A deep understanding of distinct functional differences of various defects in semiconductor materials is conducive to effectively control and rationally tune defect-induced functionalities. However, such research goals remain a substantial challenge due to great difficulties in identifying the defect types and distinguishing their own roles, especially when various defects coexist in bulk or nanoscale material. Hereby, we subtly selected a molecular-type semiconductor material as structural mode composed of supertetrahedral chalcogenide Cd-In-S nanoclusters (NCs) with intrinsic vacancy point defect at the core site and antisite point defects at the surface of supertetrahedron and successfully established the correlation of those point defects with their own electrochemiluminescence (ECL) behaviors. The multi-channel ECL properties were recorded, and the corresponding reaction mechanisms were also proposed. The predominant radiation recombination path of ECL emission peak at 585 nm was significantly distinguished from asymmetrically broad PL emission with a peak at 490 nm. In addition, the ECL performance of the coreless supertetrahedral chalcogenide nanocluster can be modulated by atomically precise doping of monomanganese ion at the core vacant site. A relatively high ECL efficiency of 2.1% was also gained. Actually, this is the first investigation of ECL behavior of semiconductor materials based on supertetrahedral chalcogenide nanoduster in aqueous solution. Current research may open up a new avenue to probe the roles of various different defects with defined composition and position in the NC. The versatile and bright ECL properties of Cd-In-S-NC combined with tunable ECL potential and ECL peak suggest that the new kind of NC-based ECL material may hold great promising for its potential applications in electrochemical analysis, sensing and imaging.