Development of a method for the identification of S alleles in Brassica oleracea based on digestion of PCR-amplified DNA with restriction endonucleases

Brace, Jennifer L.; Ockendon, D. J.; King, Graham J.

doi:10.1007/bf00227658

Cited by 53 publications

(31 citation statements)

References 18 publications

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“…All of the S-genotypes of plant materials used in this study were checked through direct cloning of SLG genes by polymerase chain reaction (PCR) using class I SLG-specific primers (Brace et al, 1993) and class II SLG-specific primers (Nishio et al, 1996).…”

Section: Plant Materialsmentioning

confidence: 99%

The Dominance of Alleles Controlling Self-Incompatibility in Brassica Pollen Is Regulated at the RNA Level

et al. 2002

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Self-incompatibility (SI) inBrassica is controlled sporophytically by the multiallelic S -locus. The SI phenotype of pollen in an S -heterozygote is determined by the relationship between the two S -haplotypes it carries, and dominant/recessive relationships often are observed between the two S -haplotypes. The S -locus protein 11 ( SP11 , also known as the S -locus cysteine-rich protein) gene has been cloned from many pollen-dominant S -haplotypes (class I) and shown to encode the pollen S -determinant. However, SP11 from pollen-recessive S -haplotypes (class II) has never been identified by homology-based cloning strategies, and how the dominant/recessive interactions between the two classes occur was not known. We report here the identification and molecular characterization of SP11 s from six class II S -haplotypes of B. rapa and B. oleracea . Phylogenetic analysis revealed that the class II SP11s form a distinct group separated from class I SP11s. The promoter sequences and expression patterns of SP11 s also were different between the two classes. The mRNA of class II SP11 , which was detected predominantly in the anther tapetum in homozygotes, was not detected in the heterozygotes of class I and class II S -haplotypes, suggesting that the dominant/recessive relationships of pollen are regulated at the mRNA level of SP11 s. INTRODUCTIONMany species of hermaphrodite plants have evolved mechanisms to prevent self-fertilization. Self-incompatibility (SI) is one physiological means of avoiding self-fertilization through recognition of self-pollen in or on the female pistil. Classic genetic analyses have revealed the presence of two major types of homomorphic SI systems, gametophytic and sporophytic (de Nettancourt, 1977). Although the recognition of self-pollen is controlled genetically by a single highly polymorphic locus called the S -locus in both of these systems, the SI phenotype of pollen (gametophyte) is determined by its own S -haplotype in the gametophytic system, whereas in the sporophytic system, the SI phenotype is controlled by the S -haplotypes of the diploid parent (sporophyte).The majority of the members of the cruciferous plant genus Brassica possess a strong sporophytic SI system. Thus, the SI phenotype of pollen as well as stigma is determined by relationships between the two S -haplotypes carried by its parent (Bateman, 1955). In other words, a codominant or a dominant/recessive relationship between the two S -haplotypes influences the ultimate SI phenotype of both pollen and stigma (Thompson and Taylor, 1966). The following observations have been made about dominance relationships among S -haplotypes: (1) codominance is common; (2) dominance/recessiveness is frequent in pollen; (3) dominance relationships among stigmas are different from those among pollen; and (4) dominance relationships are nonlinear (Thompson and Taylor, 1966;Ockendon, 1975;Visser et al., 1982; Hatakeyama et al., 1998a).Recent molecular studies have revealed that the S -locus of Brassica encodes three highly polymorphic ...

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Section: Plant Materialsmentioning

confidence: 99%

The Dominance of Alleles Controlling Self-Incompatibility in Brassica Pollen Is Regulated at the RNA Level

et al. 2002

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“…Relatively conserved regions were identified that can be used to design primers for PCR-based analysis of alleles. Combining PCR with restriction enzyme digestion has made it possible to obtain allele-specific bands, for typing plants of self-incompatible horticultural species (Brace et al, 1993;Janssens et al, 1995). This opens the way for sequencing and thus study of allelic diversity among plants in natural populations, even when, as is usual, they are heterozygotes.…”

Section: Quantitative Measures Of the Self-incompatibility Locus Polymentioning

confidence: 99%

Flowering plant self-incompatibility: the molecular population genetics of Brassica S-loci

Charlesworth

Awadalla

1998

Heredity

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Self-incompatibility systems in different angiosperm families are reviewed, and the evidence that incompatibility has arisen several times is outlined. New data on the sequence polymorphism of self-incompatibility loci from two different angiosperm families are compared with results from other highly polymorphic loci, particularly MHC loci. We discuss what molecular genetic analyses of these sequences can tell us about the nature and maintenance of the polymorphism at self-incompatibility loci. We suggest that there is evidence for recombination at the Brassica selfincompatibility loci, so that it may be possible to discern regions that are particularly functionally important in the recognition reaction, even though the long-term maintenance of polymorphisms in these amino acid residues has caused the evolution of many other sequence differences between alleles.

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“…In Brassica, Brace et aL (1993) reported the preferential amplification of specific S alleles, possibly due to the low level of homology between primer and some of the S sequences. We have made a similar observation in one S-segregating rye inbred line (not shown).…”

Section: Resultsmentioning

confidence: 99%

“…made to identify S alleles with molecular methods such as the use of S-specific oligonucleotide probes (Scutt and Croy, 1992) or PCR primers which amplify S allele-specific sequences (Brace et aL, 1993). We report on an alternative approach for S allele identification in rye by denaturing gradient gel electrophoresis (DGGE) of PCR amplification products.…”

Section: Discussionmentioning

confidence: 99%

“…After denaturing gradient gel electrophoresis (DGGE), a 280 bp PCR-fregment displays a polymorphism perfectly correlated to the underlying S-genotypes. This is the first report on S-related DNA polymorphism in a bifactorial self-incompatibility system of the Poaceae.made to identify S alleles with molecular methods such as the use of S-specific oligonucleotide probes (Scutt and Croy, 1992) or PCR primers which amplify S allele-specific sequences (Brace et aL, 1993). We report on an alternative approach for S allele identification in rye by denaturing gradient gel electrophoresis (DGGE) of PCR amplification products.…”

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confidence: 99%

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Identification of S‐locus linked PCR fragments in rye (Secale cereale L.) by denaturing gradient gel electrophoresis

Wehling¹,

Hackauf²,

Wricke³

1994

The Plant Journal

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SummaryRye inbred lines segregating at the S-locus and homozygous at the Z-locus were investigated by PCR with primers derived from Brassica SLG-sequences. After denaturing gradient gel electrophoresis (DGGE), a 280 bp PCR-fregment displays a polymorphism perfectly correlated to the underlying S-genotypes. This is the first report on S-related DNA polymorphism in a bifactorial self-incompatibility system of the Poaceae.made to identify S alleles with molecular methods such as the use of S-specific oligonucleotide probes (Scutt and Croy, 1992) or PCR primers which amplify S allele-specific sequences (Brace et aL, 1993). We report on an alternative approach for S allele identification in rye by denaturing gradient gel electrophoresis (DGGE) of PCR amplification products. The DGGE technique, often combined with PCR, provides a fast and non-radioactive method of analysing DNA fragments of similar size for allelic sequence variation and has successfully been used in plants (Dweikat et aL, 1990; Riedel et aL, 1993). The power of this method is based on altered melting properties of short double-stranded DNA fragments, even in the presence of only single base pair differences.

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Development of a method for the identification of S alleles in Brassica oleracea based on digestion of PCR-amplified DNA with restriction endonucleases

Cited by 53 publications

References 18 publications

The Dominance of Alleles Controlling Self-Incompatibility in Brassica Pollen Is Regulated at the RNA Level

The Dominance of Alleles Controlling Self-Incompatibility in Brassica Pollen Is Regulated at the RNA Level

Flowering plant self-incompatibility: the molecular population genetics of Brassica S-loci

Identification of S‐locus linked PCR fragments in rye (Secale cereale L.) by denaturing gradient gel electrophoresis

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