Evidence map›Paper›PMID 34539120›Full record

ArticlePhysiology and molecular biology of plants : an international journal of functional plant biology2021

A simple flow cytometry-based assay to study global methylation levels in onion, a non-model species.

Saravanakumar Somasundaram, Viswanathan Satheesh, Major Singh, Sivalingam Anandhan

Abstract read
In one paragraph

Article in Physiology and molecular biology of plants : an international journal of functional plant biology, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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1 · What the graph read from it

What it found

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2 · The registry

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3 · Its place in the literature

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4 · The record

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5 · Who and what money

Authors and funding

4 authors.

Saravanakumar SomasundaramICAR-Directorate of Onion and Garlic Research, Pune, 410505 India.
Viswanathan SatheeshShanghai Center for Plant Stress Biology, CAS Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai, 200032 China.
Major SinghICAR-Directorate of Onion and Garlic Research, Pune, 410505 India.
Sivalingam AnandhanICAR-Directorate of Onion and Garlic Research, Pune, 410505 India.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

DNA methylation is an important epigenetic mark and global methylation dynamics regulate plant developmental processes. Even though genome sequencing technologies have made DNA methylation studies easier, it is difficult in non-model species where genome information is not available. Therefore in this study, we developed a simple assay for analysing global methylation levels in plants by washless immunolabelling of unfixed nuclei using flow cytometry. Onion leaf tissue was used as a model system, and mean fluorescence intensity due to anti-5- methyl cytosine (5-mC) antibodies were used as a measure of global methylation levels. Among three nuclear isolation buffers evaluated, the highest nuclear yield with the low background was obtained with LB01. To maintain a balance between high DNA fluorescence value and low coefficient of variation of DNA peaks, 45 min of hydrolysis with 0.2 N hydrochloric acid was used for chromatin denaturation resulting in six-fold increase in 5-mC fluorescence compared to control. This method was used successfully to detect 5-Azacytidine induced DNA hypomethylation in onion leaf tissues. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-021-01047-6.

Indexed as

5-Azacytidine5-methylcytosineGlobal DNA methylation levelsOnionWashless staining

Identifiers

PMID34539120
PMCPMC8405793

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.