Differential gene expression in response to Papaya ringspot virus infection in Cucumis metuliferus using cDNA-amplified fragment length polymorphism analysis.

Yu-Tsung Lin,Fuh-Jyh Jan,Chia-Wei Lin,Chien-Hung Chung,Jo-Chu Chen,Hsin-Mei Ku,Shy-Dong Yeh

doi:10.1371/journal.pone.0068749

Yu-Tsung Lin, Fuh-Jyh Jan + Show 5 more

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https://doi.org/10.1371/journal.pone.0068749

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Journal: PloS one	Publication Date: Jul 9, 2013
Citations: 9	License type: CC BY 4.0

Affiliation: National Chung Hsing University

Abstract

A better understanding of virus resistance mechanisms can offer more effective strategies to control virus diseases. Papaya ringspot virus (PRSV), Potyviridae, causes severe economical losses in papaya and cucurbit production worldwide. However, no resistance gene against PRSV has been identified to date. This study aimed to identify candidate PRSV resistance genes using cDNA-AFLP analysis and offered an open architecture and transcriptomic method to study those transcripts differentially expressed after virus inoculation. The whole genome expression profile of Cucumis metuliferus inoculated with PRSV was generated using cDNA-amplified fragment length polymorphism (cDNA-AFLP) method. Transcript derived fragments (TDFs) identified from the resistant line PI 292190 may represent genes involved in the mechanism of PRSV resistance. C. metuliferus susceptible Acc. 2459 and resistant PI 292190 lines were inoculated with PRSV and subsequently total RNA was isolated for cDNA-AFLP analysis. More than 400 TDFs were expressed specifically in resistant line PI 292190. A total of 116 TDFs were cloned and their expression patterns and putative functions in the PRSV-resistance mechanism were further characterized. Subsequently, 28 out of 116 candidates which showed two-fold higher expression levels in resistant PI 292190 than those in susceptible Acc. 2459 after virus inoculation were selected from the reverse northern blot and bioinformatic analysis. Furthermore, the time point expression profiles of these candidates by northern blot analysis suggested that they might play roles in resistance against PRSV and could potentially provide valuable information for controlling PRSV disease in the future.

Highlights

Viral resistance is always a top priority to plant breeders
This provided evidence that no obvious response was seen in PI 292190 plants after virus infection, the plants has been exposed to a potent inoculum and the changed gene expression of PRSVinoculated resistant line PI 292190 was caused by the Papaya ringspot virus (PRSV) infection
Comparing gene expression patterns through cDNAAFLP analysis, more than 400 Transcript derived fragments (TDFs) were found in the resistant line PI 292190-PRSV

Summary

Introduction

Viral resistance is always a top priority to plant breeders. Numerous innate defense systems against pathogens have evolved in plants. The hypersensitive response (HR) is induced in the infected region of a leaf and restricts the spread of pathogens [1]. A substantial number of secondary metabolites such as salicylic acid (SA) or reactive oxygen species (ROSs) e.g., superoxide radical (O22), hydrogen peroxide (H2O2) and hydroxyl radical (OH) are able to be produced to trigger the whole plant systemic acquired resistance (SAR). These signals can activate specific or nonspecific defense responses which contribute to a plant’s ability to protect itself against future pathogen infection [2]

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