, Volume 187, Issue 3, pp 461–469 | Cite as

Identification and molecular cytology analysis of cold tolerance introgression lines derived from Oryza sativa L. mating with O. rufipogon Griff.

  • Xiang-Dong Luo
  • Liang-Fang Dai
  • Juan-Fang Cao
  • Shui-Rong Jian
  • Ya-Ling Chen
  • Biao-Lin Hu
  • Jian-Kun Xie


A previously constructed introgression lines (ILs) population including 239 lines was used to evaluate the cold-tolerant ability. The ILs with strong cold tolerance which were identified in the present study were used for further cytological and microsatellite (SSR) marker analyses. The results showed that the IL5243 and IL5335 had strong cold-tolerant ability. Cytological analysis showed that the rate of pollen mother cells (PMCs) with normal meiotic behavior in the IL5243 and IL5335 was to 89.93 and 90.22 %, respectively, and finally formed normal mature pollen. And meanwhile, the low frequency of abnormal chromosome behavior was observed in the IL5243 and IL5335, such as univalent, 8-shape bivalent, multivalent at diakinesis. At anaphase I, one or two lagging chromosomes were observed in some PMCs (3.95–5.15 %). The results of SSR marker analysis further confirmed that partial alien DNA of common wild rice has been transferred into the IL5243 and IL5335. These results implied that the IL5243 and IL5335 might be excellent bridging germplasm for exploring and utilizing the cold-tolerant gene of common wild rice. In addition, IL5243 and IL5335 would provide a better experimental system for understanding some epigenetic phenomenon induced by alien gene introgression.


Introgression Cold tolerance Meiosis Chromosome recombination 



This research was partially supported by the National Natural Science Foundation of China (30860120 and 30900781), the Natural Science Foundation of Jiangxi Province, China (2008GQN0059), the Development Program for Young Scientists of Jiangxi Province, China (20112BCB23007), and the Scientific Planning Project of Jiangxi Provincial Education Department (GJJ12184).


  1. Andaya VC, Mackill DJ (2003) Mapping of QTL associated with cold tolerance during the vegetative stage in rice. J Exp Bot 54:2579–2585PubMedCrossRefGoogle Scholar
  2. Andaya VC, Tai TH (2007) Fine mapping of the qCTS4 locus associated with seedling cold tolerance in rice (Oryza sativa L.). Mol Breeding 20:349–358CrossRefGoogle Scholar
  3. Ansari HA, Ellison NW, Williams WM (2008) Molecular and cytogenetic evidence for an allotetraploid origin of Trifolium dubium (Leguminosae). Chromosoma 117:159–167PubMedCrossRefGoogle Scholar
  4. Bertin P, Bouharmont J (1997) Use of somaclonal variation and in vitro selection for chilling tolerance improvement in rice. Euphytica 96:135–142CrossRefGoogle Scholar
  5. Chen JF, Luo XD, Qian CT, Molly MJ, Staub JE, Zhuang FY, Lou QF, Ren G (2004) Cucumis monosomic alien addition lines: morphological, cytological, and genotypic analyses. Theor Appl Genet 108:1343–1348PubMedCrossRefGoogle Scholar
  6. Chen LZ, Lou QF, Zhuang Y, Chen JF, Zhang XQ, Wolukau JN (2007) Cytological diploidization and rapid genome changes of the newly synthesized allotetraploids Cucumis × hytivus. Planta 225:603–614PubMedCrossRefGoogle Scholar
  7. Chen XR, Yang KS, Fu JR, Zhu CL, Peng XS, He P, He HH (2008) Identification and genetic analysis of fertility restoration ability in Dongxiang wild rice (Oryza rufipogon). Rice Sci 15:21–28CrossRefGoogle Scholar
  8. Fujino K (2004) A major gene for low temperature germinability in rice (Oryza sativa L.). Euphytica 136:63–68CrossRefGoogle Scholar
  9. Groszmann M, Greaves IK, Albertyn ZI, Scofield GN, Peacock WJ, Dennis ES (2011) Changes in 24-nt siRNA levels in Arabidopsis hybrids suggest an epigenetic contribution to hybrid vigor. Proc Natl Acad Sci USA 108:2617–2622PubMedCrossRefGoogle Scholar
  10. Guo JY, Chen JF, Cao QH, Luo XD, Chen LZ (2004) Cytological studies on microsporogenesis and male gametophyte development of a Cucumis allotriploid derived from C. hytivus × C. sativus. Cytologia 69:335–340CrossRefGoogle Scholar
  11. Han LZ, Qiao YL, Cao GL, Zhang YY, An YP, Ye JD, Koh HJ (2005) QTL analysis on cold tolerance during early growth period in rice. Chin J Rice Sci 19:122–126Google Scholar
  12. He GM, Zhu XP, Elling AA, Chen LB, Wang XF, Guo L, Liang MZ, He H, Zhang HY, Chen FF, Qi YJ, Chen RS, Deng XW (2010) Global epigenetic and transcriptional trends among two rice subspecies and their reciprocal hybrids. Plant Cell 22:17–33PubMedCrossRefGoogle Scholar
  13. Liu FX, Sun CQ, Tan LB, Fu YC, Li DJ, Wang XK (2003) Identification of QTL for cold tolerance at booting and flowering stage in Jiangxi Dongxiang wild rice. Chin Sci Bull 48:1864–1867Google Scholar
  14. Lou QJ, Chen L, Sun ZX, Xing YZ, Li J, Xu XY, Mei HW, Luo LJ (2007) A major QTL associated with cold tolerance at seedling stage in rice (Oryza sativa L.). Euphytica 158:87–94CrossRefGoogle Scholar
  15. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497Google Scholar
  16. Murray HG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucl Acids Res 8:4321–4326PubMedCrossRefGoogle Scholar
  17. Osborn TC, Pires JC, Birchler JA, Auger DL, Chen ZJ, Lee HS, Comai L, Madlung A, Doerge RW, Colot V, Martienssen RA (2003) Understanding mechanisms of novel gene expression in polyploids. Trends Genet 19:141–147PubMedCrossRefGoogle Scholar
  18. Pritchard JK, Stephens M, Donnelly P (2000) Inference of population structure using multilocus genotype data. Genetics 155:945–959PubMedGoogle Scholar
  19. Qi B, Zhong X, Zhu B, Zhao N, Xu L, Zhang H, Yu X, Liu B (2010) Generality and characteristics of genetic and epigenetic changes in newly synthesized allotetraploid wheat lines. J Genet Genomics 37:737–748PubMedCrossRefGoogle Scholar
  20. Qu TT, Chen LY, Zhang ZH, Hu ZL, Li P, Zhu LH, Zhu YG (2003) Molecular mapping of genes conferring cold tolerance at seedling stage using doubled haploid lines from an indica × japonica cross in rice. J Wuhan Bot Res 21:385–389Google Scholar
  21. Rahman L, Khanam MS, Koh HJ (2008) QTL analysis for yield related traits using populations derived from an indica-japonica hybrid in rice (Oryza sativa L.). Czech J Genet Plant Breed 44:93–104Google Scholar
  22. Sanguinetti CJ, Dias NE, Simpson AJ (1994) Rapid silver staining and recovery of PCR products separated on polyacrylamide gels. Biotechniques 17:914–921PubMedGoogle Scholar
  23. Sthapit BR, Witcombe JR (1998) Inheritance of tolerance to chilling stressing rice during germination and plumule greening. Crop Sci 38:660–665CrossRefGoogle Scholar
  24. Takesawa T, Ito M, Kanzaki H, Kameya N, Nakamura I (2002) Over-expression of ζ glutathione S-transferase in transgenic rice enhances germination and growth at low temperature. Mol Breed 9:93–101CrossRefGoogle Scholar
  25. Tan LB, Liu FX, Xue W, Wang GJ, Ye S, Zhu ZF, Fu YC, Wang XK, Sun CQ (2007) Development of Oryza rufipogon and O. sativa introgression lines and assessment for yield-related quantitative trait loci. J Integr Plant Biol 49:871–884CrossRefGoogle Scholar
  26. Tanksley SD, Nelson JC (1996) Advanced backcross QTL analysis: a method for the simultaneous discovery and transfer of valuable QTLs from unadapted germplasm into elite breeding lines. Theor Appl Genet 92:91–203Google Scholar
  27. Thomson MJ, Edwards JD, Septiningsih EM, Harrington SE, McCouch SR (2006) Substitution mapping of dth1.1, a flowering-time quantitative trait locus (QTL) associated with transgressive variation in rice, reveals multiple sub-QTL. Genetics 172:2501–2514PubMedCrossRefGoogle Scholar
  28. Tian F, Li DJ, Fu Q, Zhu ZF, Fu YC, Wang XK, Sun CQ (2006) Construction of introgression lines carrying wild rice (Oryza rufipogon Griff.) segments in cultivated rice (Oryza sativa L.) background and characterization of introgressed segments associated with yield-related traits. Theor Appl Genet 112:570–580PubMedCrossRefGoogle Scholar
  29. Wang WS, Pan YJ, Zhao XQ, Dwivedi D, Zhu LH, Ali J, Fu BY, Li ZK (2011) Drought-induced site-specific DNA methylation and its association with drought tolerance in rice (Oryza sativa L.). J Exp Bot 62:1951–1960PubMedCrossRefGoogle Scholar
  30. Xie JK, Hu BL, Wan Y, Zhang T, Li X, Liu RN, Huang YH, Dai LF, Luo XD (2010a) Comparison of the drought resistance characters at seedling stage between Dongxiang common wild rice (Oryza rufipogon Griff.) and cultivars (Oryza sativa L.). Acta Ecol Sin 30:1665–1674Google Scholar
  31. Xie JK, Agrama HA, Kong D, Zhuang J, Hu B, Wan Y, Yan W (2010b) Genetic diversity associated with conservation of endangered Dongxiang wild rice (Oryza rufipogon). Genet Resour Crop Evol 57:597–609CrossRefGoogle Scholar
  32. Xu L, He Y, Zhang DF, Dai JR, Wang SC (2009) Identification and fine-mapping of a bacterial brown spot disease resistance gene in maize. Mol Breed 23:709–718CrossRefGoogle Scholar
  33. Zeng YW, Pu XY (2006) Genetic analysis for cold tolerance at booting stage for japonica rice (Oryza sativa L.). Indian J Genet Plant Breed 66:100–102Google Scholar
  34. Zeng YW, Yang SM, Cui H, Yang XJ, Xu LM, Du J, Pu XY, Li ZC, Cheng ZQ, Huang XQ (2009) QTLs of cold tolerance-related traits at the booting stage for NIL-RILs in rice revealed by SSR. Genes Genomics 31(2):143–154CrossRefGoogle Scholar
  35. Zhan QC, Zhu KY, Chen ZW, Zeng SZ (2005) Studies on the QTL for cold tolerance related characters of rice seedling by molecular markers. Hybrid rice 20:50–55Google Scholar
  36. Zhang ZH, Li S, Wei L, Wei C, Zhu YG (2005) A major QTL conferring cold tolerance at the early seedling stage using recombinant inbred lines of rice (Oryza sativa L.). Plant Sci 168:527–534CrossRefGoogle Scholar
  37. Zhang XS, Zhou X, Fu YC, Zhen S, Wang XK, Sun CQ (2006) Identification of a drought tolerant introgression line derived from Dongxiang common wild rice (O. rufipogon Griff.). Plant Mol Biol 62:247–259PubMedCrossRefGoogle Scholar
  38. Zhang YS, Luo L, Liu T, Xu C, Xing Y (2009) Four rice QTL controlling number of spikelets per panicle expressed the characteristics of single Mendelian gene in near isogenic backgrounds. Theor Appl Genet 118:1035–1044PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Xiang-Dong Luo
    • 1
  • Liang-Fang Dai
    • 1
  • Juan-Fang Cao
    • 1
  • Shui-Rong Jian
    • 1
  • Ya-Ling Chen
    • 1
  • Biao-Lin Hu
    • 2
  • Jian-Kun Xie
    • 1
  1. 1.College of Life ScienceJiangxi Normal UniversityNanchangChina
  2. 2.Rice Research InstituteJiangxi Academy of Agricultural ScienceNanchangChina

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