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Application of SCAR markers to self-incompatibility genotyping in breeding lines of radish (Raphanus sativus L.)

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초록

Self-incompatibility (SI) prevents self-fertilization by inhibiting the pollen tube growth of self-pollen. Molecular analysis has revealed that the S locus comprises a number of genes, such as the S-locus glycoprotein (SLG), the S-locus receptor kinase (SRK), and SP11 (SCR). Although molecular markers related to those genes have been developed, a simple S-haplotype detecting method has not been reported due to the highly polymorphic and relatively small coding regions. In this study, the sequence characterized amplified region (SCAR) markers were used to establish an efficient radish genotyping method. We identified the S-haplotypes of 192 radish accessions using 19 different markers, which proved to be highly reliable. The accessions were assigned to 17 types of S-haplotypes, including 8 types of SRKs and 9 types of SLGs. Since the developed SCAR markers are based on their gene sequences, we could easily identify the S-haplotypes by a single specific band, with the highest frequencies detected for SLG 5, SRK 1, and SLG 1, in order. Among the tested markers, the SLG 1, SRK 1, and SRK 5 markers exhibited high reliability, compared to phenotypic results. Furthermore, we identified the seven types of unreported SLGs using SLG Class -I and -II specific markers. Although the developed SCAR markers still need to be improved for the genotyping of all S-haplotypes, these markers could be helpful for monitoring inbred lines, and for developing the MAS in radish breeding programs.

Abstract

Self-incompatibility (SI) prevents self-fertilization by inhibiting the pollen tube growth of self-pollen. Molecular analysis has revealed that the S locus comprises a number of genes, such as the S-locus glycoprotein (SLG), the S-locus receptor kinase (SRK), and SP11 (SCR). Although molecular markers related to those genes have been developed, a simple S-haplotype detecting method has not been reported due to the highly polymorphic and relatively small coding regions. In this study, the sequence characterized amplified region (SCAR) markers were used to establish an efficient radish genotyping method. We identified the S-haplotypes of 192 radish accessions using 19 different markers, which proved to be highly reliable. The accessions were assigned to 17 types of S-haplotypes, including 8 types of SRKs and 9 types of SLGs. Since the developed SCAR markers are based on their gene sequences, we could easily identify the S-haplotypes by a single specific band, with the highest frequencies detected for SLG 5, SRK 1, and SLG 1, in order. Among the tested markers, the SLG 1, SRK 1, and SRK 5 markers exhibited high reliability, compared to phenotypic results. Furthermore, we identified the seven types of unreported SLGs using SLG Class -I and -II specific markers. Although the developed SCAR markers still need to be improved for the genotyping of all S-haplotypes, these markers could be helpful for monitoring inbred lines, and for developing the MAS in radish breeding programs.

참고문헌 (14)

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  2. Bateman AJ, "Self-incompatibility system in angios­perms. (III)", Cruciferae. Heredity, 9, pp.52~68 (1955)
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  4. Goring DR, "The S-locus receptor kinase gene in a self-incompatibility Brassica napus line encodes a functional serine", threonine kinase, 1992, pp.4, 1273~1281
  5. Lim SH, "Identi­fication and classification of S haplotypes in Raphanus sativus by PCR-RFLP of the S locus glycoprotein (SLG) gene and the S-locus receptor kinase (SRK) gene", Theor. Appl. Genet., 104, pp.1253~1262 (2002)
  6. Nasrallah JB, "Amino-acid sequence of glycoproteins encoded by three alleles of the S locus of Brassica oleracea", Nature, 326, pp.617~619 (1987)
  7. Nasrallah JB, "The self-incompatibility genes of Brassica: Expression and use in genetic ablation of floral tissues", Annu. Rev. Plant Phy­siol. Plant Mol. Biol., 42, pp.393~420 (1991)
  8. Niikura S, "Identification of self-incom­patibility alleles (S) by PCR-RFLP in radish (Raphanus sativus L.)", Euphytica, 102, pp.379~384 (1998)
  9. Sato Y, "Diversification and Alteration of Recognition Specificity of the Pollen Ligand SP11", SCR in Self-Incompatibility, 2004, pp.16, 3230~3241
  10. 서정팔, "무 육성계통의 자가불화합성 반수형 동정 및 자가불화합성 활력과의 관계", 한국육종학회지, 35(4), pp.313~318 (2003), 한국육종학회
  11. Jeong-Pal Seo, "Identification of S-haplotypes by PCR-RFLP and DNA Sequencing of S-Locus Receptor Kinase (SRK) in Raphanus sativus Inbred Lines", Horticulture, Environment, and Biotechnology, 45(3), pp.119~126 (2004), 한국원예학회
  12. Suzuki G, "Watanabe M. Hinata K. Genomic orga­nization of the S-locus: identification and characterization of genes in SLG", SRK region of an S9 haplotype of Bra­ssica campestris, 1999, pp.152, 391~400
  13. Takayama S, "Sequences of S-glycoproteins, products of the Brassica campestris self-incompatibility locus", Nature, 326, pp.102~110 (1987)
  14. Thompson JD, "CLUS TAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position specific gap penalties and weight matrix choice", Nucleic Acids Res., 22, pp.4673~4680 (1994)

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