DNA release from plant tissue using focused ultrasound extraction (FUSE)

IF 2.7 3区 生物学 Q2 PLANT SCIENCES Applications in Plant Sciences Pub Date : 2023-01-28 DOI:10.1002/aps3.11510
Alexia Stettinius, Hal Holmes, Qian Zhang, Isabelle Mehochko, Misa Winters, Ruby Hutchison, Adam Maxwell, Jason Holliday, Eli Vlaisavljevich
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Abstract

Premise

Sample preparation in genomics is a critical step that is often overlooked in molecular workflows and impacts the success of downstream genetic applications. This study explores the use of a recently developed focused ultrasound extraction (FUSE) technique to enable the rapid release of DNA from plant tissues for genetic analysis.

Methods

FUSE generates a dense acoustic cavitation bubble cloud that pulverizes targeted tissue into acellular debris. This technique was applied to leaf samples of American chestnut (Castanea dentata), tulip poplar (Liriodendron tulipifera), red maple (Acer rubrum), and chestnut oak (Quercus montana).

Results

We observed that FUSE can extract high quantities of DNA in 9–15 min, compared to the 30 min required for control DNA extraction methods. FUSE extracted DNA quantities of 24.33 ± 6.51 ng/mg and 35.32 ± 9.21 ng/mg from American chestnut and red maple, respectively, while control methods yielded 6.22 ± 0.87 ng/mg and 11.51 ± 1.95 ng/mg, respectively. The quality of the DNA released by FUSE allowed for successful amplification and next-generation sequencing.

Discussion

These results indicate that FUSE can improve DNA extraction efficiency for leaf tissues. Continued development of this technology aims to adapt to field-deployable systems to increase the cataloging of genetic biodiversity, particularly in low-resource biodiversity hotspots.

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聚焦超声提取(FUSE)技术从植物组织中释放DNA
在基因组学中,样品制备是分子工作流程中经常被忽视的关键步骤,并影响下游遗传应用的成功。本研究探讨了使用最近开发的聚焦超声提取(FUSE)技术,使DNA从植物组织中快速释放用于遗传分析。方法FUSE产生密集的声空化气泡云,将目标组织粉碎成脱细胞碎片。该技术应用于美洲板栗(Castanea dentata)、郁金香白杨树(Liriodendron tulipifera)、红枫(Acer rubrum)和栗树栎(Quercus montana)的叶片样品。结果我们观察到FUSE可以在9-15分钟内提取大量的DNA,而对照组的DNA提取方法需要30分钟。FUSE提取美洲板栗和红枫的DNA量分别为24.33±6.51 ng/mg和35.32±9.21 ng/mg,而对照方法提取的DNA量分别为6.22±0.87 ng/mg和11.51±1.95 ng/mg。FUSE释放的DNA质量允许成功扩增和下一代测序。这些结果表明,FUSE可以提高叶片组织DNA的提取效率。该技术的持续发展旨在适应可实地部署的系统,以增加遗传生物多样性的编目,特别是在低资源生物多样性热点地区。
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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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