Skip to main content
Genetics logoLink to Genetics
. 1998 Oct;150(2):873–882. doi: 10.1093/genetics/150.2.873

Organization and expression of the mitochondrial genome in the Nicotiana sylvestris CMSII mutant.

C Lelandais 1, B Albert 1, S Gutierres 1, R De Paepe 1, B Godelle 1, F Vedel 1, P Chétrit 1
PMCID: PMC1460359  PMID: 9755215

Abstract

Previous analyses suggested that the Nicotiana sylvestris CMSII mutant carried a large deletion in its mitochondrial genome. Here, we show by cosmid mapping that the deletion is 60 kb in length and contains several mitochondrial genes or ORFs, including the complex I nad7 gene. However, due to the presence of large duplications in the progenitor mitochondrial genome, the only unique gene that appears to be deleted is nad7. RNA gel blot data confirm the absence of nad7 expression, strongly suggesting that the molecular basis for the CMSII abnormal phenotype, poor growth and male sterility, is the altered complex I structure. The CMSII mitochondrial genome appears to consist essentially of one of two subgenomes resulting from recombination between direct short repeats. In the progenitor mitochondrial genome both recombination products are detected by PCR and, reciprocally, the parental fragments are detected at the substoichiometric level in the mutant. The CMSII mtDNA organization has been maintained through six sexual generations.

Full Text

The Full Text of this article is available as a PDF (391.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Backert S., Dörfel P., Börner T. Investigation of plant organellar DNAs by pulsed-field gel electrophoresis. Curr Genet. 1995 Sep;28(4):390–399. doi: 10.1007/BF00326439. [DOI] [PubMed] [Google Scholar]
  2. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brandt P., Sünkel S., Unseld M., Brennicke A., Knoop V. The nad4L gene is encoded between exon c of nad5 and orf25 in the Arabidopsis mitochondrial genome. Mol Gen Genet. 1992 Dec;236(1):33–38. doi: 10.1007/BF00279640. [DOI] [PubMed] [Google Scholar]
  4. Chapdelaine Y., Bonen L. The wheat mitochondrial gene for subunit I of the NADH dehydrogenase complex: a trans-splicing model for this gene-in-pieces. Cell. 1991 May 3;65(3):465–472. doi: 10.1016/0092-8674(91)90464-a. [DOI] [PubMed] [Google Scholar]
  5. Chetrit P., Rios R., De Paepe R., Vitart V., Gutierres S., Vedel F. Cytoplasmic male sterility is associated with large deletions in the mitochondrial DNA of two Nicotiana sylvestris protoclones. Curr Genet. 1992 Feb;21(2):131–137. doi: 10.1007/BF00318472. [DOI] [PubMed] [Google Scholar]
  6. Dawson A. J., Jones V. P., Leaver C. J. The apocytochrome b gene in maize mitochondria does not contain introns and is preceded by a potential ribosome binding site. EMBO J. 1984 Sep;3(9):2107–2113. doi: 10.1002/j.1460-2075.1984.tb02098.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. De Paepe R., Chétrit P., Vitart V., Ambard-Bretteville F., Prat D., Vedel F. Several nuclear genes control both male sterility and mitochondrial protein synthesis in Nicotiana sylvestris protoclones. Mol Gen Genet. 1990 Jul;222(2-3):206–210. doi: 10.1007/BF00633819. [DOI] [PubMed] [Google Scholar]
  8. Dewey R. E., Levings C. S., 3rd, Timothy D. H. Novel recombinations in the maize mitochondrial genome produce a unique transcriptional unit in the Texas male-sterile cytoplasm. Cell. 1986 Feb 14;44(3):439–449. doi: 10.1016/0092-8674(86)90465-4. [DOI] [PubMed] [Google Scholar]
  9. Dewey R. E., Levings C. S., Timothy D. H. Nucleotide sequence of ATPase subunit 6 gene of maize mitochondria. Plant Physiol. 1985 Nov;79(3):914–919. doi: 10.1104/pp.79.3.914. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Falconet D., Lejeune B., Quetier F., Gray M. W. Evidence for homologous recombination between repeated sequences containing 18S and 5S ribosomal RNA genes in wheat mitochondrial DNA. EMBO J. 1984 Feb;3(2):297–302. doi: 10.1002/j.1460-2075.1984.tb01799.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fauron C. M., Havlik M., Brettell R. I. The mitochondrial genome organization of a maize fertile cmsT revertant line is generated through recombination between two sets of repeats. Genetics. 1990 Feb;124(2):423–428. doi: 10.1093/genetics/124.2.423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Folkerts O., Hanson M. R. The male sterility-associated pcf gene and the normal atp9-1 gene in Petunia are located on different mitochondrial DNA molecules. Genetics. 1991 Nov;129(3):885–895. doi: 10.1093/genetics/129.3.885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fox T. D., Leaver C. J. The Zea mays mitochondrial gene coding cytochrome oxidase subunit II has an intervening sequence and does not contain TGA codons. Cell. 1981 Nov;26(3 Pt 1):315–323. doi: 10.1016/0092-8674(81)90200-2. [DOI] [PubMed] [Google Scholar]
  14. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gualberto J. M., Wintz H., Weil J. H., Grienenberger J. M. The genes coding for subunit 3 of NADH dehydrogenase and for ribosomal protein S12 are present in the wheat and maize mitochondrial genomes and are co-transcribed. Mol Gen Genet. 1988 Dec;215(1):118–127. doi: 10.1007/BF00331312. [DOI] [PubMed] [Google Scholar]
  16. Gutierres S., Lelandais C., Paepe R. D., Vedel F., Chétrit P. A mitochondrial sub-stoichiometric orf87-nad3-nad1 exonA co-transcription unit present in solanaceae was amplified in the genus Nicotiana. Curr Genet. 1997 Jan;31(1):55–62. doi: 10.1007/s002940050176. [DOI] [PubMed] [Google Scholar]
  17. Gutierres S., Sabar M., Lelandais C., Chetrit P., Diolez P., Degand H., Boutry M., Vedel F., de Kouchkovsky Y., De Paepe R. Lack of mitochondrial and nuclear-encoded subunits of complex I and alteration of the respiratory chain in Nicotiana sylvestris mitochondrial deletion mutants. Proc Natl Acad Sci U S A. 1997 Apr 1;94(7):3436–3441. doi: 10.1073/pnas.94.7.3436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Haouazine N., Takvorian A., Jubier M. F., Michel F., Lejeune B. The nad6 gene and the exon d of nad1 are co-transcribed in wheat mitochondria. Curr Genet. 1993 Dec;24(6):533–538. doi: 10.1007/BF00351718. [DOI] [PubMed] [Google Scholar]
  19. Hartmann C., Récipon H., Jubier M. F., Valon C., Delcher-Besin E., Henry Y., De Buyser J., Lejeune B., Rode A. Mitochondrial DNA variability detected in a single wheat regenerant involves a rare recombination event across a short repeat. Curr Genet. 1994 May;25(5):456–464. doi: 10.1007/BF00351786. [DOI] [PubMed] [Google Scholar]
  20. He S., Lyznik A., Mackenzie S. Pollen fertility restoration by nuclear gene Fr in CMS bean: nuclear-directed alteration of a mitochondrial population. Genetics. 1995 Feb;139(2):955–962. doi: 10.1093/genetics/139.2.955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hiesel R., Schobel W., Schuster W., Brennicke A. The cytochrome oxidase subunit I and subunit III genes in Oenothera mitochondria are transcribed from identical promoter sequences. EMBO J. 1987 Jan;6(1):29–34. doi: 10.1002/j.1460-2075.1987.tb04714.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Janska H., Mackenzie S. A. Unusual mitochondrial genome organization in cytoplasmic male sterile common bean and the nature of cytoplasmic reversion to fertility. Genetics. 1993 Nov;135(3):869–879. doi: 10.1093/genetics/135.3.869. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kanazawa A., Tsutsumi N., Hirai A. Reversible changes in the composition of the population of mtDNAs during dedifferentiation and regeneration in tobacco. Genetics. 1994 Nov;138(3):865–870. doi: 10.1093/genetics/138.3.865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lamattina L., Gonzalez D., Gualberto J., Grienenberger J. M. Higher plant mitochondria encode an homologue of the nuclear-encoded 30-kDa subunit of bovine mitochondrial complex I. Eur J Biochem. 1993 Nov 1;217(3):831–838. doi: 10.1111/j.1432-1033.1993.tb18311.x. [DOI] [PubMed] [Google Scholar]
  25. Lamattina L., Grienenberger J. M. RNA editing of the transcript coding for subunit 4 of NADH dehydrogenase in wheat mitochondria: uneven distribution of the editing sites among the four exons. Nucleic Acids Res. 1991 Jun 25;19(12):3275–3282. doi: 10.1093/nar/19.12.3275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mackenzie S. A., Chase C. D. Fertility Restoration Is Associated with Loss of a Portion of the Mitochondrial Genome in Cytoplasmic Male-Sterile Common Bean. Plant Cell. 1990 Sep;2(9):905–912. doi: 10.1105/tpc.2.9.905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Martínez-Zapater J. M., Gil P., Capel J., Somerville C. R. Mutations at the Arabidopsis CHM locus promote rearrangements of the mitochondrial genome. Plant Cell. 1992 Aug;4(8):889–899. doi: 10.1105/tpc.4.8.889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nehls U., Friedrich T., Schmiede A., Ohnishi T., Weiss H. Characterization of assembly intermediates of NADH:ubiquinone oxidoreductase (complex I) accumulated in Neurospora mitochondria by gene disruption. J Mol Biol. 1992 Oct 20;227(4):1032–1042. doi: 10.1016/0022-2836(92)90519-p. [DOI] [PubMed] [Google Scholar]
  29. Oldenburg D. J., Bendich A. J. Size and Structure of Replicating Mitochondrial DNA in Cultured Tobacco Cells. Plant Cell. 1996 Mar;8(3):447–461. doi: 10.1105/tpc.8.3.447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pereira de Souza A., Jubier M. F., Delcher E., Lancelin D., Lejeune B. A trans-splicing model for the expression of the tripartite nad5 gene in wheat and maize mitochondria. Plant Cell. 1991 Dec;3(12):1363–1378. doi: 10.1105/tpc.3.12.1363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Pla M., Mathieu C., De Paepe R., Chétrit P., Vedel F. Deletion of the last two exons of the mitochondrial nad7 gene results in lack of the NAD7 polypeptide in a Nicotiana sylvestris CMS mutant. Mol Gen Genet. 1995 Jul 22;248(1):79–88. doi: 10.1007/BF02456616. [DOI] [PubMed] [Google Scholar]
  32. Rottmann W. H., Brears T., Hodge T. P., Lonsdale D. M. A mitochondrial gene is lost via homologous recombination during reversion of CMS T maize to fertility. EMBO J. 1987 Jun;6(6):1541–1546. doi: 10.1002/j.1460-2075.1987.tb02398.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schardl C. L., Pring D. R., Lonsdale D. M. Mitochondrial DNA rearrangements associated with fertile revertants of S-type male-sterile maize. Cell. 1985 Nov;43(1):361–368. doi: 10.1016/0092-8674(85)90041-8. [DOI] [PubMed] [Google Scholar]
  34. Schuster W., Hiesel R., Isaac P. G., Leaver C. J., Brennicke A. Transcript termini of messenger RNAs in higher plant mitochondria. Nucleic Acids Res. 1986 Aug 11;14(15):5943–5954. doi: 10.1093/nar/14.15.5943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Shirzadegan M., Palmer J. D., Christey M., Earle E. D. Patterns of mitochondrial DNA instability in Brassica campestris cultured cells. Plant Mol Biol. 1991 Jan;16(1):21–37. doi: 10.1007/BF00017914. [DOI] [PubMed] [Google Scholar]
  36. Small I. D., Isaac P. G., Leaver C. J. Stoichiometric differences in DNA molecules containing the atpA gene suggest mechanisms for the generation of mitochondrial genome diversity in maize. EMBO J. 1987 Apr;6(4):865–869. doi: 10.1002/j.1460-2075.1987.tb04832.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Stern D. B., Newton K. J. Mitochondrial gene expression in Cucurbitaceae: conserved and variable features. Curr Genet. 1985;9(5):395–404. doi: 10.1007/BF00421611. [DOI] [PubMed] [Google Scholar]
  38. Vitart V., De Paepe R., Mathieu C., Chétrit P., Vedel F. Amplification of substoichiometric recombinant mitochondrial DNA sequences in a nuclear, male sterile mutant regenerated from protoplast culture in Nicotiana sylvestris. Mol Gen Genet. 1992 May;233(1-2):193–200. doi: 10.1007/BF00587579. [DOI] [PubMed] [Google Scholar]
  39. Wallace D. C. Mitochondrial DNA mutations in diseases of energy metabolism. J Bioenerg Biomembr. 1994 Jun;26(3):241–250. doi: 10.1007/BF00763096. [DOI] [PubMed] [Google Scholar]
  40. Young E. G., Hanson M. R. A fused mitochondrial gene associated with cytoplasmic male sterility is developmentally regulated. Cell. 1987 Jul 3;50(1):41–49. doi: 10.1016/0092-8674(87)90660-x. [DOI] [PubMed] [Google Scholar]
  41. da Silva M. V., Alves P. C., Duarte M., Mota N., Lobo-da-Cunha A., Harkness T. A., Nargang F. E., Videira A. Disruption of the nuclear gene encoding the 20.8-kDa subunit of NADH: ubiquinone reductase of Neurospora mitochondria. Mol Gen Genet. 1996 Aug 27;252(1-2):177–183. doi: 10.1007/BF02173218. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES