Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Assessing hybrid sterility in Oryza glaberrima × O. sativa hybrid progenies by PCR marker analysis and crossing with wide compatibility varieties


Interspecific crossing of the African indigenous rice Oryza glaberrima with Oryza sativa cultivars is hindered by crossing barriers causing 100% spikelet sterility in F1 hybrids. Since hybrids are partially female fertile, fertility can be restored by back crossing (BC) to a recurrent male parent. Distinct genetic models on spikelet sterility have been developed predicting, e.g., the existence of a gamete eliminator and/or a pollen killer. Linkage of sterility to the waxy starch synthase gene and the chromogen gene C, both located on chromosome 6, have been demonstrated. We selected a segregating BC2F3 population of semi-sterile O. glaberrima × O. sativa indica hybrid progenies for analyses with PCR markers located at the respective chromosome-6 region. These analyses revealed that semi-sterile plants were heterozygous for a marker (OSR25) located in the waxy promoter, whereas fertile progenies were homozygous for the O. glaberrima allele. Adjacent markers showed no linkage to spikelet sterility. Semi-sterility of hybrid progenies was maintained at least until the F4 progeny generation, suggesting the existence of a pollen killer in this plant material. Monitoring of reproductive plant development showed that spikelet sterility was at least partially due to an arrest of pollen development at the microspore stage. In order to address the question whether genes responsible for F1 sterility in intraspecific hybrids (O. sativa indica × japonica) also cause spikelet sterility in interspecific hybrids, crossings with wide compatibility varieties (WCV) were performed. WCV accessions possess "neutral" S-loci (S n) improving fertility in intraspecific hybrids. This experiment showed that the tested S n-loci had no fertility restoring effect in F1 interspecific hybrids. Pollen development was completely arrested at the microspore stage and grains were never obtained after selfing. This suggests that distinct or additional S-loci are responsible for sterility of O. glaberrima × O. sativa hybrids.

This is a preview of subscription content, log in to check access.

Fig. 1.
Fig. 2.
Fig. 3a–e.


  1. Akagi H, Yokozeki Y, Inagaki A, Fujimura T (1996) Microsatellite DNA markers for rice chromosomes. Theor Appl Genet 93:1071–1077

  2. Bougerol B, Pham JL (1989) Influence of the Oryza sativa genotype on the fertility and quantitative traits of F1 hybrids between the two cultivated rice species O. sativa L. and O. glaberrima Steud. Genome 32:810–815

  3. Causse MA, Fulton TM, Gu Cho Y, Nag Ahn S, Chunwongse J, Wu K, Xiao J, Yu Z, Ronald PC, Harrington SE, Second G, McCouch SR, Tanksley SD (1994) Saturated molecular map of the rice genome based on an interspecific backcross population. Genetics 138:1251–1274

  4. Chen X, Temnykh S, Xu Y, Cho YG, McCouch SR (1997) Development of a microsatellite framework map providing genome-wide coverage in rice (Oryza sativa L.). Theor Appl Genet 95:553–567

  5. Chu YE, Morishima H, Oka HI (1969) Reproductive barriers distributed in cultivated rice species and their wild relatives. Jpn J Genet 44:207–223

  6. Dingkuhn M, Jones M, Johnson DE, Sow A (1998) Growth and yield potential of Oryza sativa and O. glaberrima upland rice cultivars and their interspecific progenies. Field Crops Res 57:57–69

  7. Haefele SM, Wopereis MCS, Wiechmann H (2002) Long-term fertility experiments for irrigated rice in the West African Sahel: agronomic results. Field Crops Res 132:31–40

  8. Harushima Y, Yano M, Shomura A, Sato M, Shimano T, Kuboki Y, Yamamoto T, Yang Lin S, Antonio BA, Parco A, Kajiya H, Huang N, Yamamoto K, Nagamura Y, Kurata N, Khush GS, Sasaki T (1998) A high-density rice genetic linkage map with 2,275 markers using a single F2 population. Genetics 148:479–494

  9. Heuer S, Loerz H, Dresselhaus T (2000) The maize MADS box gene ZmMADS2 is specifically expressed in maize pollen and during maize pollen-tube growth. Sex Plant Reprod 13:21–27

  10. Heuer S, Miézan KM, Sié M, Gaye G (2003) Increasing biodiversity of irrigated rice in Africa by interspecific crossing of Oryza glaberrima (Steud.) × O. sativa indica (L.). Euphytica 132:31–40

  11. Ikehashi H, Akari H (1986) Genetics of F1 sterility in remote crosses of rice. In: Kush GS (ed) Rice genetics. Proc Int Rice Genetics Symposium. International Rice Research Institute, Manila, The Philippines, pp 119–130

  12. Ikehashi H, Wan J (1998) The wide compatibility system: current knowledge of its genetics and use for enhanced yield heterosis. In: Virmani SS, Siddiq EA, Muralidharan K (eds) Advances in hybrid rice technology. International Rice Research Institute, Manila, The Philippines, pp 67–77

  13. Jones MP, Mande S, Aluko K (1997a) Diversity and potential of Oryza glaberrima Steud. in upland rice breeding. Breed Sci 47:395–398

  14. Jones MP, Dingkuhn M, Aluko GK, Semon M (1997b) Interspecific Oryza sativa L. × O. glaberrima Steud. progenies in upland rice improvement. Euphytica 92:237–246

  15. Kinoshito T (1993) Report of the committee on gene symbolization, nomenclature and linkage groups. RGN 10:7–39

  16. Lin SY, Ikehashi H, Yanagihara S, Kawashima A (1992) Segregation distortion via male gametes in hybrids between Indica and Japonica or wide-compatibility varieties of rice (Oryza sativa L.). Theor Appl Genet 84:812–818

  17. Lorieux M, Ndjiondjop MN, Ghesquière A (2000) A first interspecific Oryza sativa × Oryza glaberrima microsatellite-based genetic linkage map. Theor Appl Genet 100:593–601

  18. McCouch SR, Kochert G, Yu ZH, Wang ZY, Khush GS (1988) Molecular mapping of rice chromosomes. Theor Appl Genet 76:815–829

  19. Oka HI (1974) Analysis of genes controlling F1 sterility in rice by the use of isogenic lines. Genetics 77:521–534

  20. Pallotta MA, Graham RD, Langridge P, Sparrow DHB, Barker SJ (2000) RFLP mapping of manganese efficiency in barley. Theor Appl Genet 101:1100–1108

  21. Sano Y (1986) Sterility barriers between Oryza sativa and O. glaberrima. In: Kush GS (ed) Rice genetics. Proc Int Rice Genetics Symposium International Rice Research Institute, Manila, The Philippines, pp 109–118

  22. Sano Y (1990) The genetic nature of gamete eliminator in rice. Genetics 125:183–191

  23. Sano Y (1992) Genetic comparison of chromosome 6 between wild and cultivated rice. Jpn J Breed 42:561–572

  24. Sano Y (1994) Gamete eliminator adjacent to the wx locus as revealed by pollen analysis in rice. J Hered 85:310–312

  25. Sano Y, Chu YE, Oka HI (1979) Genetic studies of speciation in cultivated rice. 1. Genetic analysis for the F1 sterility between O. sativa L. and O. glaberrima STEUD. Jpn J Genet 54:121–132

  26. Tomita M (1996) The gametic lethal gene gal: activated only in the presence of the semidwarfing gene d60 in rice. In: Kush GS (ed) Rice genetics III. Proc 3rd Int Rice Genetics Symposium. International Rice Research Institute, Manila, The Philippines, pp 396–403

  27. Xu Y, Zhu L, Xiao J, Huang N, McCouch SR (1997) Chromosomal regions associated with segregation distortion of molecular markers in F2, backcross, doubled-haploid, and recombinant inbred populations in rice (Oryza sativa L.). Mol Gen Genet 253:535–545

  28. Yanagihara S, McCouch SR, Ishikawa K, Ogi Y, Maruyama K, Ikehashi H (1995) Molecular analysis of the inheritance of the S-5 locus, conferring wide compatibility in Indica/Japonica hybrids of rice (O. sativa L.). Theor Appl Genet 90:182–188

  29. Zhang G, Lu Y (1996) Genetics of F1 pollen sterility in O. sativa. In: Kush GS (ed) Rice genetics III. Proc 3rd Int Rice Genetics Symposium. International Rice Research Institute, Manila, The Philippines, pp 418–422

Download references


We thank the Deutsche Akademischer Austauschdienst (DAAD) for granting a post-doc scholarship for Dr. S. Heuer (D/99/18141). We are grateful to Dr. S. Braconnier and colleagues at the CERAAS (CIRAD) institute in Thiès (Senegal), as well as to Prof. Dr. H. Loerz, Dr. T. Dresselhaus and the team at Hamburg University (Germany), for the provision of greenhouse and laboratory facilities. Many thanks to Dr. A. Ghesquière and Dr. M. Lorieux (IRD, Montpellier, France) for the PCR primers and stimulating discussions.

Author information

Correspondence to Sigrid Heuer.

Additional information

Communicated by H.C. Becker

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Heuer, S., Miézan, K.M. Assessing hybrid sterility in Oryza glaberrima × O. sativa hybrid progenies by PCR marker analysis and crossing with wide compatibility varieties. Theor Appl Genet 107, 902–909 (2003).

Download citation


  • Crossing barriers
  • Hybrid rice
  • Oryza glaberrima
  • Oryza sativa
  • Pollen