Expression of PEI-coated gold nanoparticles carrying exogenous gene in periwinkle mesophyll cells and its practice in huanglongbing research

• Published in: iScience Vol. 25; no. 6; p. 104479
• Main Authors: Zhang, Junzhe, Lei, Wenxue, Meng, Yixuan, Zhou, Chengqian, Zhang, Baihong, Yuan, Jun, Wang, Min, Xu, Dongqing, Meng, Xiangchun, Chen, Wenli
• Format: Journal Article
• Language: English
• Published: Elsevier Inc 17.06.2022; Elsevier
• Subjects: Biological sciences; Biomaterials; Biotechnology; Materials in biotechnology; Biotechnology; Biological sciences; Materials in biotechnology; Biomaterials
• ISSN: 2589-0042; 2589-0042
• DOI: 10.1016/j.isci.2022.104479

Huanglongbing (HLB) is a devastating disease of citrus, which is mostly caused by Candidatus Liberibacter asiaticus (CLas). To realize the specific application of nano-transgenic technology in HLB, AuNPs-PEI (Gold Nanoparticles-Polyethylenimine) was used to carry foreign genes into the leaves of periwinkle (Catharanthus roseus) by infiltration. Here, we demonstrated that NPR1-GFP protein expression was observed from the 12th hour to the 10th day after infiltrating AuNPs-PEI-pNPR1 (Arabidopsis thaliana nonexpressor of pathogenesis-related gene 1)-GFP. Fluorescence of mCherry was observed 6 h after AuNPs-PEI-pNLS (nuclear localization signal sequence)-mCherry infiltration and fluorescence of FAM was observed in the nucleus 4 h after AuNPs-PEI-FAM-siRNANPR1 infiltration. In addition, NPR1-GFP expression in CLas-infected periwinkle leaves was significantly higher than that in healthy periwinkle leaves after infiltration. Our work confirmed that the expression of exogenous NPR1-GFP could reduce the CLas titers by promoting the expression of PR (pathogenesis related) genes and ICS (isochorismate synthase) gene. [Display omitted] •AuNPs-PEI-FAM-siRNANPR1 entered the nucleus within 4 h after infiltration•AuNPs-PEI-pNLS-mCherry expressed the corresponding protein within 6 h•AuNPs-PEI-pNPR1-GFP continued to express the corresponding protein for 14 days•After AuNPs-PEI-pNPR1-GFP infiltration for 2 days, CLas titer decreased significantly Biological sciences; Biomaterials; Biotechnology; Materials in biotechnology