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dc.contributor.authorAaliya, Khadija-
dc.description.abstractPotato crop is vulnerable to a number of biotic and abiotic stresses. Among abiotic stresses, frost may cause devastating effects on quantity as well as quality of yield. In Pakistan, 75% of potato is cultivated as autumn season crop sown in Sep-Oct and harvested in Jan-Feb. This crop undergoes frost during tuber formation stage resulting in significant loss of production. The aim of the present research was the expression studies of Ice Recrystallization Inhibition Protein (IRI3) from Lolium perenne, Antifreeze protein (AFP) from Daucus carota and Zinc Finger protein (ZnFP) from Gossypium hirsutum under CaMV35S Promoter in potato variety Diamant to enhance the tolerance against cold stress. Expression of these genes in potato was enhanced under the influence of 58 base pair 5’-untranslated region of Alcohol Dehydrogenase Gene (ADH1) as a Translational enhancer. To study sub-cellular Localization of these Proteins, gene cassettes were fused with GFP gene and transformed in model plant Arabidopsis thaliana. Confocal Microscopy revealed the expression of these proteins in the cytoplasm of the cells of Germinating Arabidospsis seedling under stress conditions. To achieve the objective of this study, genes were cloned in plant expression vector pCAMBIA 1301 under the control of 35S constitutive promoter and then transformed into potato plants through Agrobacterium mediated transformation system. Transgenic plants were confirmed with GUS histochemical analysis, PCR and southern Blot analysis. Quantitative expression in Positive transgenic plants was increased under stress conditions. As high as 20.41 fold increase of IRI3 gene expression was observed under stress conditions. The expression of ZnFP and AFP was also increased 16.1 folds and 10.2 folds respectively under stress conditions. Positive transgenic plants exhibited reduction in ion leakage percentage. Ion leakage percentage was observed as 59.41% for IRI3, 65.34% for ZnFP and 70.21% for AFP as compared to 83.21% of control. Plant survival rate was enhanced significantly in transgenic plants. It can be concluded from the results that these genes induced tolerance against low temperature. Thus, use of DNA recombinant technology to combat abiotic stress could be promising for increasing the production of the potato crop in Pakistan.en_US
dc.description.sponsorshipHigher Education Commission Pakistanen_US
dc.publisherUniversity of the Punjab , Lahoreen_US
dc.subjectMolecular Biologyen_US
dc.subjectBiology Molecularen_US
dc.titleTransformation and Expression Studies of Multiple Frost Tolerant Genes in Solanum tuberosum L.en_US
Appears in Collections:PhD Thesis of All Public / Private Sector Universities / DAIs.

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