Faithful editing of a tomato-specific mRNA editing site in transgenic tobacco chloroplasts

Karcher, Daniel; Kahlau, Sabine; Bock, Ralph

doi:10.1261/rna.823508

Cited by 19 publications

(15 citation statements)

References 47 publications

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“…It was stated by the authors that the trans-acting factors of cp RNA editing could recognize the groups of cis-elements that shared a moderate sequence identity. Similar results were achieved by Karsher et al (Karcher et al, 2008) concerning sites rpoB-6 and rps12 (which was artificially introduced to the tobacco genome). From this viewpoint, 28 RNA editing sites of the cucumber cp genome have been examined, searching for possible candidates recognized by the common trans-acting factors.…”

Section: Discussionsupporting

confidence: 86%

Cucumber, melon, pumpkin, and squash: Are rules of editing in flowering plants chloroplast genes so well known indeed?

Guzowska

Fiedorowicz

Pląder

2009

Gene

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Section: Discussionsupporting

confidence: 86%

Cucumber, melon, pumpkin, and squash: Are rules of editing in flowering plants chloroplast genes so well known indeed?

Guzowska

Fiedorowicz

Pląder

2009

Gene

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“…Some RNA editing sites were shown to have conserved sequences upstream of the editing sites (Karcher et al, 2008;Kobayashi et al, 2008;Sosso et al, 2012). However, these conserved sequences were derived by comparing merely two editing sites.…”

Section: Targeting Sequences Of Emp5 Are Not Conservedmentioning

confidence: 99%

Empty Pericarp5Encodes a Pentatricopeptide Repeat Protein That Is Required for Mitochondrial RNA Editing and Seed Development in Maize

et al. 2013

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In flowering plants, RNA editing is a posttranscriptional mechanism that converts specific cytidines to uridines in both mitochondrial and plastidial transcripts, altering the information encoded by these genes. Here, we report the molecular characterization of the empty pericarp5 (emp5) mutants in maize (Zea mays). Null mutation of Emp5 results in abortion of embryo and endosperm development at early stages. Emp5 encodes a mitochondrion-targeted DYW subgroup pentatricopeptide repeat (PPR) protein. Analysis of the mitochondrial transcripts revealed that loss of the EMP5 function abolishes the C-to-U editing of ribosomal protein L16 at the rpl16-458 site (100% edited in the wild type), decreases the editing at nine sites in NADH dehydrogenase9 (nad9), cytochrome c oxidase3 (cox3), and ribosomal protein S12 (rps12), and surprisingly increases the editing at five sites of ATP synthase F0 subunit a (atp6), apocytochrome b (cob), nad1, and rpl16. Mutant EMP5-4 lacking the E+ and DYW domains still retains the substrate specificity and editing function, only at reduced efficiency. This suggests that the E+ and DYW domains of EMP5 are not essential to the EMP5 editing function but are necessary for efficiency. Analysis of the ortholog in rice (Oryza sativa) indicates that rice EMP5 has a conserved function in C-to-U editing of the rice mitochondrial rpl16-458 site. EMP5 knockdown expression in transgenics resulted in slow growth and defective seeds. These results demonstrate that Emp5 encodes a PPR-DYW protein that is required for the editing of multiple transcripts in mitochondria, and the editing events, particularly the Cto-U editing at the rpl16-458 site, are critical to the mitochondrial functions and, hence, to seed development in maize.

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“…Chloroplast transformation has become an invaluable tool to study plastid gene functions (e.g., Ruf et al 1997;Hager et al 1999) and the mechanisms of plastid gene expression (Allison and Maliga 1995;Eibl et al 1999;Karcher et al 2008). In addition, the transplastomic technology has created new opportunities for plant biotechnologists.…”

Section: Introductionmentioning

confidence: 99%

Chloramphenicol acetyltransferase as selectable marker for plastid transformation

2010

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Chloroplast transformation remains a demanding technique and is still restricted to relatively few plant species. The limited availability of selectable marker genes and the lack of selection markers that would be universally applicable to all plant species represent some of the most serious technical problems involved in extending the species range of plastid transformation. Here we report the development of the chloramphenicol acetyltransferase gene cat as a new selectable marker for plastid transformation. We show that, by selecting for chloramphenicol resistance, tobacco chloroplast transformants are readily obtained. Transplastomic lines quickly reach the homoplasmic state (typically in one additional regeneration round), accumulate the chloramphenicol acetyltransferase enzyme to high levels and transmit their plastid transgenes maternally into the next generation. No spontaneous antibiotic resistance mutants appear upon chloramphenicol selection. Several lines of evidence support the assumption that plant mitochondria are also sensitive to chloramphenicol suggesting that the chloramphenicol acetyltransferase may be a good candidate selectable marker for plant mitochondrial transformation.

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Faithful editing of a tomato-specific mRNA editing site in transgenic tobacco chloroplasts

Cited by 19 publications

References 47 publications

Cucumber, melon, pumpkin, and squash: Are rules of editing in flowering plants chloroplast genes so well known indeed?

Cucumber, melon, pumpkin, and squash: Are rules of editing in flowering plants chloroplast genes so well known indeed?

Empty Pericarp5Encodes a Pentatricopeptide Repeat Protein That Is Required for Mitochondrial RNA Editing and Seed Development in Maize

Chloramphenicol acetyltransferase as selectable marker for plastid transformation

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