Browsing by Subject "S1"

To determine the role of metalloprotease EGY1 and hormone signalling pathways in PSII repair cycle, a mutant named white because of early senescence was identified having 4bp deletion in EGY1. To further characterize the growth responses in white mutant, two suppressors (white suppressor 1 and white suppressor 2), mutated in STAY GREEN1 (SGR1) which prevents chlorophyll degradation, restored the normal white phenotype was identified upon suppressor mutant screens. This study investigated the effect of chloroplast translation inhibitors (lincomycin/chloramphenicol) and MV (methyl viologen) on photosynthesis in Arabidopsis thaliana single and double white mutants. Furthermore, a second goal was to verify the correct identification of the mutations in white suppressor 1 and white suppressor 2. Western blotting and pulse amplitude modulated fluorimeter (PAM) was used to quantify the D1 protein (reaction core of PSII) levels and photochemical efficiency (Fv/Fm) respectively. Immunoblotting revealed a pronounced decrease in D1 levels for both white and egy1. PAM results showed a high tolerance of white mutant towards lincomycin/chloramphenicol. The white suppressors complemented the lincomycin/chloramphenicol tolerance of white mutant. The white mutant was highly MV sensitive. This MV response was altered in white double mutants (white ein2-1, white sr1-4D and white rcd1-4), suggesting that hormone signalling was involved in the response to MV. The decreased abundance of D1 in the white mutant suggests a role for EGY1 in PSII assembly and D1 turnover under light stress. In all assays (immunoblotting and PAM), the white mutant and egy1-2 gave the same results, this confirms the correct identification of the white mutant as a new egy1 allele. The successful restoration of lincomycin /chloramphenicol tolerance by white suppressors (S1 and S2), implicates that chlorophyll breakdown impacts on correct photosynthesis function. The suppressors S1 and S2 were transformed with wildtype SGR1, which restored the white mutant phenotype. Thus, the suppressor phenotype was caused by mutations in SGR1.