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Transcription, splicing and editing of plastid RNAs in the nonphotosynthetic plant Epifagus virginiana

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Abstract

Expression of the vestigial plastid genome of the nonphotosynthetic, parasitic flowering plant Epifagus virginiana was examined by northern analysis and by charaterization of cDNAs. Probes for each of 12 plastid genes tested hybridized to all lanes of northern blots containing total RNA prepared from stems and fruits of Epifgus and from leaves of tobacco. Certain transcript patterns in Epifagus plastids are highly complex and similar to those of tobacco operons. In contrast, genes such as rps2, which have become orphaned in Epifagus as a result of evolutionary loss of formerly cotranscribed genes, show simpler transcript patterns in Epifagus than in tobacco. Sizing and sequencing of cDNAs generated by reverse transcriptase-PCR for three genes, rps12, rpl2, and clpP, show that their transcripts are properly cis-and/or trans-spliced at the same five group II intron insertion sites used in photosynthetic plants. A single, conventional C→U edit in rps12 was found among the total of 1401 nucleotides of cDNA sequence that was determined for the three genes. An octanucleotide sequence identical to putative guide RNA of plant organelles and perfectly complementary to the rps12 edit site itself was identified just 200 bp upstream of the edit site. These data, together with previous results from the complete sequencing of the Epifagus plastid genome, provide compelling evidence that this degenerate genome is nonetheless expressed and functional. Analysis of the putative maturase MatK, encoderd by the group II intron of trnK in photosynthetic land plants but by a freestanding gene in Epifagus, leads us to hypothesize that it acts ‘in trans’ to assist the splicing of group II introns other than the one in which it is normally encoded.

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Author notes

  1. Stephanie C. Ems

    Present address: Department of Medical and Molecular Genetics, Indiana University School of Medicine, 46202, Indianapolis, IN, USA

  2. Clifford W. Morden

    Present address: Department of Botany/H.E.B.P., University of Hawaii at Manoa, 96822, Honolulu, HI, USA

  3. Colleen K. Dixon

    Present address: Eli Lilly and Company, Inc., Lilly Corporate Center, 46285, Indianapolis, IN, USA

  4. Kenneth H. Wolfe

    Present address: Department of Genetics, University of Dublin, Trinity College, Dublin 2, Ireland

  5. Claude W. de Pamphilis

    Present address: Department of Biology, Vanderbilt University, 37235, Nashville, TN, USA

Authors and Affiliations

  1. Department of Biology, Indiana University, 47405, Bloomington, IN, USA

    Stephanie C. Ems, Clifford W. Morden, Colleen K. Dixon, Kenneth H. Wolfe, Claude W. de Pamphilis & Jeffrey D. Palmer

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Ems, S.C., Morden, C.W., Dixon, C.K. et al. Transcription, splicing and editing of plastid RNAs in the nonphotosynthetic plant Epifagus virginiana . Plant Mol Biol 29, 721–733 (1995). https://doi.org/10.1007/BF00041163

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