Cytoplasmic male sterility (CMS) is a maternally inherited trait of higher plants that can be suppressed by nuclear restorers of fertility ( Rf) genes that normally down-regulate the expression of a CMS-associated mitochondrial gene. In Brassica napus, nap CMS is associated with the expression of the orf222/nad5c/orf2101 mitochondrial gene region and is suppressed by the restorer gene Rfn. I present here an extensive analysis of the expression of the orf222/nad5c/orf2101 region in nap CMS and fertility-restored plants. Using RT-PCR methodology, I mapped transcript initiation sites, processing sites, and 3' termini. I identified two processing events, one within and one immediately downstream of orf222, that are specific to fertility restored plants and I suggest possible mechanisms by which the Rfn protein may recognize cognate RNA substrates. Unexpectedly, I also found that levels of atp8 transcripts are much lower in CMS than in restored plants. / nad5c, one of the components of the nap CMS-associated region, is the small central exon of the nad5 gene. In higher plants, nad5c transcripts must be joined to exons b and d through two group II intron trans-splicing events. I found that in the dicot Brassica and the monocot wheat, proper splicing requires exon c and d joining occur prior to the splicing of c with b. Joining of c to a/b transcripts prior to c/d splicing results exclusively in mis-spliced products in which the 5' end of c is joined to cryptic sites within exon b. It is suggested that intron sequences downstream of c base-pair with exon a, leading to mis-folding of the b/c intron and mis-splicing. In Oenothera, where the c/d intron is further fragmented into a tri-partite intron, mis-splicing does not occur. I suggest that avoidance of mis-splicing may be a factor that drives fragmentation of trans -splicing group II introns and may have contributed to the eventual evolution of spliceosomal RNAs from a group II intron precursor.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.115915 |
| Date | January 2007 |
| Creators | Elina, Helen. |
| Publisher | McGill University |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | application/pdf |
| Coverage | Doctor of Philosophy (Department of Biology.) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 002840554, proquestno: AAINR66717, Theses scanned by UMI/ProQuest. |
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