These authors contributed equally to this work.
Versatile roles of Arabidopsis plastid ribosomal proteins in plant growth and development
Article first published online: 12 OCT 2012
© 2012 The Authors. The Plant Journal © 2012 Blackwell Publishing Ltd
The Plant Journal
Volume 72, Issue 6, pages 922–934, December 2012
How to Cite
Romani, I., Tadini, L., Rossi, F., Masiero, S., Pribil, M., Jahns, P., Kater, M., Leister, D. and Pesaresi, P. (2012), Versatile roles of Arabidopsis plastid ribosomal proteins in plant growth and development. The Plant Journal, 72: 922–934. doi: 10.1111/tpj.12000
- Issue published online: 6 DEC 2012
- Article first published online: 12 OCT 2012
- Accepted manuscript online: 18 AUG 2012 06:12AM EST
- Received 1 June 2012; revised 23 July 2012; accepted 14 August 2012; published online 12 October 2012.
Figure S1. Relative expression and intron-exonstructures of PRPL1, PRPL4 and PRPL24transcript variants quantified by real-time PCR. (a) To monitor theaccumulation of the two splicing variants for PRPL1(PRPL1.1, At3g63490.1; PRPL1.2, At3g63490.2) andPRPL24 (PRPL24.1, At5g54600.1; PRPL24.2,At5g54600.2) loci, as well as the four different transcriptspredicted for PRPL4 locus (PRPL4.1-to-4,At1g07320.1-to-.4), transcript variant quantification was conductedby real-time PCR on cDNA obtained from leaves and siliques of WT(Col-0) plants. cDNA amplification products were obtained by usingthe transcript variant specific primers reported in Table S6.Transcript variant expression has been normalized on PRPL4.1 showing the highest fold expression with respect to actin, employed as internal reference gene. White/black bars indicate expression fold changes in leaves/siliques.
(b) Intron-exon structures of PRPL1, PRPL4 andPRPL24 transcript variants. Exons are indicated as numbered white boxes, introns as black lines. Arrow-heads indicate the positions of translation initiation and stop codons. The beginning and end of exons is indicated.
Figure S2. Growth behaviour of prpl28-1albino plants. Progeny obtained by self-pollination of heterozygousPRPL28/prpl28-1 plants was cultivated either on soil or MS medium with or without 1% sucrose. Images were obtained at 4, 6, 8 and 10 days after germination (DAG). Albino plants grown on soil died after 10 DAG at the 2-cotyledon stage. A similar behaviour could be observed on MS medium without sucrose, whereas albino plants were able to extend roots and develop the first two true leaves on MS medium with 1% sucrose (for details on growth conditions see Experimental Procedures). Taken together, the data indicate that PRPL28 is essential for autotrophic growth in Arabidopsis. White arrow-heads indicate the position of albino plants on soil.
Table S1. Overview of the outcome ofcomplementation experiments. The number of resistant andcomplemented plants (containing the transgene in a homozygousmutant background and showing wild-type-like ΦIIvalues and leaf pigmentation) observed in T0 generationof transformed heterozygous or homozygous mutant plants isprovided. pro, 1 kbp of native promoter, together with the genomicregion of the corresponding gene. 35S-CaMV, constitutive promoterfrom Cauliflower Mosaic Virus controlling the expression of thecoding sequence of PRPS1 and PRPS17 genes. Leaf pigment content (Chl a+b) is expressed as pmol/mg leaf fresh weight. Note that all complemented plants showed a WT-like growth rate. See also Table 1 to compare the phenotype of the complemented plants with the one of the viable homozygous mutants.
Table S2. Silique length, seed set andfrequency of normal and aborted or (in case ofPRPL28/prpl28-1) albino seeds observed in WT (Col-0)and heterozygous mutant siliques at 20 DAF.
Table S3. Overview of the functions of theribosomal protein studied in chloroplasts and the prokaryote E.coli. Note that essential roles have been attributed to E.coli ribosomal proteins according to Hashimoto et al.(2005) (see PEC database,www.shigen.nig.ac.jp/ecoli/pec/index.jsp).
Table S4. Oligonucleotide sequences employed for genotyping insertion lines. Gene-specific sense (SP) and antisense (ASP) oligonucleotide combinations were used to identify the WT alleles, whereas T-DNA and gene-specific oligonucleotide combinations (either SP or ASP) were employed to detect the mutant alleles.
Table S5. Oligonucleotide sequences employed for complementation tests.
Table S6. Oligonucleotide sequences employed for gene expression analysis.
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