Abstract: Taking 36 populations of 6 wild kiwifruit species from Guizhou as experimental materials, the transcriptomic data of ‘Hongyang’ kiwifruit in response to Bactrocera dorsalis（Hendel） larval feeding was analyzed to search SSR（simple sequence repeat） loci for EST-SSR primers development through bioinformatics analysis, and genetic diversity and genetic relationships in 180 accessions of 36 populations from six kinds of wild kiwifruit were analyzed, in order to provide reference for the wild kiwifruit resources protection and utilization.The results showed that the frequency of EST-SSR was 43.07% and the repetitive number of six repeat units was 5-24.The predominant repeat motifs was dinucleotide with a proportion of 69.19%.By testing 40 pairs of EST-SSR primers, 28 pairs of EST-SSR primers produced clear polymorphic fragments in all six kiwifruit species.Using the 28 EST-SSR primers that produced during preliminary screening, we detected 348 alleles in the 180 studied accessions.Polymorphism information content（PIC） ranged from a minimum of 0.376 6（P3-22） to a maximum of 0.930 4（P4-37） with an average of 0.783 6,which revealed only 1 pairs of SSR primers with moderate polymorphism and 27 with high polymorphism.The cluster analysis showed that the six kiwifruit species were grouped into two categories at a genetic distance of 0.72,A.fulvicoma,A.eriantha and A.latifolia were clustered as a class; A.callosa var.henryi, A.callosa var. discolor and A.chinensis var.deliciosa were anothor one.