Powell Shane M, Ferguson Susan H, Bowman John P, Snape Ian
Tasmanian Institute of Agricultural Research, University of Tasmania, Hobart 7001, Australia.
Microb Ecol. 2006 Oct;52(3):523-32. doi: 10.1007/s00248-006-9131-z. Epub 2006 Aug 31.
A real-time polymerase chain reaction (PCR) method to quantify the proportion of microorganisms containing alkane monooxygenase was developed and used to follow changes in the microbial community in hydrocarbon-contaminated Antarctic soil during a bioremediation field trial. Assays for the alkB and rpoB genes were validated and found to be both sensitive and reproducible (less than 2% intrarun variation and 25-38% interrun variation). Results from the real-time PCR analysis were compared to analysis of the microbial population by a culture-based technique [most probable number (MPN) counts]. Both types of analysis indicated that fertilizer addition to hydrocarbon-contaminated soil stimulated the indigenous bacterial population within 1 year. The proportion of alkB containing microorganisms was positively correlated to the concentration of n-alkanes in the soil. After the concentration of n-alkanes in the soil decreased, the proportion of alkane-degrading microorganisms decreased, but the proportion of total hydrocarbon-degrading microorganisms increased, indicating another shift in the microbial community structure and ongoing biodegradation.
开发了一种实时聚合酶链反应（PCR）方法，用于定量含有烷烃单加氧酶的微生物比例，并在生物修复现场试验中跟踪烃污染南极土壤中微生物群落的变化。对alkB和rpoB基因的检测进行了验证，发现它们既灵敏又可重复（批内变异小于2%，批间变异为25%-38%）。将实时PCR分析的结果与基于培养技术的微生物种群分析[最可能数（MPN）计数]进行了比较。两种分析类型均表明，向烃污染土壤中添加肥料在1年内刺激了本地细菌种群。含alkB微生物的比例与土壤中正构烷烃的浓度呈正相关。土壤中正构烷烃浓度降低后，烷烃降解微生物的比例下降，但总烃降解微生物的比例增加，表明微生物群落结构发生了另一次转变，生物降解仍在进行。
