Cloning of a new S-RNase and development of a PCR-RFLP system for the determination of the S-genotypes of Japanese pear [Pyrus pyrifolia]
2007
Kim, H.(Chiba Univ., Matsudo (Japan). Faculty of Horticulture) | Kakui, H. | Koba, T. | Hirata, Y. | Sassa, H.
Japanese pear exhibits gametophytic self-incompatibility that is controlled by the multiallelic S-locus. The S-locus encodes polymorphic S-RNase for the pistil-part S determinant in Japanese pear. Information about the S-genotypes is important for the selection of pollen donors for fruit production and breeding of Japanese pear cultivars. We determined the S-genotypes of three Japanese pear cultivars, 'Kinchaku' (Ssup(4)Ssup(k)), 'Natsuhikari' (Ssup(3)Ssup(4)) and 'Wakahikari' (Ssup(3)Ssup(4)), and an Fsub(1) hybrid cultivar between a Japanese pear cultivar and a European pear cultivar, 'Oharabeni' (Ssup(9)Ssup(e)), by PCR-RFLP analysis of S-RNase. A new Ssup(k)-RNase allele was identified in 'Kinchaku', and full length cDNA encoding Ssup(k)-RNase was sequenced. The Ssup(k)-RNase was not effectively amplified by the previously published primers. We designed a new PCR primer combination for the amplification of the Ssup(k)-RNase allele in addition to the Ssup(1) to Ssup(9)-RNase alleles of Japanese pear, and developed a PCR-RFLP system for identifying the Ssup(1)-to Ssup(9)- and Ssup(k)-alleles.
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