Tissue Culture And Quality-related Gene Cloning In Carthmus Tinctorius L.

Flos Carthami, dried flower petals of Carthamus tinctorius L, is an important crude drug in traditional Chinese medicine for promoting blood circulation and removing obstruction in the channels. The chemical composition hydroxysafflor yellow A (HSYA) is the main active component in safflower. The quality of safflower is influenced by the content of the HSYA. It is important to find out the functional genes to regular pattern of heredity about the safflower quality. As a result, identification the genes linked to HSYA of Carthamus tinctorius L. at the cDNA molecular level will lay a foundation for breeding spineless cultivars at the molecular level. Meanwhile, the purpose of this study was to develop a plantlet regeneration system for a Safflower breeding program with the final objective of applying transgenic techniques to prepare for further validation of gene function. The main results were as follows:1 We found the optimal time for sterilizing of seeds in 0.1% HgCl2 was 30min.We discovered that cotyledon was the best material for induction by testing different materials. Shoot regeneration from primary was optimal MS basal salts medium containing 0.1mg/L NAA+02.0mg/L6-BA+15mg/L or KT0.1mg/L NAA+02.0mg/L6-BA+20mg/LKT2 We mensurated the content of HSYA in 09 safflower, ranged from 0.00%to 2.6%3 In the present study, the gene F3H has been firstly cloned, using the method of rapid amplification of cDNA ends (RACE). Characterization, structure prediction and biotinformatics analsysis were performed. The full length of cDNA of F3H was 1415 bp (genbank accession No. JF737995) with 1086 bp ORF (open reading frame) encoding a polypeptide of 361 amino acid residues.4 The expression level of F3H in safflower was determined by real-time PCR, and the results revealed that the expression level of F3H in safflower with yellow flowers is much higher than that with the white flowers.