Abstract
Chrysanthemum morifolium, is a well-known flowering plant worldwide, and has a high commercial, floricultural, and medicinal value. In this study, simple-sequence repeat (SSR) markers were generated from EST datasets and were applied to assess the genetic diversity among 32 cultivars. A total of 218 in silico SSR loci were identified from 7300 C. morifolium ESTs retrieved from GenBank. Of all SSR loci, 61.47% of them (134) were hexa-nucleotide repeats, followed by tri-nucleotide repeats (17.89%), di-nucleotide repeats (12.39%), tetra-nucleotide repeats (4.13%), and penta-nucleotide repeats (4.13%). In this study, 17 novel EST-SSR markers were verified. Along with 38 SSR markers reported previously, 55 C. morifolium SSR markers were selected for further genetic diversity analysis. PCR amplification of these EST-SSRs produced 1319 fragments, 1306 of which showed polymorphism. The average polymorphism information content of the SSR primer pairs was 0.972 (0.938–0.993), which showed high genetic diversity among C. morifolium cultivars. Based on SSR markers, 32 C. morifolium cultivars were separated into two main groups by partitioning of the clusters using the unweighted pair group method with arithmetic mean dendrogram, which was further supported by a principal coordinate analysis plot. Phylogenetic relationship among C. morifolium cultivars as revealed by SSR markers was highly consistent with the classification of medicinal C. morifolium populations according to their origin and ecological distribution. Our results demonstrated that SSR markers were highly reproducible and informative, and could be used to evaluate genetic diversity and relationships among medicinal C. morifolium cultivars.
Highlights
Chrysanthemum morifolium (Ramat.) Kitam is an important member of the family Asteraceae
17 novel C. morifolium EST-simple-sequence repeat (SSR) were developed successfully, and 38 C. nankingense EST-SSRs were confirmed with transferability for application in a related species
These 55 pairs of SSR primers were used for further genetic diversity analysis in C. morifolium cultivars (Table 4)
Summary
Chrysanthemum morifolium (Ramat.) Kitam is an important member of the family Asteraceae. A large number of C. morifolium cultivars are ornamental and medicinally important plants that are planted all over the world (Teixeira da Silva, 2003; Bhattacharya and da Silva, 2006). In addition to their esthetic value, some C. morifolium cultivars are used medicinally for their curative. Molecular markers have potentials to reveal the genetic diversity among medicinal C. morifolium germplasms. A few studies have been reported on genetic diversity in cultivated chrysanthemums (Bhattacharya and da Silva, 2006; Xu et al, 2006; Shao et al, 2010; Zhang et al, 2011b), preliminary genetic linkage map construction, and QTL detection (Zhang et al, 2010, 2011a) using RAPD, AFLP, ISSR, and SRAP markers
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