Multiple Transgenes Poplar Plants Obtained By Biolistic Bombardment

Populus L. is an important genus globally planted in commercial forest plantations and some species are used as model plants in the study of forest biotechnology. Up to now, there are a lot of reports of genetic transformation in poplar, but the ideal plant is still difficult to achieve because a lot of poplar important characters are usually controlled by multiple genes, and most researches into poplar genetic transformation is that one or two genes were transferred into plants. In order to obtain transgenic plants with high yields, high quality and multiple resistant characters, the transformations of three or more genes into poplar were needed and manipulate multiple genes.In this study, genes of SacB, Vgb, Bt Cry3A+OC-1, JERFs were co-transferred into poplar plants of P. ×euramericacana 'Guariento' and P.alba ×P.glandulosa via biolistic bombardment. The results were as follows:1 Kanamycin selection . 27 P.×euramericacana 'Guariento' and 41 P.alba ×P.glandulosa resistant lines were obtained.2 Dot blotting . The results showed that 18 of 27 P .×euramericacana 'Guariento' were positive (approximately 66.7%), 29 of 41 P.alba ×P.glandulosa were positive(approximately 70.7%), and most lines were negative. All five transgenes were detected in 8 P.×euramericacana 'Guariento' lines, three transgenes in 7 lines, two transgenes in 2 lines, and one transgene in 1 line. All five transgenes were detected in 8 lines P.alba ×P.glandulosa, four transgenes in 6 lines, three transgenes in 6 lines, two transgenes in 2 lines, and one transgenes in 7 lines.3 Southern blotting. Based on dot blotting, southern blotting results in P.×euramericacana 'Guariento' with all five transgenes show that all transgenes were detected in 7 lines.4 Bt-cry3A ELISA . Based on dot blotting, Bt-cry3A ELISA were carried out in P.×euramericacana 'Guariento' with all five transgenes. The results showed that Cry3A were expressed in 7 lines.