Molecular Biology Reports

, Volume 36, Issue 2, pp 273–280 | Cite as

Mitochondrial atpA gene is altered in a new orf220-type cytoplasmic male-sterile line of stem mustard (Brassica juncea)

  • Jing-Hua Yang
  • Yan Huai
  • Ming-Fang ZhangEmail author


The purpose of this research is to identify the probable mitochondrial factor associated with cytoplasmic male sterility (cms) by comparative analysis of cms and its isogenic maintainer lines in stem mustards. Dramatic variations in the morphology of floral organs were observed in cms stem mustard. Mitochondrial atpA gene was shown to be altered in cms compared with that in its maintainer line, of which mitochondrial atpA gene from its maintainer line was sequenced to encode 507 amino acids. It was indicative of high homology with mitochondrial atpA genes from other species, even as high as 94% in similarity with Oryza sativa in terms of amino acid constituents. However, only 429 amino acids were deduced in cms showing 83% similarity with atpA gene from its maintainer line. Two copies were observed in its maintainer line, but only one was found in cms. Such numerous differences of mitochondrial atpA gene between cms and its maintainer lines may not be the results of evolutionary divergence but the rearrangements of mitochondria. Expression of mitochondrial atpA gene was shown to be down-regulated in cms by using Northern blot. Consequently, mitochondrial ATP synthesis was severely decreased more than one fold in cms stem mustard indicating deficiency in mitochondrial ATP synthesis in this type of cms. Therefore, we deduced that mitochondrial atpA gene altered in cms could be associated with male-sterility in this type of cms.


Cytoplasmic male sterility Mitochondrial atpA Brassica juncea 



This work was partially supported by a grant from the National Natural Science Foundation of China (NSFC, 30571270). We thank Dr. Nakazono Mikio (The University of Tokyo) for providing critical comments on the experiment.

Supplementary material

11033_2007_9176_MOESM1_ESM.pdf (23 kb)
Alignment of the deduced amino acids of mitochondrial atpA from cms and its maintainer line of stem mustard (PDF 24 kb)


  1. 1.
    Laser KD, Lersten NR (1972) Anatomy and cytology of microsporogenesis in cytoplasmic male-sterile angiosperms. Bot Rev 38:425–454CrossRefGoogle Scholar
  2. 2.
    Kaul MLH (1988) Male sterility in higher plants. Springer, BerlinGoogle Scholar
  3. 3.
    Dewey RE, Levings CS III, Timothy DH (1986) Novel recombinations in the maize mitochondrial genome produce a unique transcriptional unit in the Texas male-sterile cytoplasm. Cell 44:439–449PubMedCrossRefGoogle Scholar
  4. 4.
    Dewey RE, Timothy DH, Levings CS III (1987) A mitochondrial protein associated with cytoplasmic male sterility in the T-cytoplasm of maize. Proc Natl Acad Sci USA 84:5374–5378PubMedCrossRefGoogle Scholar
  5. 5.
    Schnable PS, Wise RP (1998) The molecular basis of cytoplasmic male sterility and fertility restoration. Trends Plant Sci 3:175–180CrossRefGoogle Scholar
  6. 6.
    Budar F, Pelletier G (2001) Male sterility in plant: occurrence, determinism, significance and use. Life Sci 324:543–550Google Scholar
  7. 7.
    Hanson MR, Bentolila S (2004) Interaction of mitochondrial and nuclear genes that affect male gametophyte development. Plant Cell 16:S154–S169PubMedCrossRefGoogle Scholar
  8. 8.
    Linker B, Börner T (2005) Mitochondrial effects on flower and pollen development. Mitochondrion 5:389–402CrossRefGoogle Scholar
  9. 9.
    Pelletier G, Budar F (2007) The molecular biology of cytoplasmically inherited male sterility and prospects for its engineering. Curr Opin Biotechnol 18:121–125PubMedCrossRefGoogle Scholar
  10. 10.
    Small ID, Isaac PG (1987) Stoichiometric differences in DNA molecules containing the atpA gene suggest mechanisms for the generation of mitochondrial genome diversity in maize. EMBO J 6:865–869PubMedGoogle Scholar
  11. 11.
    Makaroff CA, Apel IJ, Palmer JD (1990) Characterization of radish mitochondrial atpA: influence of nuclear background on transcription of atpA-associated sequences and relationship with male sterility. Plant Mol Biol 15:735–746PubMedCrossRefGoogle Scholar
  12. 12.
    Kohler RH, Horn R, Lossl A, Zetsche K (1991) Cytoplasmic male sterility in sunflower is correlated with the co-transcription of a new open reading frame with the atpA gene. Mol Gen Genet 227:369–376PubMedCrossRefGoogle Scholar
  13. 13.
    Sakai T, Imamura J (1992) Alteration of mitochondrial genomes containing atpA genes in the sexual progeny of cybrids between Raphanus sativus cms line and Brassica napus cv. Westar. Theor Appl Genet 84:923–926CrossRefGoogle Scholar
  14. 14.
    Xue YB, Collin S, Davies DR, Thomas CM (1994) Differential screening of mitochondrial cDNA libraries from male-fertile and cytoplasmic male-sterility sugar-beet reveals genome rearrangements at atp6 and atpA loci. Plant Mol Biol 25:91–103PubMedCrossRefGoogle Scholar
  15. 15.
    Makaroff CA, Apel IJ, Palmer JD (1989) The atp6 coding region has been disrupted and a novel reading frame generated in the mitochondrial genome of cytoplasmic male-sterile radish. J Biol Chem 264:11706–11713PubMedGoogle Scholar
  16. 16.
    Singh M, Brown GG (1991) Suppression of cytoplasmic male sterility by nuclear genes alters expression of a novel mitochondrial gene region. Plant Cell 3:1349–1362PubMedCrossRefGoogle Scholar
  17. 17.
    Handa H, Nakajima K (1992) Different organization and altered transcription of the mitochondrial atp6 gene in the male-sterile cytoplasm of rapeseed (Brassica napus L.) Curr Genet 21:153–159PubMedCrossRefGoogle Scholar
  18. 18.
    Szklarczyk M, Oczkowski M, Augustyniak H (2000) Organization and expression of mitochondrial atp9 genes from cms and fertile carrots. Thero Appl Genet 100:263–270CrossRefGoogle Scholar
  19. 19.
    Canal de la L., Crouzillat D, Quetier F, Ledoigt G (2001) A transcriptional alteration on the atp9 gene is associated with a sunflower male-sterile cytoplasm. Thero Appl Genet 102:1185–1189CrossRefGoogle Scholar
  20. 20.
    Dieterich JH, Braun HP, Schmitz UK (2003) Alloplasmic male sterility in Brassica napus (cms ‘Tournefortii-Stiewe’) is associated with a special gene arrangement around a novel atp9 gene. Mol Gen Genomics 269:723–731CrossRefGoogle Scholar
  21. 21.
    Sandhu APS, Abdelnoor RV, Mackenzie SA (2007) Transgenic induction of mitochondrial rearrangements for cytoplasmic male sterility in crop plants. Proc Natl Acad Sci USA 104(6):1766–1770PubMedCrossRefGoogle Scholar
  22. 22.
    Marienfeld JR, Unseld M, Brandt P, Brennicke A (1997) Mosaic open reading frames in the Arabidopsis thaliana mitochondrial genome. Biol Chem 378:859–862PubMedCrossRefGoogle Scholar
  23. 23.
    Leino M, Landgren M, Glimelius K (2005) Alloplasmic effects on mitochondrial transcriptional activity and RNA turnover result in accumulated transcripts of Arabidopsis orfs in cytoplasmic male-sterile Brassica napus. Plant J 42:469–480PubMedCrossRefGoogle Scholar
  24. 24.
    Carlsson J, Leino M, Glimelius K (2007) Mitochondrial genotypes with variable parts of Arabidopsis thaliana DNA affect development in Brassica napus lines. Theor Appl Genet 115:627–641PubMedCrossRefGoogle Scholar
  25. 25.
    Zhang M-F, Chen L-P, Wang B-L, Yang J-H, Chen Z-J, Yutaka H (2003) Characterization of atpA and orf220 genes distinctively present in a cytoplasmic male-sterile line of tuber mustard. J Hortic Sci Biotechnol 78(6):837–841Google Scholar
  26. 26.
    Dellaporta SL, Wood J, Hicks JB (1983) A Plant DNA minipreparation: version. Plant Mol Biol Rep 1(4):19–24CrossRefGoogle Scholar
  27. 27.
    Thompson JD, Higgins DG, Gibson TJ, Clustal W (1994) Improving the sensitivity of progressive multiple sequence alignment through sequence weighing, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680PubMedCrossRefGoogle Scholar
  28. 28.
    Ames BN (1966) Assay of inorganic phosphate, total phosphate and phosphatases. Method Enzymol 8:115–118CrossRefGoogle Scholar
  29. 29.
    Bunney TD, Walraven HS, Boer AH (2001) 14-3-3 protein is a regulator of the mitochondrial and chloroplast ATP synthase. PNAS 98:4249–4254PubMedCrossRefGoogle Scholar
  30. 30.
    Yang JH, Zhang MF, Yu JQ (2007) Alterations of RNA editing for the mitochondrial atp9 gene in a new orf220-type cytoplasmic male-sterile line of stem mustard (Brassica juncea var. tumida). J Integr Plant Biol 49(5):672–677CrossRefGoogle Scholar
  31. 31.
    Bergman P, Edqvist J, Farbos I, Glimelius K (2000) Male-sterile tobacco displays abnormal mitochondrial atp1 transcript accumulation and reduced floral ATP/ADP ratio. Plant Mol Biol 42:531–544PubMedCrossRefGoogle Scholar
  32. 32.
    Sabar M, Gagliaradi D, Balk J, Leaver CJ (2003) ORFB is a subunit of F1F0-ATP synthase: insight into the basis of cytoplasmic male sterility in sunflower. EMBO Rep 4:381–386PubMedCrossRefGoogle Scholar
  33. 33.
    Nakagami H, Pitzschke A, Hirt H (2005) Emerging MAP kinase pathway in plant stress signaling. Trends Plant Sci 10(7):339–346PubMedCrossRefGoogle Scholar
  34. 34.
    Hamel LP, Nicole MC, Sritubtim S, Morency MJ, Ellis M, Ehilting J, Beaudoin N, Barbazuk B, Klessig D, Lee J, Martin G, Mundy J, Ohashi Y, Scheel D, Sheen J, Xing T, Zhang SQ, Seguin A, Ellis BE (2006) Ancient signals: comparative genomics of plant MAPK and MAPKK gene families. Trends Plant Sci 11(4):192–198PubMedCrossRefGoogle Scholar
  35. 35.
    Fujii S, Komatsu S, Toriyama K (2007) Retrograde regulation of nuclear gene expression in CW-CMS of rice. Plant Mol Biol 63(3):405–417PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  1. 1.Laboratory of Genetic Resources & Functional Improvement for Horticultural Plants, Department of HorticultureZhejiang UniversityHangzhouP.R. China
  2. 2.Laboratory of Horticultural Plant Growth, Development & BiotechnologyMinistry of AgricultureHangzhouP.R. China

Personalised recommendations