Cellular Delivery of Plasmid DNA into Wheat Microspores Using Rosette Nanotubes

Abstract

Plant genetic engineering offers promising solutions to the increasing demand for efficient, sustainable, and high-yielding crop production as well as changing environmental conditions. The main challenge for gene delivery in plants is the presence of a cell wall that limits the transportation of genes within the cells. Microspores are plant cells that are, under the right conditions, capable of generating embryos, leading to the formation of haploid plants. Here, we designed cationic and fluorescent rosette nanotubes (RNTs) that penetrate the cell walls of viable wheat microspores under mild conditions and in the absence of an external force. These nanomaterials can capture plasmid DNA to form RNT–DNA complexes and transport their DNA cargo into live microspores. The nanomaterials and the complexes formed were nontoxic to the microspores.

Plant biomacromolecule delivery methods in the 21st century

“…Another hollow nanotube platform called rosette nanotubes (RNTs) were composed via the self-assembly of complementary guanine−cytosine motifs, and these nanoparticles were complexed with plasmid DNA for plant delivery (Cho et al, 2020). RNTs entered wheat microspores and did not affect health, division, or regeneration abilities of microspores.…”

“…More recently, pollens were transformed using different nanoparticle carriers, such as rosette nanotubes (RNTs) in wheat (Cho et al, 2020), sheet-like clay nanoparticles in tomato (Yong et al, 2021), single-walled carbon nanotubes (SWNTs) in oil palm (Lew et al, 2020b), cationic cellpenetrating peptides (CPP) in wheat (Bilichak, Luu and Eudes, 2015) and triticale (Chugh, Amundsen and Eudes, 2009;Pepper et al, 2017), and iron oxide magnetic nanoparticles (MNPs) in cotton (Zhao et al, 2017) and maize . However, like many other pollen transformation methods, some of these methods remain controversial due to the inability to reproduce results or obtain similar results in a different plant system or by a different research group.…”

Chiral nanotubes self-assembled from discrete non-covalent macrocycles

“…The potential use of these systems has been undoubtedly demonstrated by the high number of publications related to applications in biomedical [31][32][33][34][35] and environmental fields 36 among others.…”

Delivery of siRNA using cationic rosette nanotubes for gene silencing

“…31 We have recently shown that cationic and fluorescent RNTs can capture plasmid DNA, penetrate the plant cell walls to deliver the cargo into live wheat microspores under mild conditions and in the absence of an external force. 39 Herein, we present the synthesis of fifteen lysine-functionalized RNTs (KnT, n = 1-15), which were used to bind siRNA. The binding was examined at various molar ratios (KnT RNTs : siRNA) by gel retardation assay and the resulting KnT RNTs/siRNA complexes were subjected to nucleases in bovine serum to determine the protective ability of KnT RNTs against serum degradation.…”