摘要
Poaching and trans-boundary trafficking of tigers and body parts are threatening the world’s last remaining wild tigers.Development of an efficient molecular genetic assay for tracing the origins of confiscated specimens will assist in law enforcement and wildlife forensics for this iconic flagship species.We developed a multiplex genotyping system“tigrisPlex”to simultaneously assess 22 short tandem repeat(STR,or microsatellite)loci and a gender-identifying SRY gene,all amplified in 4 reactions using as little as 1 ng of template DNA.With DNA samples used for between-run calibration,the system generates STR genotypes that are directly compatible with voucher tiger subspecies genetic profiles,hence making it possible to identify subspecies via bi-parentally inherited markers.We applied“tigrisPlex”to 12 confiscated specimens from Russia and identified 6 individuals(3 females and 3 males),each represented by duplicated samples and all designated as Amur tigers(Panthera tigris altaica)with high confidence.This STR multiplex system can serve as an effective and versatile approach for genetic profiling of both wild and captive tigers as well as confiscated tiger products,fulfilling various conservation needs for identifying the origins of tiger samples.
基金
This research was supported by the Peking-Tsinghua Center for Life Sciences at Peking University(PKU)and the National Natural Science Foundation of China(NSFC No.31271320,No.31471179).We thank Zhi Dong,Yan Zhuang,Yuechen Liu,Lin Miao and Galina Nemkova for technical assistance.Reference tiger samples for calibration were kindly provided by Dr Stephen O’Brien under a Material Transfer Agreement signed between PKU and the National Cancer Institute,USA,and in full compliance with the Convention on International Trade in Endangered Species.We also thank Dale Miquelle for helpful comments on an earlier version of the manuscript.
