Skip to main content
SpringerLink
Log in

Towards map-based cloning: fine mapping of a recessive genic male-sterile gene (BnMs2) in Brassica napus L. and syntenic region identification based on the Arabidopsis thaliana genome sequences

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

Abstract

S45AB, a recessive genic male sterile (RGMS) line, originated as a spontaneous mutant in Brassica napus cv. Oro. The genotypes of sterile (S45A) and fertile plants (S45B) are Bnms1ms1ms2ms2 and BnMs1ms1ms2ms2, respectively. In our previous studies, Yi et al. (Theor Appl Genet 113:643–650, 2006) mapped the BnMs1 locus to a region of 0.4 cM, candidates of which have been identified and genetic transformation is in progress. We describe the fine mapping of BnMs2 exploiting amplified fragment length polymorphism (AFLP) and amplified consensus genetic marker (ACGM) methodologies, and the identification of a collinear region probably containing BnMs2 orthologue in Arabidopsis thaliana. A near isogenic line (NIL) population S4516AB which segregated for BnMs2 locus was generated by crossing, allelism testing and repeated full-sib mating. From the survey of 1,024 AFLP primer combinations, 12 tightly linked AFLP markers were obtained and five of them were successfully converted into co-dominant or dominant sequence characterized amplified region (SCAR) markers. A population of 2,650 sterile plants was screened using these markers and a high-resolution map surrounding BnMs2 was constructed. The closest AFLP markers flanking BnMs2 were 0.038 and 0.075 cM away, respectively. Subsequently, an ACGM marker was developed to delimit the BnMs2 locus at an interval of 0.075 cM. We extended marker sequences to perform BlastN searches against the Arabidopsis genome and identified a collinear region containing 68 Arabidopsis genes, in which the orthologue of BnMs2 might be included. We further integrated BnMs2 linked AFLP or SCAR markers to two doubled-haploid (DH) populations derived from the crosses Tapidor × Ningyou7 (Qiu et al., Theor Appl Genet 114:67–80, 2006) and Quantum × No.2127-17 (available in our laboratory), and BnMs2 was mapped on N16. Molecular markers developed from these investigations will facilitate the marker-assisted selection (MAS) of RGMS lines, and the fine map and syntenic region identified will greatly hasten the process of positional cloning of BnMs2 gene.

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

  • Arumuganathan K, Earle ED (1991) Nuclear DNA content of some important plant species. Plant Mol Biol Rep 9:208–218

    Article  CAS  Google Scholar 

  • Brandle JL, Mcvetty PBE (1990) Geographical diversity parental selection and heterosis in oilseed rape. Can J Plant Sci 70:935–940

    Google Scholar 

  • Brown G, Formanova N, Jin H, Wargachuk R, Dendy C, Patil P, Laforest M, Zhang J, Cheung W, Landry B (2003) The radish Rfo restorer gene of Ogura cytoplasmic male sterility encodes a protein with multiple pentatricopeptide repeats. Plant J 35:262–272

    Article  PubMed  CAS  Google Scholar 

  • Brunel D, Froger N, Pelletier G (1999) Development of amplified consensus genetic markers (ACGMs) in Brassica napus from Arabidopsis thaliana sequences of known biological function. Genome 42:387–402

    Article  PubMed  CAS  Google Scholar 

  • Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15

    Google Scholar 

  • Foisset N, Delourme R, Barret P, Hubert N, Landry BS, Renard M (1996) Molecular-mapping analysis in Brassica napus using isozyme, RAPD and RFLP markers on a doubled-haploid progeny. Theor Appl Genet 93:1017–1025

    Article  CAS  Google Scholar 

  • Giancola S, Marhadour S, Desloire S, Clouet V, Falentin-Guyomarc’h H, Laloui W, Falentin C, Pelletier G, Renard M, Bendahmane A, Delourme R, Budar F (2003) Characterization of a radish introgression carrying the Ogura fertility restorer gene Rfo in rapeseed, using the Arabidopsis genome sequence and radish genetic mapping. Theor Appl Genet 107:1442–1451

    Article  PubMed  Google Scholar 

  • Grant I, Beversdorf WD (1985) Heterosis and combining ability estimates in spring oilseed rape (Brassica napus L.). Can J Genet Cytol 27:472–478

    Google Scholar 

  • Hong DF, Wan LL, Liu PW, Yang GS, He QB (2006) AFLP and SCAR markers linked to the suppressor gene (Rf ) of a dominant genetic male sterility in rapeseed (Brassica napus L.). Euphytica 151:401–409

    Article  CAS  Google Scholar 

  • Imai R, Koizuka N, Fujimoto H, Hayakawa T, Sakai T, Imamura J (2003) Delimitation of the fertility restorer locus Rfk1 to a 43-Kb contig in Kosena radish (Raphanus sativus L.). Mol Gen Genomics 269:388–394

    Article  CAS  Google Scholar 

  • Ke LP, Sun YQ, Liu PW, Yang GS (2004) Identification of AFLP fragments linked to one recessive genic male sterility (RGMS) in rapeseed (Brassica napus L.) and conversion to SCAR markers for marker-aided selection. Euphytica 138:163–168

    Article  CAS  Google Scholar 

  • Lander ES, Green P, Abrahamson J, Barlow A, Daly MJ, Lincoln SE, Newburg L (1987) MAPMAKER: an interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics 1:174–181

    Article  PubMed  CAS  Google Scholar 

  • Lincoln S, Daly M, Lander E (1992) Constructing genetic linkage maps with Mapmaker/exp 3.0: a tutorial and reference manual, 3rd edn. Whitehead Institute Technical Report

  • Li SL, Zhou ZJ, Zhou XR (1993) Inheritance of recessive genic male sterile line S45AB of rape (Brassica napus L.). Acta Agric Shanghai 9:1–7

    Google Scholar 

  • Li SL, Zhou ZJ, Zhou XR (1995) Three-line method of genetic male sterility for hybrid seed production in Brassica napus L. Acta Agric Shanghai 11:21–26

    Google Scholar 

  • Liu N, Shan Y, Wang FP, Xu CG, Peng KM, Li XH, Zhang Q (2001) Identification of an 85-kb DNA fragment containing pms1, a locus for photoperiod-sensitive genic male sterility in rice. Mol Genet Genomics 266:271–275

    Article  PubMed  CAS  Google Scholar 

  • Liu ZW, Fu TD, Tu JX, Chen BY (2005) Inheritance of seed colour and identification and AFLP markers linked to the seed in rapeseed (Brassica napus L.). Theor Appl Genet 110:303–310

    Article  CAS  Google Scholar 

  • Lowe AJ, Jones AE, Raybould AF, Trick M, Moule CJ, Edwards KJ (2002) Transferability and genome specificity of a new set of microsatellite primers among Brassica species of the U triangle. Mol Ecol Notes 2:7–11

    Article  CAS  Google Scholar 

  • Lukens L, Zou F, Lydiate D, Parkin I, Osborn T (2003) Comparison of a Brassica oleracea genetic map with the genome of Arabidopsis thaliana. Genetics 164:359–372

    PubMed  CAS  Google Scholar 

  • Lu GY, Yang GS, Fu TD (2004) Molecular mapping of a dominant genic male sterility gene Ms in rapeseed (Brassica napus). Plant Breed 123:262–265

    Article  CAS  Google Scholar 

  • Mayerhofer R, Wilde K, Mayerhofer M, Lydiate D, Bansal VK, Good AG, Parkin IAP (2005) Complexities of chromosome landing in a highly duplicated genome: towards map based cloning of a gene controlling blackleg resistance in Brassica napus. Genetics 171:1977–1988

    Article  PubMed  CAS  Google Scholar 

  • Michelmore RW, Paran I, Kesseli RV (1991) Identification of markers linked to disease-resistance genes by bulked segregant analysis: a rapid method to detect markers in specific genomic regions by using segregating populations. Proc Natl Acad Sci 88:9828–9832

    Article  PubMed  CAS  Google Scholar 

  • Muangprom A, Osborn TC (2004) Characterization of a dwarf gene in Brassica rapa, including the identification of a candidate gene. Theor Appl Genet 108:1378–1384

    Article  PubMed  CAS  Google Scholar 

  • Muangprom A, Thomas SG, Sun T, Osborn TC (2005) A novel dwarfing mutation in a green revolution gene from Brassica rapa. Plant Physiol 137:931–938

    Article  PubMed  CAS  Google Scholar 

  • Pan T, Zeng FY, Wu SH, Zhao Y (1988) A study on breeding and application GMS line of low eruci acid in rapeseed (B. napus). Oil Crop China 3:5–8

    Google Scholar 

  • Parkin IAP, Gulden SM, Sharpe AG, Lukens L, Trick M, et al (2005) Segmental structure of the Brassica napus genome based on comparative analysis with Arabidopsis thaliana. Genetics 171:765–781

    Article  PubMed  CAS  Google Scholar 

  • Pflieger S, Lefebvre V, Causse M (2001) The candidate gene approach in plant genetics: a review. Mol Breed 7:275–291

    Article  CAS  Google Scholar 

  • Qiu D, Morgan C, Shi J, Long Y, Liu J, Li R, Zhuang X, Wang Y, Tan X, Dietrich E, Weihmann T, Everett C, Vanstraelen S, Beckett P, Fraser F, Trick M, Barnes S, Wilmer J, Schmidt R, Li J, Li D, Meng J, Bancroft I (2006) A comparative linkage map of oilseed rape and its use for QTL analysis of seed oil and erucic acid content. Theor Appl Genet 114:67–80

    Article  PubMed  CAS  Google Scholar 

  • Schmidt R (2002) Plant genome evolution: lessons from comparative genomics at the DNA level. Plant Mol Biol 48:21–37

    Article  PubMed  CAS  Google Scholar 

  • Sernyk JL, Stefansson BR (1983) Heterosis in summer rape (Brassica napus L.). Can J Plant Sci 63:407–413

    Article  Google Scholar 

  • Shen JX, Fu TD, Yang GS (2001) Primary study on heterosis of self-incompatibility in Brassica napus. J Huazhong Agri Univ 20:528–530

    Google Scholar 

  • Siebert PD, Chenchik A, Kellogg DE, Lukyanov KA, Lukyanov SA (1995) An improved PCR method for walking in uncloned genomic DNA. Nucleic Acids Res 23:1087–1088

    Article  PubMed  CAS  Google Scholar 

  • Snowdon RJ, Friedt W (2004) Molecular markers in Brassica oilseed breeding: current status and future possibilities. Plant Breed 123:1–8

    Article  CAS  Google Scholar 

  • Tanksley SD, Ganal MW, Martin GB (1995) Chromosome langding: a paradigm for map-based gene cloning in plants with large genomes. Trends Genetics 11:63–68

    Article  CAS  Google Scholar 

  • Thomas CM, Vos P, Zabeau M, Jones D, Norcott KA, Chadwick BP, Jones JDG (1995) Identification of amplified restriction fragment polymorphism (AFLP) markers tightly linked to the tomato Cf-9 gene for resistance to Cladosprorium fulvum. Plant J 8:785–794

    Article  PubMed  CAS  Google Scholar 

  • Vos P, Hogers R, Bleeker M, Reijans M, Van de Lee T, Hornes M, Freijters A, Pot J, Peleman J, Kuiper M, Zabeau M (1995) AFLP: a new technique for DNA fingerprinting. Nucleic Acids Res 23:4407–4414

    Article  PubMed  CAS  Google Scholar 

  • Wang H, Hou GZ, Zhao JX (1993) Study on transfer of a recessive GMS for double-low Brassica napus. Oil Crop China 1:1–6

    Google Scholar 

  • Yi B, Chen YN, Lei SL, Tu JX, Fu TD (2006) Fine mapping of the recessive genic male-sterile gene (Bnms1) in Brassica napus L. Theor Appl Genet 113:643–650

    Article  PubMed  CAS  Google Scholar 

  • Zhang RM, Wang H (1996) Transferring of yellow seed canola to the recessive genic male sterile materials in Brassica napus. Southwest China J Agric Sci 9:33–37

    Google Scholar 

Download references

Acknowledgments

The authors wish to thank Prof. Meng for providing DNA of Tapidor × Ningyou 7 population. This research was financed by the funds from the High-tech program “863”(2006AA10Z146), the High-tech program “863”(2006AA10A), the National Key Basic Research Special Foundation of China (2001CB1088), the Program for Changjiang Scholar and Innovative Research Team in university (IRT0442), and the Program for “948” (2003-Q04).

Author information

Authors and Affiliations

  1. National Key Laboratory of Crop Genetic Improvement, National Sub-center of Rapeseed Improvement in Wuhan, Huazhong Agricultural University, Wuhan, 430070, China

    Shaolin Lei, Xueqin Yao, Bin Yi, Wei Chen, Chaozhi Ma, Jinxing Tu & Tingdong Fu

Authors
  1. Shaolin Lei
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xueqin Yao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Bin Yi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wei Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Chaozhi Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jinxing Tu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Tingdong Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jinxing Tu.

Additional information

Communicated by G. Wenzel.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lei, S., Yao, X., Yi, B. et al. Towards map-based cloning: fine mapping of a recessive genic male-sterile gene (BnMs2) in Brassica napus L. and syntenic region identification based on the Arabidopsis thaliana genome sequences. Theor Appl Genet 115, 643–651 (2007). https://doi.org/10.1007/s00122-007-0594-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00122-007-0594-1

Keywords

Search

Navigation