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Male-sterile tobacco displays abnormal mitochondrial atp1 transcript accumulation and reduced floral ATP/ADP ratio

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Abstract

Alloplasmic male sterility is commonly obtained in Nicotiana by combining the nucleus from N. tabacum with the cytoplasm from other Nicotiana species. Besides being male-sterile, most of these cultivars also display changes in floral organ structure. Flowers from male-sterile plants containing the nucleus from N. tabacum combined with N. repanda cytoplasm develop stamen with shortened filaments and shrivelled anthers capped with stigmatoids. Male fertility and normal floral development can be restored by introduction of a restorer chromosome fragment from the cytoplasmic donor N. repanda into the N. tabacum nucleus. A novel reading frame, orf274, located upstream of atp1 in the mitochondrial genomes of both N. tabacum and N. repanda, as well as in the male-sterile and fertility-restored plants was identified. Co-transcripts of orf274 and atp1 were detected by RT-PCR in all four cultivars, but these transcripts accumulate to levels detectable by northern hybridization only in male-sterile plants. These co-transcripts neither generate detectable levels of an ORF274 polypeptide nor lead to altered expression of the ATP synthase subunit α. Measurement of ATP and ADP steady-state levels, however, revealed that the ATP/ADP ratio is significantly lower in young floral buds of male-sterile plants than in fertile plants.

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References

  • Araya, A., Begu, D. and Litvak, S. 1994. RNA editing in plants. Physiol. Plant. 91: 543–550.

    Google Scholar 

  • Binder, S., Marchfelder, A. and Brennicke, A. 1996. Regulation of gene expression in plant mitochondria. Plant Mol. Biol. 32: 303–314.

    Google Scholar 

  • Bonnett, H.T., Kofer, W., Hå kansson, G. and Glimelius, K. 1991. Mitochondrial involvement in petal and stamen development studied by sexual and somatic hybridization of Nicotiana species. Plant Sci. 80: 119–130.

    Google Scholar 

  • Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248–254.

    Google Scholar 

  • Braun, C.J. and Levings, C.S. III. 1985. Nucleotide sequence of the F1-ATPase α subunit gene from maize mitochondria. Plant Physiol. 79: 571–577.

    Google Scholar 

  • Braun, C.J., Brown, G.G., Levings, C.S. III and Hermann, R.G. 1992. Cytoplasmic male sterility. In: R.G. Hermann (Ed.), Cell Organelles: Plant Gene Research, Springer-Verlag, Wien/New York, pp. 219–245.

    Google Scholar 

  • Burk, L.G. 1967. An interspecific bridge-cross: Nicotiana repanda through N. sylvestris to N. tabacum. J. Hered. 58: 215–218.

    Google Scholar 

  • Chanut, F.A., Grabau, E.A. and Gesteland, R.F. 1993. Complex organization of the soybean mitochondrial genome: recombination repeats and multiple transcripts at the atpA loci. Curr. Genet. 23: 234–247.

    Google Scholar 

  • Chase, C.D. 1994. Expression of CMS-unique and flanking mitochondrial DNA sequences in Phaseolus vulgaris L. Curr. Genet. 25: 245–251.

    Google Scholar 

  • Chase, C.D. and Ortega, V.M. 1992. Organization of ATPA coding and 30 flanking sequences associated with cytoplasmic male sterility in Phaseolus vulgaris L. Curr. Genet. 22: 147–153.

    Google Scholar 

  • Chaumont, F., Boutry, M., Briquet, M. and Vassarotti, A. 1988. Sequence of the gene encoding the mitochondrial F1-ATPase α subunit from Nicotiana plumbaginifolia. Nucl. Acids Res. 16: 6247.

    Google Scholar 

  • Chen, G.T. and Inouye, M. 1994. Role of the AGA/AGG codons, the rarest codons in global gene expression in Escherichia coli. Genes Dev. 8: 2641–2652.

    Google Scholar 

  • Conley, C.A. and Hanson, M.R. 1994. Tissue-specific protein expression in plant mitochondria. Plant Cell 6: 85–91.

    Google Scholar 

  • DeBlock, M. and Debrouwer, D. 1996. RNA-RNA in situ hybridization using digoxigenin-labeled probes: the use of high molecular weight polyvinyl alcohol in the alkaline phosphatase indoxyl-nitroblue tetrazolium. Anal. Biochem. 216: 88–89.

    Google Scholar 

  • Gardeström, P. and Wigge, B. 1988. Influence on ATP/ADP ratios in the chloroplasts, mitochondria, and cytosol, studied by rapid fractionation of barley (Hordeum vulgare) protoplasts. Plant Physiol. 88: 69–76.

    Google Scholar 

  • Gerstel, D.U., Burns, J.A. and Burk. L.G. 1978. Cytoplasmic male sterility in Nicotiana, restoration of fertility, and the nucleolus. Genetics 89: 157–169.

    Google Scholar 

  • Goldman, E., Rosenberg, A.H., Zubay, G. and Studier, F.W. 1995. Consecutive low-usage leucine codons block translation only when near the 50 end of a message in Escherichia coli. J. Mol. Biol. 245: 467–473.

    Google Scholar 

  • Hanson, M.R. 1991. Plant mitochondrial mutations and male sterility. Annu. Rev. Genet. 25: 461–486.

    Google Scholar 

  • He, S., Lyznik, A. and Mackenzie, S. 1995. Pollen fertility restoration by nuclear gene Fr in CMS bean: nuclear-directed alteration of a mitochondrial population. Genetics 139: 955–962.

    Google Scholar 

  • Hill, D.H. and Straka, J.G. 1988. Protein determination using bicinchoninic acid in the presence of sulfhydryl reagents. Anal. Biochem. 170: 203–208.

    Google Scholar 

  • Hå kansson, G. 1992. Nuclear-mitochondrial interactions and its relevance for male sterility in Nicotiana. Ph.D. thesis, Swedish University of Agricultural Sciences, Dept. of Plant Breeding, ISBN 91-576-4572-8.

  • Hå kansson, G. and Glimelius, K. 1991. Extensive nuclear influence on mitochondrial transcription and genome structure in malefertile and male-sterile alloplasmic Nicotiana materials. Mol. Gen. Genet. 229: 380–388.

    Google Scholar 

  • Hå kansson, G., van der Mark, F., Bonnett, H.T. and Glimelius, K. 1988. Variant mitochondrial protein and DNA patterns associated with cytoplasmic male-sterile lines of Nicotiana. Theor. Appl. Genet. 76: 431–437.

    Google Scholar 

  • Köhler, R.H., Horn, R., Lössl, A. and Zetsche, K. 1991. Cytoplasmic male sterility in sunflower is correlated with the cotranscription of a new open reading frame with the atpA gene. Mol. Gen. Genet. 227: 369–376.

    Google Scholar 

  • Koltunow, A.M., Truettner, J., Cox, K.H., Wallroth, M. and Goldberg, R.B. 1990. Different temporal and spatial gene expression patterns occur during anther development. Plant Cell 2: 1201–1224.

    Google Scholar 

  • Kyte, J. and Doolittle, R.F. 1982. A simple method for displaying the hydropathic character of a protein. J. Mol. Biol. 157: 105–132.

    Google Scholar 

  • Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.

    Google Scholar 

  • Landgren, M. and Glimelius, K. 1990. Analysis of chloroplast and mitochondrial segregation in three different combinations of somatic hybrids produced within Brassicaceae. Theor. Appl. Genet. 80: 776–784.

    Google Scholar 

  • Laser, B. and Kück, U. 1995. The mitochondrial atpA/atp9 cotranscript in wheat and triticale: RNA processing depends on the nuclear genotype. Curr. Genet. 29: 50–57.

    Google Scholar 

  • Laver, H.K., Reynolds, S.J., Monéger, F. and Leaver, C.J. 1991. Mitochondrial genome organization and gene expression associated with cytoplasmic male sterility in sunflower (Helianthus annuus). Plant J. 1: 186–193.

    Google Scholar 

  • Lee, S. and Warmke, H.E. 1979. Organelle size and number in fertile and T-cytoplasmic male-sterile corn. Am. J. Bot. 66: 141–148.

    Google Scholar 

  • Levings, C.S. III. 1990. The Texas cytoplasm of maize: cytoplasmic male sterility and disease susceptibility. Science 250: 942–947.

    Google Scholar 

  • Logemann, J., Schell, J. and Willmitzer, L. 1987. Improved method for the isolation of RNA from plant tissues. Anal. Biochem. 163: 16–20.

    Google Scholar 

  • Luethy, M.H., Horak, A. and Elthon, T.E. 1993. Monoclonal antibodies to the α-and β-subunits of the plant mitochondrial F1-ATPase. Plant Physiol. 101: 931–937.

    Google Scholar 

  • Mackenzie, S., He, S. and Lyznik, A. 1994. The elusive plant mitochondrion as a genetic system. Plant Physiol. 105: 775–780.

    Google Scholar 

  • Maier, R.M., Zeltz, P., Kössel, H., Bonnard, G., Gualberto, J.M. and Grienenberger, J.M. 1996. RNA editing in plant mitochondria and chloroplasts. Plant Mol. Biol. 32: 343–365.

    Google Scholar 

  • Nieman, R.H. and Clark, R.A. 1984. Measurement of plant nucleotides by high-performance liquid chromatography. J. Chromat. 317: 271–281.

    Google Scholar 

  • Sambrook, E., Fritsch, F. and Maniatis, T. 1989. Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  • Sand, S.A. and Christoff, G.T. 1973. Cytoplasmic-chromosomal interactions and altered differentiation in tobacco. J. Hered. 64: 24–30.

    Google Scholar 

  • Sanger, F., Nicklen, S. and Coulson, A.R. 1977. DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. USA 74: 5463–5467.

    Google Scholar 

  • Sarria, R., Lyznik, A., Vallejos, C.E. and Mackenzie, S.A. 1998. A cytoplasmic male sterility-associated mitochondrial peptide in common bean is post-translationally regulated. Plant Cell 10: 1217–1228.

    Google Scholar 

  • Sharpe, A.G., Parkin, I.A.P., Keith, D.J. and Lydiate, D.J. 1995. Frequent nonreciprocal translocations in the amphidiploid genome of oilseed rape (Brassica napus). Genome 38: 1112–1121.

    Google Scholar 

  • Small, I.D., Isaac, P.G. and Leaver, C.J. 1987. Stoichiometric differences in DNA molecules containing the atpA gene suggest mechanisms for the generation of mitochondrial genome diversity in maize. EMBO J. 6: 865–869.

    Google Scholar 

  • Smart, C.J., Monéger, F. and Leaver, C.J. 1994. Cell specific regulation of gene expression in mitochondria during anther development in sunflower. Plant Cell 6: 811–825.

    Google Scholar 

  • Tadege, M. and Kuhlemeier, C. 1997. Aerobic fermentation during tobacco pollen development. Plant. Mol. Biol. 35: 3433–3454.

    Google Scholar 

  • Thelen, J.J., Miernyk, J.A. and Randall, D.D. 1999. Molecular cloning and analysis of the mitochondrial pyruvate dehydrogenase from maize. Plant Physiol. 119: 635–643.

    Google Scholar 

  • Towbin, H., Staehelin, T. and Gordon, J. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA 76: 4350–4354.

    Google Scholar 

  • Wise, R.P., Dill, C.L. and Schnable, P.S. 1996. Mutator-induced mutations of the rf1 nuclear fertility restorer of T-cytoplasm maize alter the accumulation of T-urf13 mitochondrial transcripts. Genetics 143: 1383–1394.

    Google Scholar 

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Bergman, P., Edqvist, J., Farbos, I. et al. Male-sterile tobacco displays abnormal mitochondrial atp1 transcript accumulation and reduced floral ATP/ADP ratio. Plant Mol Biol 42, 531–544 (2000). https://doi.org/10.1023/A:1006388814458

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