Molecular Markers Related To The Quality Of Carthamus Tinctorius L. Screening With AFLP

The quality (attribute character) of the Chinese medical materials is closely related to the content of its activities. Flos Carthami, dried flower petals of safflower (Carthamus tinctorius L.), is a significant TCM for promoting blood circulation and removing obstruction in the channels. The quality of Flos Carthami depends on the content of its active compound chalcone HS YA (hydroxysafflor yellow A). Therefore, the content of HSYA was considered as an important clue to screening fine varieties.Based on the strategy of bulk segregate analysis (BSA), two gene pools were respectively constructed according to the extreme highest and lowest content of HSYA in Flos Carthami. A total of 64 primer combinations were tested on the two gene pools. Of these, the strips of 48 primer pairs in PAGE electrophoresis were clearer and polymorphisms were better. Especially, the polymorphic bands of two primer combinations were sifted from the 48 primer pairs by trial and error, which were EcoR\-ACT-MseI-CAG and EcoRI-ACC-MseI-CTC. To confirm the linkage of candidate AFLP markers to quality trait of Flos Carthami, the parents, the two gene poors and the segregating F₂ population were screened for polymorphism. Finally, we obtained four special AFLP markers tightly linked to high content of HSYA in Flos Carthami, named AFLP5, AFLP7, AFLP15 and AFLP16. Respectively, the exact lengths of which were 936bp, 852bp, 730bp and 277bp by sequence information. Furthermore, the methord of cDNA-AFLP was used and six special cDNA-AFLP markers were found to be associated with high-content HSYA in Flos Carthami.Our study established a foundation for efficient breeding of Carthamus tinctorius L. and offers an effetive way for the research of gene expression and regulation mechanism of quality in offcinal.