The optimization of ecological security pattern is an important approach to achieve regionally ecological security and maintain regionally sustainable development. The Taolai River basin is a typically arid inland river basin with fragile and sensitively ecological background and prominently ecological problems. According to the ecological environment situation of the study area, the evaluation framework of ecosystem services and ecosystem health was developed to assess the ecological security of the basin in 2000 and 2015 based on "contribution, vigor, organization, and resilience" model. On this basis, the ecological sources were identified. With the help of Linkage Mapper tool, the least cost corridors were extracted. Sub-optimal corridors were constructed based on the Circuitscape software. In addition, barriers were identified by using Barrier Mapper tool. In order to provide references for the development and construction of arid areas and ecological protection, the optimization strategies of ecological security pattern in the basin were put forward. The results showed that:(1) on the whole, the ecological security of the basin was not optimistic. In terms of area, in 2000 and 2015, the low security area accounted for 63.68% and 61.25% of the total area, respectively, while the high level security areas and higher level security areas accounted for only 13.93% and 15.80%. From the perspective of spatial distribution, there were significant differences in the ecological security. The south and the central regions were higher than the north region. (2) We constructed and optimized the ecological security pattern based on the identification results of the ecological source, the least cost corridor, the sub-optimal corridor, and the barrier. The ecological sources were mainly distributed in the mountain of Sunan Yugur Autonomous County in the south of the basin and the agricultural areas of Jinta County, Jiayuguan City and Suzhou District in the middle areas of the basin. A total of 1122 least cost paths with a total length of 3468.15 km were generated among the ecological sources. There were 242 corridors distributed among the relatively far away source areas with higher resistance, which separated by higher resistance, such as bare rock and bare land. (3) The optimized division of the ecological pattern of the basin was divided into ecologically stable areas, ecological development areas and key protected areas. The optimization strategy of the ecological security pattern of the basin is proposed to provide a reference for the ecological protection, development and construction of the arid inland river basin.