关键词: 橙色红曲菌, 原生质体, 形成, 再生, 转化

Abstract: In this research, formation and regenaration of protoplast conditions were optimized. The spores suspension up to a final concentration of 1×107～4×107/ml are inoculated in the liquid medium with MPPY, cultured at 30℃, 200r/min for 35 h. Mycelial masses were resuspended in suitable enzyme solution (1% snaliase + 0.3% lysing enzyme + 4 % cellulase, lytic enzyme dissolved in 0.6 mol/L MgSO4, adjusting pH to 6), and incubated at 30 ℃ with orbital shaking (60 r/min). The number of protoplasts were counted by a hemacytometer during the protoplasts formation, and the pesk of 8×107/ml after 3.5 h of incubation. The peak of regenerantion frequence is 18%. PEG and CaCl2-mediated protoplast transformation of strain AS3.4384 with pH4, hygromycin B as selective marker is established.

Key words: Monascus aurantiacus, protoplast, formation, regeneration, transformation