The invention discloses a preparation method of an alkynol selective hydrogenation catalyst. The preparation method comprises the following steps: (1), agitating and mixing a nitrogen-containing biomass with a surfactant, next adding an indium-containing metallic salt and acid liquor, mixing uniformly, and then performing drying and high-temperature calcination to obtain an indium oxide compoundednitrogen-doped porous carbon material; and (2), impregnating the indium oxide compounded nitrogen-doped porous carbon material in a Pd precursor solution, and then performing agitation, oven-drying,calcination and hydrogen reduction treatment in sequence to obtain the alkynol selective hydrogenation catalyst. The invention also discloses the alkynol selective hydrogenation catalyst prepared by the preparation method and application of the alkynol selective hydrogenation catalyst to an alkynol selective hydrogenation reaction in a vitamin industrial chain. The preparation method is simple; the prepared alkynol selective hydrogenation catalyst is compact in structure; nano particles of a Pd-In alloy are highly dispersed; and the alkynol selective hydrogenation catalyst has excellent catalytic performance.