High-throughput sequence analysis of small RNAs in grapevine (Vitis vinifera L.) affected by grapevine leafroll disease

Mol Plant Pathol. 2012 Dec;13(9):1060-76. doi: 10.1111/j.1364-3703.2012.00815.x. Epub 2012 Jul 24.

Abstract

Grapevine leafroll disease (GLRD) is one of the most economically important virus diseases of grapevine (Vitis spp.) worldwide. In this study, we used high-throughput sequencing of cDNA libraries made from small RNAs (sRNAs) to compare profiles of sRNA populations recovered from own-rooted Merlot grapevines with and without GLRD symptoms. The data revealed the presence of sRNAs specific to Grapevine leafroll-associated virus 3, Hop stunt viroid (HpSVd), Grapevine yellow speckle viroid 1 (GYSVd-1) and Grapevine yellow speckle viroid 2 (GYSVd-2) in symptomatic grapevines and sRNAs specific only to HpSVd, GYSVd-1 and GYSVd-2 in nonsymptomatic grapevines. In addition to 135 previously identified conserved microRNAs in grapevine (Vvi-miRs), we identified 10 novel and several candidate Vvi-miRs in both symptomatic and nonsymptomatic grapevine leaves based on the cloning of miRNA star sequences. Quantitative real-time reverse transcriptase-polymerase chain reaction (RT-PCR) of selected conserved Vvi-miRs indicated that individual members of an miRNA family are differentially expressed in symptomatic and nonsymptomatic leaves. The high-resolution mapping of sRNAs specific to an ampelovirus and three viroids in mixed infections, the identification of novel Vvi-miRs and the modulation of certain conserved Vvi-miRs offers resources for the further elucidation of compatible host-pathogen interactions and for the provision of ecologically relevant information to better understand host-pathogen-environment interactions in a perennial fruit crop.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Cloning, Molecular
  • Closteroviridae / genetics*
  • Closteroviridae / isolation & purification
  • Gene Expression Profiling
  • Gene Expression Regulation, Plant
  • Gene Library
  • Genome, Viral / genetics
  • High-Throughput Nucleotide Sequencing / methods*
  • MicroRNAs / chemistry
  • MicroRNAs / genetics
  • MicroRNAs / metabolism
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • Plant Diseases / genetics
  • Plant Diseases / virology*
  • Plant Leaves / genetics
  • Plant Leaves / virology
  • RNA, Plant / genetics*
  • RNA, Viral / classification
  • RNA, Viral / genetics*
  • Reproducibility of Results
  • Viroids / genetics
  • Viroids / isolation & purification
  • Vitis / genetics*
  • Vitis / virology*

Substances

  • MicroRNAs
  • RNA, Plant
  • RNA, Viral

Associated data

  • GENBANK/JQ989190
  • GENBANK/JQ989191
  • GENBANK/JQ989192
  • GENBANK/JQ989193
  • GENBANK/JQ989194
  • GENBANK/JQ989195
  • GENBANK/JQ989196
  • GENBANK/JQ989197
  • GENBANK/JQ989198
  • GENBANK/JQ989199
  • GENBANK/JQ989200
  • GENBANK/JQ989201
  • GENBANK/JQ989202
  • GENBANK/JQ989203
  • GENBANK/JQ989204
  • GENBANK/JQ989205
  • GENBANK/JQ989206
  • GENBANK/JQ989207
  • GENBANK/JQ989208
  • GENBANK/JQ989209
  • GENBANK/JQ989210
  • GENBANK/JQ989211
  • GENBANK/JQ989212
  • GENBANK/JQ989213
  • GENBANK/JQ989214
  • GENBANK/JQ989215
  • GENBANK/JQ989216
  • GENBANK/JQ989217
  • GENBANK/JQ989218
  • GENBANK/JQ989219
  • GENBANK/JQ989220
  • GENBANK/JQ989221
  • GENBANK/JQ989222