Cloning And Functional Analysis Of MdMIF2 Gene In Apple

Apple(Malus domestica)is a perennial woody plant of the genus Rosaceae that has been favored by many people because of its variety,color,aroma and sweet taste.In recent years,the molecular biology research using apple as a test material which has become more and more extensive.The research about in vitro regeneration and genetic transformation of apple leaves has made great progress.The mini zinc finger 2 protein is a small protein containing only the ZF finger domain,which was previously studied in Arabidopsis and tomato.This study was the first to clone the full length of Md MIF2 gene in ?GL-3? apple,construct overexpression and silencing vector of Md MIF2 gene,and transform apple ?GL-3? by Agrobacterium-mediated method.Md MIF2 gene overexpression and silencing transgenic plants were obtained,and the function of Md MIF2 gene was explored by measuring the rate of regeneration bud regeneration,leaf regeneration rate and average number of regenerated buds,which provided a theoretical basis for the study of efficient genetic transformation system of apple.The main results are as follows:1.The Md MIF2 gene coding region was cloned from ?GL-3? apple,which was 273 bp in length and encoded 90 amino acids,containing a ZF domain.The Md MIF2 gene was analyzed by spatiotemporal expression pattern and promoter online analysis,and the expression characteristics of Md MIF2 gene were analyzed.2.Overexpression vector and silencing vector of apple Md MIF2 gene were constructed.?GL-3? apple seedlings were transformed using Agrobacterium-mediated genetic transformation systems.Three Md MIF2 gene overexpression lines and three Md MIF2 gene silenced lines were obtained.3.Through the leaf regeneration test,it was found that compared with non-transgenic plants,the regeneration ability of Md MIF2 gene overexpressing plants was significantly enhanced,while the Md MIF2 gene silencing plant leaf regeneration ability was significantly weakened.This study shows that the apple Md MIF2 gene has the function of promoting leaf regeneration.