Rainwater chemistry was investigated at a semi-rural site in Ya'an, Sichuan basin with rain samples collected from May 2013 to July 2014. The rainwater pH values ranged from 3.25 to 6.86, with an annual volume-weighted mean (VWM) of 4.38, and the acid rain frequency was 74 %. Such severe acidification, 15 % of the total events showed a pH below 4.0, attributed to the deficiency of Ca(2+), significant anthropogenic pollution contribution, and rainy pattern to this area. The annual VWM of total ions concentration was 477.19 μeq/L. NH4 (+) was the most abundant ionic species, followed by SO4 (2-), NO3 (-), Ca(2+), Cl(-), Na(+), K(+), Mg(2+), and F(-) in a descending order. The total ionic concentrations presented a seasonal trend of lower values in autumn and summer but higher ones in winter and spring. Based on enrichment factor, correlation analysis and principle component analysis, three factors were identified: factor 1 (NH4 (+), SO4 (2-), NO3 (-), K(+), and Cl(-), 47.45 % of the total variance) related to anthropogenic sources (coal/fuel combustion, biomass burning and agriculture), factor 2 (Ca(2+), Mg(2+), Na(+), and Cl(-), 34.01 % of the total variance) associated with natural sources, and factor 3 (H(+), 11.78 % of the total variance) related to free acidity. Back trajectory analysis indicates that the rainwater chemistry in Ya'an was mainly affected by regional air masses from Sichuan basin. Long-range transported air masses from southwest with heavy anthropogenic pollution increased the total ion concentration and acidity of rainwater. Considering its special topography, anthropogenic emissions from regional and long-range transport (especially from southwest) must be controlled effectively to improve the acid rain condition of non-urban areas in Sichuan basin.

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Conserving aquatic ecosystems requires efficient tools to accurately assess the biodiversity of aquatic species. However, existing knowledge is insufficient in terms of the reliability and the comparability of methods measuring fish diversity. Environmental DNA (eDNA), as a promising method, was used to detect fish taxa in this study. We optimized the eDNA method in the laboratory, and applied the optimal eDNA method to survey fish diversity in a natural aquatic life reserve. We simulated necessary steps of the eDNA method in the lab to increase the confidence of the field survey. Specifically, we compared different eDNA sampling, extraction, and sequencing strategies for accurately capturing fish species of the target area. We found that 1L water samples were sufficient for sampling eDNA information of the majority taxa. The filtration was more effective than the centrifugal precipitation for the eDNA extraction. The cloning sequencing was better than the high-throughput sequencing. The field survey showed that the Shannon–Wiener diversity index of fish taxa was the highest in Huairou Reservoir. The diversity index also showed seasonal changes. The accuracy rate of detecting fish taxa was positively correlated with the eDNA concentration. This study provides a scientific reference for an application of the eDNA method in terms of surveying and estimating the biodiversity of aquatic species.