Thomsen Philip Francis, Elias Scott, Gilbert M Thomas P, Haile James, Munch Kasper, Kuzmina Svetlana, Froese Duane G, Sher Andrei, Holdaway Richard N, Willerslev Eske
Centre for Ancient Genetics and Environments, Natural History Museum and Institute of Biology, University of Copenhagen, Copenhagen, Denmark.
PLoS One. 2009;4(4):e5048. doi: 10.1371/journal.pone.0005048. Epub 2009 Apr 1.
A major challenge for ancient DNA (aDNA) studies on insect remains is that sampling procedures involve at least partial destruction of the specimens. A recent extraction protocol reveals the possibility of obtaining DNA from past insect remains without causing visual morphological damage. We test the applicability of this protocol on historic museum beetle specimens dating back to AD 1820 and on ancient beetle chitin remains from permafrost (permanently frozen soil) dating back more than 47,000 years. Finally, we test the possibility of obtaining ancient insect DNA directly from non-frozen sediments deposited 3280-1800 years ago -- an alternative approach that also does not involve destruction of valuable material.
METHODOLOGY/PRINCIPAL FINDINGS: The success of the methodological approaches are tested by PCR and sequencing of COI and 16S mitochondrial DNA (mtDNA) fragments of 77-204 base pairs (-bp) in size using species-specific and general insect primers.
CONCLUSION/SIGNIFICANCE: The applied non-destructive DNA extraction method shows promising potential on insect museum specimens of historical age as far back as AD 1820, but less so on the ancient permafrost-preserved insect fossil remains tested, where DNA was obtained from samples up to ca. 26,000 years old. The non-frozen sediment DNA approach appears to have great potential for recording the former presence of insect taxa not normally preserved as macrofossils and opens new frontiers in research on ancient biodiversity.
对昆虫遗骸进行古DNA(aDNA)研究面临的一个主要挑战是,采样过程至少会对标本造成部分破坏。最近的一种提取方案显示了从过去的昆虫遗骸中获取DNA而不造成可见形态损伤的可能性。我们测试了该方案对可追溯到公元1820年的历史博物馆甲虫标本以及对来自距今47000多年的永久冻土(永久冻结土壤)中的古代甲虫几丁质遗骸的适用性。最后,我们测试了直接从3280 - 1800年前沉积的非冷冻沉积物中获取古代昆虫DNA的可能性——这是一种同样不涉及破坏珍贵材料的替代方法。
方法/主要发现:通过使用物种特异性和通用昆虫引物对大小为77 - 204碱基对(-bp)的COI和16S线粒体DNA(mtDNA)片段进行PCR和测序,来测试这些方法的成功率。
结论/意义:所应用的非破坏性DNA提取方法在可追溯到公元1820年的历史时期昆虫博物馆标本上显示出有前景的潜力,但在测试的古代永久冻土保存的昆虫化石遗骸上效果较差,在这些遗骸中,DNA是从距今约26000年的样本中获得的。非冷冻沉积物DNA方法似乎在记录通常不作为大化石保存的昆虫类群的过去存在方面具有巨大潜力,并为古代生物多样性研究开辟了新的前沿领域。