Abstract: ObjectiveThe optimal medium for Bletilla striata(Thumb.)Reichb.f.protocorm induction and prolifera-tion were screened and effective protocorm proliferation system of B.striata was establishand to provide technical support for industrialized production of tissue culture seedlings of B. striata.MethodB. striata seeds were used as explant to select the proper methods of aspetic germination.Taking MS or 1/2MS as the basic medium,and adding plant growth sub-stance with different concentrations and additives to determine the optimum light and temperature conditions by single factor test as well as select the most suitable medium for its protocorm induction.Through orthogonal test,the most suitable medium for its proliferation was screened.ResultAfter treating B.striata capsule by 75% ethyl alcohol for 10 min,the aspetic germination rate could reach 90.0% and the seed germination was not affected.In the process of proto-corm induction,30.0μmol/m2·s illumination intensity and 25℃temperature benefited the germination of protocorm.The optimal medium for protocorm induction was 1/2MS medium+1.0 mg/L 6-BA+0.1 mg/L NAA.During the proliferation of protocorm,the addition of mashed potato,coconut juice and banana mud was beneficial to the proliferation of protocorm, and 50.0 g/L mashed potato had the best effects. The orthogonal test showed that the optimum medium for protocorm proliferation was MS+0.5 mg/L 6-BA+0.2 mg/L NAA+50.0 g/L mashed potato,and the proliferation mutiple reached 5.270.ConclusionThe established effective B.striata protocorm proliferation system can be widely used in the factory in the production of tissue culture seedlings of B.striata.