分光光度法测定黑曲霉孢子浓度的研究

Determination of Aspergillus niger spores concentration by spectrophotometry

采用分光光度计测定黑曲霉(Aspergillus niger)xj在不同生长期、不同稀释倍数下孢子悬液的OD_(600nm)值,对比血球板计数法得到的孢子悬液浓度,研究在对数期和稳定期OD_(600nm)值与孢子浓度之间的关系;采用微电影拍摄法观察黑曲霉xj产孢结构形成过程,并绘制固体发酵条件下的生长曲线。结果表明,在10~20倍稀释区间内,稀释倍数与OD_(600nm)值之间呈现良好线性关系(对数期R^2=0.984 8、稳定期R2=0.991 3),且OD_(600nm)值与孢子浓度之间也保持良好的线性关系(对数期R^2=0.995 3、稳定期R^2=0.993 6);通过微电影拍摄法观察到黑曲霉xj产孢结构形成是分阶段进行的过程;在固体发酵过程中,孢子浓度呈现"S"型增长趋势。该研究为测定丝状真菌孢子浓度提供另一种可借鉴的方法。

OD600nm values of Aspergillus niger xj spore suspension was determined by spectrophotometer under different growth stages and dilution ratios, comparing to the spore suspension concentration which was determined by the blood bat counting method, the relationship between OD600nm value and spore concentration in log phase and stable phase were investigated. The micro film shooting method was used to observe the forming process of A. niger xj conidial fi＇uctification and the growth curve of spore under a solid-state fermentation condition was drawn. The results showed that in the 10-20 times dilution range, it showed a good linear relationship between dilution ratio and OD600nm value （log phase R2 =0.984 8, stable phase R2=0.991 3）, and there also had a good linear relationship between OD600nm value and spore concentration （log phase R^2=0.995 3, stable phase R^2=0.993 6）. Moreover, it could be observed that the formation of A. niger xj conidial fructification was a stage process through the micro film shooting method, and spore concentration showed a ＂S＂ type growth trend during a solid-state fermentation. The study provided another referential method for the determination of filamentous fimgal spore concentration.