The invention discloses recombinant fluorescent dual-hybrid yeast and a detection method and the applications thereof. The recombinant fluorescent yeast is a recombinant microzyme obtained by transferring a fusion gene into a microzyme Y187; and the fusion gene is formed by sequentially connecting DNA (Deoxyribonucleic Acid) fragments shown by the 9th bit to the 1030th bit in a sequence (5) in a sequence table, a reporter gene, a terminator, a selection marker protein expression cassette and DNA fragments shown by the 4336th bit to the 5226th bit in the sequence (5) in the sequence table. Therecombinant fluorescent yeast provided by the invention can be used for rapidly detecting interaction among protein.