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A search for a dark baryon is performed for the first time in the two-body decay Ξ−→π−+invisible using (10.087 ± 0.044) × 109 J / ψ events collected at a center-of-mass energy of s=3.097GeV with the BESIII detector at the BEPCII collider. No significant signal is observed, and the 90% (95%) confidence level upper limits on the branching fraction B(Ξ−→π−+invisible) are determined to be 4.2×10−5 (5.2×10−5), 6.9×10−5 (8.4×10−5), 6.5×10−4 (7.6×10−4), 1.1×10−4 (1.3×10−4) and 4.5×10−5 (5.5×10−5), under the dark baryon mass hypotheses of 1.07 GeV/ c 2, 1.10 GeV/ c 2, mΛ (1.116 GeV/ c 2), 1.13 GeV/ c 2, and 1.16 GeV/ c 2, respectively. The constraints obtained on the Wilson coefficients Cus,sL and Cus,sR are more stringent than the previous limits derived from the LHC searches for the colored mediators.