Isolation, expression and functional analysis of a putative RNA-dependent RNA polymerase gene from maize (Zea mays L.)

Mol Biol Rep. 2010 Feb;37(2):865-74. doi: 10.1007/s11033-009-9692-2. Epub 2009 Aug 15.

Abstract

RNA-dependent RNA polymerases (RdRPs) in plants have been reported to be involved in post-transcriptional gene silencing (PTGS) and antiviral defense. In this report, an RdRP gene from maize (ZmRdRP1) was obtained by rapid amplification of cDNA ends (RACE) and RT-PCR. The mRNA of ZmRdRP1 was composed of 3785 nucleotides, including a 167 nt 5' untranslated region (UTR), a 291 nt 3'UTR and a 3327 nt open reading frame (ORF), which encodes a putative protein of 1108 amino acids with an estimated molecular mass of 126.9 kDa and a predicated isoelectric point (pI) of 8.37. Real-time quantitative RT-PCR analysis showed that ZmRdRP1 was elicited by salicylic acid (SA) treatment, methyl jasmonate (MeJA) treatment and sugarcane mosaic virus (SCMV) infection. We silenced ZmRdRP1 by constitutively expressing an inverted-repeat fragment of ZmRdRP1 (ir-RdRP1) in transgenic maize plants. Further studies revealed that the ir-RdRP1 transgenic plants were more susceptible to SCMV infection than wild type plants. Virus-infected transgenic maize plants developed more serious disease symptoms and accumulated more virus than wild type plants. These findings suggested that ZmRdRP1 was involved in antiviral defense in maize.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Cloning, Molecular
  • Gene Expression Profiling
  • Gene Expression Regulation, Plant
  • Gene Targeting
  • Immunity, Innate / genetics
  • Molecular Sequence Data
  • Mosaic Viruses / physiology
  • Phylogeny
  • Plant Diseases / genetics
  • Plant Diseases / immunology
  • Plant Diseases / virology
  • Plants, Genetically Modified
  • RNA Interference
  • RNA-Dependent RNA Polymerase / genetics*
  • RNA-Dependent RNA Polymerase / isolation & purification
  • RNA-Dependent RNA Polymerase / physiology*
  • Sequence Homology, Amino Acid
  • Viral Load / genetics
  • Zea mays / genetics*
  • Zea mays / immunology
  • Zea mays / physiology
  • Zea mays / virology

Substances

  • RNA-Dependent RNA Polymerase