黄海沉积物柱状样中有孔虫古DNA的提取和PCR扩增的方法学初探

PRELIMINARY STUDY ON THE METHODS OF THE EXTRACTION AND PCR AMPLIFICATION OF ANCIENT FORAMINIFERAL DNA IN CORE SEDIMENT FROM THE YELLOW SEA

海洋沉积物柱状样有孔虫古DNA是近年来国际新兴的研究技术,对于解释海洋全球变化可以获取到传统形态学检获不到的遗传信息,并对其进行比较和补充。目前国际上有孔虫古DNA的研究主要开展于深海和极地等有利于DNA保存的环境,但对于近海陆架海域尚无相关有效的技术研究。为了探索适合提取和扩增陆架浅海环境沉积物柱状样中的有孔虫古DNA的方法,本实验以采自山东半岛附近的黄海沉积物柱状样为研究对象,通过改进DNA提取过程的涡旋震荡时间和洗脱液体积、比较不同的PCR扩增条件和引物对,对陆架浅海地区沉积物柱状样中有孔虫古DNA提取和PCR扩增的方法进行了探索。借助ImageJ软件对PCR产物的凝胶电泳图像进行了定量分析与比较。研究结果显示,延长涡旋震荡时间和减少洗脱液体积可以提高对海洋沉积物环境总古DNA的提取效能,使用引物对s14F0和s15以及优化后的PCR条件能成功扩增陆架浅海环境沉积物中的有孔虫古DNA。本文探索了适用于陆架浅海环境沉积物柱状样的有孔虫古DNA的研究技术,可为古海洋和古环境研究提供新的研究思路。

Ancient DNA of the foraminifers in the core sediments is a hot spot of the international scientific research in recent years. For the interpretation of global ocean changes, genetic information that cannot be detected by traditional morphology can be obtained, and it can be compared and supplemented. At present, the research on ancient DNA of foraminifera in the world is mainly carried out in the deep sea and polar region, which are conducive to DNA preservation, but there is no relevant and effective technical research on the offshore shelf. In order to explore a suitable method for extracting and amplifying ancient foraminiferal DNA from the sediments in the shallow continental shelf,this experiment adopted the core sediments from the Yellow Sea near the Shandong Peninsula as the research object.The extraction and PCR amplification methods were explored by improving the eddy oscillation time and eluting volume of DNA extraction process, and comparing different PCR amplification conditions and primer pairs. The gel electrophoresis images of PCR products were quantitatively analyzed and compared by ImageJ software. The results show that the extraction efficiency of total ancient DNA in marine sediment can be improved by prolonging the eddy oscillation time and reducing the eluting volume, using the primers s14F0 and S15 and the optimized PCR conditions.