红曲霉T-DNA插入突变库中色素突变子的筛选

摘要

Monascus purpureus has been an important traditional edible and officinal microbe in China that produces Monascus pigment, Monacolin K, -γ-aminobutyric acid, ergosterol and other substances. It has been widely used in the production of food fermentation agents, food coloring agent, nutraceutical, medicine etc. The application history was no fewer than a thousand years. As materials 734 mutants from Monascus mutant library mediated by Agrobacterri-iun tumefacUns were studied. Among them nine mutants of pigment mulon with significant changes in producing Monascus pigment were selected using ethanol extraction, the color value of mutant S50 in 505 nm was decreased by 0. 12 fold than the original strain S, almost turning to an albino strain. The results of UV-VIS scanned map in 300 -600 nm indicated that the pigment mutons changed not only in color value, but also in absorption wavelength and the absorption peak. Hereafter, the features of pigment mutons, including colony morphology, microscopic morphology, hygromycin resistance, genetic stability and citrinin production capacity, were analyzed. It laid the foundation for the research of Monasaa pigment-related genes.%红曲霉(Monacus purpureus )是我国重要的传统食用和药用微生物,能分泌红曲色素、莫纳卡琳(Monacolin K)、γ-氨基丁酸、麦角固醇等物质,被广连用于生产食品发酵剂、食品着色剂、保健食品和药品等,在我国有上千年的应用历史.本实验以根癌农杆菌(Agrobacterrium tumefaciens)介导的红曲霉突变体库中754株突变子菌株为材料,通过乙醇提取法筛选出了9株产红曲色素发生显著变化的色素突变子,其中S50在505 nm波长处的色价降为原始菌株S的0.12倍,几乎变为白化株.300 ～ 600 nm波长下UV-VIS扫描图谱特征显示,色素突变子不仅色价发生了变化,而且吸收波长、吸收峰值也发生了不同程度的变化.最后,对色素突变子菌落形态、显微结构、潮霉素抗性、遗传稳定性以及产桔霉素能力等进行分析,为研究红曲霉产色素相关基因奠定了基础.