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Allelic differentiations and effects of the Rf3 and Rf4 genes on fertility restoration in rice with wild abortive cytoplasmic male sterility

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Biologia Plantarum


To reveal the allelic differentiations at the two genes for fertility restoration (Rf) on chromosomes 1 (Rf3) and 10 (Rf4), 15 chromosome single segment substitution lines (SSSLs) with the Rf3 locus and 18 SSSLs with the Rf4 locus were crossed with Bobai A (BbA), a cytoplasmic male sterility line with wild abortive type of cytoplasm (WA-CMS), respectively. Based on the pollen and seed fertility of the F1 hybrids, the Rf3 and Rf4 genes were each classified into four alleles, namely Rf3-1, Rf3-2, Rf3-3, and Rf3-4 for Rf3, and Rf4-1, Rf4-2, Rf4-3, and Rf4-4 for Rf4. Out of the 33 SSSLs, an SSSL W23-19-06-06-11 carrying the genotype Rf3-4Rf3-4/Rf4-4Rf4-4 possessed the strongest restoring ability for BbA. To determine the genetic effects of Rf3 and Rf4 for WA-CMS, one BC3F2 population possessing the genetic background of W23-19-06-06-11 was generated from the cross between W23-19-06-06-11 and BbA by backcrossing and marker-assisted selection. In the BC3F2 population, the plants carrying the Rf3Rf3/Rf4Rf4, Rf3Rf3/rf4rf4, and rf3rf3/Rf4Rf4 genotypes were selected and their phenotyping for pollen and spikelet fertility were evaluated. The result showed that under the genetic background of SSSL W23-19-06-06-11, the effect of Rf4 appeared to be slightly larger than that of Rf3 and their effects were additive for WA-CMS system. These studies will lead to the transfer of Rf genes into adapted cultivars through marker-assisted selection in active hybrid rice breeding programs.

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cytoplasmic male sterility


marker-assisted selection


polymerase chain reaction


pentatricopeptide repeat


quantitative trait loci

Rf :

fertility restorer gene


restriction fragment length polymorphism


single segment substitution lines


wild abortive


  • Bentolila, S., Alfonso, A.A., Hanson, M.R.: A pentatricopeptide repeat-containing gene restores fertility to cytoplasmic male sterile plants. — PNAS 99: 10887–10892, 2002.

    Article  PubMed  CAS  Google Scholar 

  • Chase, C., Babay-Laughnan, S.: Cytoplasmic male sterility and fertility restoration by nuclear genes. — In: Daniell, H., Chase, C. (ed.): Molecular Biology and Biotechnology of Plant Organelles. Pp. 593–622. Kluwer Academic Publishers, Dordrecht 2004.

    Chapter  Google Scholar 

  • Chen, J.J., Ding, J.H., Ouyang, Y.D., Du, H.Y., Yang, J.Y., Cheng, K., Zhao, J., Qiu, S.Q., Zhang, X.L., Yao, J.L., Liu, K.D., Wang, L., Xu, C.G., Li, X.H., Xue, Y.B., Xia, M., Ji, Q., Lu, J., Xu, M..L., Zhang, Q.F.: A triallelic system of S5 is a major regulator of the reproductive barrier and compatibility of indica-japonica hybrids in rice. — Proc. nat. Acad. Sci. USA 105: 11436–11441, 2008.

    Article  PubMed  CAS  Google Scholar 

  • Dellaporta, R.P., Wood, J., Hicks, J.D.: A plant DNA minipreparation: version II. — Plant mol. Biol. Rep. 1: 19–21, 1983.

    Article  CAS  Google Scholar 

  • Hanson, M.R., Bentolila, S.: Interactions of mitochondrial and nuclear genes that affect male gametophytic development. — Plant Cell 16 (Suppl.): S154-S169, 2004.

  • He, F.H., Zeng, R.Z., Xi, Z.Y., Talukdar, A., Zhang, G.Q.: [Genetic diversity of different waxy genotypes in rice.] — Mol. Plant Breed. 1: 179–186, 2003. [In Chin.]

    CAS  Google Scholar 

  • He, F.H., Xi, Z.Y., Zeng, R.Z., Talukdar, A., Zhang, G.Q.: [Mapping of heading date QTLs in rice (Oryza sativa L.) using single segment substitution lines.] — Sci. agr. sin. 38: 1505–1513, 2005a. [In Chin.]

    CAS  Google Scholar 

  • He, F.H., Xi, Z.Y., Zeng, R.Z., Talukdar, A., Zhang, G.Q.: [Developing single segment substitution lines (SSSLs) in rice (Oryza sativa L.) using advanced backcrosses and MAS.] — Acta genet. sin. 32: 825–831, 2005b. [In Chin.]

    PubMed  Google Scholar 

  • Ikehashi, H., Araki, H.: Genetics of F1 sterility in remote crosses of rice (Oryza sativa L.). — In: Rice Genetics. Pp. 119–130. International Rice Research Institute, Manila 1986.

    Google Scholar 

  • Jing, R., Li, X., Yi, P., Zhu, Y.: Mapping fertility restoring genes of rice WA cytopasmic male sterility using SSLP markers. — Bot. Bull. Acad. sin. 42: 167–171, 2001.

    CAS  Google Scholar 

  • Li, S., Yang, D.C., Zhu, Y.G.: Characterization and use of male sterility in hybrid rice breeding. — J. Integrative Plant Biol. 49: 791–804, 2007.

    Article  CAS  Google Scholar 

  • Lin, S.C., Yuan, L.P.: Hybrid rice breeding in China. — In: Innovative Approaches to Rice Breeding. Pp. 35–51. International Rice Research Institute, Manila 1980.

    Google Scholar 

  • Liu, G.F., Zeng, R.Z., Zhu, H.T., Zhang, Z.M., Ding, X.H., Zhao, F.M., Li, W.T., Zhang, G.Q.: Dynamic expression of nine QTLs for tiller number detected with single segment substitution lines in rice. — Theor. appl. Genet. 118: 443–453, 2009.

    Article  PubMed  Google Scholar 

  • Liu, G.F., Zhang, Z.M., Zhu, H.T., Zhao, F.M., Ding, X.H., Zeng, R.Z., Li, W.T., Zhang, G.Q.: Detection of QTLs with additive effects and additive-by-environment interaction effects on panicle number in rice (Oryza sativa L.) with single-segment substitution lines. — Theor. appl. Genet. 116: 923–931, 2008.

    Article  PubMed  CAS  Google Scholar 

  • Liu, G.F., Zhu, H.T., Liu, S., Zeng, R.Z., Zhang, Z.M., Li, W.T., Ding, X.H., Zhao, F.M., Zhang, G.Q.: Unconditional and conditional QTL mapping for the developmental behavior of tiller number in rice (Oryza sativa L.). — Genetics 138: 885–893, 2010.

    Google Scholar 

  • Long, Y.M., Zhao, L.F., Niu, B.X., Su, J., Wu, H., Chen, Y.L., Zhang, Q.Y., Guo, J.X., Zhuang, C.Q., Mei, M.T., Xia, J.X., Wang, L., Wu, H.B., Liu, Y.G.: Hybrid male sterility in rice controlled by interaction between divergent alleles of two adjacent genes. — Proc. nat. Acad. Sci. USA 105: 18871–18876, 2008.

    Article  PubMed  CAS  Google Scholar 

  • McCouch, S.R.., Teytelman, L., Xu, Y.B., Lobos, K.B., Clare, K., Walton, M., Fu, B., Maghiran, R., Li, Z., Xing, Y., Zhang, Q., Kono, I., Yano, M., Jellstrom, R.F., Declerck, G., Schneider, D., Cartinhour, S., Ware, D., Stein, L.: Development and mapping of 2240 new SSR markers for rice (Oryza sativa L.). — DNA Res. 9: 199–207, 2002.

    Article  PubMed  CAS  Google Scholar 

  • Mishra, G.P., Singh, R.K., Mohapatra, T., Singh, A.K., Prabhu, K.V., Zaman, F.U., Sharma, R.K.: Molecular mapping of a gene for fertility restoration of wild abortive (WA) cytoplasmic male sterility using a basmati rice restorer line. — J Plant Biochem. Biotechnol. 12: 37–42, 2003.

    Article  CAS  Google Scholar 

  • Oka, H.I.: Experimental studies on the origin of cultivated rice. — Genetics 78: 475–486, 1974.

    PubMed  CAS  Google Scholar 

  • Panaud, O., Chen, X., McCouch, S.R.: Development of microsatellite markers and characterization of simple sequence length polymorphism (SSPL) in rice (Oryza sativa L.). — Mol. gen. Genet. 252: 597–607, 1996.

    PubMed  CAS  Google Scholar 

  • Peleman, J.D., Van der Voort, J.R.: Breeding by design. — Trends Plant Sci. 8: 330–334, 2003.

    Article  PubMed  CAS  Google Scholar 

  • Sattari, M., Kathiresan, A., Glenn, B., Gregorio, S., Virmani, S.: Comparative genetic analysis and molecular mapping of fertility restoration genes for WA, Dissi, and Gambiaca cytoplasmic male sterility systems in rice. — Euphytica 160: 305–315, 2008.

    Article  CAS  Google Scholar 

  • Sheeba, N.K., Viraktamath, B.C., Sivaramakrishnan, S., Gangashetti, M.G,. Khera, P., Sundaram, R.M.: Validation of molecular markers linked to fertility restorer gene (s) for WA-CMS lines of rice. — Euphytica 167: 217–227, 2009.

    Article  CAS  Google Scholar 

  • Sun, Y., Gu, Y.J., Zhang, H.G., Tian, S., Tang, S.Z., Gu, M.H.: [Genetic effects of 3 different male sterile cytoplasms in rice.] — J. Yangzhou Univ. 27(2): 1–4, 2006. [In Chin.]

    Google Scholar 

  • Tan, X.L., Vanavichit, A., Amornsilpa, S., Trangoonrung, S.: Genetic analysis of rice CMS-WA fertility restoration based on QTL mapping. — Theor. appl. Genet. 96: 994–999, 1998.

    Article  Google Scholar 

  • Tan, Y.P., Li, S.Q., Wang, L., Liu, G., Hu, J. Zhu, Y.G.: Genetic analysis of fertility-restorer genes in rice. — Biol. Plant. 52: 469–474, 2008.

    Article  Google Scholar 

  • Teng, B., Zeng, R.Z., Wang, Y.C., Liu, Z.Q., Zhang, Z.M., Zhu, H.T., Ding, X.H., Li, W.T., Zhang, G.Q.: Detection of allelic variation at the Wx locus with single-segment substitution lines in rice (Oryzasativa L.). — Mol. Breed. 30: 583–595, 2012.

    Article  Google Scholar 

  • Wang, S.K., Wu, K., Yuan, Q.B., Liu, X.Y., Liu, Z.B., Lin, X.Y., Zeng, R.Z., Zhu, H.T., Dong, G.J., Qian, Q., Zhang, G.Q., Fu X.D.: Control of grain size, shape and quality by OsSPL16 in rice. — Natur. Genet. doi:10.1038/ng.2327, 2012.

  • Xi, Z.Y., He, F.H., Zeng, R.Z., Zhang, Z.M., Ding, X.H., Li, W.T., Zhang, G.Q.: Development of a wide population of chromosome single-segment substitution lines in the genetic background of an elite cultivar of rice (Oryza sativa L.). — Genome 49: 476–484, 2006.

    Article  PubMed  CAS  Google Scholar 

  • Yano, M., Sasaki, T.: Genetic and molecular dissection of quantitative traits in rice. — Plant mol. Biol. 35: 145–153, 1997.

    Article  PubMed  CAS  Google Scholar 

  • Yao, F.Y., Xu, C.G., Yu, S.B., Li, J.X., Gao, Y.J., Li, X.H., Zhang, Q.: Mapping and genetic analysis of two fertility restorer loci in the wild abortive cytoplasmic male sterility system of rice (Oryza sativa L.). — Euphytica 98: 183–187, 1997.

    Article  CAS  Google Scholar 

  • Yuan, L.P.: The execution and theory of developing hybrid rice. — Chin. agr. Sci. 1: 27–31, 1977.

    Google Scholar 

  • Zhang, G., Bharaj, T.S., Virmani, S.S., Huang, N.: Mapping of the Rf-3 nuclear fertility-restoring gene for WA cytoplasmic male sterility in rice using RAPD and RFLP markers. — Theor. appl. Genet. 94: 27–33, 1997.

    Article  PubMed  CAS  Google Scholar 

  • Zhang, G., Lu, Y., Zhang, H., Yang, J., Liu, G.: [Genetic studies on the hybrid sterility in cultivated rice (Oryza sativa L.) IV. Genotypes for F1 pollen sterility.] — Acta genet. sin. 21: 34–41, 1994. [In Chin.]

    CAS  Google Scholar 

  • Zhang, G.Q., Lu, Y.G.: [Genetic studies of the hybrid sterility in cultivated rice (Oryza sativa) II. A genic model for F1 pollen sterility.] — Acta genet. sin. 20: 222–228, 1993. [In Chin.]

    Google Scholar 

  • Zhang, G.Q., Zeng, R.Z., Zhang, Z.M., Ding, X.H., Li, W.T., Liu, G.M., He, F.H., Tulukdar, A., Huang, C.F., Xi, Z.Y., Qin, L.J., Shi, J.Q., Zhao, F.M., Feng, M.J., Shan, Z.L., Chen, L., Guo, X.Q., Zhu, H.T., Lu, Y.G.: The construction of a library of single segment substitution lines in rice (Oryza sativa L.). — Rice Genet. Newslett. 21: 85–87, 2004.

    Google Scholar 

  • Zhang, Q.Y., Liu, Y.G., Mei, M.T.: Molecular mapping of the fertility restorer gene Rf4 for WA cytoplasmic male sterility. — Acta Genet. sin. 29: 1001–1004, 2002.

    PubMed  Google Scholar 

  • Zhang, Q.Y., Lu, Y.G.: [Genetic studies of the hybrid sterility in cultivated rice (Oryza sativa L.). I. Diallel analysis of the hybrid sterility among isogenic F1 sterile lines.] — Chin. J. Rice Sci. 3: 97–101, 1989. [In Chin.]

    Google Scholar 

  • Zhang, Y.X., Yang, J.Y., Shan, Z.L., Chen, S., Qiao, W.H., Zhu, X.Y., Xie, Q.J., Zhu, H.T., Zhang, Z.M., Zeng, R.Z., Ding, X.H., Zhang, G.Q.: Substitution mapping of QTLs for blast resistance with SSSLs in rice (Oryza sativa L.). — Euphytica 184: 141–150, 2012.

    Article  Google Scholar 

  • Zhu, Y.G.: [Studies on rice male sterility and its restoration.]. — In: Zhu, Y.G., Li, Y.Q., Wang, M.Q. (ed.): Biology of Male Sterility in Rice. Pp. 398–438. Wuhan University Press, Wuhan 2000. [In Chin.]

    Google Scholar 

  • Zhu, Y.G.: [Studies on male sterile lines of rice with different cytoplasms.]. — Acta agron. sin. 5: 29–38, 1979. [In Chin.]

    Google Scholar 

  • Zhuang, J.Y., Fan, Y.Y., Wu, J.L., Rao, Z.M., Xia, Y.W., Zheng, K.L.: [Maping genes for rice CMS-WA fertility restoration.] — Acta genet. sin. 28: 129–134, 2001. [In Chin.]

    PubMed  CAS  Google Scholar 

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Correspondence to G. -Q. Zhang.

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Acknowledgements: This work was supported by National Natural Science Foundation of China (30830074) and Department of Education of Anhui Province Natural Science Foundation of China (No. KJ2010B155).

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Cai, J., Liao, Q.P., Dai, Z.J. et al. Allelic differentiations and effects of the Rf3 and Rf4 genes on fertility restoration in rice with wild abortive cytoplasmic male sterility. Biol Plant 57, 274–280 (2013).

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