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

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

doi:10.1016/j.isci.2022.104479

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

iScience. 2022 May 29;25(6):104479. doi: 10.1016/j.isci.2022.104479. eCollection 2022 Jun 17.

Authors

Affiliations

¹ MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, Guangdong Provincial Key Laboratory of Laser Life Science, Guangzhou Key Laboratory of Spectral Analysis and Functional Probes, College of Biophotonics, South China Normal University, Guangzhou 510631, China.
² Neuroscience Laboratory, Hugo Moser Research Institute at Kennedy Krieger, Baltimore MD 21205, USA.
³ College of Resources and Environmental Sciences, Nanjing Agricultural University, Weigang No.1, Nanjing 210095, China.
⁴ College of Life Science, Nanjing Agricultural University, Weigang No.1, Nanjing 210095, China.
⁵ Key Laboratory of South Subtropical Fruit Biology and Genetic Resource Utilization (MOA), Key Laboratory of Tropical and Subtropical Fruit Tree Research, Institute of Fruit Tree Research, Guangdong Academy of Agricultural Science, Guangzhou 510640, China.

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.

Keywords: Biological sciences; Biomaterials; Biotechnology; Materials in biotechnology.