The Development Of SSR Primers And Genetic Analysis In Akebia Trifoliata(Thunb.)Koidz.

Akebia trifoliate(Thunb.)Koidz.belongs to Lardizabalaceae group,Akebia Decne.,which is divided to Akebia trifoliate ssp.trifoliata,Akebia trifoliate ssp.australis,and Akebia trifoliate ssp.longisepala.Akebia trifoliate has a highly medicinal,economic and ecological value,there is a huge potential of development and broad prospects of application for it.But so far,it is still not clear for its germplasm resources,which greatly hinder the development and utilization of Akebia trifoliate.In this paper,selecting Akebia trifoliate as sample material,we tried to develop SSR primer for the systematics research of it,and laid the foundation for the research of germplasm resources.In addition,using these highly polymorphic loci,we analyzed the condition of genetic relationships and genetic structure for it.The main results obtained in this research are as follows:1)Using bead enrichment,we established the microsatellite library of Akebia trifoliate(Thunb.)Koidz.,and obtained 148 positive clones of 150 colonies tested.After being sequenced and analyzed,there were 57 sequences that could be used to design primers,which accounted for 38%of the total proportion,and 63 pairs of primers were designed;2)Using orthogonal design,we optimized the reaction system of 63 pairs of primers,and 54 pairs of primers obtained the fragment.We screened to obtain 33 pairs of polymorphic primers in 54 pairs of primers by using 7 samples.Then we screened to get 16 pairs of highly polymorphic primers of 33 pairs of primers by using 54 and 48 samples;3)Analyzing the genetic diversity of three generations of individuals by 16 pairs of highly polymorphic SSR primers,we found that these loci had higher genetic diversity among populations and can provide effective tools and instruments as the study for germplasm resources and genetic diversity of Akebia.Moreover,there was high heterozygosity of the population;4)Through clustering analysis and analyzing genetic structure of the individuals of three generations,Akebia trifoliate(Thunb.)Koidz.is cross-pollinated plant that higher heterozygosity,which provide guidance for breeding cultivation.