由于软体动物的多样性和分布广泛,监测其生物多样性是一项巨大的挑战。环境DNA(eDNA)技术越来越多地应用于生物多样性监测,但是对海洋软体动物的相关研究仍然有限。尽管以前的研究已经开发了几对用于软体动物eDNA分析的引物,他们中的大多数只针对一小群软体动物。在这项研究中,为软体动物群落设计了7个引物,并与8对已发表的引物进行了验证和比较,以选择最佳候选物。经过硅片测试,MollCOI154和MollCOI255引物显示非特异性扩增,在已发表的引物中也获得了相同的结果(COI204,Sepi,和veneroida)。Moll12S100、Moll12S195和Moll16S引物未能扩增来自选定软体动物的所有基因组DNA。除了Moll16S,在长江口的四个eDNA样品上成功扩增了所有开发的和两个已发表的(unionoida和veneroida)引物。在注释扩增的序列后,注解MollCOI253显示出比其他引物更高的扩增成果。总之,MollCOI253在扩增成功和特异性方面具有更好的性能,可以为基于eDNA的研究提供技术支持,这将有利于软体动物生物多样性的调查和保护。
Monitoring
mollusk biodiversity is a great challenge due to their large diversity and broad distribution. Environmental DNA (eDNA) technology is increasingly applied for biodiversity monitoring, but relevant studies on marine mollusks are still limited. Although previous studies have developed several pairs of primers for
mollusk eDNA analyses, most of them targeted only a small group of mollusks. In this study, seven primers were designed for the
mollusk community and validated and compared with eight pairs of published primers to select the best candidates. After in silico test, MollCOI154 and MollCOI255 primers showed non-specific amplification, and same results were also obtained in published primers (COI204, Sepi, and veneroida). Moll12S100, Moll12S195 and Moll16S primers failed to amplify across all genomic DNA from selected
mollusk. Except Moll16S, all developed and two published (unionoida and veneroida) primers were successfully amplified on four eDNA samples from Yangtze River estuary. After annotation of the amplified sequences, MollCOI253 showed higher annotation of the amplification results than the other primers. In conclusion, MollCOI253 had better performance in terms of amplification success and specificity, and can provide technical support for eDNA-based research, which will be beneficial for molluscan biodiversity investigation and conservation.