苏丹黑B染色法原位检测2株淡水小球藻胞内油脂累积  

In-situ detection of lipid accumulation in two strains of Chlorella by Sudan black B staining

作　　者：金雪洁[1] 巩三强[1] 夏金兰[1,2] 聂珍媛[1,2] 

机构地区：[1]中南大学资源加工与生物工程学院,湖南长沙410083 [2]中南大学生物冶金教育部重点实验室,湖南长沙410083

出　　处：《中南大学学报（自然科学版）》2014年第8期2547-2552,共6页Journal of Central South University:Science and Technology

基　　金：湖南省研究生创新学位论文资助课题(2010ssxt246);中南大学创新教育资助课题(2010bsxt05;CL11029;CL11038)

摘　　要：以2株淡水小球藻Chlorella pyrenoidosa XJ01和Chlorella sorokiniana XJ02为研究对象,对苏丹黑B染色法测定微藻胞内油脂质量分数的条件进行优化,建立苏丹黑B染色法原位测定微藻油脂的方法;并在通气条件下,检测微藻脂质质量分数及脂质累积过程。研究结果表明:当辅助染色破壁的盐酸浓度为1.0 mol/L,染色时间和温度分别为10 min和100℃时,苏丹黑B染色效果最好;在不同生长阶段,C.sorokiniana XJ02胞内油脂质量分数均比C.pyrenoidosa XJ01的高;此外,2株藻胞内油脂质量分数在对数生长期中期开始快速增加,到生长稳定期油脂质量分数达到最高,C.pyrenoidosa XJ01和C.sorokiniana XJ02油脂质量分数最高分别为25%和38.2%,随后油脂质量分数开始缓慢降低;利用苏丹黑B将微藻细胞染色后在显微镜下观察到的胞内油滴结果与其胞内油脂测定结果基本相符。Sudan black B staining method was optimized and applied to measure lipid content and monitor the lipid accumulation in two oil-rich freshwater microalgae C. pyrenoidosa XJ01 and C. sorokiniana XJ02, respectively. The optimal HCl concentration for disrupting cell wall of microalgal strains is 1.0 mol/L, Sudan black B staining has the highest effectiveness when staining time and temperature are 10 min and 100 ℃, respectively. The conducted comparative test with traditional gravimetric method and stability test indicates that Sudan black B staining method can be regarded as a feasible method for quantification of lipids in microalgae in situ. Lipid in microalgal cells starts to accumulate from interim of logarithmic phase and reaches highest value at interim of stationary phase and then begin to decrease, which is consistent with the results of oil-droplet images obtained by microscope.

关 键 词：苏丹黑B染色 小球藻 油脂累积 原位检测 

分 类 号：Q939.97[生物学—微生物学]