41 primers were designed from SSR clones, consisting of perfect or interrupted repeats of AG/CT, AC/GT motifs. 38 loci showed successful amplification on genotypes of three populations with number of alleles ranging from 2 to 14, observed and expected heterozygosity of 0.167 to 0.933 and 0.422 to 0.917, respectively and average polymorphic information content of 0.104 to 0.811. A high gene flow (Nm= 5.436) demonstrated high genetic diversity within the populations. 19 loci displayed significant deviations from Hardy Weinberg equilibrium. Overall no linkage disequilibrium and bottleneck was detected among sampled populations. DARwin and STRUCTURE analysis grouped the populations into two original clusters and admixed genetic stock irrespective of their geographical locations. These novel loci will support further genotyping studies in R. arboreum.
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    Sequence-based phygeography and conservation of seaweeds from indian subcontinents
    (Central University of Punjab, 2014) Rani, Pooja; Bast, Felix
    Phylogeography is the study of historical events which are responsible for evolution and current distribution of a species in different geographical area. However very less record is available about marine macroalgae of Indian subcontinent. This study investigated the DNA barcoding and phylogeographical distribution of marine algae from the Indian subcontinent. Different algae samples collected from various coasts of Indian subcontinent are amplified using ITS, COX and rbcL primers. In our results, we found the occurrence of green algae like Ulva reticulata, Ulva intestinalis, Ulva fasciata, Ulva proliifera, Ulva ohnoi and one sample with Caulerpa scalpelliformis ; Red algae, Gracilaria foliifera, Gracilaria domingensis, Gracilaria corticata, Grateloupia Sp., Ceramium Sp., Centroceras clavulatum, Erythrocladia Sp., Erythrocladia irregularis, Acanthophora Sp., Dilsea socialis, Hypnea stelullifera, Sirodotia tenuissima and Dichotomaria Sp.; Brown algae, Sargassum zhangii, Sargassum megalocystum, Sargassum aquifolium and Turbinaria ornata in Indian subcontinent. Gracilaria domingenesis, Dilsea socialis, Sargassum megalosystum were first time reported in India. On the basis of molecular studies, we found that Ceramium Sp. Nov., Erythrocladia Sp. Nov., Acanthophora Sp Nov., Grateloupia Sp. Nov. and Dichotomaria Sp. Nov. were identified as new species. Erythrocladia irregularis was identified as endophytic algae inside green algae Cladophora glomerata. Phylogenetic tree was generated to analyse the evolutionary distance between different samples. Morphological and microscopic studies were performed for each sample. This study further helps in identification and documentation of new species and cryptic species. All samples were pressed for herbarium voucher.
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    Oxidative stress and innate immune response in A549 lung carcinoma cells
    (Central University of Punjab, 2014) Upadhyay, Shishir; Dhiman, Manisha
    Cancer immunology is the study of cross-talk between the immune response and cancer cells. The immune response, including the recognition of cancer-specific antigens is of particular interest as knowledge gained drives the development of new vaccines and antibody therapies. The molecular mechanisms which are disturbed in the susceptible patients who proceed to develop cancer are very complicated and still largely unknown. It proposed that apart from the reported genetic modifications on tumor cells there are modifications due to oxidative stress (resulting in the formation of chemical adducts, e.g. 3-hydroxynonenal, 3- nitrotyrosine, carbonyl etc.) at the vicinity of tumor where the immune cells infiltrate. The central hypothesis of the present work is that respiratory burst which is host?s mechanism to kill the foreign particles (tumor cells) is used as defence mechanism by the tumor cells by forming neoantigen which in turn make them undetectable and can further help them to escape the host immune surveillance. The lung carcinoma A549 cells were treated with100μM H2O2 and using 1-D gel electrophoresis the oxidized tumor proteins in normal and treated cells were visualized. To confirm the oxidized modifications at membrane levels and at proteins levels the lipid peroxidation and protein carbonyls were detected respectively. It was observed that the oxidative stress induces the lipid peroxidation and protein carbonyls in tumor cells. To determine if neo (oxidized) tumor antigens elicit any alteration in immune responses where they show compromised phagocytosis, thus resulting in a failure to elicit effector immune functions were analyzed via phagocytosis and respiratory burst using spectrophotometry and microscopic techniques. The present study has identified a novel mechanism(s) of carcinogenesis initiation and which can further provide directions for the development of adjunct therapies to control cancer in its initial stages and at the same time it advocates for new ventures to increase the efficacy of the chemotherapeutic drugs.
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    Development and charaterization of UGMS markers for genetic diversity analysis in Rhododendron arboreum
    (Central University of Punjab, 2013) Sharma, Gagan; Bhardwaj, Pankaj
    Rhododendron arboreum is an ecologically important species growing in Himalayan regions. It is widely popular due to its medicinal potential, edible, sacred & economic value, however very little is known about its genetic diversity. The development and characterization of UGMS markers will provide a major impact on genetic diversity analysis. Total 792 (615 singletons and 177 contigs) unigenes were predicted from 1,241 publicly available EST database from NCBI (http://www.ncbi.nlm.nih.gov/) site in R.catawbiense by clustering of 2 - 33 random EST sequences. From 50 SSR contained Unigenes, 36 UGMS primers were designed for the cross amplification approach and six pairs of microsatellite primers were produced clear PCR amplification. The cross related species/genera transferability rate was 16.66% from R.catawbiense to R.arboreum. The polymorphism information content (PIC) ranged from 0.2756 to 0.9212 with an average of 0.5765.The average observed heterozygosity (Ho) was 0.8666 i.e. greater than average expected heterozygosity (He) was 0.6792. The genetic diversity was found with an average of 0.6222 within population of R.arboreum and genetic diversity found with an average of 0.3436 among the population.This study revealed the insight of abundance & distribution of microsatellite in the expressed region of the Rhododendron arboreum genome.
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    Isolation and characterization of heavy metal resistant bacteria from soil
    (Central University of Punjab, 2014) Mehra, Richa; Saini,R.G
    The exploitation of microbes to remove heavy metals from contaminated sites and industrial effluents is an extensive area of research. In this study, isolation of heavy metal resistant bacteria was done from potentially heavy metal contaminated agricultural soil of Bathinda region. The ICAP-AES analysis of soil revealed toxic concentrations of lead and nickel whereas most other heavy metals were also present in considerable amounts. The organisms showing maximum resistance to eight heavy metals tested (Pb, Cu, Ni, Cr, Co, Cd, Fe and As) were screened. The 29 potent metal resistant organisms were subjected to biochemical characterization and 16S rDNA sequencing, and tentatively identified as strains of Bacillus and Streptomyces spp. All the strains showed multiple heavy metal resistance with varied degrees of resistance. A strain of B. megaterium FE1 exhibited maximum resistance to all the heavy metals- Pb (2.5 mg/ml), Cu (1.5 mg/ml), Ni (1.0 mg/ml), Cr (3.0 mg/ml), Co (0.5 mg/ml), Cd (2.5 mg/ml) and As (2.5 mg/ml). The incidence of such extent of heavy metal resistance in native bacteria suggest that agricultural soil is undergoing stress which needs more intensive studies.
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    DNA Barcoding and phylogography of brown seaweeds of coasts of indian subcontinent
    (Central University of Punjab, 2013) Bhushan, Satej; Bast, Felix
    Algae are one of the diverse groups of ubiquitous autotrophs. Their use as food was more or less initially confined to few East Asian and South American countries like China, Japan, Korea, Chile, Argentina and Brazil, but with the increased demand and limited supply for food combined with the medicinal properties of the marine macroalgae, they started getting the attention of policy makers and researchers alike all around the world. Brown seaweeds (Phaeophyceae) are mostly marine and characterized by presence of pigment fucoxanthin which gives them its coloured appearance. The present work aims to provide detailed molecular analysis of the brown seaweeds found in Indian coastal regions to study and characterize it taxonomically which has not been done till now in Indian context. Out of all the samples processed, one invasive species was detected, Sargassum zhangii, which is the first report of this algal species outside Chinese waters. The conspecificity was confirmed by a multi-faceted approach, including comparative morphology, microscopy, genetic distance analysis and computational phylogenetics using Maximum Likelihood and Bayesian Inference methods.
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    Effect of amyloid beta (25-35) peptide on mitochondrial respiratiory function in neuroral cells over expressing ape1
    (Central University of Punjab, 2014) Kaur, Navrattam; Mantha, Anil K.
    Alzheimer's disease (AD) is an important public health problem which affects millions of people worldwide. The major pathological hallmarks associated with AD are the accumulation of amyloid beta (A?) in senile plaques and neurofibrillary tangles (NFTs) made up of hyperphosphorylated tau proteins. Accumulating evidences point towards the role of oxidative stress and mitochondrial dysfunction in the pathogenesis of AD. Aging is considered as one of the greatest risk factor for AD. In order to maintain genome integrity, base excision repair (BER) pathway is the predominant pathway for repairing oxidized base lesions in neuronal cells. APE1 is the central enzyme of the BER-pathway, having both repair and redox activities and shown to enhance neuronal survival after oxidative stress. In my study, effect of A?(25-35) on mitochondrial ROS/RNS levels and activities of respiratory complexes (I, III, & IV) in neuronal cells was studied with and without ectopic APE1 expression and the neuro- modulatory role of Ginkgolide B (from leaves of G. biloba) was evaluated. It was seen that A?(25-35) increases the ROS/RNS levels in these cells which was decreased when pre-treated with Ginkgolide B (G.B) before treating with A?(25-35). APE1 levels were found to be decreased on treating with A?(25-35) and were increased on pre- treatment with G.B and subsequent treatment with A?(25-35). These results indicate that ectopic APE1 expression in the mitochondria of the neuronal cells might overcome the oxidative damage caused by A?(25-35). Also, phytochemical G.B has shown to modulate the mitochondrial complex activity upon A?(25-35)-induced oxidative stress and modulate the ROS/RNS levels in the presence of APE1. Further studies are needed to understand the mechanism of action of APE1 in relation to the above results, which will be carried out during my Ph.D. work.