Environmental DNA (eDNA) can exist in both intracellular and extracellular forms in natural ecosystems. When targeting harmful cyanobacteria, extracellular eDNA indicates the presence of traces of cyanobacteria, while intracellular eDNA indicates the potential for cyanobacteria to occur. However, identifying the “actual” potential for harmful cyanobacteria to occur is difficult using the existing sediment eDNA analysis method, which uses silica beads and cannot distinguish between these two forms of eDNA. This study analyzes the applicability of a density gradient centrifugation method (Ludox method) that can selectively analyze intracellular eDNA in sediment to overcome the limitations of conventional sediment eDNA analysis. PCR was used to amplify the extracted eDNA based on the two different methods, and the relative amount of gene amplification was compared using electrophoresis and Image J application. While the conventional bead beating method uses sediment as it is to extract eDNA, it is unknown whether the mic gene amplified from eDNA exists in the cyanobacterial cell or only outside of the cell. However, since the Ludox method concentrates the intracellular eDNA of the sediment through filtration and density gradient, only the mic gene present in the cyanobacteria cells could be amplified. Furthermore, the bead beating method can analyze up to 1 g of sediment at a time, whereas the Ludox method can analyze 5 g to 30 g at a time. This gram of sediments makes it possible to search for even a small amount of mic gene that cannot be searched by conventional bead beating method. In this study, the Ludox method secured sufficient intracellular gene concentration and clearly distinguished intracellular and extracellular eDNA, enabling more accurate and detailed potential analysis. By using the Ludox method for environmental RNA expression and next-generation sequencing (NGS) of harmful cyanobacteria in the sediment, it will be possible to analyze the potential more realistically.
연안 준설퇴석물을 열처리, 생물침출공정을 이용한 중금속처리, 중화처리 등의 다양한 방법으로 전처리한 재료를 이용하여 연안해수에 함유된 질산염의 흡착특성에 대한 실험실적 연구를 수행하였다. 전처리된 퇴적물의 흡착평형시간은 생물침출-열처리 퇴적물이 17분으로 가장 빨랐으며 생물침출-중화-열처리 퇴적물과 생물침출퇴적물이 각각 25분. 27분으로 황토와 열처리 황토에서의 3,3분, 32분 보다 빠르게 흡착되는 것으로 평가되었다. 시험된 재료들에 대한 흡착공정은 Freundlich 등온흡착식으로 해석할 수 있었으며, 전처리한 퇴적물의 흡착속도가 황토에 비해 빠름을 알 수 있었다. 질산염의 흡착량은 퇴적물과 열처리 퇴적물에서 각각 2.l2, 2.19 mg NO3-N/g 으로 가장 많았으며 열처리 등으로 전처리된 퇴적물은 연안해수의 수질개선을 위한 흡착재로서 활용 가능한 것으로 평가되었다.