The preparation of enantiomerically pure 1,4-oxazines remains a continuous challenge in synthetic chemistry because of their potential application in the total synthesis of morpholines. Herein, a one-pot asymmetric transfer hydrogenation/cyclization enantio-relay process catalyzed by Ru and Au bimetallic catalysts was developed. This enantio-relay process firstly allows the asymmetric transfer hydrogenation of alkynones to form chiral alkynols in the presence of (S,S)-mesityleneRuClTsDPEN (TsDPEN = N-(p-toluenesulfonyl)-1,2-diphenylethylenediamine) and then the chiral alkynols could be converted into 3,4-dihydro-2H-1,4-oxazines in moderate to good yields with excellent enantioselectivity retention accompanied by AuCl(PPh₃)/AgNTf₂ and HCOOH. A series of sulfonamide-tethered alkynones were well tolerated in this process. Mechanistic studies indicated that the alkynols formed in the first step were initially transformed into a methylenemorpholine intermediate when AuCl(PPh₃)/AgNTf₂ and HCOOH were added, which can be further isomerized into chiral 3,4-dihydro-2H-1,4-oxazines with the addition of a second HCOOH. This protocol offers not only a practical method to access 3,4-dihydro-2H-1,4-oxazines from alkynones in an enantioselective version, but also enriches the arsenal of gold chemistry from a mechanistic point of view.