Visualization of extracellular DNA released during border cell separation from the root cap

Fushi Wen, Gilberto Curlango-Rivera, David A. Huskey, Zhongguo Xiong, Martha C Hawes

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

PREMISE OF THE STUDY: Root border cells are programmed to separate from the root cap as it penetrates the soil environment, where the cells actively secrete >100 extracellular proteins into the surrounding mucilage. The detached cells function in defense of the root tip by an extracellular trapping process that also requires DNA, as in mammalian white blood cells. Trapping in animals and plants is reversed by treatment with DNase, which results in increased infection. The goal of this study was to evaluate the role of DNA in the structural integrity of extracellular structures released as border cells disperse from the root tip upon contact with water. METHODS: DNA stains including crystal violet, toluidine blue, Hoechst 33342, DAPI, and SYTOX green were added to root tips to visualize the extracellular mucilage as it absorbed water and border cell populations dispersed. DNase I was used to assess structural changes occurring when extracellular DNA was degraded. KEY RESULTS: Complex masses associated with living border cells were immediately evident in response to each stain, including those that are specific for DNA. Treating with DNase I dramatically altered the appearance of the extracellular structures and their association with border cells. No extracellular DNA was found in association with border cells killed by freezing or high-speed centrifugation. This observation is consistent with the hypothesis that, as with border cell extracellular proteins, DNA is secreted by living cells. CONCLUSION: DNA is an integral component of border cell extracellular traps.

Original languageEnglish (US)
Pages (from-to)970-978
Number of pages9
JournalAmerican Journal of Botany
Volume104
Issue number7
DOIs
StatePublished - 2017

Fingerprint

root cap
Cell Separation
visualization
DNA
cells
Meristem
mucilage
root tips
deoxyribonuclease I
Deoxyribonuclease I
trapping
mucilages
dyes
Coloring Agents
protein
border
Gentian Violet
Tolonium Chloride
structural change
gentian violet

Keywords

  • Corn
  • ExDNA
  • Extracellular traps
  • Fabaceae
  • Pea
  • Pisum sativum
  • Poaceae
  • Rhizosphere
  • Root border cells
  • Root caps
  • Zea mays

ASJC Scopus subject areas

  • Ecology, Evolution, Behavior and Systematics
  • Genetics
  • Plant Science

Cite this

Visualization of extracellular DNA released during border cell separation from the root cap. / Wen, Fushi; Curlango-Rivera, Gilberto; Huskey, David A.; Xiong, Zhongguo; Hawes, Martha C.

In: American Journal of Botany, Vol. 104, No. 7, 2017, p. 970-978.

Research output: Contribution to journalArticle

Wen, Fushi ; Curlango-Rivera, Gilberto ; Huskey, David A. ; Xiong, Zhongguo ; Hawes, Martha C. / Visualization of extracellular DNA released during border cell separation from the root cap. In: American Journal of Botany. 2017 ; Vol. 104, No. 7. pp. 970-978.
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abstract = "PREMISE OF THE STUDY: Root border cells are programmed to separate from the root cap as it penetrates the soil environment, where the cells actively secrete >100 extracellular proteins into the surrounding mucilage. The detached cells function in defense of the root tip by an extracellular trapping process that also requires DNA, as in mammalian white blood cells. Trapping in animals and plants is reversed by treatment with DNase, which results in increased infection. The goal of this study was to evaluate the role of DNA in the structural integrity of extracellular structures released as border cells disperse from the root tip upon contact with water. METHODS: DNA stains including crystal violet, toluidine blue, Hoechst 33342, DAPI, and SYTOX green were added to root tips to visualize the extracellular mucilage as it absorbed water and border cell populations dispersed. DNase I was used to assess structural changes occurring when extracellular DNA was degraded. KEY RESULTS: Complex masses associated with living border cells were immediately evident in response to each stain, including those that are specific for DNA. Treating with DNase I dramatically altered the appearance of the extracellular structures and their association with border cells. No extracellular DNA was found in association with border cells killed by freezing or high-speed centrifugation. This observation is consistent with the hypothesis that, as with border cell extracellular proteins, DNA is secreted by living cells. CONCLUSION: DNA is an integral component of border cell extracellular traps.",
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