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Analysis of metabolome effects of hop (H. lupulus) transcription factors in heterologous of Petunia hybrida
MORAVCOVÁ, Vendula
Plant transformation is now a key research in plant biology, and also is a unique practical tool in the improvement of varieties of cultivated plants. The aim of this work was to obtain transgenic plants Petunia x hybrida containing transgene nptII (kanamycin resistance gene) using the indirect transformation by Agrobacterium tumefaciens. In terms of in-vivo was grown from seeds experimental material in the form of plant Petunia x hybrida cv. Andrea. For the experiments three different constructs HLWD40 3278-80 and HLbHLH 3577 and HLbHLH 3677 GFP were used. The leaf explants from Petunia plants were prepared. Explants were transferred using tweezers into the crucible containing liquid ? MS and prepared bacteria were added. Thus prepared explants remained in bacteria the next day, when they were transferred to the regenerating solid ground. The explants formed callus gradually (clusters of cells) which regenerate a new plant. Transformation should satisfy the condition that the transformed plants should be fertile. For this reason, an attempt was made crossing the transformed plants were crossed Pap1 1527/2 and Pap1 1572/4 with plants crossed HLWD 40 3278-80. The grown plants were analysed by a PCR reaction. The explants began during the first 2-4 weeks after transfromation to form callus and during the next week to remove the first regenerants. At one exulant fell averaged around 7 regenerants and the explants regenerated by construct HLbHLH 3577 regenerate better than regenerants transformed with construct HLWD 40 3278-80. During the transformation we obtained regenerants. With "tissue" PCR, it was found that 15 plants carried the gene for kanamycin resistance. Transformation efficiency was 3.75%.
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GFP and GUS reporters in plant trangenesis
HRABÁKOVÁ, Lenka
The main goal of this bachelor thesis was the confrontation of two oftently used reporter genes gfp and gus, their structure, utility and methods of screening their activity. The experimental part of this bachelor thesis was based on transformation of tobacco with Agrobacterium tumefacies strains.
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Verification of transformation of rhododendron leaf segments
SKOTNICOVÁ, Petra
The selection procedure on kanamycin for rhododendron leaf segments of two species after transformation by Agrobacterium tumefaciens was examined in this thesis. GUS activity and presence of gene gus by polymerase chain reaction (PCR) was determined with plants regenerated on medium with concentration of kanamycin 100mg/l.
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A comparison of efficacy of direct and indirect methods of genetic transformation of potato (Solanum tuberosum L.)
PŘIBYLOVÁ, Marie
Potato is one of the main targets for genetic improvement by gene transfer. The aim of this study was to compare the efficacy of genetic transformation of potato, cultivar Bintje, using two methods: Agrobacterium tumefaciens mediated transformation and microprojectile bombardment. The same plasmid p35SGUSint, which cosists of 35S CaMV promoter, gus and nptII genes, was used for both transformations of internodal potato explants. Kamamycin selection, transient and stable expressions of {$\beta$}-glucuronidase and PCR amplification of gus and nptII transgenes were used for transgenic plant selection, identification and analysis.
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