摘要: 16S rRNA gene sequencing is a widely used method to estimate the diversity of bacterial species in environmental samples. However, the reliability of results is often severely compromised owing to PCR artifacts such as chimeric 16S rRNA gene sequences. Not all researchers have taken these limitations of polymerase chain reaction (PCR) into account in their experimental designs. In order to evaluate the effect of chimeras on bacterial symbionts surveying studies in insects, we identified chimeras among the pooled 16S rRNA gene sequences obtained from wheat aphid Sitobion miscanthi bacterial microbiota in this study. Chimeras were observed to occur at a high frequency; in which 18 of the 39 sequences tested were found to be chimeric. Most of these chimeras were composed of the 16S rRNA gene sequences of aphid obligate symbiont Buchnera aphidicola and species of Pseudomonas. Furthermore, we found that the widely used chimera identification algorithms (Pintail, Decipher, Uchime and Chimera Slayer) had differing capacities for the identification of chimeras, which suggested that results obtained from these algorithms may not be compared directly and should be combined to produce a comprehensive final result. Our study highlights that the identification and exclusion of chimeras is necessary in surveys of insect bacterial symbionts that use 16S rRNA gene sequencing methods.