HACC-Based Nanoscale Delivery of the NbMLP28 Plasmid as a Crop Protection Strategy for Viral Diseases

Abstract

Resistant genes as an effective strategy to antivirus of plants are at the core of sustainability efforts. We use the antiviral protein major latex protein 28 (NbMLP28 plasmid) and N -2-hydroxypropyl trimethyl ammonium chloride chitosan (HACC) designated as the HACC/NbMLP28 complex as protective gene delivery vectors to prepare nanonucleic acid drugs. The maximum drug loading capacity of HACC was 4. The particle size of HACC/NbMLP28 was measured by transmission electron microscopy and found to be approximately 40–120 nm, the particle dispersion index (PDI) was 0.448, and the ζ-potential was 22.3 mV. This facilitates its ability to deliver particles. Different controls of laser scanning confocal experiments verified the effective expression of NbMLP28 and the feasibility of nanodelivery. The optimal ratio of HACC/plasmid was 2:1. Finally, the nanoparticle/plasmid complex was tested for its ability to control diseases and was found to significantly improve resistance to three viruses. The enhanced resistance was particularly notable 4 days after inoculation. Taken together, these results indicate that HACC/NbMLP28 is a promising tool to treat plant viruses. To the best of our knowledge, this is the first study that successfully delivered and expressed antiviral protein particles in plants. This gene delivery system can effectively load antiviral plasmids and express them in plant leaves, significantly affecting the plant resistance of three RNA viruses.

Nanoplatforms for the Delivery of Nucleic Acids into Plant Cells

“…Despite the numerous beneficial properties of chitosan and a great number of studies on chitosan applications, only a few examples of chitosan-mediated delivery of nucleic acids into plant cells have been described currently [51,57,[59][60][61][62].…”

“…Moreover, the same TPP-chitosan nanoparticles were demonstrated to efficiently deliver a ribonucleoprotein complex containing Cas9 endonuclease and guide RNA into the cells of the potato apical meristem via vacuum infiltration, which was confirmed via the sequencing of edited coilin [60] and phytoene desaturase (PDS) [59] genes in regenerated cell lines. Zhang et al [61] used chitosan quaternary ammonium salt to obtain nanoparticles with incorporated pDNA. The chitosan/pDNA nanocomplexes were infiltrated into N. benthamiana leaves.…”

Non-transgenic, PAMAM co-delivery DNA of interactive proteins NbCRVP and NbCalB endows Nicotiana benthamiana with a stronger antiviral effect to RNA viruses

“…Furthermore, nanocarriers have also been used to protect these biomolecules from various degradation processes in plant cells [ 6 ]. For example, N -2-hydroxypropyl trimethyl ammonium chloride chitosan can protect the plasmid carrying the resistance protein NbMLP28 from nuclease degradation and successfully delivery it to achieve the antiviral effect [ 7 ].…”