Molecular Typing of S-alleles through Identification, Characterization and cDNA Cloning for S-RNases in Sweet Cherry

in Journal of the American Society for Horticultural Science
Authors:
Ryutaro TaoLaboratory of Pomology, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan

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Hisayo YamaneLaboratory of Pomology, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan

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Akira SugiuraLaboratory of Pomology, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan

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Hideki MurayamaFaculty of Agriculture, Yamagata University, Tsuruoka 997-8555, Japan

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Hidenori SassaKihara Institute for Biological Research, Yokohama City University, Yokohama 244-0813, Japan

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Hitoshi MoriGraduate School of Bioagricultural Science, Nagoya University, Nagoya 464-8601, Japan

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This report identifies S-RNases of sweet cherry (Prunus avium L.) and presents information about cDNA sequences encoding the S-RNases, which leads to the development of a molecular typing system for S-alleles in this fruit tree species. Stylar proteins of sweet cherry were surveyed by two dimensional polyaclylamide gel electrophoresis (2D-PAGE) to identify S-proteins associated with gametophytic self-incompatibility. Glycoprotein spots linked to S-alleles were found in a group of proteins which had Mr and pI similar to those of other rosaceous S-RNases. These glycoproteins were present at highest concentration in the upper segment of the mature style and shared immunological characteristics and N-terminal sequences with those of S-RNases of other plant species. cDNAs encoding these glycoproteins were cloned based on the N-terminal sequences. Genomic DNA and RNA blot analyses and deduced amino acid sequences indicated that the cDNAs encode S-RNases; thus the S-proteins identified by 2D-PAGE are S-RNases. Although S1 to S6-alleles of sweet cherry cultivars could be distinguished from each other with the genomic DNA blot analysis, a much simpler method of PCR-based typing system was developed for the six S-alleles based on the DNA sequence data obtained from the cDNAs encoding S-RNases.

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