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Use of immunochemical and SSCP analyses to test homozygosity at theS locus ofBrassica oleracea genotypes
Authors:Valérie Delorme  Thierry Gaude  Philippe Heizmann  Christian Dumas
Institution:(1) Reconnaissance Cellulaire et Amélioration des Plantes, UMR CNRS-INRA, Ecole Normale Supérieure de Lyon, 46 Allée d'Italie, 69364 Lyon Cedex 07, France
Abstract:The self-incompatibility reaction of cruciferous plants prevents self-fertilization and has been shown to be controlled by at least two genes situated at a single multiallelic locus, theS locus. One of these two genes, theS locus glycoprotein (SLG) gene, encodes an abundant glycoprotein secreted to the cell wall of stigma papillae. Identification of thoseS alleles present at theS locus is of prime interest when studying the self-incompatibility response and can be achieved by identifying the SLG of the stigma. Here, we show that using anti-SLG antibodies in an immunochemical analysis, combined with a SSCP (single-strand conformation polymorphism) approach to characterize the corresponding stigma-specific, SLG mRNA, allowed the identification of plants heterogeneous at theS locus among populations of plants that were thought to be homozygous for known SLG alleles. This analysis stresses the importance of testing the homozygosity at theS locus of lines considered inbred for a knownS allele as mix-up of seeds may occur during the breeding programme.
Keywords:Brassica oleracea  immunochemical analysis  self-incompatibility  S locus glycoprotein  SSCP  stigma
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