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冷冻保存的DNA与腊叶标本组织样本用于叶绿体组装和基因组浅层测序的比较。

A comparison of freezer-stored DNA and herbarium tissue samples for chloroplast assembly and genome skimming.

作者信息

McAssey Edward V, Downs Cassidy, Yorkston Mitsuko, Morden Clifford, Heyduk Karolina

机构信息

School of Life Sciences University of Hawai'i at Mānoa Honolulu Hawai'i USA.

Department of Ecology and Evolutionary Biology University of Connecticut Storrs Connecticut USA.

出版信息

Appl Plant Sci. 2023 Jun 5;11(3):e11527. doi: 10.1002/aps3.11527. eCollection 2023 May-Jun.

Abstract

PREMISE

The use of DNA from herbarium specimens is an increasingly important source for evolutionary studies in plant biology, particularly in cases where species are rare or difficult to obtain. Here we compare the utility of DNA from herbarium tissues to their freezer-stored DNA counterparts via the Hawaiian Plant DNA Library.

METHODS

Plants collected for the Hawaiian Plant DNA Library were simultaneously accessioned as herbarium specimens at the time of collection, from 1994-2019. Paired samples were sequenced using short-read sequencing and assessed for chloroplast assembly and nuclear gene recovery.

RESULTS

Herbarium specimen-derived DNA was statistically more fragmented than freezer-stored DNA derived from fresh tissue, leading to poorer chloroplast assembly and overall lower coverage. The number of nuclear targets recovered varied mostly by total sequencing reads per library and age of specimen, but not by storage method (herbarium or long-term freezer). Although there was evidence of DNA damage in the samples, there was no evidence that it was related to the length of time in storage, whether frozen or as herbarium specimens.

DISCUSSION

DNA extracted from herbarium tissues will continue to be invaluable, despite being highly fragmented and degraded. Rare floras would benefit from both traditional herbarium storage methods and extracted DNA freezer banks.

摘要

前提

植物标本馆标本中的DNA是植物生物学进化研究中日益重要的来源,特别是在物种稀有或难以获取的情况下。在这里,我们通过夏威夷植物DNA文库比较了植物标本馆组织中的DNA与其冷冻保存的DNA对应物的效用。

方法

1994年至2019年收集用于夏威夷植物DNA文库的植物在采集时同时作为植物标本馆标本登记。使用短读测序对配对样本进行测序,并评估叶绿体组装和核基因回收情况。

结果

统计学上,植物标本馆标本来源的DNA比新鲜组织冷冻保存的DNA片段化程度更高,导致叶绿体组装较差且总体覆盖率较低。回收的核靶标数量主要因每个文库的总测序读数和标本年龄而异,而不是因储存方法(植物标本馆或长期冷冻库)而异。尽管有证据表明样本中存在DNA损伤,但没有证据表明其与储存时间长短有关,无论是冷冻保存还是作为植物标本馆标本保存。

讨论

尽管从植物标本馆组织中提取的DNA高度片段化和降解,但它仍将具有极高的价值。珍稀植物群落将从传统的植物标本馆储存方法和提取的DNA冷冻库中受益。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/93c6/10278930/cefc9231b2da/APS3-11-e11527-g001.jpg

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