Polyacrylamide gel electrophoresis of S-RNase fragments for identification of S-genotypes of Japanese pear (Pyrus pyrifolia)

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  • Kato Masaki
    Graduate School of Horticulture, Chiba University
  • Kato Shu
    Chiba Prefectural Agriculture and Forestry Research Center
  • Sassa Hidenori
    Graduate School of Horticulture, Chiba University

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  • Polyacrylamide gel electrophoresis of <i>S-RNase</i> fragments for identification of <i>S</i>-genotypes of Japanese pear (<i>Pyrus pyrifolia</i>)

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Japanese pear (Pyrus pyrifolia) exhibits gametophytic self-incompatibility (GSI) controlled by a complex and multiallelic S locus. The pistil-part product of the S locus is the polymorphic ribonuclease, S-RNase. Information on S-genotypes is important for the production and breeding of Japanese pears. Molecular analyses of S-genotypes of Japanese pear have been conducted with the CAPS (cleaved amplified polymorphic sequence) system; PCR amplification of S-RNase fragments by a common primer pair followed by digestion with restriction enzymes each of which cleaves a specific S haplotype. Here, we show that the separation of S-RNase fragments by polyacrylamide gel electrophoresis (PAGE) distinguishes four out of nine S haplotypes of Japanese pear without restriction digestion. S3-, S5-, S6- and S8-RNases were identified as distinct bands by PAGE. S3- and S5-RNases were separated by PAGE despite their identical fragment sizes. Using this system, three Japanese pear lines with unknown S-genotypes were analyzed. The newly determined S-genotypes of the lines were confirmed by CAPS analysis.

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