Exploring fish diversity inhabiting Lake Dianchi by metabarcording multiple universal primer defined markers

MENG Han; XU Shan; LV Jiacheng; ZHAO Kaisong; ZHAO Zheng; LIN Yuanyuan

doi:10.13984/j.cnki.cn37-1141.2025.06.017

Abstract：The selection of universal primers is very important for by environmental DNA(eDNA)me-tabarcoding in monitoring fish biodiversity.In this study,the eDNA samples were taken from Lake Di-anchi,the amplification performance of six pairs of universal primers(Teleo,Meta,CY45M,1634M,MiFish,FishCB)were compared.The results showed that 1634M,Meta and Teleo amplify the eDNAs well,yielding bright,clear and single target bands suitable for subsequent high-throughput sequen-cing.With 1634M,Meta and Teleo,eDNA samples from Lake Dianchi in dry and wet periods were amplified and sequenced.A total of 37 species of fish belonging to 8 orders,13 families and 36 genera were annotated,which include common fish in Lake Dianchi like Misgurnus anguillicaudatus,Caras-sius auratus,Monopterus albus,Cyprinus carpio,Pseudorasbora parva and endangered rare Sinocy-clocheilus grahami.There was a significant difference in the composition of fish species annotated with the data from the three primers,24(1634M),9(Meta)and 10(Teleo)species,respectively.In total,33 fish species including Sinocyclocheilus grahami,Cultrichthys erythropterus,Ctenopharyngodon idella among others were specifically annotated by one of the three primers.The fish annotated by all three primers were Hyporhamphus intermedius and Cyprinus carpio.The fish annotated by 1634M and Meta is Rhinogobius similis.The fish annotated by Teleo and Meta is Carassius auratus.The mo-nitoring results based on all the three primers could clearly reflect the seasonal variations of fish diversi-ty in Lake Dianchi,and the difference of fish species composition in the wet period was smaller than that in the dry period.Combination of multiple universal primers would improve coverage rate of fish species composition and the effect of diversity monitoring.