Expression Vector Construction And Soybean Transformation Of Low-Phosphorous Tolerance Transcription Factor GmPTF1

Phosphorus is one of the most important nutrients that significantly affect soybean growth and metabolism. Phosphorus deficiency will limit vegetative organ of plant. Based on the cloning and identification of GmPTF1, the eukaryotic expression vector pGN-GmPTF1 and pX6-GmPTF1 were constructed in our study and further introduced into Arabidopsis to research the biological function of the gene; Meanwhile, To creates new transgenic materials and provide an important genetic resources for the cultivation of low phosphorus tolerant soybean varieties, GmPTF1 was transformed into soybean varieties via Agrobacterium-mediated cotyledonary-node and pollen tube pathway transformation methods. The main results were as follows:1. Construction the eukaryotic expression vector of GmPTF1: The over-expression vector pGN-GmPTF1 and marker-free expression vector pX6-GmPTF1 were constructed completely in this study by using pGN and pX6 vector.2. Study the biological function of transcription factor gene GmPTF1 in Arabidopsis: The vector pGN-GmPTF1 and pX6-GmPTF1 were transformed into Arabidopsis with Agrobacterium tumefaciens GV3101. The T2 seed of transgenic Arabidopsis was also obtained. The T2 transgenic Arabidopsis plants with pGN-GmPTF1 grew better than that of the wide type in low phosphate condition. Therefore, the results showed that the function of GmPTF1 was association with the tolerance to inorganic phosphate starvation in plant.3. Transformation low-phosphorous tolerance transcription factor GmPTF1 into soybean: Expression vector pGN-GmPTF1 and pX6-GmPTF1 were introduced into jidou12, jidou16, wuxing1 and luguandou1 by Agrobacterium-mediated and pollen tube pathway transformation methods. PCR analysis results of transformation plants showed that total sixty-five PCR positive plants of T0 generation, thirteen T1, thirty-eight T2 and three T3 were obtained.4. Obtained six new genetically modified soybean materials: Results of PCR, RT-PCR and seqence analysis showed that GmPTF1 had integrated into and transcripted successfully in the soybean genome. Now we have obtained six genetically modified materials (JPF-1, JPF-2, JPF-3, JPF-4, JPF-5, WPF-1).