Using Vertically Aligned Carbon Nanofiber Arrays on Rigid or Flexible Substrates for Delivery of Biomolecules and Dyes to Plants

Jessica M. Morgan; Joanna Jelenska; Dale K. Hensley; Pengju Li; Bernadeta R. Srijanto; Scott T. Retterer; Robert F. Standaert; Jennifer L. Morrell-Falvey; Jean T. Greenberg

doi:10.3791/65602

Using Vertically Aligned Carbon Nanofiber Arrays on Rigid or Flexible Substrates for Delivery of Biomolecules and Dyes to Plants

Jessica M. Morgan, Joanna Jelenska, Dale K. Hensley, Pengju Li, Bernadeta R. Srijanto, Scott T. Retterer, Robert F. Standaert, Jennifer L. Morrell-Falvey, Jean T. Greenberg

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

Abstract

The delivery of biomolecules and impermeable dyes to intact plants is a major challenge. Nanomaterials are up-and-coming tools for the delivery of DNA to plants. As exciting as these new tools are, they have yet to be widely applied. Nanomaterials fabricated on rigid substrate (backing) are particularly difficult to successfully apply to curved plant structures. This study describes the process for microfabricating vertically aligned carbon nanofiber arrays and transferring them from a rigid to a flexible substrate. We detail and demonstrate how these fibers (on either rigid or flexible substrates) can be used for transient transformation or dye (e.g., fluorescein) delivery to plants. We show how VACNFs can be transferred from rigid silicon substrate to a flexible SU-8 epoxy substrate to form flexible VACNF arrays. To overcome the hydrophobic nature of SU-8, fibers in the flexible film were coated with a thin silicon oxide layer (2-3 nm). To use these fibers for delivery to curved plant organs, we deposit a 1 µL droplet of dye or DNA solution on the fiber side of VACNF films, wait 10 min, place the films on the plant organ and employ a swab with a rolling motion to drive fibers into plant cells. With this method, we have achieved dye and DNA delivery in plant organs with curved surfaces.

Original language	English
Article number	e65602
Journal	Journal of Visualized Experiments
Volume	2023
Issue number	197
DOIs	https://doi.org/10.3791/65602
State	Published - Jul 2023

Funding

Nanofiber arrays were fabricated at the Center for Nanophase Materials Sciences, which is a Department of Energy Office of Science User Facility (Proposal ID: CNMS2019-103 and CNMS2022-A-1182). Support from CNMS is awarded through a peer-reviewed proposal system and is provided at no cost to successful applicants who intend to publish their results (http://www.cnms.ornl.gov/user/ becoming_a_user.shtml). We thank Kevin Lester and CNMS for assistance with the production of nanofiber arrays. We thank Dr. John Caughmen, Dr. Timothy McKnight, Dr. Amber Webb, Daryl Briggs, and Travis Bee for critical discussions on experimental design. We thank Dr. Adam Rondinone for the schematic of the PECVD machine. We thank Leslie Carol for the scientific illustrations. This work was funded by the Bioimaging Science Program, U.S. Department of Energy, Office of Science, Biological and Environmental Research, DE-SC0019104, and United States Department of Agriculture, 2021-67013-34835. JMM was supported by the United States Department of Agriculture: National Institute of Food and Agriculture: Agriculture and Food Research Initiative Predoctoral Fellowship 2021-67034-35167.

Funders	Funder number
Bioimaging Science Program
Center for Nanophase Materials Sciences
U.S. Department of Energy
U.S. Department of Agriculture	2021-67013-34835
National Institute of Food and Agriculture	2021-67034-35167
Office of Science	CNMS2019-103, CNMS2022-A-1182
Biological and Environmental Research	DE-SC0019104

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Morgan, J. M., Jelenska, J., Hensley, D. K., Li, P., Srijanto, B. R., Retterer, S. T., Standaert, R. F., Morrell-Falvey, J. L., & Greenberg, J. T. (2023). Using Vertically Aligned Carbon Nanofiber Arrays on Rigid or Flexible Substrates for Delivery of Biomolecules and Dyes to Plants. Journal of Visualized Experiments, 2023(197), Article e65602. https://doi.org/10.3791/65602

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abstract = "The delivery of biomolecules and impermeable dyes to intact plants is a major challenge. Nanomaterials are up-and-coming tools for the delivery of DNA to plants. As exciting as these new tools are, they have yet to be widely applied. Nanomaterials fabricated on rigid substrate (backing) are particularly difficult to successfully apply to curved plant structures. This study describes the process for microfabricating vertically aligned carbon nanofiber arrays and transferring them from a rigid to a flexible substrate. We detail and demonstrate how these fibers (on either rigid or flexible substrates) can be used for transient transformation or dye (e.g., fluorescein) delivery to plants. We show how VACNFs can be transferred from rigid silicon substrate to a flexible SU-8 epoxy substrate to form flexible VACNF arrays. To overcome the hydrophobic nature of SU-8, fibers in the flexible film were coated with a thin silicon oxide layer (2-3 nm). To use these fibers for delivery to curved plant organs, we deposit a 1 µL droplet of dye or DNA solution on the fiber side of VACNF films, wait 10 min, place the films on the plant organ and employ a swab with a rolling motion to drive fibers into plant cells. With this method, we have achieved dye and DNA delivery in plant organs with curved surfaces.",

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Using Vertically Aligned Carbon Nanofiber Arrays on Rigid or Flexible Substrates for Delivery of Biomolecules and Dyes to Plants

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