High-throughput sequencing has dramatically expanded our understanding of bacterial communities based on the information of the species types and their relative abundances. Recently, researchers have also become aware of a deficiency in not considering the absolute abundance in this technique. Combining two or more different methods has typically been used to achieve absolute quantification of microbial communities. However, making a combination of different methods not only is time-consuming but also involves potential uncertainty due to variations in the experimental conditions. To simplify the experimental procedure and improve the high-throughput absolute abundance quantification (HAAQ) of a soil bacterial community, we propose an HAAQ method that uses an internal standard strain (ISS) HAAQ-GFP to simultaneously obtain both the relative and absolute abundances in the soil bacterial community. The results showed that a soil bacterial community and its dynamics can be better characterized by the HAAQ method when the optimal concentrations of ISS HAAQ-GFP (10⁵ to 10⁷ cells g⁻¹) were used, and a 16S rRNA gene copy number adjustment was applied. Based on the HAAQ method, we first found that soil bacterial absolute abundances at the genus level fitted well to the partial log-normal distribution function, and most genera concentrations were in the range of 10^3.5 to 10^6.5 cells g⁻¹ in the test soils. Our case studies also indicated that more comprehensive descriptions of soil bacterial communities and their dynamics can be achieved by both the relative and absolute abundances than by the relative abundance alone. The improved HAAQ method can be potentially applied to other microbial ecological studies and to stimulating the development of quantitative bacterial ecology studies.

基于物种类型及其相对丰度的信息，高通量测序极大地扩展了我们对细菌群落的了解。最近，研究人员也意识到没有考虑这种技术的绝对丰度的缺陷。通常已使用两种或多种不同方法的组合来实现微生物群落的绝对定量。然而，将不同方法组合在一起不仅费时，而且由于实验条件的变化还可能带来不确定性。为了简化实验过程并提高土壤细菌群落的高通量绝对丰度定量（HAAQ），我们提出一种使用内标菌株（ISS）HAAQ-GFP的HAAQ方法，以同时获得土壤细菌群落中的相对和绝对丰度。结果表明，当ISS HAAQ-GFP的最佳浓度为10时，用HAAQ方法可以更好地表征土壤细菌群落及其动力学。^使用5至10 ^7个 细胞g ^-1），并进行16S rRNA基因拷贝数调节。基于HAAQ方法，我们首先发现在属水平上的土壤细菌绝对丰度非常适合部分对数正态分布函数，并且大多数属浓度在10 ^3.5至10 ^6.5个 细胞g ^-1之间在测试土壤中。我们的案例研究还表明，相对和绝对丰度比单独的相对丰度可以更全面地描述土壤细菌群落及其动态。改进的HAAQ方法可以潜在地应用于其他微生物生态学研究，并刺激定量细菌生态学研究的发展。