Study On The System For Agrobacterium-Mediated Genetic Transformation Of Lilium With PeACC Gene In RNAi Vector

Lily is well-known for the characters of the big flower ,graceful posture,bright color and strong fragrant. The scales volume of fresh cut flower is considerable billions in the whole world. Lily is also suitable for use in the garden, shrubborderand potted plant. But while lily flower blooming, it was the same as the other ones senescening rapidly. How to prolong the longevity of lily flower was subject for studying recent 20 years. With the limit of common hybridization and negative effection of chemical on fresh cut flowers, many studies are engaged in transgenetic plant.In this study, we established the acceptor system of Agrobacteruim-mediated transfotmation of"Sorbonne". Then by using the seedling leaf as the gene transformation acceptor, the foreign gene PeACC which cloned from peony was built as a dsRNA-mediated gene silencing vector and was transformed by Agrobacterium EHA105.The conclusions are as followed.1 The medium suitable for inducing the scales to differentiate adventitious bud was MS+6-BA1.0 mg·L–1+NAA0.1 mg·L–1.2 High-efficiency acceptor system of direct differentiation for the gene transformation of"Sorboone"leaves was conducted. (1) The medium suitable for the petioles of the test-tube plantlets to differentiate adventitious bud was MS+6-BA2.0mg·L–1+NAA0.3mg·L–1;(2) The medium suitable for the scales leaves of the test-tube plantlets to differentiate adventitious bud was MS+6-BA1.0mg·L–1 +2,4-D0.3mg·L–1;(3) The optimum medium for inducing root is MS+NAA0.5mg/l.;(4) The screening concentration of kanamycin for the petioles of the test-tube plantlets was 80mg·L–1 and the scales leaves of the test-tube plantlets was 120mg·L–1 ;(5) The proper concentration of Carbenicillin for restraining bacteria was 300mg·L–1.3 The study on influcing factors of transformation showed that:(1) It was suitable for instrusion with OD600=0.8 Agrobactium;(2) The efficiency of intrusion could be increase by supplying 100 umol·L–1 AS;(3) The petioles and scales leaves of plantlets in vitro showed a higher transformtion rate with preculture 3 days.4 40 kanamycin resistant seedlings were achieved, 6 kanamycin resistant seedlings were at random selected for PCR and got the strip. It proved preliminarily that foreign gene had been integrated into these 6 plants genome.