Molecular characterization and pathogenicity analysis of prunus necrotic ringspot virus isolates from China rose ( Rosa

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ORIGINAL ARTICLE

Molecular characterization and pathogenicity analysis of prunus necrotic ringspot virus isolates from China rose (Rosa chinensis Jacq.) Fei Xing1,4 · Dehang Gao2 · Huan Liu2 · Hongqing Wang2 · Nuredin Habili3 · Shifang Li1,4  Received: 6 March 2020 / Accepted: 15 June 2020 © Springer-Verlag GmbH Austria, part of Springer Nature 2020

Abstract Prunus necrotic ringspot virus (PNRSV) is a viral pathogen with worldwide distribution, infecting many commercial fruit trees and ornamental plants. So far, the correlation between PNRSV infection and China rose mosaic disease has not been studied. Rose mosaic disease is characterized by severe symptoms, including mosaic, line pattern, and ringspot. Six viruses that were potentially associated with mosaic disease, including PNRSV, were tested in China roses. Only PNRSV was detected in China roses showing mosaic disease, and asymptomatic samples tested negative for this virus. This result was confirmed by small RNA sequencing, but rose leaf rosette-associated virus and rose spring dwarf-associated virus were also identified in both samples with mosaic disease and asymptomatic samples. This implied that PNRSV might be associated with China rose mosaic disease. Full genome sequences of two PNRSV isolates were determined, and the RNA1, 2 and 3 segments were found to be 3,332, 2,594 and 1,951 nucleotides (nt) in length, respectively. The three RNA segments shared 88.7–89.1% nt sequence identity in the 3′UTR, while RNA2 and RNA3 shared 98.2–99.4% identity. The higher variability in RNA1 suggests that it might have been under greater selection pressure. Phylogenetic analysis showed that the two PNRSV isolates clustered in group PV-32. Full-length infectious cDNA clones of PNRSV from China rose were constructed and used to agroinfiltrate cucumber seedlings. The inoculated cucumber leaves showed yellowing, chlorotic spots, necrosis, dwarfing, and decline at 23 to 39 days post-inoculation, demonstrating the virulence of the PNRSV isolate from China rose. These data lay a foundation for determining the molecular mechanism of rose mosaic disease caused by PNRSV.

Introduction

Handling editor: Jesús Navas-Castillo. Electronic supplementary material  The online version of this article (https​://doi.org/10.1007/s0070​5-020-04739​-8) contains supplementary material, which is available to authorized users. * Shifang Li [email protected] 1



State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China

2



Department of Fruit Science, College of Horticulture, China Agricultural University, Beijing 100193, China

3

The Australian Wine Research Institute Waite Precinct, University of Adelaide, Adelaide 5000, Australia

4

Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, China



Prunus necrotic ringspot virus (PNRSV) is a viral pathogen with worldwide distribution that infects many commercial fruit trees, such as