, 214:23 | Cite as

Production and genetic analyses of novel Brassica rapa L. introgressions from interspecific crosses with Brassica juncea L. landraces native to the Qinghai-Tibet Plateau

  • Changcai Teng
  • Yan Niu
  • Dezhi Du
  • Qinglan Yu
  • Zhigang Zhao


Interspecific hybrids between related species have long been used for transferring desirable genes, broadening genetic diversity and utilizing intersubgenomic heterosis. In this study, we developed a novel Brassica rapa type (AA, 2n = 20) exhibiting certain features derived from interspecific hybridization between natural B. rapa and Brassica juncea (AABB, 2n = 36). In pollen mother cells (PMCs) of the novel B. rapa type, normal chromosome pairing with 10 bivalents and 10:10 segregation was observed, and the novel B. rapa lines were completely fertile. However, GISH showed that certain B chromosomes or fragments were introgressed into B. rapa. Genetic components of the novel B. rapa lines were investigated by GISH, AFLP and SSR analyses. GISH analysis of F1, BC1F1, and BC1F2 plants confirmed the identities of three addition lines and seven translocation lines. AFLP and SSR analyses of 60 hybrid progenies from BC1F4 plants, their parents, and some B. juncea and B. rapa resources indicated that the AJ and B chromosome(s) or fragment(s) introgressed to the novel B. rapa. AFLP revealed that 60 BC1F4 plants contained B chromosomes or fragments, which evidenced introgression into the hybrid progeny. SSR analysis indicated that the A-genome (A1–A10) of B. juncea was introgressed into the hybrid progeny at 1.0 to 42.7%. Lastly, we obtained some yellow-seed and early-flowering B. rapa resources. The novel B. rapa lines can be used to genetically improve B. rapa in the Qinghai-Tibet Plateau and to study the origin and evolution of the A- and B-genomes.


Amplified fragment length polymorphism (AFLP) B. juncea B. rapa Genetic introgression Genomic in situ hybridization (GISH) Simple sequence repeat (SSR) 



The authors are grateful to Dr. Bin Zhu for his critical reading of the manuscript.

Authors’ contribution

ZZ and DD designed and managed this study. CT and YN performed the experiments and analyzed the data. CT wrote the manuscript. QY designed and executed the artificial synthesis of B. rapa.


This study was financially supported by funds from the National Key Research and Development Plan of China (2016YFD0100202), the Key Laboratory of Spring Rape Genetic Improvement of Qinghai Province (2017-ZJ-Y09) and the Industry Technology Systems for Rapeseed in China (CARS-13).

Compliance with ethical standards

Conflict of interest

We declare that we have no financial or personal relationships with other people or organizations that can inappropriately influence our work. There are no professional or other personal interests of any nature in any product, service and/or company that could be construed as influencing the position presented in, or the review of, the manuscript entitled, “Production and genetic analyses of novel Brassica rapa L. introgressions from interspecific crosses with Brassica juncea L. landraces native to the Qinghai-Tibet Plateau”.

Ethical approval

This article does not describe any studies involving human participants or animals.

Supplementary material

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  1. Anamthawat-Jonsson K (2001) Molecular cytogenetics of introgression hybridization in plants. Methods Cell Sci 23:139–148. CrossRefPubMedGoogle Scholar
  2. Attia T, Robbelen G (1986) Cytogenetic relationship within cultivated Brassica analyzed in amphihaploids from the three diploid ancestor. Can J Genet Cytol 28:323–329. CrossRefGoogle Scholar
  3. Attia T, Busso C, Robbelen G (1987) Digenomic triploids for an assessment of chromosome relationships in the cultivated diploid Brassica species. Genome 29:326–330. CrossRefGoogle Scholar
  4. Barba-Gonzalez R, Van Silfhout AA, Visser RGF, Ramanna MS, Van Tuyl JM (2006) Progenies of allotriploids of Oriental × Asiatic lilies (lilium) examined by GISH analysis. Euphytica 151:243–250. CrossRefGoogle Scholar
  5. Bennett RA, Séguin-Swartz G, Rahman H (2012) Broadening genetic diversity in Canola using the C-genome species Brassica oleracea L. Crop Sci 52:2030–2039. CrossRefGoogle Scholar
  6. Chen HF, Wang H, Li ZY (2007) Production and genetic analysis of partial hybrids in intertribal crosses between Brassica species (B. rapa, B. napus) and Capsella bursa-pastoris. Plant Cell Rep 26:1791–1800. CrossRefPubMedGoogle Scholar
  7. Choudhary BR, Joshi P, Rao SR (2002) Cytogenetics of Brassica juncea × Brassica rapa hybrids and patterns of variation in the hybrid derivatives. Plant Breed 121:292–296. CrossRefGoogle Scholar
  8. Cui C, Ge X, Gautam M, Kang L, Li Z (2012) Cytoplasmic and genomic effects on meiotic pairing in Brassica hybrids and allotetraploids from pair crosses of three cultivated diploids. Genetics 191:725–738. CrossRefPubMedPubMedCentralGoogle Scholar
  9. Dhaka N, Rout K, Yadava SK, Sodhi YS, Gupta V, Pental D, Pradhan AK (2017) Genetic dissection of seed weight by QTL analysis and detection of allelic variation in Indian and East European gene pool lines of Brassica juncea. Theor Appl Genet 130:293–307. CrossRefPubMedGoogle Scholar
  10. Du XZ, Ge XH, Yao XC, Zhao ZG, Li ZY (2009) Production and cytogenetic characterization of intertribal somatic hybrids between Brassica napus and Isatis indigotica and backcross progenies. Plant Cell Rep 28:1105–1113. CrossRefPubMedGoogle Scholar
  11. Fredua-Agyeman R, Coriton O, Huteau V, Parkin IA, Chèvre AM, Rahman H (2014) Molecular cytogenetic identification of B genome chromosomes linked to blackleg disease resistance in Brassica napus × B. carinata interspecific hybrids. Theor Appl Genet 127:1305–1318. CrossRefPubMedGoogle Scholar
  12. Fu CH, Chen CL, Guo WW, Deng XX (2004) GISH, AFLP and PCR-RFLP analysis of an intergeneric somatic hybrid combining Goutou sour orange and Poncirus trifoliata. Plant Cell Rep 23:391–396. CrossRefPubMedGoogle Scholar
  13. González G, Comas C, Confalonieri V, Naranjo CA, Poggio L (2006) Genomic affinities between maize and Zea perennis using classical and molecular cytogenetic methods (GISH–FISH). Chromosome Res 14:629–635. CrossRefPubMedGoogle Scholar
  14. Guttman B (2001) Evolution. In: Brenner S, Miller JH (eds) Encyclopedia of genetics, vol 2. Academic, San Diego, pp 663–666CrossRefGoogle Scholar
  15. Hua YW, Li ZY (2006) Genomic in situ hybridization analysis of Brassica napus × Orychophragmus violaceus hybrids and production of B. napus aneuploids. Plant Breed 125:144–149. CrossRefGoogle Scholar
  16. Hua YW, Liu M, Li ZY (2006) Parental genome separation and elimination of cells and chromosomes revealed by AFLP and GISH analyses in a Brassica carinata × Orychophragmus violaceus cross. Ann Bot 97:993–998. CrossRefPubMedPubMedCentralGoogle Scholar
  17. Jiang J, Gill BS (1994) Nonisotopic in situ hybridization and plant genome mapping: the first 10 years. Genome 37:717–725. CrossRefPubMedGoogle Scholar
  18. Kang L, Du X, Zhou Y, Zhu B, Ge X, Li Z (2014) Development of a complete set of monosomic alien addition lines between Brassica napus and Isatis indigotica (Chinese woad). Plant Cell Rep 33:1355–1364. CrossRefPubMedGoogle Scholar
  19. Leflon M, Eber F, Letanneur JC, Chelysheva L, Coriton O, Huteau V, Ryder CD, Barker G, Jenczewski E, Chèvre AM (2006) Pairing and recombination at meiosis of Brassica rapa (AA) × Brassica napus (AACC) hybrids. Theor Appl Genet 113:1467–1480. CrossRefPubMedGoogle Scholar
  20. Li Z, Liu HL, Luo P (1995) Production and cytogenetics of intergeneric hybrids between Brassica napus and Orychophragmus violaceus. Theor Appl Genet 91:131–136. PubMedGoogle Scholar
  21. Li MT, Li ZY, Zhang CY, Qian W, Meng JL (2005) Reproduction and cytogenetic characterization of interspecific hybrids derived from crosses between Brassica carinata and B. rapa. Theor Appl Genet 110:1284–1289. CrossRefPubMedGoogle Scholar
  22. Li M, Chen X, Meng J (2006) Intersubgenomic heterosis in rapeseed production with a partial new-typed Brassica napus containing subgenome Ar from B. rapa and Cc from Brassica carinata. Crop Sci 46:234–242. CrossRefGoogle Scholar
  23. Li Q, Mei J, Zhang Y, Li J, Ge X, Li Z, Qian W (2013) A large-scale introgression of genomic components of Brassica rapa into B. napus by the bridge of hexaploid derived from hybridization between B. napus and B. oleracea. Theor Appl Genet 126:2073–2080. CrossRefPubMedGoogle Scholar
  24. Li G, Lang T, Dai G, Li D, Li C, Song X, Yang Z (2015) Precise identification of two wheat–Thinopyrum intermedium substitutions reveals the compensation and rearrangement between wheat and Thinopyrum chromosomes. Mol Breed 35:1–10. CrossRefGoogle Scholar
  25. Ma N, Li ZY, Cartagena JA, Fukui K (2006) GISH and AFLP analyses of novel Brassica napus lines derived from one hybrid between B. napus and Orychophragmus violaceus. Plant Cell Rep 25:1089–1093. CrossRefPubMedGoogle Scholar
  26. Mason AS, Huteau V, Eber F, Coriton O, Yan G, Nelson MN, Cowling W, Chèvre A (2010) Genome structure affects the rate of autosyndesis and allosyndesis in AABC, BBAC and CCAB Brassica interspecific hybrids. Chromosome Res 18:655–666. CrossRefPubMedGoogle Scholar
  27. Molnár I, Molnár-Láng M (2010) GISH reveals different levels of meiotic pairing with wheat for individual Aegilops biuncialis chromosomes. Biol Plant 54:259–264. CrossRefGoogle Scholar
  28. Nagaharu U (1935) Genome analysis in Brassica with special reference to the experimental formation of B. napus and peculiar mode of fertilization. Jpn J Bot 7:389–452Google Scholar
  29. Navabi ZK, Parkin IA, Pires JC, Xiong Z, Thiagarajah MR, Good AG, Rahman MH (2010) Introgression of B-genome chromosomes in a doubled haploid population of Brassica napus × B. carinata. Genome 53:619–629. CrossRefPubMedGoogle Scholar
  30. Navabi ZK, Stead KE, Pires JC, Xiong Z, Sharpe AG, Parkin IAP, Rahman MH, Good AG (2011) Analysis of B-Genome chromosome introgression in interspecific hybrids of Brassica napus × B. carinata. Genetics 187:659–673. CrossRefPubMedPubMedCentralGoogle Scholar
  31. Nei M, Li WH (1979) Mathematical model for studying genetic variation in terms of restriction endouncleases. Proc Natl Acad Sci 76:5269–5273CrossRefPubMedPubMedCentralGoogle Scholar
  32. Rahman H, Bennett RA, Séguin-Swartz G (2015) Broadening genetic diversity in Brassica napus canola: development of canola-quality spring B. napus from B. napus × B oleracea var. alboglabra interspecific crosses. Can J Plant Sci 95:29–41. CrossRefGoogle Scholar
  33. Röbbelen G (1960) Beitrage zur analyse des Brassica-genoms. Chromosoma 11:205–228. CrossRefGoogle Scholar
  34. Rohlf FJ (1997) NTSYS-pc 2.1. Numerical taxonomy and multivariate analysis system. Exeter Software, SetauketGoogle Scholar
  35. Tan C, Cui C, Xiang Y, Ge X, Li Z (2017) Development of Brassica oleracea-nigra monosomic alien addition lines: genotypic, cytological and morphological analyses. Theor Appl Genet. PubMedCentralGoogle Scholar
  36. Tang X, Shi D, Xu J, Li Y, Li W, Ren Z, Fu T (2014) Molecular cytogenetic characteristics of a translocation line between common wheat and Thinopyrum intermedium with resistance to powdery mildew. Euphytica 197:201–210. CrossRefGoogle Scholar
  37. Vos P, Hogers R, Bleeker M, Reijans M, van de Lee T, Hornes M, Frijters A, Pot J, Peleman J, Kuiper M, Zabeau M (1995) AFLP: a new technique for DNA fingerprinting. Nucleic Acids Res 23:4407–4414. CrossRefPubMedPubMedCentralGoogle Scholar
  38. Waara S, Glimelius K (1995) The potential of somatic hybridization in crop breeding. Euphytica 85:217–233. CrossRefGoogle Scholar
  39. Wang YP, Snowdon RJ, Rudloff E, Wehling P, Friedt W, Sonntag K (2004) Cytogenetic characterization and fae1 gene variation in progenies from asymmetric somatic hybrids between Brassica napus and Crambe abyssinica. Genome 47:724–731. CrossRefPubMedGoogle Scholar
  40. Warude D, Chavan P, Joshi K, Patwardhan B (2003) DNA isolation from fresh, dry plant samples with highly acidic tissue extracts. Plant Mol Biol Rep 21:467. CrossRefGoogle Scholar
  41. Wen J, Tu J, Li Z, Fu T, Ma C, Shen J (2008) Improving ovary and embryo culture techniques for efficient resynthesis of Brassica napus from reciprocal crosses between yellow-seeded diploids B. rapa and B. oleracea. Euphytica 162:81–89. CrossRefGoogle Scholar
  42. Woods DL, Capcara JJ, Downey RK (1991) The potential of mustard (Brassica juncea (L.) Coss) as an edible oil crop on the Canadian Prairies. Can J Plant Sci 71:195–198. CrossRefGoogle Scholar
  43. Xie S, Khan N, Ramanna MS, Niu L, Marasek-Ciolakowska A, Arens P, van Tuyl JM (2010) An assessment of chromosomal rearrangements in neopolyploids of lilium hybrids. Genome 53:439–446. CrossRefPubMedGoogle Scholar
  44. Yamashita K, Takatori Y, Tashiro Y (2005) Chromosomal location of a pollen fertility-restoring gene, Rf, for CMS in Japanese bunching onion (Allium fistulosum L.) possessing the cytoplasm of A. galanthum Kar. et Kir. revealed by genomic in situ hybridization. Theor Appl Genet 111:15–22. CrossRefPubMedGoogle Scholar
  45. Yao XC, Ge XH, Chen JP, Li ZY (2010) Intra- and intergenomic relationships in interspecific hybrids between Brassica (B. rapa, B. napus) and a wild species B. maurorum as revealed by genomic in situ hybridization (GISH). Euphytica 173:113–120. CrossRefGoogle Scholar
  46. Younis A, Ramzan F, Hwang YJ, Lim KB (2015) FISH and GISH: molecular cytogenetic tools and their applications in ornamental plants. Plant Cell Rep 34:1477–1488. CrossRefPubMedGoogle Scholar
  47. Zhong XB, Hans de Jong JH, Zabel P (1996) Preparation of tomato meiotic pachytene and mitotic metaphase chromosomes suitable for fluorescence in situ hybridization (FISH). Chromosome Res 4:24–28. CrossRefPubMedGoogle Scholar
  48. Zhou J, Yang Z, Li G, Liu C, Tang Z, Zhang Y, Ren Z (2010) Diversified chromosomal distribution of tandemly repeated sequences revealed evolutionary trends in secale (Poaceae). Plant Syst Evol 287:49–56. CrossRefGoogle Scholar
  49. Zhu JS, Struss D, Röbbelen G (1993) Studies on resistance to Phoma lingam in Brassies napus-Brassica nigra addition lines. Plant Breed 111:192–197. CrossRefGoogle Scholar

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© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.State Key Laboratory of Plateau Ecology and Agriculture of Qinghai UniversityQinghai UniversityXiningChina
  2. 2.Key Laboratory of Qinghai Province for Spring Rapeseed Genetic Improvement, National Key Laboratory Breeding Base-Key Laboratory of Qinghai Province for Plateau Crop Germplasm Innovation and Utilization, Academy of Agriculture and Forestry SciencesQinghai UniversityXiningChina

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