, Volume 164, Issue 2, pp 309–316 | Cite as

Identification of ‘Sib’ plants in hybrid cauliflowers using microsatellite markers

  • Ida A. Astarini
  • Julie A. Plummer
  • Rachel A. Lancaster
  • Guijun Yan


Hybrid cauliflowers have been developed to exploit heterosis and to improve uniformity of production. Two breeding systems are commonly employed, self-incompatibility (SI) and cytoplasmic male sterility (CMS). Sibs, assumed to be self-inbred, often contaminate hybrid seed lots in the SI system and whilst self-inbreeding is not possible in the CMS system, plants that look like sibs occur. The objective of this study was to develop microsatellite markers for male and female cauliflower parent lines of both SI and CMS systems and to use them to screen sibs and aberrant plants in F1 hybrids. Fifty six pairs of microsatellite primers were screened and 8 primer pairs produced co-dominant markers in parent plants and two pairs of markers were chosen for purity testing of F1 hybrid seeds. Controlled pollinations were conducted in the glasshouse to produce hybrid and selfed-seeds. These seeds were grown in a field trial to identify morphologically normal and sib plants and to assess the reliability of microsatellite markers in detecting sib plants. Microsatellite analysis of morphological sib plants from the SI system revealed that these were not always self-inbred, in contrast, most self-inbred plants showed normal growth. Similarly, all morphological sibs from the CMS system showed hybrid bands. This suggests that morphological sibs were not always due to selfing but possibly to an interaction between genetic and environmental factors and this requires further investigation.


Brassica oleracea var. botrytis Cytoplasmic male sterility Self-inbred Self-incompatibility SSR 


  1. Astarini IA, Yan G, Plummer JA (1999) Interspecific hybridisation of Boronias. Aust J Bot 47:851–864CrossRefGoogle Scholar
  2. Astarini IA, Plummer JA, Lancaster RA, Yan G (2004) Fingerprinting of cauliflower cultivars using RAPD markers. Aust J Agric Res 55:117–124CrossRefGoogle Scholar
  3. Bond JM, Mogg RJ, Squire GR, Johnstone C (2004) Microsatellite amplification in Brassica napus cultivars: cultivar variability and relationship to a long-term feral population. Euphytica 139:173–178CrossRefGoogle Scholar
  4. Boury S, Lutz I, Gavalda M-C, Guidet F, Schlesser A (1992) Genetic fingerprinting in cauliflower by the RAPD method and determination of the level of inbreeding in a set of F1 hybrid seeds. Agronomie 12:669–681CrossRefGoogle Scholar
  5. Cardi T, Earle ED (1997) Production of new CMS Brassica oleracea by transfer of ‘Anand’ cytoplasm from B. rapa through protoplast fusion. Theor Appl Genet 94:204–212CrossRefGoogle Scholar
  6. Crisp P, Tapsell CR (1993) Cauliflower, Brassica oleracea L. In: G. Kalloo B.O. Bergh (eds) Genetic improvement of vegetables crops. Pergamon Press, OxfordGoogle Scholar
  7. Crockett PA, Bhalla PL, Lee CK, Singh MB (2000) RAPD analysis of seed purity in a commercial hybrid cabbage (Brassica oleraceae var. capitata) cultivar. Genome 43:317–321PubMedCrossRefGoogle Scholar
  8. Crockett PA, Singh MB, Lee CK, Bhalla PL (2002) Genetic purity analysis of hybrid broccoli (Brassica oleracea var. italica) seeds using RAPD PCR. Aust J Agric Res 53:51–54CrossRefGoogle Scholar
  9. Fitzgerald DM, Barry D, Dawson PR, Cassells AC (1997) The application of image analysis in determining sib proportion and aberrant characterization in F1 hybrid Brassica population. Seed Sci Tech 25:503–509Google Scholar
  10. Fu T, Ping S, Xiaoniu Y, Guangsheng Y (1992) Overcoming self-incompatibility of Brassica napus by salt (NaCl) spray. Plant Breed 109:255–258CrossRefGoogle Scholar
  11. Fujime Y, Okuda N (1996) The physiology of flowering in Brassicas, especially about cauliflower and broccoli. Acta Hort 407:247–254Google Scholar
  12. Hallidri M, Pertena D (2002) Self-incompatibility test in cabbage (B. oleracea var capitata). Acta Hort 579:117–122Google Scholar
  13. Harvey E, Smith BM (1987) A recent survey of sib content in F1 hybrid Brussel sprout varieties. Cruciferae Newsl 12:122–123Google Scholar
  14. Heslop-Harrison J, Heslop-Harrison Y (1970) Evaluation of pollen viability by enzymatically-induced fluorescence; intracellular hydrolysis of fluorescein diacetate. Stain Tech 45:115–120Google Scholar
  15. Holland RL, McNeilly T (1985) Genotype environment interaction and sib content in F1 hybrid Brussels sprouts. Euphytica 34:371–376CrossRefGoogle Scholar
  16. International Seed Testing Association (ISTA) (2003) International rules for seed testing. Bassersdorf, CH-SwitzerlandGoogle Scholar
  17. Makaroff CA (1995) Cytoplasmic male sterility in Brassica species. In: Levings CS III, Vasil IF (eds) The molecular biology of plant mitochondria. Kluwer Academic Publishers, LondonGoogle Scholar
  18. Malik M, Vyas P, Rangaswamy NS, Shivanna KR (1999) Development of two new cytoplasmic male-sterile lines in B. juncea through wide hybridization. Plant Breed 118:75–78CrossRefGoogle Scholar
  19. Meng X, Ma H, Zhang W, Wang D (1998) A fast procedure for genetic purity determination of head Chinese cabbage purity seed based on RAPD markers. Seed Scie Tech 26:829–833Google Scholar
  20. Monteiro AA, Gabelman WH, William PH (1988) Use of sodium chloride solution to overcome self-incompatibility in Brassica campestris. HortScience 23:876–877Google Scholar
  21. Ruffio-Chable V, Chatelet P, Thomas G (2000) Developmentally “Aberrant” plants in F1 hybrids of Brassica oleracea. Acta Hort 539:89–94Google Scholar
  22. Saal B, Plieske J, Quiros C, Struss D (2001) Microsatellite markers for genome analysis in Brassica. II. Assignment of rapeseed microsatellites to the A and C genomes and genetic mapping in Brassica oleracea L. Theor Appl Genet 102:695–699CrossRefGoogle Scholar
  23. Williams JGK, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18:6531–6535PubMedCrossRefGoogle Scholar
  24. Zheng XY, Liu Y (1994) Inbred testing of Chinese cabbage F1 varieties by peroxidase and esterase isozyme analysis. Acta Hort Sin 21:65–70Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Ida A. Astarini
    • 1
    • 2
  • Julie A. Plummer
    • 1
  • Rachel A. Lancaster
    • 3
  • Guijun Yan
    • 1
  1. 1.Plant Biology, Faculty of Natural and Agricultural SciencesThe University of Western AustraliaCrawleyAustralia
  2. 2.Jurusan Biology, Fakultas Matematika dan Ilmu Pengetahuan AlamUniversitas UdayanaDenpasarIndonesia
  3. 3.Department of Agriculture, Western Australia BunburyAustralia

Personalised recommendations