Skip to main content
SpringerLink
Log in

Identification of cytoplasm types in rapeseed (Brassica napus L.) accessions by a multiplex PCR assay

  • Original Paper
  • Published:
Theoretical and Applied Genetics Aims and scope Submit manuscript

Abstract

Cytoplasmic male sterility (CMS) has widely been used as an efficient pollination control system in rapeseed hybrid production. Identification of cytoplasm type of rapeseed accessions is becoming the most important basic work for hybrid-rapeseed breeding. In this study, we report a simple multiplex PCR method to distinguish the existing common cytoplasm resources, Pol, Nap, Cam, Ogu and Ogu-NWSUAF cytoplasm, in rapeseed. Cytoplasm type of 35 F1 hybrids and 140 rapeseed open pollinated varieties or breeding lines in our rapeseed breeding programme were tested by this method. The results indicated that 10 of 35 F1 hybrids are the Nap, and 25 the Pol cytoplasm type, which is consistent with the information provided by the breeders. Out of 140 accessions tested, 100 (71.4%), 21 (15%) and 19 (13.6%) accessions possess Nap, Cam and Pol cytoplasm, respectively. All 19 accessions with Pol cytoplasm are from China. Pedigree analysis indicated that these accessions with Pol cytoplasm were either restorers for Pol CMS, including Shaan 2C, Huiyehui, 220, etc. or derived from hybrids with Pol CMS as female parent. Our molecular results are consistent with those of the classical testcross, suggesting the reliability of this method. The multiplex PCR assay method can be applied to CMS “three-line” breeding, selection and validation of hybrid rapeseed.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Ahmad M (2000) Molecular marker-assisted selection of HMW glutenin alleles related to wheat bread quality by PCR-generated DNA markers. Theor Appl Genet 101:892–896

    Article  CAS  Google Scholar 

  • Bonhomme S, Budar F, Lancelin D, Small L, Defrance MC, Pelletier G (1992) Sequence and transcript analysis of the Nco2.5 Ogura-specific fragment correlated with cytoplasmic male sterility in Brassica cybrids. Mol Gen Genet 235:340–348

    Article  PubMed  CAS  Google Scholar 

  • Chamberlain JS, Gibbs RA, Ranier JE, Nguyen PN, Caskey CT (1998) Deletion screening of the Duchenne muscular dystrophy locus via multiplex DNA amplification. Nucl Acids Res 16:11141–11156

    Article  Google Scholar 

  • Chang JJ, Hu SW, Zhao HX, Li ZJ (2010) Characterization of an improved Ogu-NWSUAF CMS in Brassica napus L. J Northwest A & F Univ 38(4):71–78

    Google Scholar 

  • Cheng D, Kitazaki K, Xu D, Mikami T, Kubo T (2009) The distribution of normal and male-sterile cytoplasms in Chinese sugar-beet germplasm. Euphytica 165:345–351

    Article  Google Scholar 

  • Edwards K, Johnstone C, Thompson C (1991) A simple and rapid method for the preparation of plant genomic DNA for PCR analysis. Nucl Acids Res 19:1349

    Article  PubMed  CAS  Google Scholar 

  • Engelke T, Tatlioglu T (2002) A PCR-marker for the CMS1 inducing cytoplasm in chives derived from recombination events affecting the mitochondrial gene atp9. Theor Appl Genet 104:698–702

    Article  PubMed  CAS  Google Scholar 

  • Engelke T, Terefe D, Tatlioglu T (2003) A PCR-based marker system monitoring CMS-(S). CMS-(T) and (N)-cytoplasm in the onion (Allium cepa L.). Theor Appl Genet 107:162–167

    PubMed  CAS  Google Scholar 

  • Fu TD (2008) On research and application of heterosis in rapeseed. Proceeding of oil crops scientific and technological advantage and edible oils supply safety. Chin J Oil Crop Sci 30(S):1–5

    Google Scholar 

  • Fu TD, Yang GS, Yang XN (1990) Studies on “three line” polima cytoplasmic male sterility developed in Brassica napus L. Plant Breed 104:115–120

    Article  Google Scholar 

  • Grelon M, Budar F, Bonhomme S, Pelletier G (1994) Ogura cytoplasmic male-sterility (CMS)-associated orf138 is translated into a mitochondrial membrane polypeptide in male-sterility Brassica cybrids. Mol Gen Genet 243:540–547

    Article  PubMed  CAS  Google Scholar 

  • Handa H (2003) The complete nucleotide sequence and RNA editing content of the mitochondrial genome of rapeseed (Brassica napus L.): comparative analysis of the mitochondrial genomes of rapeseed and Arabidopsis thaliana. Nucl Acids Res 31:5907–5916

    Article  PubMed  CAS  Google Scholar 

  • Handa H (2007) Investigation of the origin and transmission of linear mitochondrial plasmid based on phylogenetic analysis in Japanese rapeseed varieties. Genome 50:234–240

    Article  PubMed  CAS  Google Scholar 

  • Hanson MR, Bentolila S (2004) Interactions of mitochondrial and nuclear genes that affect male gametophyte development. Plant Cell 16:S154–S169

    Article  PubMed  CAS  Google Scholar 

  • Hu Q, Qian XZ, Wu XM (1992) A preliminary investigation on fertility of cytoplasm in rapeseed. Chinese Oil Crops 14:12–14

    Google Scholar 

  • Hu SW, Fan YF, Zhao HX, Guo XL, Yu CY, Sun GL, Liu SY, Wang HZ (2006) Analysis of MS2Bnap genomic DNA homologous to MS2 gene from Arabidopsis thaliana in two dominant digenic male sterile accessions of oilseed rape (Brassica napus L.). Theor Appl Genet 113:397–406

    Article  PubMed  CAS  Google Scholar 

  • Kemble RJ (1987) A rapid, single leaf, nucleic acid assay for determining the cytoplasmic organelle complement of rapeseed and related Brassica species. Theor Appl Genet 73:364–370

    Article  CAS  Google Scholar 

  • Kim S, Lim H, Park S, Cho K, Sung S, Oh D, Kim K (2007) Identification of a novel mitochondrial genome type and development of molecular makers for cytoplasm classification in radish (Raphanus sativus L.). Theor Appl Genet 115:1137–1145

    Article  PubMed  CAS  Google Scholar 

  • Kim S, Lee ET, Cho DY, Han T, Bang H, Patil BS, Ahn YK, Yoon MK (2009) Identification of a novel chimeric gene, orf725, and its use in development of a molecular marker for distinguishing among three cytoplasm types in onion (Allium cepa L.). Theor Appl Genet 118:433–441

    Article  PubMed  CAS  Google Scholar 

  • L’Homme Y, Brown GG (1993) Organizational differences between cytoplasmic male sterile and male fertile Brassica mitochondrial genomes are confines to a single transposed locus. Nucl Acids Res 21:1903–1909

    Article  PubMed  Google Scholar 

  • L’Homme Y, Stahl RJ, Li XQ, Hameed A, Brown GG (1997) Brassica nap cytoplasmic male sterility is associated with expression of a mtDNA region containing a chimeric gene similar to the pol CMS-associated orf224 gene. Curr Genet 31:325–335

    Article  PubMed  Google Scholar 

  • Lee YP, Kim S, Lim H, Ahn YS, Sun SK (2009) Identification of mitochondrial genome rearrangements unique to novel cytoplasmic male sterility in radish (Raphanus sativus L.). Theor Appl Genet 118:719–728

    Article  PubMed  CAS  Google Scholar 

  • Ma W, Zhang W, Gale KR (2003) Multiplex-PCR typing of high molecular weight glutenin alleles in wheat. Euphytica 134:51–60

    Article  CAS  Google Scholar 

  • Nakamura T, Vrinten P, Saito M, Konda M (2002) Rapid classification of partial waxy wheat using PCR-based markers. Genome 45:1150–1156

    Article  PubMed  CAS  Google Scholar 

  • Rousselle P, Renard M (1978) Study of a cytoplasmic male sterility in rapeseed (B. napus). Eucarpia Crucif Newslett 3:40–41

    Google Scholar 

  • Shiga T, Takayanagi K, Ohkawa Y (1976) Fertility restoration of European rapeseed cultivars to cytoplasmic male sterility. Eucarpia Crucif Newslett 1:17–18

    Google Scholar 

  • Shiga T, Ohkawa Y, Takayanagi K (1978) Segregation of fertility restoration in F2 and testcross progenies from crosses between male sterile line and European cultivars of oilseed rape, B. napus L. Eucarpia Crucif Newslett 3:44

    Google Scholar 

  • Singh M, Brown GG (1991) Suppression of cytoplasmic male sterility by nuclear genes alters expression of a novel mitochondrial gene region. Plant cell 3:1349–1362

    Article  PubMed  CAS  Google Scholar 

  • Wang WM, Li YC, Hu Q, Cheng JH, Mei SDS, Hao JY (2008) An efficient protocol for pure mitochondrial DNA extraction from oilseed rape. Chin J Oil Crop Sci 30:357–360

    Google Scholar 

  • Wei WL, Wang HZ, Liu GH (2005) Molecular Identification of the sterile cytoplasm of NCa of a cytoplasmic male sterile line in rapeseed (Brassica napus L.). Sci Agric Sin 38:1965–1972

    CAS  Google Scholar 

  • Wen YC, Zhang SF, Tian BM, Wang JP, Zhu JC, Liu JM, Zhao L (2003) Determination of cytoplasmic fertility types and nucleic fertility genotypes of rapeseed germplasm resources. Chin J Oil Crop Sci 25:1–4

    Google Scholar 

  • Yang GS (1987) Genetic investigation of cytoplasmic male sterility in Brassica napus L. Master’s thesis, Huazhong Agricultural University

  • Yang GS, Fu TD, Brown GG (1998) The genetic classification of cytoplasmic male sterility systems in Brassica napus L. Sci Agric Sin 31:27–31

    CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the earmarked fund for Modern Agro-industry Technology Research System (nycytx-00505), a grant of Northwest A&F University for SW Hu and Shaanxi Scientific and Technological Key Project (2007K01-05). Thanks to Kucera V at the Crop Research Institute, Czech Republic, Michael J. Stamm at Kansas State University, USA and XM Wu at the Oil Crops Research Institute, Chinese Academy of Agricultural Sciences for providing some rapeseed accessions, and two anonymous reviewers for their useful comments.

Author information

Authors and Affiliations

  1. College of Life Sciences, Northwest A&F University, Yangling, 712100, Shaanxi, People’s Republic of China

    H. X. Zhao, Z. J. Li & Z. H. Zhang

  2. College of Agronomy, Northwest A&F University, Yangling, 712100, Shaanxi, People’s Republic of China

    S. W. Hu & J. J. Chang

  3. Department of Biology, Saint Mary’s University, Halifax, NS, B3H 3C3, Canada

    G. L. Sun

Authors
  1. H. X. Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Z. J. Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S. W. Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. G. L. Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. J. J. Chang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Z. H. Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to S. W. Hu.

Additional information

Communicated by C. Quiros.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 1013 kb)

Supplementary material 2 (DOC 1035 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhao, H.X., Li, Z.J., Hu, S.W. et al. Identification of cytoplasm types in rapeseed (Brassica napus L.) accessions by a multiplex PCR assay. Theor Appl Genet 121, 643–650 (2010). https://doi.org/10.1007/s00122-010-1336-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00122-010-1336-3

Keywords

Search

Navigation