Functional analysis of a mirna putatively involved in powdery mildew disease susceptibility in barley

Dağdaş, Gülay
Barley is one of the most important crop species in Turkey and powdery mildew is one of the most common pathogen decreasing yield in barley. For this problem, agricultural biologists apply breeding technologies in order to select and propagate resistant barley cultivars. However, this is not a permanent solution since pathogens evolve rapidly to overcome plant resistance mechanisms. On the other hand, molecular plant pathologists are trying to understand basic mechanisms underlying plant-pathogen interactions by using molecular tools in order to develop long term solutions for preventing yield loss. In this thesis, miR159 mediated regulation of barley GAMyb transcription factor is studied. According to microRNA microarray results regarding to infection with powdery mildew pathogen Blumeria graminis f.spp hordei (Bgh) at different time points, miR159 expression level showed significant differences. Bioinformatics analysis revealed that miRNA159 targets GAMyb gene in barley. In order to investigate this relationsh‟p, both miRNA and miRNA target were cloned into GFP containing expression vectors through Gateway cloning and resulting vectors were transformed into Nicotiana benthamiana through Agrobacterium mediated gene transfer. Observations based on GFP expression showed that miRNA159 targets and decreases the expression of GAMyb in vivo. v To conclude, this study can be evaluated as a distinctive study for two aspects; (i) it is the first study assessing a “putative” barley miRNA function biologically and (ii) developed a practical and effective functional assay for miRNA studies in plants.


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Citation Formats
G. Dağdaş, “Functional analysis of a mirna putatively involved in powdery mildew disease susceptibility in barley,” M.S. - Master of Science, Middle East Technical University, 2009.