Control of plant viral diseases through cross-protection conferred by an attenuated vaccine is an important strategy for plant protection. However, the mutated site of an attenuated vaccine may not be stably inherited, while viruses have evolved efficient repair mechanisms for the maintenance of genomic integrity. Here, the wide host range and broad selection of mutation sites in cucumber mosaic virus (CMV) enabled construction of an attenuated vaccine through insertional mutation of the CMV 2b protein. CMV-R2E was stably inherited in tobacco for more than 10 generations and had a high relative control efficacy of CMV. Then, the use of polyetherimide (PEI)-modified functionalized carboxylated single-walled carbon nanotubes (PSWNTs) was investigated for vaccine delivery to address the problems of poor stability, complex procedure on field application, and exacting storage conditions with Agrobacterium inoculation. After co-incubating at a 1:300 ratio for 30 min, the vaccine and PSWNTs combined to form pCMV-R2E@PSWNTs, which resulted in a significant increase in the average height of the nanoparticles from 6.56 to 72.34 nm. The relative control efficacy of pCMV-R2E@PSWNTs to CMV was found to be 90.37%. Furthermore, the protective effect of PSWNTs on plasmids was investigated under various environmental conditions and the potential plant toxicity of pCMV-R2E@PSWNTs was assessed, providing a theoretical basis for field application of the vaccine nano-delivery system. A highly effective, stable viral vaccine for plants was thus developed and combined with nanocarriers to address the problems of field application. This approach has the potential to enable wider use of attenuated vaccines for sustainable prevention against plant viral disease in the field.
Keywords: attenuated vaccine; carbon nanotubes; cross-protection; cucumber mosaic virus; nano-delivery system; plant viral disease.