The invention discloses yarns with non-uniform fiber distribution, a fabric and a preparation method thereof. The preparation method of the yarns with non-uniform fiber distribution comprises the following steps of: (1) first opening and mixing: arranging polyester fibers and cellulose fibers on a bale plucker not uniformly, and opening and mixing the fibers; (2) first carding: carding the fibersprepared in the step (1) to obtain first carded slivers; (3) second opening and mixing: arranging natural fibers in the bale plucker, and opening and mixing the fibers; (4) second carding: carding the fibers prepared in the step (3) to obtain second carded slivers; (5) drawing: arranging the first carded slivers and the second carded slivers on an elevated platform of a drawing frame not uniformly, and drawing the carded slivers; (6) roving; (7) spinning; and (8) spooling. The surface color textures of the fabric woven by adopting the yarns are arranged transversely and non-uniformly, and thewhole fabric is very attractive.