Development and validation of SSR markers related to flower color based on full-length transcriptome sequencing in Chrysanthemum

Abstract

Chrysanthemum (Chrysanthemum moriforlium Ramat.) is one of the most popular flowers worldwide, with very high ornamental and economic values. However, the limitations of available DNA molecular markers and the lack of full genomic sequences hinder the study of genetic diversity and the molecular breeding of chrysanthemum. Here, we developed simple sequence repeat (SSR) from the full-length transcriptome sequences of chrysanthemum cultivar 'Hechengxinghuo'. A total of 11,699 SSRs with mono-, di-, tri-, tetra-, penta- and hexanucleotide repeats were identified, of which eight out of eighteen SSR loci identified based on sixteen transcripts participated in carotenoid metabolism or anthocyanin synthesis were validated as polymorphic SSR markers. These SSRs were used to classify 117 chrysanthemum accessions with different flower colors at the DNA and cDNA levels. The results showed that four SSR markers of carotenoid metabolic pathway divided 117 chrysanthemum accessions into five groups at cDNA level and all purple chrysanthemum accessions were in the group III. Furthermore, the SSR marker CHS-3, LCYE-1 and 3MaT may be related to green color and the PSY-1b marker may be related to yellow color. Overall, our work may be provide a novel method for mining SSR markers associated with specific traits.

Figure 1

Numbers of mono-, di-, and trinucleotide repeat SSRs.

Figure 2

Electrophoretic map of the PSY-1a marker in some chrysanthemum accessions at the DNA and cDNA levels. Lanes 1 to 30 correspond to chrysanthemum accessions 1 to 30 in Supplementary Table 1; M: 100 bp DNA ladder marker. The images were cropped and the original images are presented in Supplementary Fig. 10.

Figure 3

Cluster analysis of 117 chrysanthemum accessions with different colors using LCYE-1, LCYE-4, PSY-1a, and PSY-1b SSR markers at DNA (a) and cDNA (b) levels.