4.6 Article

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

Journal

VIRUSES-BASEL
Volume 13, Issue 6, Pages -

Publisher

MDPI
DOI: 10.3390/v13061050

Keywords

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

Categories

Funding

  1. National Natural Science Foundation of China [32072506, 31770168]
  2. China Agriculture Research System [CARS-24-C-01]
  3. Jiangsu Agriculture Science and Technology Innovation Fund [CX (19)3108]
  4. Priority Academic Program Development of Jiangsu Higher Education Institutions (2020 PAPD)

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The full-length genome of a Chinese isolate of Tomato mottle mosaic virus (ToMMV) was successfully isolated and cloned, leading to the development of an infectious cDNA clone that was able to infect both model and natural host plants. The infectious clone induced high infection rates in tomato plants, providing a valuable tool for further studies on the virus's gene function, pathogenicity, and virus-host interaction.
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.

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