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In vivo fluorescence correlation microscopy (FCM) reveals accumulation and immobilization of Nod factors in root hair cell walls

In vivo fluorescence correlation microscopy (FCM) reveals accumulation and immobilization of Nod... Fluorescence correlation microscopy (FCM) is a new single‐molecule detection technique based on the confocal principle to quantify molecular diffusion and concentration of fluorescent molecules (particles) with sub‐micron resolution. In this study, FCM is applied to examine the diffusional behaviour of fluorescent Nod factor analogues on living Vicia sativa root hairs. Three recently described Nod factors with a fluorescent acyl chain (Goedhart et al. Biochemistry 1999, 38, 10898–10907) were used. Plasmolysis of fluorescently labelled root hairs showed that the Nod factors are predominantly located in the cell wall, as hardly any fluorescence could be detected in the plasma membrane. After Nod factor‐induced root hair deformation, the new outgrowth was not labelled, indicating a lack of migration of Nod factors to the newly synthesized cell wall. In agreement, FCM showed a > 1000‐fold reduction of molecular mobility of the fluorescence Nod factors upon binding to the cell wall. In addition, FCM demonstrated that Nod factors, when exogenously applied in aqueous solution at 10 nM, markedly concentrate in the cell wall of root hairs (up to 50‐fold). The feasibility of applying FCM for the study of living plant cells as well as the implications of our results for the perception of Nod factors are discussed. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png The Plant Journal Wiley

In vivo fluorescence correlation microscopy (FCM) reveals accumulation and immobilization of Nod factors in root hair cell walls

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References (61)

Publisher
Wiley
Copyright
Copyright © 2000 Wiley Subscription Services, Inc., A Wiley Company
ISSN
0960-7412
eISSN
1365-313X
DOI
10.1046/j.1365-313x.2000.00656.x
Publisher site
See Article on Publisher Site

Abstract

Fluorescence correlation microscopy (FCM) is a new single‐molecule detection technique based on the confocal principle to quantify molecular diffusion and concentration of fluorescent molecules (particles) with sub‐micron resolution. In this study, FCM is applied to examine the diffusional behaviour of fluorescent Nod factor analogues on living Vicia sativa root hairs. Three recently described Nod factors with a fluorescent acyl chain (Goedhart et al. Biochemistry 1999, 38, 10898–10907) were used. Plasmolysis of fluorescently labelled root hairs showed that the Nod factors are predominantly located in the cell wall, as hardly any fluorescence could be detected in the plasma membrane. After Nod factor‐induced root hair deformation, the new outgrowth was not labelled, indicating a lack of migration of Nod factors to the newly synthesized cell wall. In agreement, FCM showed a > 1000‐fold reduction of molecular mobility of the fluorescence Nod factors upon binding to the cell wall. In addition, FCM demonstrated that Nod factors, when exogenously applied in aqueous solution at 10 nM, markedly concentrate in the cell wall of root hairs (up to 50‐fold). The feasibility of applying FCM for the study of living plant cells as well as the implications of our results for the perception of Nod factors are discussed.

Journal

The Plant JournalWiley

Published: Jan 1, 2000

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