Nest construction is one of the most interesting behavior in paper wasps however, little information is available on the plants which are being used by wasps. We evaluated the possibility of using DNA metabarcoding approach using rbcL and trnL molecular markers in qualitative and quantitative detection of plants in mixed origin woody samples with artificially mixture of bark samples. A threshold of 0.01 proportion of reads were applied based on a presence of false positives due to possible contamination of bark samples by airborne pollen however this threshold for median calculation was only 0.0025. In case of taxa richness, the median analysis showed better performance with higher taxa detection power however, rbcL marker outperformed in quantitative analysis. Then we applied DNA metabarcoding to identify the plant species used in nest construction by two Polistes species, P. mandarinus and P. rothneyi. The results revealed that woody plants were the most commonly used material in nest construction, with Quercus and Robinia being the two most dominant plant taxa for nest construction in both species. More exclusive wasp plant interaction was observed in P. rothneyi demonstrating their ability to use diverse taxa in their nest construction and perhaps their higher adaptation abilities in adverse climate condition. Overall, this study highlights the importance of using molecular methods for plant identification in nest construction studies and provides insights into the diverse range of plant taxa used by these wasp species.