Molecular Breeding

, 35:83 | Cite as

Molecular marker-assisted pyramiding of broad-spectrum disease resistance genes, Pi2 and Xa23, into GZ63-4S, an elite thermo-sensitive genic male-sterile line in rice

  • Jiefeng Jiang
  • Dabing Yang
  • Jauhar AliEmail author
  • Tongmin MouEmail author


The two-line system of hybrid rice seed production based on thermo-sensitive genic male-sterile (TGMS) lines is much more cost-effective, simple and efficient than the three-line system with cytoplasmic male sterility. In this study, we report the introgression of a rice blast resistance gene Pi2 from VE6219 and a bacterial blight (BB) resistance gene Xa23 from HBQ810 into Guangzhan63-4S (GZ63-4S), an elite TGMS rice line, through marker-assisted pyramiding, leading to the development of Hua1015S, a blast- and BB-resistant pyramided line. The newly developed TGMS line Hua1015S and its derived hybrids showed resistance to rice blast and BB. Hua1015S had a similar critical temperature point of fertility–sterility alteration to GZ63-4S and similar key agronomic and grain quality traits. However, Hua1015S gave higher hybrid seed yields on account of increased outcrossing rates due to better floral traits, e.g. spikelet dehiscence, angle of glume opening, and stigma exsertion. The newly developed TGMS parental line Hua1015S with Pi2 and Xa23 is currently being utilized for heterosis breeding of broad-spectrum blast- and BB-resistant two-line rice hybrids and as an improved disease donor source for further TGMS parental line improvement.


Thermo-sensitive genic male sterile line Blast resistance Bacterial blight resistance Hybrid rice Marker-assisted pyramiding GZ63-4S Pi2 and Xa23 



The research work was supported by Grants from the National High Technology (863) Plan of China (2014AA10A604), Key Technology Research Project of Wuhan City, China (2014020202010136) and the Bill & Melinda Gates Foundation “Green Super Rice for the Resources-Poor of Africa and Asia”.

Supplementary material

11032_2015_282_MOESM1_ESM.pdf (318 kb)
Supplementary material 1 (PDF 318 kb)
11032_2015_282_MOESM2_ESM.pdf (115 kb)
Supplementary material 2 (PDF 114 kb)


  1. Chen S, Li XH, Xu CG, Zhang QF (2000) Improvement of bacterial blight resistance of ‘Minghui 63’, an elite restorer line of hybrid rice, by molecular marker-assisted selection. Crop Sci 40:239–244CrossRefGoogle Scholar
  2. Chen HQ, Chen ZX, Ni S, Zuo SM, Pan XB, Zhu XD (2008) Pyramiding three genes with resistance to blast by marker assisted selection to improve rice blast resistance of Jin 23B. Chin J Rice Sci 22:23–27Google Scholar
  3. Chu Z, Yuan M, Yao J, Ge X, Yuan B, Xu C, Li X, Fu B, Li Z, Bennetzen JL, Zhang Q, Wang S (2006) Promoter mutations of an essential gene for pollen development result in disease resistance in rice. Gene Dev 20:1250–1255CrossRefPubMedCentralPubMedGoogle Scholar
  4. Dai Z, Liu G, Li A, Xu M, Liu X, Zhou C, Zhang H (2005) Breeding of two-line indica hybrid rice combination, “Yangliangyou 6”, and studying on its culture characteristics. Chin Agric Sci Bull 21:114–116Google Scholar
  5. Dellaporta SL, Woo J, Hicks JB (1983) A plant DNA mini preparation: version II. Plant Mol Biol Rep 1:19–21CrossRefGoogle Scholar
  6. Deng YW, Zhu XD, Shen Y, He ZH (2006) Genetic characterization and fine mapping of the blast resistance locus Pigm(t) tightly linked to Pi2 and Pi9 in a broad-spectrum resistant Chinese variety. Theor Appl Genet 113:705–713CrossRefPubMedGoogle Scholar
  7. Fan YR, Zhang QF (2014) Understanding a key gene for thermosensitive genic male sterility in rice. Sci China Life Sci 57:1241–1242CrossRefPubMedGoogle Scholar
  8. Fukuoka S, Saka N, Koga H, Ono K, Shimizu T, Ebana K, Hayashi N, Takahashi A, Hirochika H, Okuno K, Yano M (2009) Loss of function of a proline-containing protein confers durable disease resistance in rice. Science 325:998–1001CrossRefPubMedGoogle Scholar
  9. Gao LG, Gao HL, Li RB, Li DY, Zhou M, Yan Q, Zhou WY, Zhang J, Deng GF (2010) Optimization and verification of molecular for rice bacterial blight resistance gene Xa23. Mol Plant Breed 8:660–664Google Scholar
  10. Hayashi K, Hashimoto N, Daigen M, Ashikawa I (2004) Development of PCR-based SNP markers for rice blast resistance genes at the Piz locus. Theor Appl Genet 108:1212–1220CrossRefPubMedGoogle Scholar
  11. Hittalmani S, Parco A, Mew TV, Zeigler RS, Huang N (2000) Fine mapping and DNA marker-assisted pyramiding of the three major genes for blast resistance in rice. Theor Appl Genet 100:1121–1128CrossRefGoogle Scholar
  12. Huang B, Xu JY, Hou MS, Ali J, Mou TM (2012) Introgression of bacterial blight resistance genes Xa7, Xa21, Xa22 and Xa23 into hybrid rice restorer lines by molecular marker-assisted selection. Euphytica 187:449–459CrossRefGoogle Scholar
  13. IRRI (2002) Standard evaluation system for rice (SES). International Rice Research Institute, Los BanosGoogle Scholar
  14. Ji GH, Zhang SG, Qian J (2003) Preliminary analysis on races of Xanthomonas oryzae pv. oryzae in Yunnan Province. Plant Prot 29:19–21Google Scholar
  15. Jiang HC, Feng YT, Bao L, Li X, Gao GJ, Zhang QL, Xiao JH, Xu CG, He YQ (2012) Improving blast resistance of Jin 23B and its hybrid rice by marker-assisted gene pyramiding. Mol Breed 30:1679–1688CrossRefGoogle Scholar
  16. Khush GS (2001) Green revolution: the way forward. Nat Rev Genet 2:815–822CrossRefPubMedGoogle Scholar
  17. Li CY, Wei J, Lin YJ, Chen H (2012) Gene silencing using the recessive rice bacterial blight resistance gene xa13 as a new paradigm in plant breeding. Plant Cell Rep 31:851–862CrossRefPubMedGoogle Scholar
  18. Liu G, Lu G, Zeng L, Wang GL (2002) Two broad-spectrum blast resistance genes, Pi9(t) and Pi2(t), are physically linked on rice chromosome 6. Mol Genet Genomics 267:472–480CrossRefPubMedGoogle Scholar
  19. Liu H, Yang W, Hu B, Liu F (2007) Virulence analysis and race classification of Xanthomonas oryzae pv. oryzae in China. J Phytopathol 155:129–135CrossRefGoogle Scholar
  20. Liu J, Wang X, Mitchell T, Hu Y, Liu X, Dai L, Wang GL (2010) Recent progress and understanding of the molecular mechanisms of the rice–Magnaporthe oryzae interaction. Mol Plant Pathol 11:419–427CrossRefPubMedGoogle Scholar
  21. Luo YC, Yin ZC (2013) Marker-assisted breeding of Thai fragrance rice for semidwarf phenotype, submergence tolerance and disease resistance to rice blast and bacterial blight. Mol Breed 32:709–721CrossRefGoogle Scholar
  22. Luo YC, Sangha JS, Wang SH, Li ZF, Yang JB, Yin ZC (2012) Marker-assisted breeding of Xa4, Xa21 and Xa27 in the restorer lines of hybrid rice for broad-spectrum and enhanced disease resistance to bacterial blight. Mol Breed 30:1601–1610CrossRefGoogle Scholar
  23. Mackill DJ, Bonman JM (1992) Inheritance of blast resistance in near-isogenic lines of rice. Phytopathology 82:746–749CrossRefGoogle Scholar
  24. Maruthasalam S, Kalpana K, Kumar KK, Loganathan M, Poovannan K, Raja JAJ, Kokiladevi E, Samiyappan R, Sudhakar D, Balasubramanian P (2007) Pyramiding transgenic resistance in elite indica rice cultivars against the sheath blight and bacterial blight. Plant Cell Rep 26:791–804CrossRefPubMedGoogle Scholar
  25. Ou SH (1985) Rice diseases, 2nd edn. Commonwealth Mycological Institute, RichmondGoogle Scholar
  26. Skamnioti P, Gurr SJ (2009) Against the grain: safeguarding rice from rice blast disease. Trends Biotechnol 27:141–150CrossRefPubMedGoogle Scholar
  27. Song WY, Wang GL, Chen LL, Kim HS, Pi LY, Holsten T, Gardner J, Wang B, Zhai WX, Zhu LH, Fauquet C, Ronald P (1995) A receptor kinase-like protein encoded by the rice disease resistance gene, Xa21. Science 270:1804–1806CrossRefPubMedGoogle Scholar
  28. Sundaram RM, Vishnupriya RM, Biradar SK, Laha GS, Ashok Reddy G, Shobha Rani N, Sarma P, Sonti RV (2008) Marker assisted introgression of bacterial blight resistance in Samba Mahsuri, an elite indica rice variety. Euphytica 160:411–422CrossRefGoogle Scholar
  29. Virmani SS, Sun ZX, Mou TM, Ali J, Mao CX (2003) Two-line hybrid rice breeding manual. International Rice Research Institute, Los BanosGoogle Scholar
  30. Wang ZH, Zou YJ, Li XY, Zhang QY, Chen L, Wu H, Su DH, Chen YL, Guo JX, Luo D, Long YM, Zhong Y, Liu YG (2006) Cytoplasmic male sterility of rice with boro II cytoplasm is caused by a cytotoxic peptide and is restored by two related PPR motif genes via distinct modes of mRNA silencing. Plant Cell 18:676–687CrossRefPubMedCentralPubMedGoogle Scholar
  31. Wang CL, Zhang XP, Fan YL, Gao Y, Zhu QL, Zheng CK, Qin TF, Li YQ, Che JY, Zhang MW, Yang B, Liu YG, Zhao KJ (2015) XA23 is an executor R protein and confers broad-spectrum disease resistance in rice. Mol Plant 8:290–302. doi: 10.1093/mp/ssu132 CrossRefPubMedGoogle Scholar
  32. Xu JJ, Wang BH, Wu YH, Du PN, Wang J, Wang M, Yi CD, Gu MH, Liang GH (2011) Fine mapping and candidate gene analysis of ptgms2-1, the photoperiod-thermo-sensitive genic male sterile gene in rice (Oryza sativa L.). Theor Appl Genet 122:365–372CrossRefPubMedGoogle Scholar
  33. Yang QK, Liang CY, Zhuang W, Li J, Deng HB, Deng QY, Wang B (2007) Characterization and identification of the candidate gene of rice thermo-sensitive genic male sterile gene tms5 by mapping. Planta 225:321–330CrossRefPubMedGoogle Scholar
  34. Yi G, Lee SK, Hong YK, Cho YC, Nam MH, Kim SC, Han SS, Wang GL, Hahn TR, Ronald PC, Jeon JS (2004) Use of Pi5(t) markers in marker-assisted selection to screen for cultivars with resistance to Magnaporthe grisea. Theor Appl Genet 109:978–985CrossRefPubMedGoogle Scholar
  35. Yuan L (2004) Hybrid rice technology for food security in the world. Crop Res 18:185–186Google Scholar
  36. Zeng LX, Huang SH, Wu SZ (2002) The resistance of IRBB21 (Xa21) against 5 races of Guangdong Province. Acta Phytopathol Sin 29:97–100Google Scholar
  37. Zhang Q (2009) Genetics and improvement of bacterial blight resistance of hybrid rice in China. Rice Sci 16:83–92CrossRefGoogle Scholar
  38. Zhang Q, Lin SC, Zhao BY, Wang CL, Yang WC, Zhou YL, Li DY, Chen CB, Zhu LH (1998) Identification and tagging a new gene for resistance to bacterial blight (Xanthomonas oryzae pv. oryzae) from O. rufipogon. Rice Genet Newsl 15:138–142Google Scholar
  39. Zhang Q, Wang CL, Zhao KJ, Zhao YL, Caslana VC, Zhu XD, Li DY, Jiang QX (2001) The effectiveness of advanced rice lines with new resistance gene Xa23 to rice bacterial blight. Rice Genet Newsl 18:71–72Google Scholar
  40. Zhou B, Qu S, Liu G, Dolan M, Sakai H, Lu G, Bellizzi M, Wang GL (2006) The eight amino-acid differences within three leucine-rich repeats between Pi2 and Piz-t resistance proteins determine the resistance specificity to Magnaporthe grisea. Mol Plant Microbe Interact 19:1216–1228CrossRefPubMedGoogle Scholar
  41. Zhou YL, Xu JL, Zhou SC, Yu J, Xie XW, Xu MR, Sun Y, Zhu LH, Fu BY, Gao YM, Li ZK (2009) Pyramiding Xa23 and Rxo1 for resistance to two bacterial diseases into an elite indica rice variety using molecular approaches. Mol Breed 23:279–287CrossRefGoogle Scholar
  42. Zhou YL, Uzokwe VNE, Zhang CH, Cheng LR, Wang L, Chen K, Gao XQ, Sun Y, Chen JJ, Zhu LH, Zhang Q, Ali J, Xu JL, Li ZK (2011) Improvement of bacterial blight resistance of hybrid rice in China using the Xa23 gene derived from wild rice (Oryza rufipogon). Crop Prot 30:637–644CrossRefGoogle Scholar
  43. Zhou H, Zhou M, Yang YZ, Li J, Zhu LY, Jiang DG, Dong JF, Liu QJ, Gu LF, Zhou LY, Feng MJ, Qin P, Hu XC, Song CL, Shi JF, Song XW, Ni ED, Wu XJ, Deng QY, Liu ZL, Chen MS, Liu YG, Cao XF, Zhuang CX (2014) RNase ZS1 processes Ub L40 mRNAs and controls thermosensitive genic male sterility in rice. Nat Commun 5:4884–4892CrossRefPubMedGoogle Scholar
  44. Zhu X, Chen S, Yang J, Zhou S, Zeng L, Han J, Su J, Wang L, Pan Q (2012) The identification of Pi50(t), a new member of the rice blast resistance Pi2/Pi9 multigene family. Theor Appl Genet 124:1295–1304CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  1. 1.National Key Laboratory of Crop Genetic ImprovementHuazhong Agricultural UniversityWuhanChina
  2. 2.International Rice Research InstituteMetro ManilaPhilippines

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