Πλοήγηση ανά Συγγραφέας "Manousoudaki, Kleanthi"

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    Control of ERULUS and PERSEUS genes’ expression in Arabidopsis thaliana.
    (Τ.Ε.Ι. Κρήτης, Σχολή Τεχνολογίας Γεωπονίας και Τεχνολογίας Τροφίμων (Σ.Τε.Γ.Τε.Τ), Τμήμα Τεχνολόγων Γεωπόνων, 2018-07-12) Manousoudaki, Kleanthi; Μανουσουδάκη, Κλεάνθη
    Previous studies have confirmed that the RSL4 transcription factor is expressed in root hairs. In this work it was attempted to answer the question whether RSL4 may be responsible for the expression of many trichoblasts-specific genes, such as the two receptor-like kinase genes, namely At5g61350 (given the name ERULUS) and At3g07070 (given the name PERSEUS) using reverse genetics. These two kinases were studied in the present thesis. Expression of both kinase genes was tested by evaluating the activity of their promoter when each gene product was constructed in fusion with a GFP expressed in mutant Arabidopsis plants that did not express RSL4 (named as rsl4-1, knock out plants). Expression of ERULUS PERSEUS genes fused with GFP was clearly observed and resulted in new phenotype that grew root hairs. Thus, various tests were carried out to record the characteristics of this new mutant phenotype, by checking the size of shoots and siliques of rsl4-1 plants, the phenotype of the root hairs on different media, and finally comparing significant differences root hair growth by alterating the constitution of these media. Expression of ERULUS and PERSEUS kinases genes was observed in the rsl4-1 plants which were operating as knock outs (i.e. did not produce) to the RSL4 protein. It is concluded that the RSL4 protein is not the only transcription factor that regulates the expression of these two genes ERULUS and PERSEUS, but it appears that something else must be involved as well. In the second part of the experiments, wild type Arabidopsis was transformed with the promPER::PER-GFP construct to express the PERSEUS protein fused with GFP. This produced many independent lines of these transformants whose identification was based on phenotyping checks and PCR analysis. In the last part of this thesis, it was verified whether the promPER::PER-GFP construct results in production of an active GFP-tagged PER-proteins. From previous work from IMPRES group University of Antwerp the promPER::PER-GFP fusion was inserted into plants that had inactivated this gene (Perseus). Comparing these plants with perseus mutant plants that do not produced PERSEUS protein and with wild type plants, it was observed that perseus plants with the introduced GFP-tagged protein had the same root hair phenotype as the wild type, leading to the conclusion that PERSEUS-GFP protein works and can restore wild type root hair phenotype.