, Volume 217, Issue 4, pp 559–565 | Cite as

Male-sterility of thermosensitive genic male-sterile rice is associated with premature programmed cell death of the tapetum

  • Sujin Ku
  • Hyejin Yoon
  • Hak Soo Suh
  • Yong-Yoon Chung
Original Article


The tapetum plays a crucial role in pollen development. This secretory tissue produces numerous nutritive proteins necessary for pollen maturation. The tapetum, whose cells undergo programmed cell death (PCD), is completely diminished by the time the pollen is fully mature. Our previous studies on a thermosensitive genic male-sterile (TGMS) rice (Oryza sativa L.) suggested that male-sterility was due to failure in pollen development. In this paper we describe how further analysis of the TGMS rice revealed that male-sterility is associated with premature PCD of the tapetum. Cytological observations of TGMS rice anthers at various developmental stages indicated that PCD initiates at an early stage of pollen development and continues until the tapetal cells are completely degraded, resulting in pollen collapse. Transmission electron microscopy showed the morphologically distinct hallmarks of apoptosis, including cytoplasmic shrinkage, membrane blebbing, and vacuolation. Identification of DNA fragmentation using the TUNEL assay supports the hypothesis that premature PCD is associated with male-sterility in the rice. The tissue-specific feature of the thermosensitive genic male-sterile phenotype is discussed with regard to PCD during anther development.


Anther Male-sterility Oryza Programmed cell death Tapetum 



cytoplasmic male-sterility


environmental genic male-sterility


programmed cell death


thermo-sensitive genic male-sterility


TdT-mediated dUTP nick-end labelling



This work was supported by the Center for Plant Molecular Genetics and Breeding Research, Korea Science and Engineering Foundation. We acknowledge the Wild Crop Germplasm Bank of Yeungnam University for providing the TGMS rice used in this study. We also thank Dr. Martin B. Dickman of the University of Nebraska for his comments regarding this manuscript and Sanghyun Lee for his helps in preparing figures. S. Ku and H. Yoon contributed equally to this work.


  1. Balk J, Leaver CJ (2001) The PET1-CMS mitochondrial mutation in sunflower is associated with premature programmed cell death and cytochrome c release. Plant Cell 13:1803–1818PubMedGoogle Scholar
  2. Bethke PC, Lonsdale JE, Fath A, Jones, RL (1999) Hormonally regulated programmed cell death in barley aleurone cells. Plant Cell 11:1033–1045CrossRefPubMedGoogle Scholar
  3. Goetz M, Godt DE, Guivarc'h A, Kahmann U, Chriqui D, Roitsch T (2001) Induction of male sterility in plants by metabolic engineering of the carbohydrate supply. Proc Natl Acad Sci USA 98:6522–6527CrossRefPubMedGoogle Scholar
  4. Hernould M, Suharsono, Zabaleta E, Carde JP, Litvak S, Araya A, Mouras A (1998) Impairment of tapetum and mitochondria in engineered male-sterile tobacco plants. Plant Mol Biol 36:499–508CrossRefPubMedGoogle Scholar
  5. Heslop-Harrison J (1976) The adaptive significance of the exine. In: Ferguson IK, Muller J (eds) The evolutionary significance of the exine. Academic Press, New York, pp 27–38Google Scholar
  6. Kapoor S, Kobayashi, A, Takatsuji, H (2002) Silencing of the tapetum-specific zinc finger gene TAZ1 causes premature degeneration of tapetum and pollen abortion in petunia. Plant Cell 14:2353–2367CrossRefPubMedGoogle Scholar
  7. Kaul M (1988) Male sterility in higher plants, vol 10. Springer, Berlin Heidelberg New YorkGoogle Scholar
  8. Kofer W, Glimelius K, Bonnett HT (1991) Modifications of mitochondrial DNA cause changes in floral development in homeotic-like mutants of tobacco. Plant Cell 3:759–769CrossRefPubMedGoogle Scholar
  9. Ku S-J, Cho K-H, Choi Y-J, Baek W-K, Kim S, Suh H-S, Chung Y-Y (2001) Cytological observation of two environmental genic male-sterile lines of rice. Mol Cells 12:403–406PubMedGoogle Scholar
  10. Laser KD, Lersten NR (1972) Anatomy and cytology of microsporogenesis in cytoplasmic male sterile angiosperms. Bot Rev 38:425–454Google Scholar
  11. Mackenzie S, Shichuan H, Lyznik A (1994) The elusive plant mitochondrion as a genetic system. Plant Physiol 105:775–780PubMedGoogle Scholar
  12. Mariani C, Beuckeleer MD, Truettner J, Leemans J, Goldberg RB (1990) Induction of male sterility in plants by a chimaeric ribonuclease gene. Nature 347:737–741Google Scholar
  13. Meuter-Gerhards A, Riegart S, Wiermann R (1999) Studies on sporopollenin biosynthesis in Cucurbita maxima (DUCH-II): the involvement of aliphatic metabolism. J. Plant Physiol 154:431–436Google Scholar
  14. Mittler R, Lam E (1996) Sacrifice in the face of foes: pathogen-induced programmed cell death. Trends Microbiol 4:10–15CrossRefPubMedGoogle Scholar
  15. Mittler R, Feng X, Cohen M (1998) Post-transcriptional suppression of cytosolic ascorbate peroxidase expression during pathogen-induced programmed cell death in tobacco. Plant Cell 10:461–473Google Scholar
  16. Paxson-Sowders DJ, Dodrill CJ, Owen HA, Makaroff CA (2001) DEX1, a novel plant protein, is required for exine pattern formation during pollen development in Arabidopsis. Plant Physiol 127:1739–1749Google Scholar
  17. Rozema J, Croekman RA, Blokker P, Meijkamp BB, de Bakker N, van de Staaij J, van Beem A, Ariese F, Kars SM (2001) UV-B absorbance and UV-B absorbing compounds (para-coumaric acid) in pollen and sporopollenin: the perspective to track historic UV-B levels. J Photochem Photobiol B 62:108–117CrossRefPubMedGoogle Scholar
  18. Schnabel PS, Wise RP (1998) The molecular basis of cytoplasmic male-sterility and fertility restoration. Trends Plant Sci 3:175–180CrossRefGoogle Scholar
  19. Stanley RG, Linskens HF (1974) Pollen: biology, biochemistry, management. Springer, Berlin Heidelberg New York, pp 13–28Google Scholar
  20. Van der Meer IM, Stam ME, van Tunen AJ, Mol JNM, Stuitje AR (1992) Antisense inhibition of flavonoid biosynthesis in petunia anthers results in male sterility. Plant Cell 4:253–262CrossRefPubMedGoogle Scholar
  21. Wang M, Hoekstra S, van Bergen S, Lamers GEM, Oppedijk BJ, van der Heijden MW, de Priester W, Schilperoort RA (1999) Apoptosis in developing anthers and the role of ABA in this process during androgenesis in Hordeum vulgare L. Plant Mol Biol 39:489–501PubMedGoogle Scholar
  22. Xu Y, Hanson MR (2000) Programmed cell death during pollination-induced petal senescence in petunia. Plant Physiol 122:1323–1333CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • Sujin Ku
    • 1
  • Hyejin Yoon
    • 1
  • Hak Soo Suh
    • 2
  • Yong-Yoon Chung
    • 1
  1. 1.School of Life Sciences and BiotechnologyKorea UniversitySeoulKorea
  2. 2.School of Biological Resources, College of Natural ResourcesYeungnam UniversityKyongsanKorea

Personalised recommendations