Waters Christopher, Hurt Carla, Krosnick Shawn
School of Environmental Studies Tennessee Technological University 200 W. 10th Street, Box 5152 Cookeville 38505 Tennessee USA.
Department of Biology Tennessee Technological University 1100 North Dixie Avenue Cookeville 38505 Tennessee USA.
Appl Plant Sci. 2025 Feb 5;13(2):e11633. doi: 10.1002/aps3.11633. eCollection 2025 Mar-Apr.
The importance of natural history collections in modern ecological and genetic research cannot be overstated. Herbarium specimens provide historical information that can be used to investigate community ecology, phenology, and population genetics. In this study, environmental DNA (eDNA) metabarcoding and next-generation sequencing were used to test the efficacy of detecting historical plant-animal interactions from herbarium specimen flowers.
A modified eDNA isolation method and standard Illumina sequencing protocols were used. Animal eDNA was amplified using both cytochrome c oxidase subunit I (COI) and 16S primers to increase detection probability. The relationship between specimen age (0-69 years) and target taxa read depth was also investigated.
We generated and identified over 1.5 million sequences of animal taxa belonging to 29 clades (families or orders). These methods enabled the detection of taxa including birds, mammals, hymenopterans, lepidopterans, coleopterans, and taxa belonging to "intrafloral" communities. While herbarium specimens overall yielded less identifiable eDNA compared to fresh material, the age of the herbarium specimen negligibly affected the amount of target and/or non-target eDNA detected in flowers.
With careful consideration of the types of data that may be obtained through sampling eDNA from herbarium specimens, these methods could prove valuable to future research on plant-animal interactions.
自然历史藏品在现代生态学和遗传学研究中的重要性怎么强调都不为过。植物标本提供的历史信息可用于调查群落生态学、物候学和种群遗传学。在本研究中，利用环境DNA（eDNA）宏条形码技术和下一代测序技术来测试从植物标本花中检测历史上动植物相互作用的有效性。
使用了改良的eDNA分离方法和标准的Illumina测序方案。使用细胞色素c氧化酶亚基I（COI）和16S引物扩增动物eDNA，以提高检测概率。还研究了标本年龄（0至69年）与目标分类单元读取深度之间的关系。
我们生成并鉴定了超过150万个属于29个分支（科或目）的动物分类单元序列。这些方法能够检测到包括鸟类、哺乳动物、膜翅目昆虫、鳞翅目昆虫、鞘翅目昆虫以及属于“花内”群落的分类单元。虽然与新鲜材料相比，植物标本总体上产生的可识别eDNA较少，但植物标本的年龄对花中检测到的目标和/或非目标eDNA的量影响可忽略不计。
在仔细考虑通过从植物标本中采集eDNA可能获得的数据类型后，这些方法可能对未来动植物相互作用的研究具有重要价值。
