%0 Journal Article
%T Rapid identification of apple (Malus¡Ádomestica Borkh.) S alleles using sequencing-based DNA marker APPLid
%A Ichiro Kasajima
%A Nobuyuki Yoshikawa
%A Teppei Kikuchi
%J Archive of "Plant Biotechnology".
%D 2017
%R 10.5511/plantbiotechnology.17.0503a
%X All apple cultivars harbor the trait called self-incompatibility. Self-incompatibility represents that the pistils of the flowers are not successfully fertilized with own, the same cultivar¡¯s pollens. Compatibility or incompatibility of apple flowers are determined by S alleles. For example, the most popular apple cultivar ¡®Fuji¡¯ possesses the S1 and S9 alleles (S1S9). Thus, ¡®Fuji¡¯ is incompatible with S1S9 cultivars, but is compatible with the cultivars possessing different combinations of S alleles such as S2S7 and S1S7. Apple S alleles have been identified by performing a series of allele-specific PCR amplifications, to detect more than ten different S alleles separately. Here, we developed a new type of sequencing-based DNA marker of the apple S-RNase gene, which identifies S alleles. This DNA marker was named APPLid (apple S-allele identifier). A 53-base region in the first coding sequence of S-RNase is the target of APPLid sequencing. Variation in nucleotide sequences in this APPLid sequence enables allele identifications. This region is amplified from apple genomic DNA by using a pair of degenerate primers. The forward primer is attached with ¡®DS5 adaptor.¡¯ After PCR amplification, electrophoresis and gel extraction of 177-bp DNA fragments, APPLid sequence is determined by direct sequencing with a sequencing primer. The APPLid sequences of 20 apple cultivars completely matched their S alleles, which include triploid cultivars. In conclusion, APPLid identifies apple S alleles (S1, S2, S3, S4, S5, S7, S9, S10, S20, S24, S25, S26, S27 and S28, so far) just by a single sequencing analysis
%K apple
%K degenerate primer
%K DNA marker
%K DNA sequencing
%K S allele
%U https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6543762/