Simultaneous transformation of three biphenyl dioxygenase bacterial genes into Arabidopsis plants

Polychlorinated biphenyls (PCBs) are chlorinated biphenyl rings that due to their properties such as heat resistance and stability were widely used in different industries from 1920s to 1980s. Due to their characteristics such as stability, resistance to decomposition and harmful effects on human health, their production was banned in 1980s., Worldwide contamination of water and soil with PCBs is one of the important environmental problems at present time. A way to reduce PCBs contamination is transformation and expression of biphenyl dioxygenase (BPDO) genes having catabolic degradation activity of polychlorinated biphenyls into plants. The aim of present study was developing a method for simultaneous transformation of bphA, bphE, and bphG genes, coding for the components of BPDO enzyme, into Arabidopsis plants. According to obtained results, three bphA, bphE, and bphG genes cloned into pGreen vector were transformed into E.coli, Agrobacterium and as well as Arabidopsis plants. There were differences in transformation efficiency of two strains LBA4404 and C58C1 of Agrobacterium, used in this study. The highest number of transgenic plants 0.85%% was obtained by LBA4404 strain. The transgenic nature of Arabidopsis plantlets was confirmed by selecting fully green plants on 50 mg/l Kanamycin as well as PCR analysis. Transgenic plants were successfully transferred into soil and continued their growth.