The typical farmland soil of Wuxue in Hubei Province was collected.Based on the phospholipid fatty acid (PLFA) extraction method existed,we set different methanol extraction time,soil sample weight and hexane dissolved volume.The results showed that the phospholipid fatty acids were extracted from the start of the methanol elution.The optimal weight of the soil in Wuxue typical farmland system was 2.0 g.The phospholipid fatty acid methyl esters dissolved in 250 μL hexane were proved to provide the most complete result.The further analysis of the microbial community structure in soils of different crops in Wuxue were conducted to verify the effect of the adjusted PLFA extraction method.The result showed that this method was of good stability and repeatability,reflecting the diversity of soil microbial community structure in typical farmland of Wuxue.