用于叶绿体组装和基因组脱脂的冷冻储存DNA和植物标本组织样本的比较

IF 2.7 3区 生物学 Q2 PLANT SCIENCES
Edward V. McAssey, Cassidy Downs, Mitsuko Yorkston, Clifford Morden, Karolina Heyduk
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引用次数: 2

摘要

从植物标本室标本中提取DNA是植物生物学进化研究中越来越重要的来源,特别是在物种稀少或难以获得的情况下。在这里,我们通过夏威夷植物DNA库比较了从植物标本组织中提取的DNA与冷冻储存的DNA的效用。方法1994-2019年,夏威夷植物DNA文库收集的植物在收集时同时作为标本馆标本加入。使用短读测序对配对样品进行测序,并评估叶绿体组装和核基因恢复。结果植物标本来源的DNA比冷冻保存的新鲜组织来源的DNA碎片化程度更高,导致叶绿体组装更差,总体覆盖率更低。核靶的数量主要受每个文库的总测序读数和标本年龄的影响,而不受保存方法(植物标本馆或长期冷冻)的影响。尽管有证据表明样本中存在DNA损伤,但没有证据表明这与储存时间长短有关,无论是冷冻还是作为植物标本馆标本。从植物标本组织中提取的DNA尽管高度破碎和退化,但仍将是无价的。稀有植物群将受益于传统的植物标本馆储存方法和提取的DNA冷冻库。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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

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

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.

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来源期刊
CiteScore
7.30
自引率
0.00%
发文量
50
审稿时长
12 weeks
期刊介绍: Applications in Plant Sciences (APPS) is a monthly, peer-reviewed, open access journal promoting the rapid dissemination of newly developed, innovative tools and protocols in all areas of the plant sciences, including genetics, structure, function, development, evolution, systematics, and ecology. Given the rapid progress today in technology and its application in the plant sciences, the goal of APPS is to foster communication within the plant science community to advance scientific research. APPS is a publication of the Botanical Society of America, originating in 2009 as the American Journal of Botany''s online-only section, AJB Primer Notes & Protocols in the Plant Sciences. APPS publishes the following types of articles: (1) Protocol Notes describe new methods and technological advancements; (2) Genomic Resources Articles characterize the development and demonstrate the usefulness of newly developed genomic resources, including transcriptomes; (3) Software Notes detail new software applications; (4) Application Articles illustrate the application of a new protocol, method, or software application within the context of a larger study; (5) Review Articles evaluate available techniques, methods, or protocols; (6) Primer Notes report novel genetic markers with evidence of wide applicability.
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