Genetic Transformation Of Populus Tomentosa And Arabidopsis With Sterility Constructs Of PtAP3, PtAG And PtAP1

Populus tomentosa Carr. (Chinese white poplar) is a native specie in section Leuce with f ast-growing and high wood quality in northern China. However, its catkin and pollen pollution bring a lot of inconvenience to people's life and production activity. To obtain sterile poplar, s even sterility counstructs including 35S-PtAP3-IR-NOS,35S-PtAG-IR-NOS,35S-PtAG-IR-NOS(MA R),35S-PtAP1-IR-NOS,35S-PtAP1::PtAG-IR-NOS,35S-AP1M2-E9 and 35S-AP1M3-E9 were transf ormed into Populus tomentosa. The positive transgenic poplar individuals were primarily verifie d by PCR. Sterility counstructs were also transformed into Arabidopsis thaliana considering the long juvenile period of poplar. Molecular detection and phenotypic analysis of transgenic A rabidopsis thaliana were conducted to provide a reference for transformation of poplars. Major results and conclusuions are described as follows.The Populus tomentosa clone TC1521 was used for genetic transformation.The positive res ults verified by PCR include nine 35S-PtAP3-IR-NOS transgenic poplar plants,five 35S-PtAG-IR-NOS transgenic poplar plants, six 35S-PtAG-IR-NOS (MAR) transgenic poplar plants,four 35S-PtAP1-IR-NOS transgenic poplar plants, six 35S-PtAP1::PtAG-IR-NOS transgenic poplar plants,s even 35S-AP1M2-E9 transgenic poplar plants and eight 35S-AP1M3-E9 transgenic poplar plants. The conversions rates were 1.6%,0.9%,1.1%,0.7%,1.1%,1.2% and 1.5% respectively.RT-qPCR showed little difference in expression among transgenic lines. The hightest level of expression is 4.96 times more than that of the reference gene. The lowest level of expression is 3.02 times. The expression of upstream gene LFY and PI' in transgenic plants relatively declined compared with the wild type, there is no dramatic changes of downstream gene Sep2,Sep3 and Sep4 between transgenic poplar plants and wide type.Through the phenotypic observation of transgenic arabidopsis phenotype,there are great differences between transgenic 35S-AP1M2-E9 plants and wide type. Some transgenic arabidopsis lines flowered 10.7 days later than wild type during the observation.Some Transgenic arabidopsis lines did not even get flowered during the observation. Besides, several floral abnormal variations were found on sepal, petal and stamens of transgenic arabidopsis.