Site directed mutations of one of the plant pathogen effector gene for functional analysis

Mustafa, Zemran
Wheat (Triticum aestivum L.) is one of the most important crops planted in the world and used as major food resource. Yet, there are major losses of productivity due to different biotic and abiotic destructive agents. Among the biotic factors especially the yellow rust is one of the most frequent. Yellow rust or also called “stripe rust”, due to stripe like appearances on the leaves, disease is caused by Puccinia striiformis f. sp. tritici, which is an obligate plant fungal pathogen. To cope with high yield losses caused by these pathogens, investigation of host-pathogen interaction at the molecular level can empower the scientists in fight against the pathogen. In this thesis research, one of the Puccinia striiformis f. sp. tritici candidate effector genes (PstHa2a5 / Yin et al., 2009) was studied for functional analysis by conducting site directed mutagenesis (SDM) on the gene. The amino acid sequence of this effector vi indicates the presence of a conserved FXC motif. The function of this motif is currently unknown but it is suspected that it is needed for intracellular targeting of effector proteins. Mutated versions of the gene were cloned into pK7WGF2 and pK7FWG2 gateway destination vectors to be transformed first into Agrobacterium tumefaciens by electroporation and then delivered into Nicotiana benthamiana leaves via agrobacterium-mediated gene transformation. The distorted motif of PstHa2a5 by SDM was tested for its role in intracellular localization of the protein. By expressing the mutant protein in N. benthamiana leave cells and tracking green fluorescent protein (GFP) expressed as a fusion to the modified gene, under microscope functional analysis were conducted.


Phylogenetic relationships among Triticum L. and Aegilops L. species as genome progenitors of bread wheat based on sequence diversity in trnT-F region of chloroplast DNA
Dizkirici, Ayten; Kansu, Cigdem; Önde, Sertaç; AVCI BİRSİN, MELAHAT; Ozgen, Murat; Kaya, Zeki (Springer Science and Business Media LLC, 2013-12-01)
Cultivated wheat, (Triticum aestivum L.), is one of the most important food crops in the world. The Aegilops L. genus is frequently utilized by plant breeders for improving the current wheat cultivars due to their close relationships. Therefore, understanding the phylogenetic relationships among the species of these genera is not only valuable for plant taxonomy, but also for plant breeding efforts. The presented phylogenetic analysis was based on the sequences of trnT-F chloroplast DNA containing three non...
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Boylu, Barış; Akkaya, Mahinur S.; Hakkı, Erdoğan Eşref; Department of Biotechnology (2011)
Wheat diseases such as rusts and powdery mildews are among the most important and ancient diseases that affect wheat cultivation worldwide. The pathogen race specific resistance genes cannot maintain long lasting resistance. On the other hand, the presence of genes confers the non-race specific resistance last much longer. The durable resistance phenotypes in wheat against various rust and powdery mildew diseases were reported as Lr34, Yr18, and Pm38 separately; nevertheless, they were known to locate very ...
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Baloğlu, Mehmet Cengiz; Yücel, Ayşe Meral; Öktem, Hüseyin Avni; Department of Biology (2011)
Wheat is the most important grain crop grown in our country providing greatest part of the daily nutritional requirement. Abiotic factors including salinity, drought, cold and heat stresses affect quality and yield of wheat varieties used for the production of both bread and pasta flour. NAC proteins form one of the widest families of plant specific transcription factors. Members of this family are related with development, defense and abiotic stress responses. TaNAC69-1 and TtNAM-B2 genes were isolated fro...
Particle bombardment transformation of some Turkish wheat cultivars with TaNAC69-1 and TtNAMB2 genes
Battal, A.; BALOĞLU, MEHMET CENGİZ; Kavas, M.; Yücel, Mustafa; Öktem, Hüseyin Avni (2012-09-23)
Wheat is one of the most important crop plants in the world.The production of wheat is seriously affected by biotic (pests,bacterial and fungal diseases) and abiotic (drought, salinity andfreezing) stresses. NAC family proteins (NAM/ATAF1-2/CUC2) areplant specific transcription factors and related with development,defense and abiotic stress responses.TaNAC69-1andTtNAMB2genes are the transcription factors from this protein family. Theaim of this study was to transform of some Turkish ...
Bacterial expression of an effector protein of yellow rust pathogen and a resistance protein of wheat and characterization of the effector protein
Erdoğan, Sayıt Mahmut; Akkaya, Mahinur S.; Özçubukçu, Salih; Department of Biotechnology (2019)
Yellow rust is one of the most important wheat disease encountered in large parts of the world and in our country caused by a pathogen which is called as Puccinia striiformis f. sp. tritici (Pst). Rapid alterations in the pathogen virulency can make previously resistant varieties susceptible to the disease. Finding out common or species-specific genes, participated in the plant-pathogen interactions will provide an understanding of the biological mechanisms of the disease. Within the scope of this thesis fo...
Citation Formats
Z. Mustafa, “Site directed mutations of one of the plant pathogen effector gene for functional analysis,” M.S. - Master of Science, Middle East Technical University, 2015.