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

聚乙烯亚胺包覆金纳米粒子携外源基因在长春花叶肉细胞中的表达及在黄龙病研究中的实践

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作者:Junzhe Zhang, Wenxue Lei, Yixuan Meng, Chengqian Zhou, Baihong Zhang, Jun Yuan, Min Wang, Dongqing Xu, Xiangchun Meng, Wenli Chen

Abstract

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-siRNA NPR1 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.

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