玉米丝轴黑粉菌原生质体制备与再生条件的研究

Preparation and Regeneration of Protoplasts of Sporisorium reilianum f.

以玉米丝轴黑粉菌(Sporisorium reilianum f.)为供试菌株,对其孢子原生质体的制备和再生条件进行研究。结果表明,交配型菌株SR1加入5 mg/mL溶壁酶(Lysing Enzymes)、50 mg/mL崩溃酶(Drise-lase)与50 mg/mL蜗牛酶(Snailase)混合酶液,置于28℃100 r/min摇床上酶解10 min,原生质体得率为99.36%、产量1.35×108个/mL,以山梨醇为稳渗剂,原生质体涂布于含有1.0 mol/L山梨醇的CM再生培养基上,再生率可达35.63%。

To establish protoplast mediated genetic transformation,preparation and regeneration of protoplasts from Sporisorium reilianum f.were studied.The results showed that the optional condition for the protoplast preparation was that,mating type strains SR1 were incubated in the mixed enzyme liquid consisting of 5 mg / mL lysing enzymes,50 mg / mL driselase,50 mg / mL snailase at 28 ℃ with shaking at 100 r / min for 10 min,and the yield of protoplast reached 1.35×108mL and the highest production rate of protoplast reached 99.36%.The protoplast was regenerated on the CM plates containing 1.0 mol / L D-sorbitol and the highest regeneration rate could reach 35.63%.