| Lilies(Lilium spp.)not only have special beautiful flowers, but also have officinal and edible value. They are very popular for many people in the world. With the planting areas expanded gradually, the flowers often bring lots of disease on the field which are sensitive for illness. So they produced hugeness lost. But tradition preventions increase labour cost,meanwhile they could effect the environment. It is more effective and direct method for preventing the plant diseases to use using the disease-resistant varieties. Because the tradition breeding method need a long time for breeding and is effected by F1 generation greatly, the demand of the Lilies production couldn't be meted. On the background of biology technique fast development, we could transfer new disease-resistant gene of other sources into the Lilies by using genetic engineering means .It will break a new path in the new disease-resistant breeding of Lilies.This study took the less-disease resistant oriental lily'Sorbonne'as material. We establish regeneration system of direct shoot method, introduce callus from Lilies bulbs。We observe the transformation from callus to somatic embryo. We compare the cases of shoot on different medias in the process of direct regeneration. We use the bulbs of Lilies as the explants of genetic transformation, The broad-spectrum disease resistance gene (NPR1 gene) transferred into Lilies mediated by the agrobacterium. we establish genetic transformation system of Lilies. Finally, we get positive plants after PCR. The main content and result of experiment are as follows:1.The regeneration system though bulbs: we use the slice of Lilies bulbs as explants, MS basal medium supplemented with 6-BA0.4mg/L and NAA1.0mg/L was advantageous to shoot induction, the frequency of shoot induction is 96.7%; MS basal medium supplemented with IBA0.5 mg/L could develop their roots, the rate of robust plantlets is 95.2%.2.The callus regeneration of Lilies by somatic embryogenesis: we use the bulbs of Lilies as explants, the good method for induce callus is MS basal medium supplemented with 2,4-D2.0mg/L and 6-BA0.5 mg/L and LH 300mg/L, the rate of produce callus is 68.3%. MS+2,4-D 2.0 mg/L +6-BA 0.5mg/L+LH150mg/L+Gln150mg/L is the best culture medium that the induction of embryogenesis callus ,and the rate of produce embryogenesis callus is 75.0% .Then transfer the embryogenesis callus into the culture media MS+2,4-D 0.5 mg/L +KT 0.3 mg/L + Sucrose 40g/L, which rate is 60.0%. Somatic embryo could germinate on the MS basal medium supplemented with 6-BA 0.5mg/L. The appearance and growth of somatic embryo were observed through paraffin slices. Histological observations showed that surface cells first experienced dedifferentiation and followed with callus-forming, then the top of callus appear a tubercular structure. These embryogenic cell masses experienced globular, heart stages and developed into whole plantlets .3.The establishment of genetic transformation system: Taking the slice of Lilies bulbs as explants, pre-culture of callus for 2d;the agrobacterium tumefaciens solution was OD600=0.5,the time of infect was 20min,the time of co-culture was 2d at 20℃,the step up the concentration of Km 30mg/L from the first day,80mg/L from the third day,130mg/L from the sixth day. Concentration of Cef was 400mg/L in the process of sterilization, until the plantlet formed. MS added 100mg/L trans-Rerulic acid and subtracted NH4NO3 can improve the transformation ratio.4.Detection of the transformed plant:For shoot regeneration,we gained 56 resistant plants.Three of them is PCR positive plant, positive percentage is 5.4% and transformation percentage is 0.33% in 900 explants.
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