Construction And Transformation Of Artificial Expression Vectors Of MiR398 In Eucalyptus Grandi×E.urophylla

Eucalyptus is belonged to Myrtaceae Eucalyptus,and originating from Australia.It is widely cultivated around the world because of its fast-growing,high yield and barren tolerance,Eucalyptus is one of the fast reproducing tree species(Eucalyptus,populus,Pinus Linn and bamboo)over the world.miR398 is the first reported miRNA which is down regulated in oxidative stress.AmiRNAs(artificial miRNAs)are exogenous small RNAs generated by endogenous miRNA precursors,which substituted core miRNA and its complementary sequence miRNA*,Construction of plant amiRNA expression vector and importing into plants is an important method to study the function of miRNA.In this article we take adventage of sequence of Euphoria longan and pRS300 vector which contains Arabidopsis thaliana miR319 to build amiR398 plant binary-vector system by overlap PCR,as the result,we get four amiRNA expression vectors named Z-pBI398a?F-pBI398a?Z-pBI398b/c and F-pBI398b/c,after this,we import these expression vectors into Eucalyptus by Agrobacterium,finally we got Eucalyptus seedlings.