Study On The Transformation Of Lotus Corniculatus With Agrobacterium

Lotus corniculatus L,which is one of the important perennial legume forage grass, widely distributes in Midwest area and all over the world. Lotus can resist to abiotic and biotic stress, as well as with many excellent quality traits of forage. For instance, the quality include resistance for trampling, re-growth, grazing and enduring, furthermore, Lotus live for longer green stage, for good forage quality on foods, cattle will not be inflated by eating. During the life process, Lotus often undergoes the threatening of low temperature, drought and high salinity environments. So, it is very important for improving the resistance to harmful environments and other factor in Lotus. This experiment aimed to construct the method of agrobacterium-mediated genetic transformation system in Lotus. Our work will build on the foundation of transgenic research in the future. By the agrobacterium-mediated method, the DREB gene of alfalfa was transferred into lotus.The results show that:Cytyledons of Lotus corniculatus as the explants for infection, pre-incubated for2days, when the concentration of agrobacteria reach to OD6oo=0.2-0.4the selected explants was infected for5minutes, and then begin to co-cultured with infected explants for2days, after that, transferred explants to callus induction culture containing350mg/L Cef for callus induction, using the20mg/L Kan+350mg/L Cef to delay selection finally, the genetic transformation system were carried out for the differentiation and rooting. The seedlings were selected for transgenic checking by PCR method, by this way,21strains show positive.