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Front. Microbiol. | doi: 10.3389/fmicb.2018.03034

Exploration of an actin promoter-based transient expression vector to trace the cellular localization of nucleorhabdovirus proteins in leafhopper culture cells

 Taiyun Wei1*,  Xiao-Feng Zhang1, Yunjie Xie1,  Haitao Wang1, Juan Wang1, Tianbao Zeng1, Yibing Zhao1 and Hongyan Chen1
  • 1State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, Institute of Plant Virology, Fujian Agriculture and Forestry University, China

Abstract:
Continuously cultured cell lines derived from planthopper and leafhopper have greatly facilitated the investigation of rice viruses transmitted by these insects. However, the lack of a suitable transient expression vector has limited their utility. Here, by cloning and analyzing the promoter sequence of the gene encoding cytoplasmic actin from the leafhopper Nephotettix cincticeps, we successfully developed the first efficient transient expression vector for cultured leafhopper cells, which can also be used to express exogenous proteins in other insect culture cell lines, including those derived from Recilia dorsalis leafhopper and Spodoptera frugiperda (Sf9). Furthermore, insertion of the Hr5 viral enhancer element and knockdown of the endogenous Dicer2 gene notably improved the vector’s expression efficiency in leafhopper cells. Using the optimized vector, we have for the first time traced the cellular localization of the proteins encoded by rice yellow stunt virus (RYSV) in cells of its insect vector and demonstrated that P6 protein is a component of the viroplasm.

Keywords: actin promoter, Transient expression vector, Insect cultured cell, RYSV P6, viroplasm

Received: 29 Sep 2018; Accepted: 23 Nov 2018.

Edited by:

Heiko Ziebell, Julius Kühn-Institut, Germany

Reviewed by:

Nicolas Bejerman, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina
Ralf G. Dietzgen, The University of Queensland, Australia  

Copyright: © 2018 Wei, Zhang, Xie, Wang, Wang, Zeng, Zhao and Chen. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: Dr. Taiyun Wei, Fujian Agriculture and Forestry University, State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, Institute of Plant Virology, Fuzhou, China, weitaiyun2000@yahoo.com