The Research On Genetic Transformation Of Soybean With Arabidopsis Salt Tolerance Relative Gene AtNHX5

Soybean is one of the most important foods and oil crop of the world, but its production and quality is restricted by some factors like environment partly. Since the increase of the area of arid lands, saline lands and secondary salinization of cultivated land year by year, and the gradual decrease of cultivable area, it is a big restriction for the production of field crops in our country. For this reason, the incubation of new antireversional variety is becoming a hot spot and an emphasis as well for the past few years. In this study, established a new pathway for incubating new antireversional variety of Soybean with transporting Stress-resistant gene into Soybean by agroinfection. In this study, used high-quailty Soybean variety of Hebei Province as material, and undertook Arabidopsis thaliana salt-tolerant gene-AtNHX5(Arabidopsis Na~~+/H~~+ antiporter gene in Arabidopsis thaliana CPA1 family) genetic transformation by embryo tip method and entirety cotylcdonary nodes method.1. In this study, first determined the hygromycin- sensitivity of high-quailty Soybean variety of Hebei province such as Jidou 17, Wuxing 2, nf 58, Jihuang 13 by morphology methods to determine the best concentration for plant transformation. The result showed that the final hygromycin screening concentration of Wuxing 2 and Jidou 17 were 10 mg/L, and the final screening concentration of Jihuang 13 and nf 58 were 8 mg/L.2. In this study, used Wuxing 2 maturate Soybean embryo tips as material, and optimized the embryo tip transformation system of soybean with the supersonic assisted Agrobacterium-mediated transformation. On this basis, We used the embryo tip explant of wuxing 2, Wuxing 3, Jidou 15, Jidou 16, Jihuang 13 as receptor systems, undertaked Arabidopsis thaliana salt-tolerant gene AtNHX5 genetic transformation by 80 W supersonic assisted Agrobacterium-mediated transformation.We demonstrated the AtNHX5 had already integrated to the genome of Wuxing 2 soybean initially by regeneration Plants PCR verification, and the transformation efficiency was 0.16 %.3. In this study, investigated the AtNHX5 transformation by new entirety cotylcdonary nodes method. Used the Hypocotyl involved one seed leaf as explantation, and transformed with pCAMBIA3301 vector plasmid contained Basta screening mark and AtNHX5. Demonstrated the AtNHX5 had already integrated to the genome of Wuxing 2 soybean initially by regeneration Plants PCR verification, and the transformation efficiency was 1.47 %. The method cancelled the tedious and time-consuming tissue culture procedure after transformation of explantation, and the Anagen cycle was decurtated obviously. This study provided an effective technique platform for soybean transformation by Agrobacterium-mediated.