Experiment: | 336 |
Title: | Determination of the transcriptome of gun1, a mutant defective in plastid-to-nucleus signalling, in |
Date: | 2007-05-30 |
Description: | Background The photosynthetic apparatus in chloroplasts of higher plants is encoded by genes distributed between the plastid and nuclear genomes. Chloroplast biogenesis and the assembly of a functional photosynthetic apparatus are therefore dependent on co-ordinated expression of genes in the plastid and nuclear genomes. There is evidence to suggest that this co-ordination is achieved by retrograde signalling from the plastid to the nucleus. Treatment with norflurazon, an inhibitor of plastid carotenoid biosynthesis, or lincomycin, an inhibitor of plastid translation results in reduced nuclear photosynthetic gene transcription. gun (genomes uncoupled) mutants express nuclear photosynthetic genes in the presence of norflurazon. Five gun mutants have been identified from screens of EMS-mutated Arabidopsis thaliana seed harbouring an Lhcb1 promoter-uidA fusion (Susek et al., 1993). The gun mutants fall into two groups; gun2-gun5 have defects in genes involved in tetrapyrrole biosynthesis (Mochizuki et al., 2001; Larkin et al., 2003) and gun1 has an uncharacterised mutation but is distinct from the gun2-gun5 group (Vinti et al., 2000; Mochizuki et al., 2001). gun1 has been shown to express nuclear photosynthetic genes in the presence of lincomycin, whereas gun2-gun5 do not.Experiment Description RNA will be prepared in triplicate. Two Arabidopsis thaliana lines, Col-0 and the mutant gun1, will be subjected to four experimental conditions as follows: To determine differences between the transcriptomes of Col-0 and gun1 in light the two lines will be grown on 1x Murashige and Skoog basal salts solidified with agar (MS) and containing 2% sucrose. Seedlings will be subjected to a regime of 5 days darkness and two days continuous 200 ?mole m-2 s-1 white light. To determine differences between the transcriptomes of Col-0 and gun1 in the absence of light the two lines will be grown on 1x MS containing 2% sucrose. These seedlings will be subjected to a regime of 7 days darkness. The effect of lincomycin, at a concentration of 0.5mM, on the transcriptomes of both Col-0 and gun1, grown under both light and dark regimes, will be determined. Comparisons between the transcriptomes of Col-0 grown with and without lincomycin, under both dark and light regimes, will reveal which nuclear transcripts are affected by loss of chloroplast translation. Comparisons between the transcriptomes of Col-0 and gun1 grown with lincomycin, under both dark and light regimes, will reveal the transcripts affected by the gun1 mutation.ReferencesLarkin, R.M., Alonso, J.M., Ecker, J.R. and Chory, J. 2003. GUN4, a regulator of chlorophyll synthesis and intracellular signaling. Science 299: 902-906.Mochizuki, N., Brusslan, J.A., Larkin, R., Nagatani, A. and Chory, J. 2001. Arabidopsis genomes uncoupled 5 (GUN5) mutant reveals the involvement of Mg-chelatase H subunit in plastid-to-nucleus signal transduction. Proc Natl Acad Sci U S A 98: 2053-2058.Susek, R.E., Ausubel, F.M.and Chory, J. 1993. Signal transduction mutants of Arabidopsis uncouple nuclear CAB and RBCS gene expression from chloroplast development. Cell 74: 787-799.Vinti, G., Hills, A., Campbell, S., Bowyer, J.R., Mochizuki, N., Chory, J. and Lopez-Juez, E. 2000. .Interactions between hy1 and gun mutants of Arabidopsis, and their implications for plastid/nuclear signalling. Plant J. 24: 883-894. |
ftp Link: | ftp Link |
Slide ID | Slide Name | Genetic Background | Tissue | Stock Code | Cel File |
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Cottage_1-10_wt+Linc-D_Rep1_ATH1 | 3896 | Whole plant | N1092 | Cottage_1-10_wt+Linc-D_Rep1_ATH1.CEL | |
Cottage_1-11_wt+Linc-D_Rep2_ATH1 | 3897 | Whole plant | N1092 | Cottage_1-11_wt+Linc-D_Rep2_ATH1.CEL | |
Cottage_1-12_wt+Linc-D_Rep3_ATH1 | 3898 | Whole plant | N1092 | Cottage_1-12_wt+Linc-D_Rep3_ATH1.CEL | |
Cottage_1-13_g1-Linc-L_Rep1_ATH1 | 3899 | Whole plant | Cottage_1-13_g1-Linc-L_Rep1_ATH1.CEL | ||
Cottage_1-14_g1-Linc-L_Rep2_ATH1 | 3900 | Whole plant | Cottage_1-14_g1-Linc-L_Rep2_ATH1.CEL | ||
Cottage_1-15_g1-Linc-L_Rep3_ATH1 | 3901 | Whole plant | Cottage_1-15_g1-Linc-L_Rep3_ATH1.CEL | ||
Cottage_1-16_g1+Linc-L_Rep1_ATH1 | 3902 | Whole plant | Cottage_1-16_g1+Linc-L_Rep1_ATH1.CEL | ||
Cottage_1-17_g1+Linc-L_Rep2_ATH1 | 3903 | Whole plant | Cottage_1-17_g1+Linc-L_Rep2_ATH1.CEL | ||
Cottage_1-18_g1+Linc-L_Rep3_ATH1 | 3904 | Whole plant | Cottage_1-18_g1+Linc-L_Rep3_ATH1.CEL | ||
Cottage_1-19_g1-Linc-D_Rep1_ATH1 | 3905 | Whole plant | Cottage_1-19_g1-Linc-D_Rep1_ATH1.CEL | ||
Cottage_1-1_wt-Linc-L_Rep1_ATH1 | 3887 | Whole plant | N1092 | Cottage_1-1_wt-Linc-L_Rep1_ATH1.CEL | |
Cottage_1-20_g1-Linc-D_Rep2_ATH1 | 3906 | Whole plant | Cottage_1-20_g1-Linc-D_Rep2_ATH1.CEL | ||
Cottage_1-21_g1-Linc-D_Rep3_ATH1 | 3907 | Whole plant | Cottage_1-21_g1-Linc-D_Rep3_ATH1.CEL | ||
Cottage_1-22_g1+Linc-D_Rep1_ATH1 | 3908 | Whole plant | Cottage_1-22_g1+Linc-D_Rep1_ATH1.CEL | ||
Cottage_1-23_g1+Linc-D_Rep2_ATH1 | 3909 | Whole plant | Cottage_1-23_g1+Linc-D_Rep2_ATH1.CEL | ||
Cottage_1-24_g1+Linc-D_Rep3_ATH1 | 3910 | Whole plant | Cottage_1-24_g1+Linc-D_Rep3_ATH1.CEL | ||
Cottage_1-2_wt-Linc-L_Rep2_ATH1 | 3888 | Whole plant | N1092 | Cottage_1-2_wt-Linc-L_Rep2_ATH1.CEL | |
Cottage_1-3_wt-Linc-L_Rep3_ATH1 | 3889 | Whole plant | N1092 | Cottage_1-3_wt-Linc-L_Rep3_ATH1.CEL | |
Cottage_1-4_wt+Linc-L_Rep1_ATH1 | 3890 | Whole plant | N1092 | Cottage_1-4_wt+Linc-L_Rep1_ATH1.CEL | |
Cottage_1-5_wt+Linc-L_Rep2_ATH1 | 3891 | Whole plant | N1092 | Cottage_1-5_wt+Linc-L_Rep2_ATH1.CEL | |
Cottage_1-6_wt+Linc-L_Rep3_ATH1 | 3892 | Whole plant | N1092 | Cottage_1-6_wt+Linc-L_Rep3_ATH1.CEL | |
Cottage_1-7_wt-Linc-D_Rep1_ATH1 | 3893 | Whole plant | N1092 | Cottage_1-7_wt-Linc-D_Rep1_ATH1.CEL | |
Cottage_1-8_wt-Linc-D_Rep2_ATH1 | 3894 | Whole plant | N1092 | Cottage_1-8_wt-Linc-D_Rep2_ATH1.CEL | |
Cottage_1-9_wt-Linc-D_Rep3_ATH1 | 3895 | Whole plant | N1092 | Cottage_1-9_wt-Linc-D_Rep3_ATH1.CEL |