Research Of Soybean Transformation Of Agrobacterium-mediated LePT1 Gene

Soybean which is rich of plant protein and grease is one of important oil crops and food crops,and it is not only the main source of plant protein and edible oil in the world,also raw materials for industrial production and important sources of feed for animals and poultry.In the study,LePT1 gene cloned from tomato genome belongs to the high affinity plant phosphate transporters Pht 1 family genes.Under conditions of Pi starvation,plants absorb Pi by roots via the high affinity system to meet their own growth and development requirements.In this experiment,with purpose of improving soybean quality,we translated the exogenous LePT1 gene into soybean genome,in order to increase phosphorus uptake and utilization,and improve the resistance of soybean,and enhance its ability to adapt to the environment.It can greatly accelerate the soybean varieties improvement.There will be some new varieties of soybeans with excellent characters,such as resilience and high yield.Designed a pair of specificity amplification primers that containing Xba I and Kpn I specific enzyme cutting sites,then connected PCR amplification products of tomato genome to the pMD19-Vector to construct pMD19-Vector-LePT1 carrier,then with the intermediate vector pROK219’s help,finally constructed the genetic transformation recombinant vector pCAMBIA2300-LePTl after digestion and connection of vector pCAMBIA2300.In this study,the experiment material is Liaodou 17.Taking embryo tip as explants and basing on the Liaodou 17 soybean regeneration system identified by former studies,we translated LePT1 gene into soybean genome via Agrobacterium-mediation to obtained transgenic plants,in this process there are a series of steps,including obtaining germ explants,pre-training,infection,coculture,recovery culture,screening culture and rooting culture,ea tl.As a result,5 survival positive plants with LePT1 gene were detected by PCR in the first generation,and then obtained second filial generation,and the PCR result detected the second filial generation shows that they are positive plants,moreover,the sequencing result blasted online shows that there is 99%similarity with the tomato LePTl gene sequence.