Characterization and reverse genetic establishment of cattle derived Akabane virus in China

Dongjie Chen,Di Wang,Junhua Deng,Xiangmei Lin,Fang Wei,Shaoqiang Wu,Yufang Kong

doi:10.1186/s12917-021-03054-x

Dongjie Chen, Di Wang + Show 5 more

Open Access

https://doi.org/10.1186/s12917-021-03054-x

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Abstract

BackgroundAkabane virus (AKAV) is an important insect-borne virus which is widely distributed throughout the world except the Europe and is considered as a great threat to herbivore health.ResultsAn AKAV strain defined as TJ2016 was firstly isolated from the bovine sera in China in 2016. Sequence analysis of the S and M segments suggested that the isolated AKAV strain was closely related to the AKAV strains JaGAr39 and JaLAB39, which belonged to AKAV genogroup II. To further study the pathogenic mechanism of AKAV, the full-length cDNA clone of TJ2016 S, M, and L segment was constructed separately into the TVT7R plasmid at the downsteam of T7 promoter and named as TVT7R-S, TVT7R-M, and TVT7R-L, respectively. The above three plasmids were further transfected into the BSR-T7/5 cells simultaneously with a ratio of 1:1:1 to produce the rescued virus AKAV. Compared with the parental wild type AKAV (wtAKAV), the rescued virus (rAKAV) was proved to be with similar cytopathic effects (CPE), plaque sizes and growth kinetics in BHK-21 cells.ConclusionWe successfully isolated a AKAV strain TJ2016 from the sera of cattle and established a reverse genetic platform for AKAV genome manipulation. The established reverse genetic system is also a powerful tool for further research on AKAV pathogenesis and even vaccine studies.

Highlights

Akabane virus (AKAV) is an important insect-borne virus which is widely distributed throughout the world except the Europe and is considered as a great threat to herbivore health
We described the first cattle-derived AKAV isolate TJ2016 in China and investigated the genetic diversities of the current isolate with the previously reported AKAV isolates
Phylogenetic analysis of the S and M segments For the phylogenetic analysis of the isolated AKAV TJ2016, nine pairs of primers were designed based on the published genome sequence of AKAV strain DHL10M110 (KY284023.1, KY284022.1, KY284021.1) and were used to amplify the whole genome of AKAV isolate (Table 1, Number 1 to 18)

Summary

Introduction

Akabane virus (AKAV) is an important insect-borne virus which is widely distributed throughout the world except the Europe and is considered as a great threat to herbivore health. AKA is widely distributed throughout the world except the Europe and has caused serious economic losses to animal husbandry, which poses a great threat to the cattle and sheep breeding industry [3,4,5]. In China, a serological survey of AKAVs in Xinjiang province indicated that the neutralizing antibody positive rate of AKAV in cattle and sheep was 20.32 and 18.15%, respectively [9]. A largescale serological survey of AKAV from 2006 to 2015 in 24 provinces of China indicated that the overall seroprevalence rate for AKAV antibodies was 21.3% in cattle and 12.0% in sheep or goats [10]. We established the reverse genetic system based on cattle-derived AKAV isolate, which was a useful tool for studying the basic mechanism of AKAV pathogenesis

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Results

Conclusion