Studies On Transformation Of ACC Oxide Antisense Gene Into Lily By Agrobacterium Tumfacience

Lily flower is one of the five world famous cut flowers, and it is widely favored by people for the characters of the big flower, varied flower color, graceful posture, and strong fragrance. The sales of fresh cut flower can be counted in billions around the world. But like other fresh cut flowers, lily will decay rapidly while blooming. How to prolong the longevity of lily flower has become an urgent problem being settled on practice and consumption and one of the main subjects for studying recently. According to results, lily is a kind of flower of ethylene releases.It is a concomitance between blossom and ethylene releases and decay. On the basic research of senescent mechanism in lily cut flower, through transferring ACC oxidase antisense gene to the cells of lily, and controlling the senescence of lily by manipulating the expression of ethylene, the purpose of prolonging the longevity of lily flower can be attained.On the basis of the ACC oxidase antisense expression vectors built, the system of callus recipient and the system of direct differentiation is built in oriental hybrid lily'Sorbonne'.Then tresferring ACC oxidase antisense gene to the lily by Agrobacterium tumefaciens-mediated. For the purpose of achieving transgenic lily of ACC oxide antisense gene and provide some beneficial foundation in lily in the future. The results were as following:1. A regeneration system of callus was initiated from Lilium oriental'Sorbonne'A regeneration system of callus was initiated from Lilium oriental'Sorbonne'with the scale leaf, the regenerated scale leaf and petioles as explants. Abundant calluses could be induced from the three explants by the same media.The combination of MS+NAA 0.5 mg/L+6-BA 0.4 mg/L+sucrose 90 g/L+VB1 4.0 mg/L was the optimum medium for the induction of calluses; the medium of MS+6-BA 0.5 mg/L was the best medium for regeneration adventitions buds from callus, and the sensibility of callus to hygromycin was 20 mg/L.2. A regeneration system of direct differentiation was initiated from Lilium oriental'Sorbonne'A regeneration system of direct differentiation was initiated from Lilium oriental'Sorbonne'with the regenerated scale leaf and petioles as explants. Adventitious buds could be induced from the regenerated scale leaf and petioles in media MS+NAA 1.5 mg/L and MS+NAA 0.5 mg/L respectively. The sensibility of regenerated scale leaf and petioles to hygromycin was 15 mg/L.3. An Agrobacterium tumefaciens-mediated transformation protocol was established for Lilium oriental'Sorbonne'An Agrobacterium tumefaciens-mediated transformation protocol was established by using the recipient system of callus and the recipient system of direct differentiation as transformation recipient...