Abstract
SummaryClementines are an important group of citrus cultivars. However, little is known about their genetic diversity. In this study, methylation-sensitive amplification polymorphism (MSAP), based on the application of isoschizomers (Hpa II and Msp I), was used to analyse cytosine methylation patterns in 18 Clementine cultivars. Conventional AFLP analysis showed only two polymorphic bands out of a total of 1,822 AFLP bands generated using 28 primer pairs. Three types of bands were generated by MSAP using 27 pairs of primers. Type I were present in both Eco RI + Hpa II and Eco RI + Msp I gels. Type II or Type III were present only in Eco RI + Hpa II, or Eco RI + Msp I gels, respectively. The total numbers of these three Types of bands were 1,377, 98, and 93, respectively. Among these three Types of bands, the number of polymorphic bands were 76 (5.5%), 30 (30.6%), and 15 (16.1%), respectively. Of these, non-reproducible polymorphic bands in duplicated samples were 27 (35.5%), 18 (60.0%), and 9 (60.0%). Each Clementine cultivar had specific and relatively fixed methylated CCGG sites, which could be revealed from their specific MSAP patterns. The reproducible polymorphic bands in duplicated samples were used to analyse the extent of variation in methylation. The diversity of the cultivars revealed by the resulting dendrogram suggests that DNA methylation may be one of the main factors accounting for the difference in agronomic traits between Clementine cultivars.
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