Establishment Of High Efficient Regeneration System And Study On Genetic Transformation Of Two Insect-resistant Genes In Gerbera Jamesonii

Gerbera jamesonii Bolus is a famous cut flower with big gorgeous flower, which have long vase life and great field in the world. In addition, people enjoy its color, appearance and artistic conception. It is worthy worldly planting this ornamental plant. Now the most of the good varieties of Gerbera jamesonii are come from foreign countries. It is necessary to cultivate the good varieties in China.This work includes: Establishing the regeneration system and preliminarily establishing transformation system by transforming two insect–resistant genes of Gerbera jamesonii. The intention is providing material and technology for application of the good varieties. The receptacle,younger leaves,stalk,footstalk of Gerbera jamesonii were used as explants to establish the regeneration system and two insect–resistant genes were transfered by Agrabacterium tumefaciens mediated. The transgenetic plants were detected by PCR and Southern Blot. The factors which affect transformation frequency and preliminarily established transforming system have be studied. The main results were as follows:1. The regeneration system of Gerbera jamesonii has been established. the medium MS+BA10mg/L+NAA0.5mg/Lwas used for inducing the calli and the medium containing MS+BA2.0mg/L+NAA1.0mg/L+AD5.0mg/L+PVP1.0mg/L+Vc1.0mg/L was used for inducing the bud. Multiplicating the bud by using MS+BA2.0 mg/L +NAA0.5 mg/L. The plant has the fine rooting capacity on 1/2MS basic medium containing 0.2mg/LIBA.2. The transformation system has been preliminarily established. The procedure: The leafstalk tissue pieces were pre-cultivated for 2 days. The leafstalk tissue pieces were immersed in the bacterial suspension with OD600 of 0.4 for 15 minutes and then co-cultivated on MS basic medium for 2 days. After washing the explants to remove excessive bacterium, the leafstalk tissue pieces was induced on MS basic medium supplemented with 400mg/LCarb+30mg/Kan. 8 plants had been gained in second culture on above medium.3. PCR analysis demonstrated the transgenic plants were positive. Southern blotting show that the DNA of transgenic plants has markedness signal, but not in the negative...