Expression of a Self-Incompatibility Gene in a Self-Compatible Line of Brassica oleracea

Gaude, Thierry; Friry, Aline; Heizmann, Philippe; Mariac, Chantal; Rougier, M.; Fobis, Isabelle; Dumas, Christian

doi:10.2307/3869430

Cited by 15 publications

(41 citation statements)

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“…acephala, previously denoted P57 Sc [9] and here simply designated Sc, was kindly provided by Dr V6ronique Ruffio from the INRA centre at Rennes, France. The genetics and self-compatibility phenotype of this line have already been described [9]. The class I lines homozygous for the $3 and S16 haplotypes, respectively, were B. oleracea var.…”

Section: Plant Materialsmentioning

confidence: 99%

“…Details on obtaining MAb 85-36-71 and its reactivity towards SLGs have already been described [9]. For the production of polyclonal anti-class I SLG antibodies, rabbits were immunized with the peptide IN-TLSSTESLY cross-linked to ovalbumin by bisdiazobenzidine [37].…”

Section: Antibody Productionmentioning

confidence: 99%

“…On the other hand, in a recent study devoted to the expression of the SLG gene in a self-compatible (Sc) line of B. oleracea, we have shown that the SLG-Sc product was highly homologous to the SLG glycoproteins of the pollen recessive $2 and $5 haplotypes [9]. The present study was undertaken to analyse comparatively the self-incompatibility phenotype and the expression of the SLG gene in the Sc line and in the three class II S haplotypes of Brassica oleracea.…”

Section: Introductionmentioning

confidence: 99%

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Expression level of the SLG gene is not correlated with the self-incompatibility phenotype in the class II S haplotypes of Brassica oleracea

et al. 1995

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In Brassica, the S-locus glycoprotein (SLG) gene has been strongly implicated in the self-incompatibility reaction. Several alleles of this locus have been sequenced, and accordingly grouped as class I (corresponding to dominant S-alleles) and class II (recessive). We recently showed that a self-compatible (Sc) line of Brassica oleracea expressed a class II-like SLG (SLG-Sc) gene. Here, we report that the SLG-Sc glycoprotein is electrophoretically and immunochemically very similar to the recessive SLG-S15 glycoprotein, and is similarly expressed in stigmatic papillae. Moreover, by seed yield analysis, we observed that both alleles are associated with a self-compatibility response, in contrast with the other known recessive S haplotypes (S2 and S5). By genomic DNA blot analysis, we show the existence of molecular homologies between the Sc and S15 haplotypes, but demonstrate that they are not identical. On the other hand, we also report that the S2 haplotype expresses very low amounts of SLG glycoproteins, although it exhibits a self-incompatible phenotype. These results strongly question the precise role of the SLG gene in the molecular mechanisms that control the self-incompatibility reaction of Brassica.

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Section: Plant Materialsmentioning

confidence: 99%

Section: Antibody Productionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Expression level of the SLG gene is not correlated with the self-incompatibility phenotype in the class II S haplotypes of Brassica oleracea

et al. 1995

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“…Proteins were electrophoresed through a 7.5% SDS͞PAGE gel, electroblotted, and immunodetected as described (5,29).…”

Section: Phosphorylation In Vitro Of Microsomal Membrane Preparationsmentioning

confidence: 99%

The integral membrane S-locus receptor kinase of Brassica has serine/threonine kinase activity in a membranous environment and spontaneously forms oligomers in planta

Giranton¹

2000

Proceedings of the National Academy of Sciences

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To gain further insight into the mode of action of S-locus receptor kinase (SRK), a receptor-like kinase involved in the self-incompatibility response in Brassica, different recombinant SRK proteins have been expressed in a membranous environment using the insect cell͞baculovirus system. Recombinant SRK proteins exhibited properties close to those of the endogenous stigmatic SRK protein and were found to autophosphorylate on serine and threonine residues in insect cell microsomes. Autophosphorylation was constitutive because it did not require the presence of pollen or stigma extracts in the phosphorylation buffer. Phosphorylation was shown to occur in trans, suggesting the existence of constitutive homooligomers of membrane-anchored recombinant SRK. To investigate the physiological relevance of these results, we have examined the oligomeric status of SRK in planta in cross-linking experiments and by velocity sedimentation on sucrose gradients. Our data strongly suggest that SRK is associated both with other SRK molecules and other stigma proteins in nonpollinated flowers. These findings may have important implications for our understanding of self-pollen signaling. baculovirus ͉ oligomerization ͉ sporophytic self-incompatibility I n the Brassica family, the cell-cell interaction that leads to the rejection of self-pollen at the stigmatic surface [the selfincompatibility (SI) reaction] is controlled genetically by the multiallelic S-locus (1). When the pollen parent and the stigma share common S haplotypes, pollen germination or pollen tube growth is inhibited, thereby preventing self-fertilization. Several genes have been localized at the S locus but only two, the S-locus glycoprotein (SLG) and the S-locus receptor kinase (SRK) genes, exhibit the polymorphic nature expected for genes involved in the SI response (2, 3). SLG encodes a secreted glycoprotein that accumulates in the cell wall of papillae (4), whereas SRK encodes a plasmalemma-anchored glycoprotein (5, 6). SRK is structurally analogous to animal receptor kinases and belongs to the plant receptor-like kinase (RLK) family (7). Although its membrane topology has not been experimentally defined, DNA sequence analysis predicts that SRK consists of three domains: an extracellular glycosylated N-terminal domain (the S-domain) that shares extensive homology with SLG, a membrane-spanning domain, and a cytoplasmic domain (3). The cytoplasmic domain has been shown to have serine͞threonine kinase activity when expressed in bacteria (8). In the S 3 haplotype, SRK has been shown to encode, in addition to the integral membrane SRK protein, a soluble truncated form corresponding to the S-domain of SRK, the eSRK protein (9).Although there is some controversy as to whether SLG is required for the SI response (10, 11), there is strong evidence that SRK is necessary. For example, SRK was shown to be mutated in two Brassica lines that exhibited a self-compatible phenotype (12, 13). Moreover, Stahl and coworkers (14) recently have shown that the SI phenotype was altered i...

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“…An identical situation was found when recessive probe (SLG-P57SC) was used on S recessive genotypes. In that set of experiments (not shown) the immunolabelling methodology [ 8 ] involving a monoclonal antibody against the B peptide, characteristic of the recessive S-alleles [5] could be tested, since in that case one of the two membranes we used for tissue printing was nitrocellulose. Therefore direct comparison between RNA and protein expression could be carried out on the very same plant material.…”

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confidence: 99%

Tissue printing and its applications in self-incompatibility studies

1993

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In this study, the tissue printing technique has been used to rapidly localize in female tissues the presence of specific mRNA representing the products (or some of the products) of the self-incompatibility S-locus gene(s). The methodology, initially developed for Brassica oleracea (sporophytic self-incompatibility) has been successfully employed on Solanum chacoense (gametophytic self-incompatibility). In the Brassica system tissue printing has allowed rapid discrimination between S alleles belonging to class 1 (dominant types) vs. class 2 (recessive types), and thus parallels findings obtained by restriction analyses. In the Solanum system the level of the S-RNase messages was analysed by scanning laser densitometry, and it was found that the message levels of the allele S14 declined faster than those coming from S13 in mature flowers.

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Expression of a Self-Incompatibility Gene in a Self-Compatible Line of Brassica oleracea

Cited by 15 publications

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Expression level of the SLG gene is not correlated with the self-incompatibility phenotype in the class II S haplotypes of Brassica oleracea

Expression level of the SLG gene is not correlated with the self-incompatibility phenotype in the class II S haplotypes of Brassica oleracea

The integral membrane S-locus receptor kinase of Brassica has serine/threonine kinase activity in a membranous environment and spontaneously forms oligomers in planta

Tissue printing and its applications in self-incompatibility studies

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