Agrobacterium-Mediated Transformation Of Chrysanthemum×morifolium With Lc Gene

Chrysanthemum(Chrysanthemum×morifolium Ramat.) is one of the most famous pot flower and cut flower in the world.But there are two faults in chrysanthemum flower color breeding:1. Chrysanthemum have no blue-flowering cultivars.A main reason for the lack of blue flowers is the absence of 3′,5′-hydroxylated anthocyanins,which are synthesized by a key enzyme,flavonoid 3′, 5′-hydroxylase(F3′5′H);2.Few colorful red cultivars are found in Chrysanthemum.The application of genetic engineering including introducing transcription factor gene and enhancing coordinated expression of a number of genes can achieve a better plant transformation result.The results were as follows:1.The one new restriction enzyme SalI site was added to terminator behind Gus gene of pBI121 vector by PCR.Gus histochemical assay indicated that Gus gene can express as same as original pBI121 vector and the function of terminator was normal.The expression vector pBI121N-Lc was constructed by Lc gene using reconstruct vector pBI121N.2.The vector pBI121N-Lc was transferred into Chrysanthemum seven cultivars by Agrobacterium-mediated using leaf disk and tTCLs as the explants.Kanamycin-resistant shoot have been obtained in the six cultivars;only in three of the six cultivars kanamycin-resistant root were obtained.Results indicated that different Chrysanthemum cultivars have genotypic differences in the transformed and regenerated.3.The 48 Chrysanthemum 'riqietaohong' generation resistance plants were detected by PCR assay, 7 of them have the same positive band as Lc gene and 35S.The results indicated that Lc gene had been successfully introduced and integrated into the genome.And some transgenic lines roots become red.