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Characterization and phenotyping of GDP-L-fucose biosynthesis mutants in Arabidopsis thaliana

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Title: Characterization and phenotyping of GDP-L-fucose biosynthesis mutants in Arabidopsis thaliana
Author(s): Shehata, Reham
Contributor: University of Helsinki, Faculty of Agriculture and Forestry
Degree program: Erasmus-Mundus Master's Programme in Plant Breeding (EMPLANT)
Specialisation: Plant production sciences
Language: English
Acceptance year: 2022
Cell walls (CWs) are the safeguards of plant cells and have a crucial role in controlling cell integrity and plant-environment interactions. Perception of cell wall damage (CWD) results in activation of signaling pathways leading to activation of plant defiance responses. Previous research utilizing murus1 GDP-L-fucose deficient mutant has shown the role of GDP-L-fucose biosynthesis in controlling the structure and integrity of the CWs. Here, we investigated whether phenotypical consequences provoked by the lack of GDP-L-fucose, observed in mutants lacking MURUS1 can be suppressed by blocking signal transduction pathways involving THESEUS1 (THE1) receptor-like kinase or WALL-ASSOCIATED KINASE 2 (WAK2)- MAP KINASE 6 (MPK6) signaling module involved in CWD responses. For this, mur1 the1 and mur1 mpk6 double mutants were generated and analyzed. We found that the phenotypes of double mutants closely resemble that of mur1 mutants, indicating that the morphological consequences of GDP-L-fucose deficiency develop independently of THE1 and MPK6 signaling. Further, we aimed to investigate whether GDP-4-keto-6-deoxymannose-3,5-epimerase-4-reductases GER1 and GER2 controlling the final step of GDP-L-fucose synthesis are functionally redundant. Due to the inability of generating ger1 ger2 double mutants, we have applied the artificial microRNA (amiRNA) strategy to generate ger1 ger2 amiRNA knockdown plants. We found that these knockdown plants exhibit severe developmental growth defects similar to those observed in plants impaired in import of GDP-L-fucose into the Golgi apparatus and these growth defects exceeded those observed in plants lacking MUR1. Therefore, we have concluded that GER1 and GER2 are functionally redundant.
Keyword(s): Cell wall plant water relations GDP-L-fucose cell wall integrity cell adhesion pollen germination amiRNA

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