Botany - Mphil Thesis

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

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Cytotoxic activity of saragassum wightii on pc-3 cancer cell-line
(Central University of Punjab, 2012) Jahid, Mohd.; Bast, Felix
Finding novel antitumor compounds with low side effects could be an interesting proposal. Antioxidant and anticancer potentials of seaweed extracts can possibly be explored for developing the new anticancer drugs. Polyphenols are extensively distributed in seaweeds and these are reported to be free radical scavengers. This study was designed to check the cytotoxic potential of the methanolic and hydromethanolic extracts from the brown algae S. wightii. The total phenolic content in the hydromethanolic extract and the methanolic extract of S. wightii was determined by the Folin-Ciocalteu method. The total phenolic content in methanolic extract was found to be greater than hydromethanolic extract. The percentage inhibition or scavenging activity of both the extracts was calculated by using the DPPH assay and was more for the methanolic extract. PC-3 cancer cell-line was used as an experimental model. The methanolic crude extract is significantly cytotoxic against the PC-3 cancer cells. On the other hand the hydromethanolic crude extract was not found to be the significantly cytotoxic against the PC-3 cancer cells.
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.
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
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.
Diversity of green and red macroalgee distributed in indian west-coast using morphometry and DNA barcoding
(Central University of Punjab, 2013) Aijaz Ahmad John; Bast, Felix
Modern algal systematists exploit genetic tools for molecular assisted alpha taxonomy and DNA barcoding is one such molecular tool that relies on the use of a standardized DNA region as a tag for rapid and accurate species identification. In this study Nuclear Internal Transcribed Spacer (ITS) was used as a marker for identification and phylogenetic analysis of marine green and red macroalgae in Indian subcontinent. Using Bayesian Inference for phylogenetic reconstruction with T3P model of molecular evolution and gamma distribution (T3P+G) using ITS dataset revealed that the bloom forming Ulva intestinalis found in west coast of Indian subcontinent showed strong endemism, supporting the view that that the genus Ulva encompasses a number of endemic cryptic species in addition to cosmopolitan species. Although there were two morphotypes present in Indian isolates, they constituted a single clade with robust Bayesian Posterior Probability support, confirming conspecificity of these morphotypes. Our results also indicate latitudinal gradients in the distribution of tubular Ulva, with a clade encompassing all nontropical isolates. Higher genetic heterogeneity of tropical isolates as evidenced by highest within-group T3P (Tamura-3-Parameter) distances comparing with that of non-tropical isolates is suggestive of tropics being the geographic origin of these species. While U. compressa and U. intestinalis were monophyletic within nontropical superclade, these morphotypes were polyphyletic within the tropical clade. Due to the polyphyly of currently accepted morphospecies concept and formation of distinct phylogenetic clade among Indian isolates forces us to propose a new bloom forming species of Ulva paschima. Further molecular assessment of invasive Carrageenophyte Kappaphycus alvarezii using ITS-1 region showed affinity to phylogenetic clade of mixed geographical origin confirming that the species was introduced in the subcontinent by human intervention. Surprising result of our study was an endophytic green algae Ulvella leptochaete that was found growing inside Caldophora glomerata, a first report of its kind from India.
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.
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.
Effect of metform on insuln mediated proliferation of lung cancer cell lines
(Central University of Punjab, 2012) Seema; Bast, Felix
Background: The incidence of cancer in diabetes mellitus (DM) patients is a matter of concern. Lung cancer is the most commonly diagnosed cancer and leading cause of death in males. Smoking is the main risk factor contributing to lung cancer progression. The occurrence of cancer is more with the type 2 DM. Besides, hyperglycemia and endogenous insulinaemia exist together for a long duration as a result of insulin resistance. As a consequence of this, the mitogenic activity of insulin is amplified. Moreover, other growth factors, and hormones are activated under the influence of insulin that further enhances this effect. It is also related to obesity, central fat accumulation, physical inactivity and smoking. The nicotine of smoke induces oxidative stress and endothelial malfunction creating metabolic abnormalities in lung cancer. In this perspective, role of insulin sensitizing drug, metformin in inhibiting the growth proliferation of lung cancer cells is hereby explored. Objective: The present study was aimed to evaluate the growth proliferation effect of insulin on non small cell lung carcinoma cell lines. It also proposed to evaluate role of metformin in preventing insulin mediated proliferation in p53 and liver kinase B1 (LKB1) mutant and wild type cell lines. Materials and methods: Two non small cell lung carcinoma cell lines, A549 and H1299 (p53 and LKB1 wild type and mutant) were used to analyze the mitogenic role of insulin by incubating for 24 hours with human recombinant insulin at a range of concentrations from 1nM to 10'M. This was followed by the metformin (concentrations from 1'M to 50mM) treatment for 24 hours along with insulin (500'M for A549 and 1mM for H1299). The proliferations were assessed by MTT dye reduction test and the percentage of the survival of the treated cells was compared with the control. One way ANOVA was used for the data analysis and the proliferation between cell lines were evaluated by student's t-test and two way analysis of variance (Two way ANOV). Results: Both the cell lines exhibited a significant proliferation (p<0.001) with the concentrations of insulin. Insulin stimulated the proliferation approximately by two fold and 1.78 times for A549 cells and H1299 cell line respectively compared to control cells. The growth of two lung carcinoma cell lines were significantly (p<0.001) inhibited by metformin treatment for 24 hours. The maximum reduction in growth was 73% and 67% for A549 and H1299 respectively for a concentration of 50mM of metformin compared to the control. The results followed a dose dependant response pattern for insulin as well as metformin treatment. Concentration at which 50% inhibition of growth observed (IC50) was comparable for both the cell lines. Conclusions: Insulin in high circulating concentrations can augment the growth proliferation of lung cancer cells. Metformin can inhibit this insulin mediated proliferation of lung cancer cells in a multifaceted way. The mechanism of action is independent of p53 and liver kinase B1.
Effect of natrium fluoride on growth and physiology of oryza sativa L.
(Central University of Punjab, 2012) Das, Anamika; Mittal, Sunil
The present study was undertaken to investigate the effect of natrium fluoride on growth and physiology of Oryza sativa L. Two varieties of the O. sativa (rice) viz. Basmati 1121 and PR147 were selected for the study. PR147 is a local variety of the region. The effect was studied on growth (root length and shoot length), physiological parameters (chlorophyll content and percent respiration), macromolecules and their hydrolyzing enzymes (carbohydrate and protein content, ?-amylase and protease activity) and antioxidant enzymes (peroxidase and superoxide dismutase) under controlled conditions in the laboratory. Studies revealed that NaF (in different concentrations ranging from 16, 32 and 64 mg/l) affected the growth and physiology of both varieties as compared to control. When the seed of both varieties of rice were exposed to 64 mg/l NaF concentration, the seedling growth of test plants was highly affected as compared to control. Basmati 1121 variety was found to be more sensitive than PR147. In chlorophyll content, the percent reduction at 64 mg/l of NaF was 62% in Basmati 1121 and 30% in PR147. Similarly, reduction in percent cellular respiratory ability on exposure to 64 mg/l of NaF was 75% and 22% in Basmati 1121 and PR147 variety respectively. At 64 mg/l the carbohydrate content was increased 2.9 folds in Basmati 1121 and 1.35 folds in PR147. However the protein content was variably affected as it was increased 2 folds in PR147 and decreased to nearly 55% in Basmati 1121. The macromolecular contents were affected as the activity of their hydrolyzing enzymes was also affected. The increase in oxidative stress due to high concentrations of fluoride exposure has lead to an increase in antioxidant enzyme activity of both peroxidase and superoxide dismutase enzymes. The study concludes that high content of fluoride affects the growth and physiology of O. sativa L. by inducing oxidative stress.
Evaluation of breast cancer cell lines (MCF-7 & T47D) : Influence of insulin and metformin on growth parameters
(Central University of Punjab, 2016) Sharma, Prateek; Thakur, Sanjeev
Breast cancer is the complex and heterogenous malignancy caused as a result of interaction of hormonal, environmental and genetic factors. Major advances made in the field of breast cancer research have emanated from studies involving human breast cancer cell lines MCF-7 and T47D. Insulin act as a potent mitogen in normal mammary tissue and breast cancer cells in culture. Insulin receptor is overexpressed in several human breast cancers and this overexpression results in transformed phenotype in human mammary epithelial cells. Experimental study has shown that Insulin induces significant mitogenic effect in MCF-7 and T47D cells in dose-dependent manner. 100nM concentration of Insulin enhances the growth of MCF-7 and T47D cells by ? 7.3 and ? 5 folds respectively in comparison to control. The antidiabetic drug metformin lowers risk of breast cancer via direct action (Insulin independent) and indirect action (Insulin dependent). The treatment of the hyperinsulinemia with the metformin will lower the circulating levels of insulin and improve insulin sensitivity, thus results into potential decrease in the incidence of breast cancer. The results show that MCF-7 and T47D cells growth inhibited over control at high doses (10 mM concentration) of metformin. MCF-7 and T47D cells also show decrease in insulin induced cell proliferation at higher doses of metformin.
Genetic diversity and polpulation analysis of melia azaderach L. by Rapd markers in bathinda and adjoing regions
(Central University of Punjab, 2013) Navgeet; Bhardwaj, Pankaj
Melia azedarach L. is an ecologically imperative species growing in Thar Desert. Biological effects such as antiviral, anthelminthic, antibacterial etc. makes it important, yet a little is known about its genetic diversity and structure. In this study, 30 RAPD primers were employed for DNA profiling of 47 individuals representing 6 populations from different geographic locales. A total of 87 bands were scored with an average of 2.9 bands per primer. The PIC ranged from 0.1195 to 0.4998 with an average of 0.4160. Nei's genetic diversity (h) and Shannon's information index (I) ranged from 0 to 0.5 and 0 to 0.6931 with an average Nei's genetic diversity of 0.2422. The gene flow (Nm) 1.4381 and the genetic diversity of 0.2381 at species level demonstrated overall high level of genetic diversity. Cladistics analysis using DARwin and Bayesian cluster analysis using STRUCTURE placed 47 individuals into two main clusters or original genetic stocks (K=2) which showed little or no association with the geographic origin. The lack of clear assignment of individuals to geographic regions of sampling and consideration of different populations in the same genetic cluster suggests a recent common evolutionary history.
Histopathological staging in putative prostate cancer tissues and reviewing litearture of correlation between prostate specific antigen levels and prostate cancer inci. 2012.
(Central University of Punjab, 2012) Thakur, Shweta; Thakur, Sanjeev
Prostate cancer (PCa) remains the most significant cause of cancer specific mortality in elderly men. Asymptomatic behavior, non-modifiable risk factors and metastatic nature is the main problem of PCa. It remains clinically silent and presents itself only at advanced stage. Thus, diagnosing PCa at an early stage can result in increased chances of better treatment and hence, increased survival rate. An accurate biomarker can detect the cancer at an early stage and hence, at curable stage. Clinical parameters can only suspect prostate cancer. Whereas, histopathological examination can establish definite diagnosis of PCa. Various histological patterns are associated with cancer aggressiveness. Therefore, better understanding of clinical relevance of these histopathological findings can help to evaluate a robust biomarker for early detection of PCa. Present study was divided into two parts. First part was aimed to study the histopathology of putative prostate cancer tissue specimens. In second part, the literature of association of pre-operative serum prostate specific antigen levels with cancer detection and aggressiveness was reviewed. Protocol for histopathology of prostate tissues was established. Results of histopathological findings in putative PCa specimens were evaluated. Prevalence of histological PCa was not found in putative PCa tissues. Image library of results of the study was prepared for future analysis. Review of literature of correlation of serum PSA levels with PCa incidence suggests that PSA screening for PCa is a two-sided debate. No doubt that PSA holds the probability of detecting early PCa before development of symptoms; certain v limitations are associated with it. First, it is not reported to be 100% accurate. Second, it generates false positive and false negative results. A false positive result leads to over-treatment whereas a false negative result generates false sense of security against PCa in patient, both affects quality of life. Another main concern with PSA screening is its inability to differentiate between indolent and aggressive cancer. Therefore; accurate and economical molecular biomarkers for early detection and distinction of indolent versus aggressive cancer are urgently required. Until such biomarkers are developed and more convincing evidences regarding efficacy of PSA screening becomes available, research should focus on improving the diagnostic clinical utility of PSA by utilizing novel PSA isoforms. Identifying and validating correlation of serum PSA with tissue PSA and histological grade would be beneficial in terms of requirement of less invasive diagnostic methods to be used to measure PSA expression level as well as to confirm PCa. Future research may focus on evaluating the histological expression of other putative biomarkers and comparative serum proteomic profiling in different PCa stages.
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.
MicroRNA target prediction and comparative micromics; Study different cancers- special focus on breast cancer metastasis
(Central University of Punjab, 2012) Sarkar, Bibekananda; Bast, Felix
Cancer or carcinoma is uncontrolled growth of abnormal cells. The transition of normal cell to cancerous cell is very complicated process and the cause of transition varies with situation. Metastasis is the main cause of death in cancer. There are genes which not only promote metastasis but also maintain microenvironment of tumor cells, and initiate the process of epithelium' mesenchymal transition (EMT). MicroRNA (micro RNA) is small, highly conserved noncoding RNAs that control gene expression post-transcriptionally. MicroRNA controls various cellular events like division, differentiation and apoptosis. Their deregulation may result in to cancerous growth. Most (50%) of the microRNA genes are located in the fragile chromosomal regions, which are more susceptible to amplification, deletion or translocation during tumor development. It is predicted that 30% of the all mRNA are directly or indirectly controlled by the microRNA. MetastamiR are that microRNA which have role in regulation of metastasis. MicroRNA expression profile in different cancers showed that it can act as Oncogene as well as Tumor suppressor gene. Till date there are 1921 mature human microRNA sequences registered in miRBase. MicroRNA target prediction is the first step in functional analysis of microRNA. Target prediction is complicated due to partial complementarity between microRNA and its target. There are many target prediction programs available, but the efficiency and sensitivity of these programs are not known. To enhance its efficiency, we need to know problems during prediction. Comparative analysis of different microRNA prediction tools provides an insight into the above parameter. In this study comparative analysis of seven prediction tools is carried out with help of validated microRNA targets of metastatic breast cancer.
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.
Phytogenetic analysis of bacterial diversity of heavy metal affected
(Central University of Punjab, 2012) Kumar, Rajeev; Kumar, Sanjeev
The soil of Bathinda region is affected by heavy metals due to anthropogenic activity, industrial effluent and fly ash. Three soil samples in triplicate were collected from three different places of Bathinda region, abandoned soil, agricultural soil, thermal power plant fly ash soil. Based on 16S rRNA gene cloning and sequence analysis, phylogentic analysis of bacterial diversity was performed. 366 clones were picked and through colony PCR 128 positive clones were chosen. Out of 128 clones, 126 clones were sequenced in three fragments and all three fragments aligned using SeqMan Lasergene ver.10.0 (DNA STAR) software. BLAST analysis of sequenced and aligned samples was carried out at NCBI to find out the homology with different groups of bacteria. All 126 clones aligned for phylogenetic analysis by MEGA 5 (Neighbor Joining method). 32 genus belong to 8 phyla were observed in all three samples. Most dominant phylum is Proteobacteria followed by Bacteriodates. Pseudomonas spp. is dominant in agricultural and thermal power plant soil. Abandoned soil showed maximum diversity followed by thermal power plant soil. In agricultural soil minimum diversity was observed. So with this investigation it can be concluded that lands without anthropogenic activities are rich in bacterial diversity as compared to more anthropogenic active areas.
Population analysis of media azesarach L. by RAPD markers in the region of rajasthan
(Central University of Punjab, 2013) Ahmad, Kamal; Bhardwaj, Pankaj
Melia azedarach L is ecologically imperative species growing in Thar Desert. Biological effects such as antiviral, anthelminthic, antibacterial, etc. makes it important, yet a little is known about its genetic diversity and structure. In this study, we employed 30 RAPD primers for DNA profiling of 91 individuals representing 8 geographically isolated populations. A total of 98 bands were scored with an average of 3.2 bands per primer. The PIC ranged from 0.0681 to 0.5351 with an average of 0.4103. Nei's genetic diversity (h) and Shannon's information index (I) ranged from 0.1237 to 0.2375 and 0.1845 to 0.3500 respectively with average Nei'sgenetic diversity of 0.2570. The gene flow (Nm) 0.9295 and the genetic diversity of 0.3498 at species level demonstrated overall high level of genetic diversity. Cladistics analysis using DARwin and Bayesian cluster analysis using STRUCTURE placed 44 individuals into two main clusters or original genetic stocks (K = 2) which show little or no association with the geographic origin. The lack of clear assignment of individuals to geographical regions of sampling and consideration of different populations in the same genetic cluster suggests a recent common evolutionary history. M. azedarach is a good reforestation tree because of its fast growth and drought hardy nature. This study adds a foundation for more precise inference about the biogeography and management in the reforestation projects in the Thar Desert.
Preconditiong chickpea for chilling stress: Understanding associated mechanisms and role of FADs in membrane protection
(Central University of Punjab, 2014) Saini, Rashmi; Kumar, Sanjeev
Chickpea has evolved as a spring crop in warmer climate of West Asia. Therefore, it is prone to chilling injury as in North India, chickpea is cultivated in winter season. Preconditioning the plants with mild drought stress might triggers the signaling pathways that will prepare the plants to combat lethal cold stress. In this study, one chilling tolerant genotype PDG3 and one chilling sensitive genotype GPF2 of chickpea were preconditioned with mild drought stress and then exposed to lethal cold stress at 40C. To see the effects of preconditioning various cell responses were monitored by measuring ELI, MDA content, LOX activity, antioxidative enzymes SOD, CAT, APX, GPOX and GR, and mRNA expression of fatty acid desaturase (FAD-2) and small heat shock proteins (sHsp18.5 & sHsp22.7) at different phases. Both the genotypes behave differently to preconditioning. This study shows that preconditioning increases the tolerance capacity of chilling-sensitive genotype towards chilling stress. Whereas, preconditioning brings about adaptable changes in chilling tolerant plant PDG3 so, they can cope up with chilling stress. To conclude whole study, preconditioning with mild drought stress has ability to improve the tolerance against cold stress in chickpea.
Preconditioning of chickpea seedlings for terminal heat stress: understanding associated mechanism and HSP's expression
(Central University of Punjab, 2014) Yadav, Renu; Kumar, Sanjeev
Chickpea (Cicer arietinium L.) is the second most important pulse crop grown worldwide. Changes in the cropping system, competition from other cash crops (wheat) and global warming are pushing chickpea to relatively warmer growing environment. In northern part of country chickpea come across with terminal high temperature stress during reproductive stage which lead to reduced grain yield. Therefore to prevent the plant from incoming heat stress, 11 day chickpea seedling were preconditioned with mild drought stress, then put on recovery for six days and then recovered seedlings were exposed to lethal stress (where temperature was increased step wise from 30?C to 36?C). This study revealed that % EL, Lipid peroxidation increased with the increase in temperature while percent TTC reduction and total protein content decreased with the increase in temperature. Antioxidative enzymes provide the major defence against the ROS generated during the abiotic stress, and it was found that activity of SOD, CAT and APX enzyme increased proportionately with the rising temperature. HSP's act as molecular chaperons and are over expressed at both mRNA and protein level in preconditioned seedlings exposed to high temperature stress as compared to non- preconditioned ones. To conclude the whole study, results obtained clearly reveal that preconditioning with drought stress has the ability to improve tolerance above ambient temperature (27?C 7?C), thereafter preconditioning did not have any influence in terms of the improvement in membrane damage and level of antioxidants. Higher expression of sHSP's is corroborated with the low expression of antioxidants.
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.