Botany - Master Dissertation

Permanent URI for this collectionhttps://kr.cup.edu.in/handle/32116/29

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Charaterization of G-SSR markers in rhododendron arbreum
(Central University of Punjab, 2014) Kaur, Harmeen; Bhardwaj, Pankaj
Himalayan region is one of the important hotspot, having Rhododendron arboreum as major species. Anthropogenic activities and unsustainable use of the plant for its fruits, flower and wood by local people has raised questions about the sustainability of the species. In order to answer these questions, it is important to first know the genetic structure of the population. This study is focused on the characterization of the G-SSR markers for Rhododendrons and to study the population genetic structure. A set of 38 primers out of 41 are characterized on three population of Rhododendron arboreum, a total of 30 individuals. Allele number at different loci came to be in range of 2 to 14 with an average of 5.2 alleles per locus. Level of heterozygosity observed was high which varies from 0.422 to 0.917 (He) and 0.167 to 0.933 (Ho) with a polymorphic information content 0.104 to 0.811. Out of 38, about 19 loci deviated from the Hardy Weinberg Equilibrium. Genetic diversity was high in these populations with an average gene flow of 5.436. Based on the information from the loci, there was no observed linkage disequilibrium and bottleneck effect. Software DARwin and STRUCTURE, irrespective of geographical location, revels the presence of two original populations with admixed genetic stock. These studied loci will help in the further genetic mapping of the genes and genotyping studies in Rhododendron arboreum.
Development And Characterization Of SSR Markers In Calotropis Procera
(Central University of Punjab, 2018) Vandana; Bhardwaj, Pankaj
Calotropis procera is a weed growing around the arid and semi-arid areas of the tropical and subtropical regions. The species has the potential to replace cotton and resolve the high rates of cotton-based products and at the same time reduce the use of synthetic non-biodegradable fibres. It has been used in various medicinal formulations from a long time. There is a need for developing molecular markers to facilitate breeding and genetic improvement of its varieties. Out of 12.6 million raw reads, 84636 reads were obtained after quality check and filtering. 93.7% of alignment rate and 76.31% of core genes were detected in Bowtie and BUSCO assessment, respectively. An average of 5.5 SSRs/Mb was obtained in 72.349 Mb transcriptome. 25 SSR markers were selected for characterization on 30 individuals representing 3 populations. 13 SSR loci were found polymorphic. In the population structure analysis a moderate level of genetic diversity (Na=3.9231, He=0.633) was predicted. The genetic variation among the population was found to be only 7% whereas within the population, it was 93%. The mean Shannon?s information index obtained was 1.1, showing significant diversity richness in the populations. The Dendrogram showed that the population from Bathinda and Ambala are closely related to each other than those from Barnala. The gene flow between Bathinda and Ambala population was found significantly high (6.439). The present set of loci had a high PIC value of 0.506 that makes them very informative to be further used for large scale studies.
Development of genomic microsatellite markers in commiphora wightii
(Central University of Punjab, 2014) Kaur, Gaganpreet; Bhardwaj, Pankaj
Commiphora wightii is an important medicinal plant, growing in arid to semi- arid conditions. It is widely distributed in tropical regions of Africa, Madagascar and Asia. In India, the specie is found in South- Western India and parts of Central India. The plant on incision secretes an oleo gum resin, which is used to treat various ailments since ancient times. The oleo gum resins of plant contain guggulsterones which have great medicinal value. The other uses of gum are in perfumery, calicoprinting, dyeing silk and cotton, fumigation and incense for which the plant is getting exploited by unfair tapping methods. This plant has been listed in red data book so conservation of this plant is the need of present. For this it is necessary to know about the genetic diversity and population structure of the plant. Microsatellite markers are markers of choice for such studied because they are reproducible, co- dominant and show high levels of polymorphism. In the present study genomic SSR markers were developed. A total of 22 primer pairs were designed for which, 1913 clones were analysed for fragments containing microsatellite regions and 338 clones were found positive and are selected for sequencing. Sequencing results show a total of 212 fragments with perfect repeats, interrupted repeats and compound repeats (> 5 repeats). These primer pairs can be used in genetic diversity analysis of naturally growing populations of Commiphora wightii. They can also be used in MAS, genome selection during gene introgression in plant breeding, genome mapping and gene tagging. This will help in planning the conservation strategies as well as rational utilization of this endangered plant.
Differential Alternative Splicing Analysis Under Abiotic Stress Conditions In Barley (Hordeum Vulgare)
(Central University of Punjab, 2018) Bisht, Diksha; Bhardwaj, Pankaj
Biotic and abiotic stress factors adversely affect the growth and development of plants, living in constantly changing environment. Plant abiotic stress responses seem to be frequently associated with the process of alternative splicing. Alternative splicing generates multiple products from a single gene by variable splicing of exons. RNA-seq utilizes NGS platforms to produce massive amount of sequence data. Using transcriptomic data, we tried to identify the differentially spliced genes in barley under heat and cadmium stress. Raw RNA seq data was pre-processed and aligned with reference barley genome. Then differential alternative splicing was analyzed on the basis of intron excision. In this study, a total of 196 significant differentially spliced clusters were found under two abiotic stress conditions, among them 47 and 149 clusters were associated with cadmium and heat stress condition, respectively. Overall, 133 fully annotated clusters and 63 unannotated clusters were observed. Further, annotation was performed for the significant genes to determine their functions. Some of these significant genes encode for splicing factors, stress tolerance protein, non snRNPs and antioxidants. From this study, it was concluded that barley undergo differential spliced events to maintain its homeostasis under different abiotic stress conditions.
Evaluation of chemical composition and groundwater and anticancer properties of citrullus colocynthis
(Central University of Punjab, 2014) Kaur, Jatinder; Bhardwaj, Pankaj
Colocynth (Citrullus colocynthis (L.)) is medicinal plant of family Cucurbitaceae which is the native of Turkey. This plant has been used as anti-inflammatory agent, for the treatment of jaundice, skin infections, and urinary diseases from ancient times. It is anti-diabetic, anti-microbial, anti-parasitic, antifungal and anti-oxidant in nature and thought to possess anti-cancer properties also. There is no proper study available regarding its anticancer nature. The aim of the study was to estimate the chemical composition of C. colocynthis fruit and the effect of different extracts prepared from C. colocynthis pulp and seeds on various cancer cell lines. These extracts were found to contain number of secondary metabolites which may further have useful and healing properties. When cancer cells were treated with different prepared extracts, the cell viability of various cancer cells was reduced effectively. Effect of p53 gene was also analysed by using intact p53 as well as p53 mutant cell lines. Antioxidant activity of C. colocynthis was also determined by performing H2DCFDA. Assessment of various antioxidant enzymes such as Catalase, SOD, and Glutathione reductase was also done and found to be present in this fruit. We also analysed the protective effect of C. colocynthis by first treating the cells with pesticides and then with colocynthis extracts. Results indicate that C. colocynthis neutralized the harmful effect of pesticides. The results obtained are encouraging and confirm the value of the use of C. colocynthis as an anticancer agent for fighting against various cancers. It may be helpful to find the exact component responsible for the protective action of C. colocynthis.
Impact of mitochondrial transplantation on cancer cells
(Central University of Punjab, 2014) Aggarwal, Alza; Bhardwaj, Pankaj
Mitochondria, the powerhouse of the cell, are small granular or filamentous bodies associated significantly with cellular respiration and are the main sources of energy, due to which they are present in maximum number in the organs that require large amounts of energy for doing their function like muscle cells, neural cells, etc. In case of any dysfunction of mitochondria, these organs are most affected culminating in a number of serious multi organs diseases, irrespective of age such as neurogenic weakness with ataxia and retinitis pigmentosa (NARP), or Leigh syndrome (LS), Cancer, etc. Although mutations in mitochondrial genes are common in cancer cells, they do not inactivate mitochondrial energy metabolism, but rather alter the mitochondrial bioenergetics and biosynthetic state. Literature survey also revealed that owing to mitochondrial dysfunction the clinical trial of many anticancer drugs has failed in patients. This study is focused on the impact of mitochondrial transplantation on cancer cells and their drug sensitivity against four human cancer cell lines HCT116 (WT & P53mutated), HepG2 and MCF7. The normal cell's Mitochondria was transplanted into cancer cells and then evaluated the Impact of transplantation of mitochondria from healthy cells into cancer cell upon their growth, ROS production and their drug sensitivity. The results of this study revealed that the healthy mitochondria transplanted to cancer cells decrease carcinogenesis and have drug sensitivity. So, it may be used as futuristic cancer remedy.
Investigation of combined genotoxic effect of pesticides on cancer cell lines
(Central University of Punjab, 2013) Anju; Bhardwaj, Pankaj
The use of pesticides in agriculture sector is increasing widely to alleviate crop loss due to pests. To combat problem of pest, various types of pesticides are widely used, out of which insecticide are majorly used. In Malwa region of Punjab lots of insecticides are used. These pesticides when present together they may result in combined response. This combined response can be of three types such as independent, dose addition and interaction (synergism or antagonism). This combined response is very big trouble, such as sometimes even low dose of pesticide which is not toxic individually can result in toxic response in the presence of another pesticide. In the present study, Malathion and Monocrotophos (widely used organophosphorous insecticides) selected for investigation of combined effect. Out of selected pesticide Malathion also detected in water, in present study. Various parameters for genotoxicity evaluation was carried out with exposure of pesticides on HCT-116 WT/ p53 mutant (colon cancer) cell lines. Results of present study suggested that combination of pesticides resulted in more cell death, oxidative stress, membrane instability and imbalanced antioxidative system. It is also evaluated that low dose exposure of pesticides individually and in combination resulted in increased cell survival, may be due to activation of defense mechanism with prior treatment. All parameters evaluated was with short term exposure of 1-2 days, so evaluated long term exposure of ten days and found that this resulted in very less cell number and changed cell morphology. It is observed that both pesticides showed genotoxicity individually as well as in combination. The combined exposure of both pesticides deviate from there expected effect for dose addition towards more toxicity. So it is concluded that there may be some interaction between these pesticides, which may be synergistic in nature, because observed more toxicity.