Sexual Plant Reproduction

, Volume 7, Issue 4, pp 203–208 | Cite as

A molecular approach to the identification of S-alleles in Brassica oleracea

  • J. Brace
  • G. J. King
  • D. J. Ockendon
Original Paper


A molecular technique for the identification of S-alleles involved in self-incompatibility has been used to analyse the S-allele reference collection of Brassica oleracea. The reference collection contains nearly 50 different lines each with a different S-allele present in the homozygous state. The technique consists of amplifying by the polymerase chain reaction (PCR) sequences belonging to the S multigene sequence family using a single pair of conserved primers. PCR products are then analysed further by digestion with six restriction enzymes followed by gel electrophoresis of the digestion products. A simple method of estimating the band sizes of the digestion products is described. The S-locus-related sequences can be distinguished from S-locus glycoprotein and S-receptor kinase genes by the restriction patterns. Furthermore, with any one restriction enzyme, several alleles showed the same restriction pattern. Alleles could therefore be grouped together. With two exceptions, each member of the S-allele reference collection showed a unique set of restriction patterns. Investigation of the exceptions using pollen tube growth tests showed that these accessions represented duplications within the collection. This technique therefore provides a simple and useful method for identifying different S-alleles.

Key words

Brassica oleracea Self-incompatibility S alleles PCR identification 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Boyes DC, Chen C-H, Tantikanjana T, Esch JJ, Nasrallah JB (1991) Isolation of a second S-locus-related cDNA from Brassica oleracea: genetic relationships between the S locus and two related loci. Genetics 27:211–228Google Scholar
  2. Brace J, Ockendon DJ, King GJ (1993) Development of a method for the identification of S alleles in Brassica oleracea based on digestion of PCR-amplified DNA with restriction endonucleases. Sex Plant Reprod 6:133–138Google Scholar
  3. Chen C-H, Nasrallah JB (1990) A new class of S sequences defined by a pollen recessive self-incompatibility allele of Brassica oleracea. Mol Gen Genet 222:241–248Google Scholar
  4. Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version II. Plant Mol Biol Rep 1:19–21Google Scholar
  5. Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programmes for the Vax. Nucleic Acids Res 12:387–395PubMedGoogle Scholar
  6. Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13Google Scholar
  7. Dwyer KG, Balent MA, Nasrallah JB, Nasrallah ME (1991) DNA sequences of self-incompatibility genes from Brassica campestris and B. oleracea: polymorphism predating speciation. Plant Mol Biol 16:481–486Google Scholar
  8. Hinata K, Nishio T (1978) S-allele specificity of stigma proteins in Brassica oleracea and B. campestris. Heredity 41:93–100Google Scholar
  9. Kandasamy MK, Paolillo DJ, Faraday CD, Nasrallah JB, Nasrallah ME (1989) The S-locus specific glycoproteins of Brassica accumulate in the cell wall of developing stigma papillae. Exp Biol 134:462–472Google Scholar
  10. Kho YO, Baer J (1968) Observing pollen tubes by means of fluorescence. Euphytica 17:298–302Google Scholar
  11. Lalonde BA, Nasrallah ME, Dwyer KG, Chen C-H, Barlow B, Nasrallah JB (1989) A highly conserved Brassica gene with homology to the S-locus-specific glycoprotein structural gene. The Plant Cell 1:249–258Google Scholar
  12. Nasrallah JB, Kao T-H, Goldberg ML, Nasrallah ME (1985) A cDNA clone encoding an S-locus-specific glycoprotein from Brassica oleracea. Nature 318:263–267Google Scholar
  13. Nasrallah JB, Kao T-H, Chen C-H, Goldberg ML, Nasrallah ME (1987) Amino acid sequence of glycoproteins encoded by three alleles of the S locus of Brassica oleracea. Nature 326:617–619Google Scholar
  14. Ockendon DJ (1980) Distribution of S-alleles and breeding structure of Cape broccoli (Brassica oleracea var italica). Theor Appl Genet 58:11–15Google Scholar
  15. Oerter KE, Munson PJ, McBride WO, Rodbard D (1990) Computerized estimation of size of nucleic acid fragments using the four-parameter logistic model. Analytical Biochem 189:235–243Google Scholar
  16. Scutt CP, Croy RRD (1992) An S5 self-incompatibility allele-specific cDNA sequence from Brassica oleracea shows high homology to the SLR2 gene. Mol Gen Genet 232:240–246Google Scholar
  17. Scutt CP, Gates PJ, Gatehouse JA, Boulter O, Croy RRD (1990) A cDNA encoding an S-locus-specific glycoprotein from Brassica oleracea plants encoding the S5 self-incompatibility allele. Mol Gen Genet 220:409–413Google Scholar
  18. Trick M, Flavell RB (1989) A homozygous S genotype of Brassica oleracea expresses two S-like genes. Mol Gen Genet 218:112–117Google Scholar
  19. Trick M, Heizmann P (1992) Sporophytic self-incompatibility systems: Brassica S-gene family. Int Rev Cytol 140:485–524Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • J. Brace
    • 1
  • G. J. King
    • 1
  • D. J. Ockendon
    • 1
  1. 1.Department of Breeding and GeneticsHorticulture Research InternationalWellesbourneUK

Personalised recommendations