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20225
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Subject: Biotechnology and molecular Biology × End date: 2024 × Start date: 2020 × Type: Thesis ×
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Now showing 1 - 5 of 5
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    ThesisItemOpen Access
    Genetic diversity, phytochemical and anticarcinogenic studies in cichorium intybus from Himalayan elevations of Uttarakhand
    (G. B. Pant University of Agriculture and Technology, Pantnagar, 2022-08) Arya, Mamta; Gohar Taj
    Chicory (Cichorium intybus L.) comprise of intense therapeutic and medicinal properties due to presence of phytochemical and secondary metabolites. It is often used as a medication in India to treat a number of illnesses such as fever, diarrhea, spleen enlargement, gout, and rheumatism. The purpose of this study was to determine the genetic diversity of chicory from Himalayan regions of Uttarakhand using SSR markers. So, it will be entrancing to know about the genetic diversity of chicory and if any genotypes are deficient in medicinal content then it could be enhanced through various molecular interventions. The cluster analysis of 100 genotypes grouped it into two clusters A and B, in which cluster A comprised of 94 genotypes. The other B cluster consists of 6 genotypes. Additionally, to investigate the genetic constitution of diverse 100 chicory genotypes genotyping by sequencing was performed, population structure and genetic diversity analysis were also done. Cluster analysis using neighbor joining method grouped the population into 3 clusters ‘A’, ‘B’ and ‘C’ depending on their geographical location. The main cluster ‘C’ consists of 61 genotypes, cluster B comprised of 28 genotypes and cluster A comprised of 11 genotypes. From GBS technique thousands of SNPs identified were used for the association mapping to recognize authentic markers linked to the agro-morphological traits of Cichorium intybus plant. There were 178 SNPs markers showed associations with the eight agromorphological traits at significant threshold level. Further, to study the difference in the phytochemical constituents quantity of chicory, qualitative and quantitative analysis were performed among 10 genotypes and identified that genotypes showed considerable amount of variation. Lactucine the phytochemical constituents of chicory also have anti-cancer properties and can treat various types of cancers such as human leukemia cancer. In present investigation, lactucine and its derivatives are docked with the apoptosis inducing and cell cycle progression proteins for the prediction of its anticancer effect. Among 17 isoforms of lactucine, Lactucin 15- oxalate has shown highest binding affinity with target protein with appropriate Glide and Dock score. These in silico results are valuable to proceed for the in vitro and in vivo studies related toanti-cancer role of lactucin 15-oxalate. Therefore, chicory plant has high in its diversity can be considered as a high value herbs for the medicinal purposes and even its phytochemical content are also enhancing nutritional values. The identified SNPs with the agro-morphological trait indicate its high rate of association which is useful in improving the medicinal properties. Even, due to anticancer effect of lactucinee can be used for treatment of leukemia cancer.
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    ThesisItemOpen Access
    Identification and characterization of candidate genes involved in drought tolerance in finger millet using genome wide association mapping
    (G. B. Pant University of Agriculture and Technology, Pantnagar, 2022-09) Chauhan, Ankita; Chauhan, Ankita; Lohani, Pushpa; Lohani, Pushpa
    In the last two decades, drought stress has become one of the largest risks to agriculture. Drought stress negatively affects growth, development and productivity of crops. Therefore, development of climate- resilient crops is need of the hour. The identification and characterization of important candidate genes for drought tolerance and their markers will be an important step in this direction. Finger millet is a climate-resilient crop, making it the perfect crop for discovering the genes involved in drought tolerance. Morpho-physiological and biochemical parameters for 80 genotypes of finger millet from different parts of the world were phenotyped in both irrigated and drought conditions. Highly significant differences between accessions were found in the analysis of variance for both conditions for the majority of the characteristics, demonstrating that these traits are involve in drought stress. Additionally, the extensive variance in the accessions' agronomic performance suggested that these qualities would make strong candidates for marker-trait associations. Further, a analysis of genetic diversity and population structure was done and on the basis of their geographic location and drought resistance, the population was divided into two clusters, A and B, using neighbour joining cluster analysis. The major cluster A consisted broadly of all the Indian and drought tolerant genotypes whereas cluster B comprised of exotic and drought sensitive genotypes. Thus, there was good congruence between the phylogenetic tree and the population structure. Principal component analysis showed the same results as of phylogenetic and population structure which further validated that the SSR markers are related to drought tolerance. SSRs marker data along with morpho-physiological traits under both irrigated and a drought condition was used for association mapping to identify reliable marker(s) linked to drought tolerance in finger millet. A total of eleven SSR markers,SSR2, SSR3, SSR4, SSR5, SSR6, SSR9, SSR11, SSR12, SSR13, SSR14 and SSR16 with high r2 values for different morpho-physiological and biochemical traits are used for the identification of candidate genes. A total of eleven genes (ECERF1, ECERF2, ECERF3, ECCBF4, ECWRKY19, ECMYB44, ECBZIP1, ECZEP1, ECNFYB2, ECDREB1and ECNAC2) orthologous to CGs of Oryza sativa and Setaria italica putatively involved in drought tolerance homeostasis in finger millet were identified through Genome- Transcriptome transition approaches. The drought-tolerant finger millet variety PRM 6107 was utilized to study the expression of identified genes under various abiotic stress conditions, including heat, salt, and drought. In comparison to control, the genes expression increased from 10 to 14 fold under various abiotic stresses. Which further validate the role of these genes in drought stress. In silico structural and functional annotation of identified genes highlighted their significance in drought tolerance in finger millet. In light of the lack of information on the genetic and molecular basis of drought tolerance in the crop , the findings of the current investigations have opened up new opportunities for the successful use of finger millet in future crop improvement programs.
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    ThesisItemOpen Access
    Molecular docking of Alternaria brassicae fungal proteins with phytoalexins of Brassica juncea and comparison with standard fungicides
    (G.B. Pant University of Agriculture and Technology, Pantnagar, District Udham Singh Nagar, Uttarakhand. PIN - 263145, 2022-08) Shivakumar R; Singh, B.R.
    Brassica juncea being one of the important oil seed crops in India as well as around the globe faces a major problem in imbalance between the production and consumption due to yield loss. This yield loss is accounted by several diseases and pests that infect the crop during its growth stages. The most important factor causing a reportable yield loss is due to a necrotrophic fungi Alternaria brassicae, of an average yield reduction of 30-40%. Therefore, mitigation strategies must be developed to resist the infection of this pathogen at molecular level. Though the fungicides do their role to an extent in controlling the infection, certain strategies must be developed to replace the fungicides, keeping in mind the harmful effects of fungicides towards environment, biodiversity and human well-being. In this study we attempt to use the molecular mechanism during the pathogenesis. Where the Alternaria brassicae aims in producing the proteins that actively take part in causing the disease and the intricate defense of the plants to combat the infection. Plant defense system involves the production of Pathogenesis Related proteins that inhibits the fungi and also it produces a wide range of secondary metabolites known as phytoalexins to neutralize the fungal proteins. Hence the interaction between the phytoalexins and fungal proteins are studied at in-silico level by performing molecular docking. The interactions of the selected proteins ABC transporter, Calmodulin and NRPS are analyzed by obtaining their docking scores in terms of binding energies against the Brassica’s phytoalexins such as Brassilexin, Cyclobrassinin, Indole-3-carboxyaldehyde, Methoxybrassitin and Rutalexin. The same docking analysis is also done with the standard fungicides that are recommended towards Alternaria blight infection to understand the advantages of phytoalexins over standard fungicides. The fungicides Azoxystrobin and Cyprodinil shows an effective performance against Alternaria brassicae proteins. Hence the results of docking exhibits that the phytoalexins such as Cyclobrassinin, Indole-3-carboxyaldehyde exhibits docking scores ranging from -5 Kcal/mol to -7 Kcal/mol, which is considered ideal and could be suggested as an alternative to fungicides selected in the study.
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    ThesisItemOpen Access
    Virtual screening of phytoalexin(s) of Brassica juncea as defense inducing molecule(s) against alternaria blight
    (G. B. Pant University of Agriculture and Technology, Pantnagar, 2022-08) Kurma, Devakrishna; Gohar Taj
    Brassica juncea is an important Rapeseed-mustard crop in India, accounting for more than 90% of the total area. It is used as sources of oil, vegetable and fodder. Brassica has a high potential yield in India, but there is a large gap between potential and actual yields, which is mostly due to biotic and abiotic stress. Alternaria blight is a serious disease among the biotic stresses. It causes huge economic losses in terms of both production and oil quality. Alternaria brassicae is the causal organism of Alternaria blight and can infect the host at any stage of development. Alternaria brassicae proteins MAT1, BETA-TUBULIN, and ALTA1 are involved in mating, motility, and spore germination activities respectively during the pathogenesis of Alternaria blight on Brassica spp. Physiochemical analysis through high Aliphatic Index (AI), Index of Instability and Grand Average of Hydropathy (GRAVY) reveals that all the three proteins were heat stable as well as hydrophobic proteins. In this study molecular docking-based approach was used to predict the above proteins of Alternaria brassicae as possible molecular target of phytoalexins during pathogenesis or defense response. I-TASSER was used to predict 3-D structures for above proteins which were subsequently docked with phytoalexins which included Camalexin, Glucobrassicin, Methoxybrassinin, Spirobrassinin, Indole- 3acetaldoxime N-oxide and also with the five standard fungicides. Among the phytoalexins, the results of docking of Glucobrassicin with the above target proteins showed good affinity as well as low docking score in the range of -8.57 to -3.97 whereas the fungicide Tebuconazole had docking score in the range of -5.47 to -1.24. Therefore, the phytoalexin Glucobrassicin could be used for the protection of Brassica plants against Alternaria blight. Further In-vitro studies and downstream validation would give way to use the Glucobrassicin phytoalexin(s) as a substitute for hazardous fungicides to control plant diseases.
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    ThesisItemOpen Access
    In vitro evaluation of antioxidative and immunomodulatory potential of the selected medicinal plants via cytokines expression analysis in chicken splenocytes culture system
    (G.B. Pant University of Agriculture and Technology, Pantnagar, District Udham Singh Nagar, Uttarakhand. PIN - 263145, 2022-04) Singh, Himani; Ambwani, Sonu
    Herbal immunomodulation is being traditionally practiced in India and is gaining added importance nowadays throughout the world. Due to lack of knowledge regarding the molecular targets and underlying mechanisms involved, there is pressing need to scientifically validate the molecular mechanism behind herbal immunomodulation. Considering the above facts, four plant extracts, viz. aqueous leaf extract of Nyctanthes arbor-tristis (Night-flowering Jasmine) (NAE), hydromethanolic rhizome extract, bark extract and seeds extract of Asparagus racemosus (Shatavari) (ARE), Terminalia arjuna (Arjun) (TAE) and Trigonella foenum-graecum (Fenugreek) (TFE), respectively; were explored for their antioxidative and immunomodulatory potential in chicken splenocytes culture system. These were evaluated for the presence of various phytoconstituents through GC-MS analysis. The antioxidative potential of all the four plant extracts was evaluated by various in vitro extracts based and cell based assays. The anti-inflammatory activity of the plant extracts was determined by protein denaturation method and proteinase enzyme inhibitory assay. The MNCD of NAE, ARE, TAE, TFE and DEXA were used to give in vitro exposure in chicken splenocytes to further explore the transcriptional level modulation of the selected genes through qRT-PCR. GC-MS data analysis of NAE, ARE, TAE and TFE revealed the presence of several active phytochemicals which may be responsible for their antioxidant and immunomodulatory properties. All the four plant extracts displayed significant in vitro antioxidant and anti-inflammatory properties in various assays conducted. Expression analysis displayed significant alterations in expression of the selected genes due to in vitro exposure of NAE, ARE, TAE, TFE and DEXA. The pro-inflammatory genes were found to be downregulated while the anti-inflammatory genes were found to be significantly upregulated in the plant extracts and DEXA treated splenocytes. There was significant increase in immunostimulatory genes in the plant extracts treated splenocytes while same were found to be decreased in DEXA treated cells. There was marked reduction in the transcript level expression of APAF-1, caspase-3 and caspase-9 when treated with NAE, ARE, TAE, TFE while these genes were found to be upregulated in DEXA treated cells. From the findings of the present study, it may be concluded that NAE, ARE, TAE and TFE showed potent anti-oxidative, anti-inflammatory and immunostimulatory potential while DEXA showed anti-oxidative, anti-inflammatory and immunosuppressive property. Due to the significant immunopotentiating and antioxidative potential of the plant extracts, these could be further used for development of immunoenhancing herbal preparations for poultry. However, exhaustive research is needed to characterize phytoconstituents and the findings need to be validated at protein level by employing suitable biochemical and molecular biology techniques.