Abstract
Effective methods for delivering bioprobes into the cells of intact plants are essential for investigating diverse biological processes. Increasing research on trees, such as Populus spp., for bioenergy applications is driving the need for techniques that work well with tree species. This report introduces vertically aligned carbon nanofiber (VACNF) arrays as a new tool for microdelivery of labeled molecules to Populus leaf tissue and whole plants. We demonstrated that VACNFs penetrate the leaf surface to deliver sub-microliter quantities of solution containing fluorescent or radiolabeled molecules into Populus leaf cells. Importantly, VACNFs proved to be gentler than abrasion with carborundum, a common way to introduce material into leaves. Unlike carborundum, VACNFs did not disrupt cell or tissue integrity, nor did they induce production of hydrogen peroxide, a typical wound response. We show that femtomole to picomole quantities of labeled molecules (fluorescent dyes, small proteins and dextran), ranging from 0.5-500 kDa, can be introduced by VACNFs, and we demonstrate the use of the approach to track delivered probes from their site of introduction on the leaf to distal plant regions. VACNF arrays thus offer an attractive microdelivery method for the introduction of biomolecules and other probes into trees and potentially other types of plants.
Original language | English |
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Article number | e0153621 |
Journal | PLoS ONE |
Volume | 11 |
Issue number | 4 |
DOIs | |
State | Published - Apr 2016 |
Funding
We thank Dr. David Weston and Dr. Abhijit Karve for the CmPP16-1 DNA. We acknowledge Dr. Tim Tschaplinski and Dr. Mitch Doktycz for critical review of the manuscript. We thank Trish Lankford for technical assistance, and Zackary Moore for assistance with plant growth and maintenance. Nanofiber arrays were produced at the Center for Nanophase Materials Sciences (CNMS), which is a DOE Office of Science User Facility. We thank Dale Hensley and the CNMS for assistance with the production of nanofiber arrays. This manuscript has been authored by UT-Battelle, LLC under Contract No. DE-AC05-00OR22725 with the U.S. Department of Energy. The United States Government retains and the publisher, by accepting the article for publication, acknowledges that the United States Government retains a non-exclusive, paid-up, irrevocable, world-wide license to publish or reproduce the published form of this manuscript, or allow others to do so, for United States Government purposes. The Department of Energy will provide public access to these results of federally sponsored research in accordance with the DOE Public Access Plan ( http://energy.gov/downloads/doe-public-access-plan ).