人参皂苷F1转化菌株的原生质体诱变育种

菌核青霉2246(Penicillium sclerotiorum)能够将人参皂苷Rg1转化为人参皂苷F1.以此菌为出发菌株,进行原生质体制备和再生的研究,确定原生质体的最佳形成条件:菌丝体培养24 h,用5 mg/mL溶壁酶、5mg/mL纤维素酶和5 mg/mL蜗牛酶的混合酶液进行酶解,以0.8 mol/L的KCI作为渗透压稳定剂,31℃水浴振摇2h.并对形成的原生质体进行亚硝基胍复合紫外线照射诱变,结果得到1株转化率显著提高、遗传性能稳定的诱变株( NU-1),其转化率由16.7％提高到30.5％.

Protoplast Mutation Breeding of the Biotransformation Strain for Ginsenoside F1

This study focused on the optimal conditions of protoplast preparation and regeneration of Penicillium sclerotiorum 2246, which could transform ginsenoside Rg1 into ginsenoside F1. The optimal conditions were as follows ： the mycelia were cultured for 24 hours, and then they were zymolysed for 2 hours by enzyme mixture of 5 mg/mL lywallzyme, 5 mg/mL cellulase, and 5 mg/mL snail enzyme at 31℃ when 0. 8 mol/L KCl was used as osmotic pressure stabilizer. Then protoplasts were treated with ultraviolet ray and NTG together to get the high mutation percentage strains. Finally obtained a mutation strain （NU-1） with high transformation rate and the conversion rate with increment from 16.7% to 30.5%.