Synthesis and Characterization of a Full-Length Infectious cDNA Clone of Tomato Mottle Mosaic Virus

Viruses. 2021 Jun 1;13(6):1050. doi: 10.3390/v13061050.

Abstract

Tomato mottle mosaic virus (ToMMV) is a noteworthy virus which belongs to the Virgaviridae family and causes serious economic losses in tomato. Here, we isolated and cloned the full-length genome of a ToMMV Chinese isolate (ToMMV-LN) from a naturally infected tomato (Solanum lycopersicum L.). Sequence analysis showed that ToMMV-LN contains 6399 nucleotides (nts) and is most closely related to a ToMMV Mexican isolate with a sequence identity of 99.48%. Next, an infectious cDNA clone of ToMMV was constructed by a homologous recombination approach. Both the model host N. benthamiana and the natural hosts tomato and pepper developed severe symptoms upon agroinfiltration with pToMMV, which had a strong infectivity. Electron micrographs indicated that a large number of rigid rod-shaped ToMMV virions were observed from the agroinfiltrated N. benthamiana leaves. Finally, our results also confirmed that tomato plants inoculated with pToMMV led to a high infection rate of 100% in 4-5 weeks post-infiltration (wpi), while pepper plants inoculated with pToMMV led to an infection rate of 40-47% in 4-5 wpi. This is the first report of the development of a full-length infectious cDNA clone of ToMMV. We believe that this infectious clone will enable further studies of ToMMV genes function, pathogenicity and virus-host interaction.

Keywords: Agrobacterium-mediated inoculation; full-length genome sequence; infectious cDNA clone; infectivity; tomato mottle mosaic virus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular*
  • DNA, Complementary*
  • Disease Susceptibility
  • Genome, Viral*
  • Genomics / methods
  • Phenotype
  • Phylogeny
  • Plant Diseases / virology
  • Sequence Analysis, DNA
  • Tobamovirus / genetics*
  • Tobamovirus / isolation & purification

Substances

  • DNA, Complementary