Studies On Establishment Of Genetic Transformation System Of Dendranthema Indicum Var. Aromaticum And Preliminary Study Of Epidermal Hair

Dendranthema indicum var. aromaticum is one of endemic species in China.Since it has been found,widespread attention and research on it by the various scholars because of its rare rich aroma in Asteraceae, and the efficient antibacterial antioxidant activity of its essential oils. In this study,by manual adjustments of different media components and biological modifier concentration, the transformation system of Dendranthema indicum var. aromaticum has been preliminary established.And laid a theoretical foundation for gene expression studies and variety improvement work of Dendranthema indicum var. aromaticum. At the same time,the study of structure, density and glandular hairs secretions of Dendranthema indicum var. aromaticum trichomes was also conducted. In order lay a good foundation for the study of Dendranthema indicum var. aromaticum aroma mechanism. Following is the main results:(1)The genetic transformation receptor system of Dendranthema indicum var. aromaticum was established.MS medium was regarded at the basic medium for inducting adventitious buds.While additional6-BA1.0mg/L+NAA1.0mg/L medium induced the best results.The kanamycin optimal critical tolerance concentration is45mg/L,which is most suitable the Dendranthema indicum var. aromaticum leaves differentiation.And the optimum rooting critical tolerance concentration is60mg/L. Inhibitory concentration of cefotaxime in the culture of the delay stage selection200mg/L, and selection of100mg/L in the screening culture stages.(2) The genetic transformation system of Dendranthema indicum var. aromaticum was initially established.The leaves by tissue culture was considered as infection materials,and experiment with Agrobacterium EHA105.The genetic transformation system is that, pre-incubation for3d,infection for30min,co-culture for3d,and delay cultur for3d.Under this system,we could get the highest conversion rate. The transformed plants was19,while the number of experimental explants was1520.The conversion rate was1.25%.The result of PCR detection showed that the exogenous DNA has been successfully transferred into Dendranthema indicum var. aromaticum plant genome.(3) We also carried out a preliminary study on Dendranthema indicum var. aromaticum trichomes by microscopic examination.The result of its leaves trichome distribution is that young leaves, lower epidermis,and veins were respectivele greater than mature leaves,upper epidermis,and the edge of the leaf.Main types of trichomes were single row hair and T-wool. The main components of the blade trichomes secretions by GC-MS analysis contain Bicyclo[3.1.0]hexane,4-methylene-1-(1-methylethyl)-, Eucalyptol, Bicyclo[3.1.0]hexan-3-one,[1S-(1.alpha.,4.beta.,5.alpha.)]-, Bicyclo[2.2.1]heptan-2-ol,1,7,7-trimethyl-,(1S-endo)- ,.gamma.-Elemene,and.alpha.-Bisabolol,4-methyl-l-(1-methylethyl)-.In all components,the terpenes accounted for22.29%, alcohol for31.43%,and acids for11.56%.In addition,we also identified benzene for7.05%and alkanes for6.53%.