Studies On LjCYC1 Transforming Saintpaulia Ionantha And CFL Transforming Cyclamen Persicum And Cymbidium Goeringii × Cymbidium Hybidium

Flowertypes and flowering time are important quality characteristics in ornamental plant. Saintpaulia ionantha,Cyclamen persicum and orchid are the three kinds flowers which people like deeply.Although Saintpaulia ionantha is rich in flower color, but has single flowertypes;while Cyclamen persicum, especially orchid has long vegetative period,if we can obtain the new species which has different flowertypes and flowering time,the market prospect will be very wide.CYC gene is mainly involved in controlling dorsal-ventral asymmetry of flowes,while LFY gene took part in flower transition and is related to flowering time,furthermore, LFY gene also plays a vital role in each stage of flower development.In order to research their influence on flower development,we transformed LjCYCl gene (the homologous of Lotus japonicus) to Saintpaulia ionantha,CFL gene (the homologous of Cucumber) to Cyclamen persicum and Cymbidium goeringii×Cymbidium hybridium,hope to gain the new cultivars which has changed in flowertypes and flowering time.The main results obtained were as following:1 The genetic transformation system of Saintpaulia ionantha was established and optimized.2 PCR detects showed that the transformation efficiency of LjCYC1 gene was 16.67% after optimization,it's 3 times more than control.Further detection by GFP and RT-PCR proved that the LjCYC1 gene was integrated into Saintpaulia ionantha's genome.3 The main new flower phenotypes in transgetic plants are as follows:(1) The flower symmetry was obviously changed from actinomorphy to belateral symmetry, the frequency of occurrences is 14.3%; (2) Partial sepal wre changed to petal, the frequency of occurrences is 7.1%; (4) The amount of sepals in one flower were reduced from five or six to four, its frequency is 7.1%; (4) the petals were deeply crack to the base of corolla and shape and number of petals were changed, the frequency of occurrences is 7.1%; (5) The stamens were changed into petals or degenerated, the frequency of occurrences is 57.1%; (6)The pistil degenerated or divided, the frequency of occurrences is 11.4%; (7) The partial flowers colour in two transgetic strains were changed from purple to pink.4 The genetic transformation system of Cyclamen persicum was established with leaf,petiole and callus. Detecting by PCR,GUS and PT-PCR,The resistance plants showed postive, and the transformation efficiency were up to 3.14%,0.35% and 1.67% respectively.5 To explore the genetic transformation methods of Cymbidium goeringii×Cymbidium hybridium, the protocorms transformated with Agrobacterium tumefaciens (OD600=0.3) cultured the screen medium supplemented with hygromycin.Detecting the resisent protocorms and plants by PCR indicated that the CFL gene was integrated into Cymbidium goeringii×Cymbidium hybridium genome preliminary After culturing four months on medium without hygromycin, the positive protocorms were transfered to inducing flower medium.The flower buds formated gradually after culturing 50 days, the frequency of flower bud formation can reach to 75% according to the amount of bottles or 5% according to the amount of in vitro seedlings after culturing 150 days,while the flower buds frequencies of wild type control were 5% and 0.7% respectively in the same culture condition.