# Investigating Apple Rubbery Wood Virus 2: HTS-Based Detection in Hungary and Involvement of sRNA-Based Regulation Processes During Its Infection

## TL;DR

Researchers studied apple rubbery wood virus 2 in Hungary, finding it in multiple tree types without symptoms and analyzing its impact on lignin production.

## Contribution

The study provides new insights into ARWV2 detection and its sRNA-based regulation during infection.

## Key findings

- ARWV2 was detected in 17 trees, including apples, pears, quince, and a rootstock, without showing symptoms.
- sRNA analysis revealed differential expression patterns of vsiRNAs, sRNAs, and miRNAs targeting lignin biosynthesis.
- Gene expression changes in lignification-related genes were not directly linked to virus presence, explaining latent infections.

## Abstract

Pomme fruits are propagated vegetatively, thereby facilitating frequent viral transmission. The causative agent of apple rubbery wood disease, apple rubbery wood virus 2 (ARWV2), can infect apple and pear. The branches of ARWV2-infected, symptomatic trees are flexible due to the decreased lignification of the xylem. In this research, we reanalysed our small RNA (sRNA) HTS datasets to survey the presence of ARWV2 in Hungary. Validation of HTS using RT-PCR revealed infection in several cultivars. The following RT-PCR-based survey revealed the infection of 17 trees, including not only apple, but also pears, one quince, and a rootstock, without showing any rubbery wood symptoms. Analysis of the sRNA datasets allowed us to profile the sRNA pattern of ARWV2-infected and non-infected trees, and characterise the differential expression pattern of vsiRNAs, sRNAs, and miRNAs targeting the lignin biosynthetic pathway. The results confirmed that the gene-expression changes in the genes that regulate lignification cannot be directly correlated with the presence of the virus, which can explain its frequent latent presence. The variable titre and sequence of the virus, and mixed-infection status of the trees, make its reliable diagnostics challenging, which could be achieved as a result of further research.