Gloxinia And Violet Transformation With Ccfaldh Gene

Formaldehyde (HCHO), one of the major indoor air pollutants, attracts worldwide attention because the exposure to formaldehyde may cause nasopharyngeal cancer and possibly leukemia. In the present work, We have generated transgenic flowering plants overexpressing a formaldehyde dehydrogenase-encoding gene from Chlorophytum comosum to improve the ability of plants to uptaken formaldehyde. The main results are as follows:1. The establishment of tissue culture system of gloxinia The effects of different culture mediums on the status of adventitious buds induction, proliferation, and radication of violet were tested by using leaves as explants. The results showed that the optimal medium for induction of adventitious buds was MS medium plus with1.0mg/L6-BA and0.1mg/L NAA; the optimal medium for radication was1/2MS medium with0.5mg/L NAA.2The establishment of tissue culture system of violetThe effects of different culture mediums on the status of adventitious buds induction, proliferation, and root of violet were tested by using leaves as explants. The results showed that the optimal medium for induction of adventitious buds was MS medium plus with1.0mg/L6-BA and0.2mg/L NAA; the optimal medium for proliferation was MS medium plus with1.0mg/L6-BA; the optimal medium for root was1/2MS medium with0.2mg/L NAA.3Agrobacterium-mediated genetic transformation of gloxinia and violetGenetic transformation of gloxinia:Leaves was pre-incubated in darkness for2days. The inoculation was carried out for6min when the agrobacterium cell density was adjusted to give an OD600of0.5-0.6. Following the inoculation period, excess bacteria was removed and the explants were transferred for co-cultivation. Co-cultivation was carried out in the dark for3days. After co-cultivation, the explants were transferred to the medium containing150mg/L kanamycin and500mg/L cefotaxime. After adventitious buds the explants were transferred to medium containing the same antibiotic. When sprout, they were transferred to rooting medium containing50mg/L kanamycin. Genetic transformation of violet:Leaves was pre-incubated in darkness for2days. The inoculation was carried out for10min when the agrobacterium cell density was adjusted to give an OD600of0.5-0.6. Following the inoculation period, excess bacteria was removed and the explants were transferred for co-cultivation. Co-cultivation was carried out in the dark for3days. After co-cultivation, the explants were transferred to the medium containing200mg/L kanamycin and500mg/L cefotaxime. After adventitious buds the explants were transferred to medium containing the same antibiotic. When sprout, they were transferred to rooting medium containing50mg/L kanamycin.4. Molecular detection of transgenic planWe had developed21PCR-positive independent transgenic lines by Agrobacterium-mediated transformation system for gloxinia. The expressions of foreign genes were determined by Real-time PCR methods, and the results shown that Ccfaldh gene was expressing at higher level in gloxinia lines6than wildtype. Through the analysis of formaldehyde absorption capacity, gloxinia formaldehyde absorption lines6levels are highest.