The Konqi River is in the lower reaches of the Bosten Lake Basin in Xinjiang, China, and its ecological environment is relatively fragile. Recently, water resources shortage, water ecological imbalance, and other issues have become increasingly prominent under the dual influence of human and natural factors. This paper studies the ecological baseflow and natural vegetation water requirement of the Konqi River Basin, which could provide a decision-making reference for planning and using the Konqi River resources to curb the ecological environmental degradation caused by river interruption or flow reduction. First, based on the distribution of vegetation types and annual runoff conditions in the Konqi River Basin, the river channel is divided into Parts A and B. Section A’s river channel has not been cut off all year round and reaches from the Tashidian Hydrological Station on the upper reaches of the Konqi River to the third water diversion project. Section B is the main distribution area of natural vegetation under the third water diversion project of the Konqi River. Second, based on the hydrological data of the Tashidian Hydrological Station of the Konqi River of the last 50 years, the ecological baseflow of Section A is estimated using the Tennant, annual classification, annual average value of the lowest monthly average flow, and 90% assurance rate methods. The phreatic water evaporation and quota methods are selected to calculate the water requirement of natural vegetation within the radiation range of Section B’s river channel. The results show that the annual average ecological baseflow estimated using the Tennant method is 9.13 m³·s^-1, and the corresponding basic ecological water volume is 2.88×10⁸ m³·a^-1. These values can meet the river loss value of Section A from 2000 to 2018, and the actual annual average runoff from 2009 to 2018 can also meet the ecological baseflow standard. For calculating Section B’s natural vegetation water requirement using the change in groundwater levels around the river after ecological water transfer and analyzing field survey data, all natural vegetation within one kilometer from the river is defined as the main ecological protection area, with an area of 4.66×10⁴ hm² and ecological water requirement of 0.95×10⁸ m³·a^-1. Finally, taking the ecological water conveyance project of the Konqi River as an example, the scientific regulation of water resources in the basin is conducted using the calculation results of ecological baseflow and vegetation water requirement, which has significance for realizing river channel restoration and ecological water supply in different level years.