Susceptibility Of Soybean Cotyledonary Node To LiCl And GeneticTransformation Of GmDof11Gene In Soybean

Dof transcription factor gene plays an important role in plant growth and development stage. In the plant genome, there includes many genes encoding Dof proteins. It is reported that there are36genes in arabidopsis Dof transcription factor family;30genes in rice Dof transcription facto family; and28genes in soybean Dof transcription factor family.The present study shows that:expression of the soybean’s GmDof4and GmDofll gene can increase the total fat content in transgenic arabidopsis seeds, and also can increase the grain weight.CAX gene family was shown to have the potential of improving plant resistance to heavy metal ions. Studies have shown that:the GmCAXl gene in Arabidopsis plants with lithium plasma of the antiporter, it ion balance by regulating plants, and thus the regulation of plant salt tolerance.Obtained in this experiment pROK2-GmCAX1-GmDof11vector through Agrobacterium-mediated method the GmDofll gene into soybean, on the one hand, use GmCAXl gene resistant to lithium, lithium chloride as a selective agent as an alternative to traditional antibiotics, to resolve theantibiotics as a soybean genetically modified to filter the drawbacks of the agent; the other hand, selected GmDofll gene as a target gene into the soybean hopes to improve the total fat content of soybean seeds. In addition, this study also explored the sensitivity of soybean cotyledon node of plants resistant to lithium carrier and the soybean genetic transformation system in order to improve the conversion rate, and the system the GmDofl1gene into soybean and preliminary identification.In this study, the main results are as follows:1. Transgenic system optimization:In this study, we use two varieties of soybean as the material, study the impact of seed sterilization conditions, germination conditions, the induction medium, hormone concentration on the regeneration of soybean cotyledonary node, and establish an efficient regeneration system of soybean cotyledonary node for the soybean’s genetic transformation. The results show that soybean seed’s sterilization time is5-7h appropriately; the soybean seed’s germination time has an impact on buds differentiation, and is5d appropriately; it is appropriate to add ordinary agar into germination medium and place the soybean hilum in the same direction; it is appropriate to use BAP of2mg/L when inducting soybean cotyledonary node buds, and use IB A of1.5mg/L when buds rooting.2. The findings of the sensitivity of the cotyledon section to lithium chloride:the critical concentration for browning wholly of the two soybeans88-1and monkey hair is1.0mM and1.4mM respectively; the difference of browning between different concentration of lithium chloride or different genotypesvery is significant, that is to say, we should pay attention to different genotypes of soybean’s sensitivity differences on the reaction of lithium chloride; the chloride lithium’s critical screening concentration of88-1is1.OmM, and monkey hair is1.2mM, when soybean cotyledonary node differentiation; the lithium chloride’s critical screening concentration of88-1and monkey hair is0.3mM and0.2mM respectively, when soybeans clustered buds rooting.3. With plants’ carriers resistant to lithium, we transfer GmDofll into soybean88-1, through the method of mediating cotyledonary node by agrobacterium using lithium chloride as a selective agent. We do experiments repeatedly, get9soybean transgenic plants, do preliminary test, and get16seeds.