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唐菖蒲(Gladiolus hybridus Hort)根腐病是由尖孢镰刀菌唐菖蒲专化型(Fusarium oxysporum f.sp.Gladioli)引起的真菌性病害。原生质体是研究尖孢镰刀菌唐菖蒲专化型与唐菖蒲之间的互作关系的重要工具,试验以尖孢镰刀菌唐菖蒲专化型为供试菌株,进行原生质体制备条件的优化与再生,明确培养基、菌龄、β-巯基乙醇预处理、酶选择以及酶解时间对原生质体产生的影响。试验结果表明:在PDB培养基中原生质体制备的最优条件为15μLβ-巯基乙醇预处理1 h,在5%崩溃酶溶液酶解3 h。在此条件下,12~14 h菌丝的原生质体产量可达到1.4×107个/m L,再生率为57.4%。
Gladiolus hybridus Hort root rot is a fungal disease caused by Fusarium oxysporum f.sp. Gladioli. Protoplast is an important tool for studying the interaction between Gladiolus hybridus and Gladiolus hybrids. In this study, the optimization of protoplast preparation conditions was carried out using Fusarium oxysporum Regeneration, clear media, bacterial age, β-mercaptoethanol pretreatment, enzyme selection and enzymatic hydrolysis time protoplast production. The results showed that the optimal conditions for preparation of protoplasts in PDB medium were as follows: pretreatment with β-mercaptoethanol 15μL for 1 h, enzymolysis in 5% disintegration enzyme solution for 3 h. Under these conditions, the protoplast production of mycelia of 12-14 h reached 1.4 × 107 cells / m L, and the regeneration rate was 57.4%.