A flow cytometric method for estimating S-phase duration in plants

Mickelson-Young, Leigh; Wear, Emily E.; Mulvaney, Patrick; Lee, Tae-Jin; Szymanski, Eric S.; Allen, George C.; Hanley‐Bowdoin, Linda; Thompson, William F.

doi:10.1093/jxb/erw367

Cited by 23 publications

(15 citation statements)

References 57 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Our observations also show that the main fraction of dividing cells finish mitosis between 5 and 8 h, the minimum time frame required for cell divisions in plants and other eukaryotes (Mickelson-Young et al., 2016). Therefore, signaling from the wound to initiate cell-cycle transitions in inner adjacent cells must happen very rapidly after tissue disruption and likely relies on non-genomic factors, such as mechanical properties and local geometry of disrupted tissues.…”

Section: Discussionsupporting

confidence: 64%

Re-activation of Stem Cell Pathways for Pattern Restoration in Plant Wound Healing

Marhavá

Hoermayer

Yoshida

et al. 2019

Cell

103

125

View full text Add to dashboard Cite

Summary Patterning in plants relies on oriented cell divisions and acquisition of specific cell identities. Plants regularly endure wounds caused by abiotic or biotic environmental stimuli and have developed extraordinary abilities to restore their tissues after injuries. Here, we provide insight into a mechanism of restorative patterning that repairs tissues after wounding. Laser-assisted elimination of different cells in Arabidopsis root combined with live-imaging tracking during vertical growth allowed analysis of the regeneration processes in vivo . Specifically, the cells adjacent to the inner side of the injury re-activated their stem cell transcriptional programs. They accelerated their progression through cell cycle, coordinately changed the cell division orientation, and ultimately acquired de novo the correct cell fates to replace missing cells. These observations highlight existence of unknown intercellular positional signaling and demonstrate the capability of specified cells to re-acquire stem cell programs as a crucial part of the plant-specific mechanism of wound healing.

show abstract

Section: Discussionsupporting

confidence: 64%

Re-activation of Stem Cell Pathways for Pattern Restoration in Plant Wound Healing

Marhavá

Hoermayer

Yoshida

et al. 2019

Cell

103

125

View full text Add to dashboard Cite

show abstract

“…Over the 20-min labeling period, EdU uniformly penetrates the root and labels cells in the multiple emerging cell lineages present in the meristematic region ( Figure 1B). Under the same growth conditions, the average length of S phase of mitotic cells in the terminal 1-mm region of the root was estimated to be between 2.7 and 3.9 h (Mickelson-Young et al, 2016). Hence, a 20-min labeling period represents ;10% of the length of S phase.…”

Section: Whole-genome Profiling Of Dna Replication Timing In Maize Romentioning

confidence: 96%

Genomic Analysis of the DNA Replication Timing Program during Mitotic S Phase in Maize (Zea mays) Root Tips

Wear

Song²,

Zynda³

et al. 2017

Plant Cell

Self Cite

View full text Add to dashboard Cite

All plants and animals must replicate their DNA, using a regulated process to ensure that their genomes are completely and accurately replicated. DNA replication timing programs have been extensively studied in yeast and animal systems, but much less is known about the replication programs of plants. We report a novel adaptation of the "Repli-seq" assay for use in intact root tips of maize (Zea mays) that includes several different cell lineages and present whole-genome replication timing profiles from cells in early, mid, and late S phase of the mitotic cell cycle. Maize root tips have a complex replication timing program, including regions of distinct early, mid, and late S replication that each constitute between 20 and 24% of the genome, as well as other loci corresponding to ;32% of the genome that exhibit replication activity in two different time windows. Analyses of genomic, transcriptional, and chromatin features of the euchromatic portion of the maize genome provide evidence for a gradient of early replicating, open chromatin that transitions gradually to less open and less transcriptionally active chromatin replicating in mid S phase. Our genomic level analysis also demonstrated that the centromere core replicates in mid S, before heavily compacted classical heterochromatin, including pericentromeres and knobs, which replicate during late S phase.

show abstract

“…The click iT chemistry is nowadays widely used to label replication sites by ethynyl deoxy uridine (EdU). EdU was already successfully applied in Arabidopsis , first to visualize the DNA replication phase in cultured cells ( Kotogany et al, 2010 ; Mickelson-Young et al, 2016 ), and later to track S phase progression in root meristems ( Hayashi et al, 2013 ; Yokoyama et al, 2016 ; Dvorackova et al, 2018 ) or to detect proliferation capacity ( Kazda et al, 2016 ). It was also demonstrated that EdU persists in plant material during growth and that it could be used to track cell fate ( Watson et al, 2016 ).…”

Section: Introductionmentioning

confidence: 99%

Visualization of the Nucleolus Using Ethynyl Uridine

Dvořáčková

Fajkus

2018

Front. Plant Sci.

View full text Add to dashboard Cite

Thanks to recent innovative methodologies, key cellular processes such as replication or transcription can be visualized directly in situ in intact tissues. Many studies use so-called click iT chemistry where nascent DNA can be tracked by 5-ethynyl-2′-deoxyuridine (EdU), and nascent RNA by 5-ethynyl uridine (EU). While the labeling of replicating DNA by EdU has already been well established and further exploited in plants, the use of EU to reveal nascent RNA has not been developed to such an extent. In this article, we present a protocol for labeling of nucleolar RNA transcripts using EU and show that EU effectively highlights the nucleolus. The method is advantageous, because the need to prepare transgenic plants expressing fluorescently tagged nucleolar components when the nucleolus has to be visualized can be avoided.

show abstract

A flow cytometric method for estimating S-phase duration in plants

Abstract: HighlightWe estimated S-phase duration for several plant species by following EdU-labeled nuclei from G1 to G2 using bivariate flow cytometry. S-phase duration is relatively consistent over a range of genome sizes.

Cited by 23 publications

References 57 publications

Re-activation of Stem Cell Pathways for Pattern Restoration in Plant Wound Healing

Re-activation of Stem Cell Pathways for Pattern Restoration in Plant Wound Healing

Genomic Analysis of the DNA Replication Timing Program during Mitotic S Phase in Maize (Zea mays) Root Tips

Visualization of the Nucleolus Using Ethynyl Uridine

Contact Info

Product

Resources

About