Molecular mapping of vernalization requirement and fertility restoration genes in carrot

Abstract

Carrot (Daucus carota L.) is a cool-season vegetable normally classified as a biennial species, requiring vernalization to induce flowering. Nevertheless, some cultivars adapted to warmer climates require less vernalization and can be classified as annual. Most modern carrot cultivars are hybrids which rely upon cytoplasmic male-sterility for commercial production. One major gene controlling floral initiation and several genes restoring male fertility have been reported but none have been mapped. The objective of the present work was to develop the first linkage map of carrot locating the genomic regions that control vernalization response and fertility restoration. Using an F2 progeny, derived from the intercross between the annual cultivar ‘Criolla INTA’ and a petaloid male sterile biennial carrot evaluated over 2 years, both early flowering habit, which we name Vrn1, and restoration of petaloid cytoplasmic male sterility, which we name Rf1, were found to be dominant traits conditioned by single genes. On a map of 355 markers covering all 9 chromosomes with a total map length of 669 cM and an average marker-to-marker distance of 1.88 cM, Vrn1 mapped to chromosome 2 with flanking markers at 0.70 and 0.46 cM, and Rf1 mapped to chromosome 9 with flanking markers at 4.38 and 1.12 cM. These are the first two reproductive traits mapped in the carrot genome, and their map location and flanking markers provide valuable tools for studying traits important for carrot domestication and reproductive biology, as well as facilitating carrot breeding.

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

Fig. 1

References

  1. Alessandro MS (2011) Estudio genético y molecular de la respuesta a la vernalización en zanahoria (Daucus carota L.). Ph.D. thesis, PROBIOL, Universidad Nacional de Cuyo, Argentina

  2. Alessandro MS, Galmarini CR (2007) Inheritance of vernalization requirement in carrot. J Am Soc Hort Sci 132:525–529

    Google Scholar 

  3. Atherton JG, Craigon J, Basher EA (1990) Flowering and bolting in carrot. I. Juvenility, cardinal temperatures and thermal times for vernalization. J Hort Sci 65:423–429

    Google Scholar 

  4. Bach IC, Olesen A, Simon PW (2002) PCR-based markers to differentiate the mitochondrial genomes of petaloid and male fertile carrot (Daucus carota L.). Euphytica 127:353–365

    Article  CAS  Google Scholar 

  5. Boiteux LS (2000) Characterization of the Meloidogyne javanica resistance locus employing molecular markers and isolation of candidate disease resistance loci in the carrot (Daucus carota L.) genome. Ph.D. thesis, The University of Wisconsin, Madison

  6. Boiteux LS, Belter JG, Roberts PA, Simon PW (2000) RAPD linkage map of the genomic region encompassing the root-knot nematode (Meloidogyne javanica) resistance locus in carrot. Theor Appl Genet 100:439–446

    Article  CAS  Google Scholar 

  7. Bradeen JM, Simon PW (1998) Conversion of an AFLP fragment linked to the carrot Y 2 locus to a simple, codominant PCR-based marker. Theor Appl Genet 97:960–967

    Article  CAS  Google Scholar 

  8. Bradeen JM, Simon PW (2007) Carrot. In: Genome mapping and molecular breeding in plants. In: Kole C (ed), vol 5, Vegetables, Chapter 4. Springer, Berlin

  9. Bradeen JM, Naess SK, Song J, Haberlach GT, Wielgus SM, Buell CR, Jiang J, Helgeson JP (2003) Concomitant reiterative BAC walking and fine genetic mapping enable physical map development for the broad-spectrum late blight resistance region, RB. Mol Gen Genomics 269:603–611

    Article  CAS  Google Scholar 

  10. Briard M, Le Clerc V, Grzebelus D, Senalik D, Simon PW (2000) Modified protocols for rapid carrot genomic DNA extraction and AFLP analysis using silver stain or radioisotopes. Plant Mol Biol Report 18:235–241

    Article  CAS  Google Scholar 

  11. Carlsson J, Glimelius K (2011) Cytoplasmic male-sterility and nuclear encoded fertility restoration. In: Kempken F (ed) Plant mitochondria. Springer, New York, pp 469–491

  12. Cavagnaro PF, Chung S, Szklarczyk M, Grzebelus D, Senalik D, Atkins AE, Simon PW (2009) Characterization of a deep-coverage carrot (Daucus carota L.) BAC library and initial analysis of BAC-end sequences. Mol Genet Genomics 281:273–288

    Google Scholar 

  13. Cavagnaro PF, Chung S-M, Manin S, Yildiz M, Ali A, Alessandro MS, Iorizzo M, Senalik DA, Simon PW (2011) Microsatellite isolation and marker development in carrot—genomic distribution, linkage mapping, genetic diversity analysis and marker transferability across Apiaceae. BCM Genomics 12:386

    Article  CAS  Google Scholar 

  14. Chahal A, Sidhu HS, Wolyn DJ (1998) A fertile revertant from petaloid cytoplasmic male-sterile carrot has a rearranged mitochondrial genome. Theor Appl Genet 97:450–455

    Article  CAS  Google Scholar 

  15. Dias Tagliacozzo GM, Valio IFM (1994) Effect of vernalization on flowering of Daucus carota (Cvs Nantes and Brasilia). Revista Brasileira de Fisiologia Vegetal. 6:71–73

    Google Scholar 

  16. Eckardt NA (2006) Cytoplasmic male sterility and fertility restoration. Plant Cell 18:515–517

    Article  CAS  Google Scholar 

  17. Ferreira ME, Satagopan J, Yandell BS, Williams PH, Osborn TC (1995) Mapping loci controlling vernalization requirement and flowering time in Brassica napus. Theor Appl Genet 90:727–732

    Article  Google Scholar 

  18. Galmarini CR, Della Gaspera P (1996) Determinación de requerimientos de pre-vernalización en zanahorias (Daucus carota L.) anuales. Actas de la XXI Reunión Argentina de Fisiología Vegetal, Mendoza, p 82

  19. Hansche PE, Gabelman WH (1963) Digenic control of male sterility in carrots, Daucus carota L. Crop Sci 3:383–386

    Article  Google Scholar 

  20. Iorizzo M, Senalik DA, Grzebelus D, Bowman M, Cavagnaro PF, Matvienko M, Ashrafi H, Van Deynze A, Simon PW (2011) De novo assembly and characterization of the carrot transcriptome reveals novel genes, new markers, and genetic diversity. BMC Genomics 12:389

    PubMed  Article  CAS  Google Scholar 

  21. Iovene M, Cavagnaro PF, Senalik D, Buell CR, Jiang J, Simon PW (2011) Comparative FISH mapping of Daucus species (Apiaceae family). Chromosome Res 19(4):493–506

    PubMed  Article  CAS  Google Scholar 

  22. Just BJ, Santos CAF, Fonseca MEN, Boiteux LS, Oloizia BB, Simon PW (2007) Carotenoid biosynthesis structural genes in carrot (Daucus carota): isolation, sequence-characterization, single nucleotide polymorphism (SNP) markers and genome mapping. Theor Appl Genet 114:693–704

    PubMed  Article  CAS  Google Scholar 

  23. Kole C, Quijada P, Michaels SD, Amasino RM, Osborn TC (2001) Evidence for homology of flowering-time genes VFR2 from Brassica rapa and FLC from Arabidopsis thaliana. Theor Appl Genet 102:425–430

    Article  CAS  Google Scholar 

  24. Lan TH, Paterson AH (2000) Comparative mapping of quantitative trait loci sculpting the curd of Brassica oleraceae. Genetics 155:1927–1954

    PubMed  CAS  Google Scholar 

  25. Linke B, Nothnagel T, Borner T (2003) Flower development in carrot CMS plants: mitochondria affect the expression of MADS-box genes homologous to GLOBOSA and DEFICIENS. Plant J 34:27–37

    PubMed  Article  CAS  Google Scholar 

  26. Martinez-Castilla LP, Alvarez-Buylla ER (2003) Adaptive evolution in the Arabidopsis MADS-box gene family inferred from its complete resolved phylogeny. Proc Natl Acad Sci USA 100:13407–13412

    PubMed  Article  CAS  Google Scholar 

  27. Michaels SD, Amasino RM (1999) FLOWERING LOCUS C encodes a novel MADS domain protein that acts as a repressor of flowering. Plant Cell 11:949–956

    PubMed  CAS  Google Scholar 

  28. Morelock TE (1974) Influence of cytoplasmic source on expression of male sterility in carrot (Daucus carota L.). Ph.D. thesis, The University of Wisconsin, Madison

  29. Morelock TE, Simon PW, Peterson CE (1996) Wisconsin wild: another petaloid male-sterile cytoplasm for carrot. HortScience 31:887–888

    Google Scholar 

  30. Osborn TC, Kole C, Parkin IAP, Sharpe AG, Kuiper M, Lydiate DJ, Trick M (1997) Comparison of flowering time genes in Brassica rapa, B. napus and Arabidopsis thaliana. Genetics 146:1123–1129

    PubMed  CAS  Google Scholar 

  31. Reeves PA, He Y, Schmitz RJ, Amasino RM, Panella LW, Richards CM (2007) Evolutionary conservation of the FLOWERING LOCUS C-mediated vernalization response: evidence from the sugar beet. Genetics 176:295–307

    Google Scholar 

  32. Rubatzky VE, Quiros CF, Simon PW (1999) Carrots and related vegetable umbelliferae. Crop production science in horticultural series: 10. CABI Publishers, New York

  33. Santos CAF, Simon PW (2002) Some AFLP amplicons are highly conserved DNA sequences mapping to the same linkage groups in two F2 populations of carrot. Genet Mol Biol 25(2):195–201

    CAS  Google Scholar 

  34. Santos CAF, Simon PW (2004) Merging carrot linkage groups based on conserved dominant AFLP markers in F2 populations. J Am Soc Hort Sci 129:211–217

    CAS  Google Scholar 

  35. Schnable PS, Wise RP (1998) The molecular basis of cytoplasmic male sterility and fertility restoration. Trends Plant Sci 3:175–180

    Article  Google Scholar 

  36. Schuelke M (2000) An economic method for fluorescent labeling of PCR fragments. Nat Biotechnol 18:233–234

    PubMed  Article  CAS  Google Scholar 

  37. Schulz B, Westphal L, Wricke G (1993) Linkage groups of isozymes, RFLP and RAPD markers in carrot (Daucus carota L. sativus). Euphytica 74:67–76

    Article  CAS  Google Scholar 

  38. Simon PW, Freeman RE, Vieira JV, Boiteux LS, Briard M, Nothnagel T, Michalik B, Young-Seok Kwon (2008) Carrot. In: Prohens J, Nuez F (eds) Handbook of Plant Breeding, vol 2, Vegetables II: Fabaceae, Liliaceae, Solanaceae and Umbelliferae. Springer, Heidelberg, pp 327–357

  39. Thompson DJ (1961) Studies on the inheritance of male sterility in the carrot (Daucus carota L. var. sativa). Proc Am Soc Hort Sci 78:332–338

    Google Scholar 

  40. Van Ooijen JW, Voorrips RE (2001) JoinMap® Version 3.0, software for the calculation of genetic linkage maps. Plant Research International, Wageningen, The Netherlands

  41. Vivek BS, Simon PW (1999) Linkage relationships among molecular markers and storage root traits of carrot (Daucus carota L. ssp. sativus). Theor Appl Genet 99:58–64

    Article  CAS  Google Scholar 

  42. Voorrips RE (2002) MapChart: software for the graphical presentation of linkage maps and QTLs. J Hered 93(1):77–78

    PubMed  Article  CAS  Google Scholar 

  43. Vos P, Hogers R, Bleeker M (1995) AFLP a new technique for DNA fingerprinting. Nucleic Acids Res 23:4407–4414

    PubMed  Article  CAS  Google Scholar 

  44. Welch JE, Grimball EL (1947) Male sterility in carrot. Science 106:594

    PubMed  Article  CAS  Google Scholar 

  45. Westphal L, Wricke G (1997) Construction of a linkage map of Daucus carota L. sativus and its application for the mapping of disease resistance and restorer genes. J Appl Genet 38A:13–19

    Google Scholar 

  46. Wolyn DJ, Chahal A (1998) Nuclear and cytoplasmic interactions for petaloid male-sterile accessions of wild carrot (Daucus carota L.). J Am Soc Hort Sci 123(5):849–853

    Google Scholar 

  47. Yan L, Loukoianov A, Blechl A, Tranquilli G, Ramakrishna W, San Miguel P, Bennetzen JL, Echenique V, Dubcovsky J (2004) The wheat VRN2 gene is a flowering repressor down-regulated by vernalization. Science 303:640–1644

    Google Scholar 

Download references

Acknowledgments

The authors acknowledge Dr. Aamir Ali (Department of Biological Sciences, University of Sargodha, Sargodha, Pakistan) for valuable assistance on experiments concerning marker evaluations.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Claudio R. Galmarini.

Additional information

Communicated by Y. Xue.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 12 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Alessandro, M.S., Galmarini, C.R., Iorizzo, M. et al. Molecular mapping of vernalization requirement and fertility restoration genes in carrot. Theor Appl Genet 126, 415–423 (2013). https://doi.org/10.1007/s00122-012-1989-1

Download citation

Keywords

  • Linkage Group
  • Cytoplasmic Male Sterility
  • AFLP Marker
  • Fertility Restoration
  • Vernalization Requirement