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Generation of Transgenic <i>Medicago Sativa</i> Overexpressing "<i>Osmotin-Chitinase</i>" Gene ChimeraKancharla, Jahnavi Reddy 01 May 2011 (has links)
Medicago is widely used as a forage crop. It is often susceptible to various pathogenic infections and exhibits low growth in drought and extreme climatic conditions. In the current study, a strategy was developed for over-expressing an “Osmotin-Chitinase” gene chimera in transgenic Medicago that could potentially confer resistance to different biotic and abiotic stresses. Seed germination of several cultivars of Medicago (M. sativa ssp. sativa, M. sativa ssp. falcata, M. sativa ssp. caerulea, M. truncatula, and M. Rugosa) was tested to determine the cultivars with good germination rates. Among these, M. sativa ssp. sativa showed an average of 80% germination over a period of one week and was subsequently selected for regeneration and transformation experiments. Different explants (cotyledons, hypocotyls, petioles) were tested for regeneration. Among these, hypocotyl explants showed highest (46.17 %) percent regeneration. Escherichia coli harboring Osmotin-Chitinase (OSM-CHI) gene chimera cloned into binary vector pBTEX with nptII as a selection marker was mobilized in Agrobacterium tumefaciens strain EHA105 which was employed in the transformation of hypocotyl explants of Medicago. Transformed calli were grown on callus inducing medium containing kanamycin for screening. Further screening of the positive transgenics was performed using PCR. Southern hybridization was carried out for further confirmation of successful transformation. Transformed shoots will be grown on the root inducing medium for developing into plantlets which would then be transferred to the green house and later tested for their degree of resistance to various biotic and abiotic stresses.
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Molekulární diferenciace pohlavních chromosomů předivek rodu \kur{Yponomeuta} / Molecular differentiation of sex chromosomes in ermine moths of genus \kur{ Yponomeuta}VESELÁ, Jana January 2012 (has links)
Structure of the karyotype of three species in ermine moths (Yponomeuta: Yponomeutidae, Lepidoptera) was investigated by means of several cytogenetic methods (DAPI staining, orcein staining, CGH and GISH with telomeric probe) focusing on their sex chromosomes. Additionally, the origin of the Z2 chromosome in the WZ1Z2/Z1Z1Z2Z2 sex chromosome system (specific for this family of butterflies) was tested using southern hybridization.
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