The present invention provides a mushroom cultivation unit having an inoculation hole retainer and a strain inoculation method. The method includes the steps of: first preparing a mushroom cultivation unit filled with culture medium； inserting a tool to form an inoculation hole in the culture medium； next, inserting an inoculation hole retainer in the inoculation hole, wherein the inoculation hole retainer is a hollow tube and the tube wall is opened with a plurality of through holes； finally, performing a sterilization process on the mushroom cultivation unit inserted with the inoculation hole retainer, and then inoculating the strains in the inoculation hole retainer. The inoculation hole retainer may effectively support the inoculation holes to prevent from collapsing. Thus, during inoculating strains, the strains may be firmly implanted into the culture medium to rapidly grow, so as to effectively shorten the following mycelium growth period and achieve the effect for producing mushrooms simultaneously for the whole pack.一種具接種孔保持器之菇類栽培單元及其菌種接種方法，首先取一裝填有培養基質之菇類栽培單元，並以一工具插入於培養基質中以形成有一接種孔，接著將一接種孔保持器插設於接種孔中，接種孔保持器呈中空管狀，並於其管壁上開設有數個貫穿之透孔，最後對插設有接種孔保持器之菇類栽培單元進行一滅菌處理後，將菌種接種於接種孔保持器中。藉由接種孔保持器使接種孔可獲得有效支撐，進而確保其接種孔不會崩落，使後續接種菌種時，菌種可確實被植入培養基質中快速同時生長，而可有效縮短其後續之走菌時程，並可達成整包同時出菇之功效。
