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Fang J.-G.,Nanjing Agricultural University | Song C.-N.,Nanjing Agricultural University | Qian J.-L.,Suzhou Agricultural Science and Technology College | Zhang X.-Y.,Nanjing Agricultural University | And 3 more authors.
Acta Physiologiae Plantarum

Sweet orange is an important group of citrus cultivars, which includes a number of bud sport cultivars. Little is known about the CpG methylation status of the CCGG sequences in the orange genome. In this study, methylation-sensitive amplification polymorphism (MSAP), based on the application of isoschizomers (Hpa II and Msp I), was first used to analyze cytosine methylation patterns in 57 orange cultivars that were not fully differentiated by regular DNA molecular markers. Three types of bands were generated from ten primer pairs. Type I bands were present following restriction with Eco RI + Hpa II and Eco RI + Msp I; type II or type III were present only following restriction with either Eco RI + Hpa II or with Eco RI + Msp I. The total number of these three types of bands was 802, 72, and 157, respectively. Among these, the number of polymorphic bands were 244 (30.2%), 23 (31.9%), and 32 (20.4%), in type I, II and III, respectively. The methylation patterns of these 57 cultivars are discussed and assessed by dendrograms derived from the analysis of polymorphic MSAP bands. The distribution of polymorphic bands of the above three types demonstrate the methylation patterns and frequency at the cytosine loci. We suggest that methylation events could be more frequent than demethylation events, and that the methylation patterns maybe associated with phenotypic traits. © 2010 Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków. Source

Xiaoying Z.,Nanjing Agricultural University | Xiaoying Z.,Suzhou Agricultural Science and Technology College | Qian J.,Suzhou Agricultural Science and Technology College | Wang H.,Suzhou Agricultural Science and Technology College | And 2 more authors.

DNA fingerprinting is both a popular and important technique with great advantages in plant cultivar identification. Despite this fact, this technique has never been used widely and efficiently in practical plant identification since analysis and recording of data generated from fingerprinting and genotyping for future reference is tedious and difficult. We report a novel strategy developed for efficient recording of DNA molecular fingerprints of genotyped plant individuals, which could be readily used as efficient reference information for quick plant identification. 25 orange- or white-fleshed loquat cultivars that are commercially important in China, were identified using this strategy which employed random amplified polymorphic DNA (RAPD) marker. Utilization of the strategy was verified by identification of three randomly chosen groups of loquat cultivars among the 25. The advantages of this approach include use of fewer primers and separation of all cultivars by the corresponding primers marked on the diagram. Importantly, the diagram could be easily referred to in identification of loquat cultivars in commerial practice. Source

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