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作为测定微生物群落功能多样性主要方法之一的biolog法,由于操作简单、灵敏度高、分辨力强、获得数据量丰富,受到众多研究者的青睐,但文献中报道的有关实验预处理方法却不尽相同。为了筛选最佳的预处理方法,通过比较文献中3种较为常用的方法,研究不同预处理对biolog法测定微生物群落功能多样性的影响。结果表明:AWCD值总体变化趋势B方法优于A、C;微生物对六类碳源的利用率中B方法对大部分碳源利用较高,且结果较稳定;两种处理的Simpson指数、Shannon丰富度指数及碳源利用丰富度指数均B>C>A,而Mc Intosh指数、Shannon均匀度结果不太稳定,但经方差分析,其差异不显著,且B方法的方差最小;主成分分析的结果 B方法优于A与C。总之,在250 r/min的转速下震荡30 min,并于28℃条件下培养接种平板的处理(B法)效果较好,其测定结果重现性好,且操作简便快捷,是比较理想的预处理方法。
Biolog method, one of the main methods to determine the functional diversity of microbial community, is widely used by many researchers due to its simple operation, high sensitivity, strong resolution and rich data volume. However, the biolog method reported in the literature does not include the experimental pretreatment method Do the same. In order to screen the best pretreatment methods, the effects of different pretreatments on the functional diversity of microbial communities by biolog method were studied by comparing the three commonly used methods in the literature. The results showed that the overall trend of AWCD value was better than that of A and C. The utilization rate of microorganisms in six types of carbon sources was higher for most carbon sources and the results were more stable. The Simpson index, Shannon The richness index and the carbon source utilization index of abundance B> C> A, while the Mc Intosh index and Shannon evenness result are not stable, but the variance is not significant, and the variance of B method is the least. The principal component analysis The result of B method is superior to A and C. In conclusion, the method of shaking at 30 rpm for 30 min at 250 r / min and culturing the inoculated plate at 28 ° C (Method B) is better, and the results are good, reproducible and easy to operate, which is ideal Pretreatment methods.