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

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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Construction and biological characterization of an infectious full-length cDNA clone of a Chinese isolate of Wheat yellow mosaic virus

Virology ◽

10.1016/j.virol.2021.01.018 ◽

2021 ◽

Vol 556 ◽

pp. 101-109

Author(s):

Fan Zhang ◽

Shuang Liu ◽

Tianye Zhang ◽

Zhuangxin Ye ◽

Xiaolei Han ◽

...

Keyword(s):

Mosaic Virus ◽

Cdna Clone ◽

Full Length ◽

Yellow Mosaic Virus ◽

Chinese Isolate ◽

Biological Characterization ◽

Wheat Yellow Mosaic Virus ◽

Full Length Cdna

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Construction of an infectious full-length cDNA clone of potato aucuba mosaic virus

Archives of Virology ◽

10.1007/s00705-021-05018-w ◽

2021 ◽

Vol 166 (5) ◽

pp. 1427-1431

Author(s):

Buyang Chen ◽

Qi Lin ◽

Yueyan Yin ◽

Liangliang Jiang ◽

Fang Wang ◽

...

Keyword(s):

Mosaic Virus ◽

Cdna Clone ◽

Full Length ◽

Full Length Cdna ◽

Aucuba Mosaic

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Infectious Transcripts from Cloned Genome-Length cDNA of Porcine Reproductive and Respiratory Syndrome Virus

Journal of Virology ◽

10.1128/jvi.72.1.380-387.1998 ◽

1998 ◽

Vol 72 (1) ◽

pp. 380-387 ◽

Cited By ~ 82

Author(s):

J. J. M. Meulenberg ◽

J. N. A. Bos-de Ruijter ◽

R. van de Graaf ◽

G. Wensvoort ◽

R. J. M. Moormann

Keyword(s):

Cdna Clone ◽

De Novo ◽

Infectious Clone ◽

Full Length ◽

Respiratory Syndrome Virus ◽

Cdna Clones ◽

Genome Length ◽

Full Length Cdna ◽

Lung Macrophages ◽

Growth Properties

ABSTRACT The 5′-terminal end of the genomic RNA of the Lelystad virus isolate (LV) of porcine reproductive and respiratory syndrome virus was determined. To construct full-length cDNA clones, the 5′-terminal sequence was ligated to cDNA clones covering the complete genome of LV. When RNA that was transcribed in vitro from these full-length cDNA clones was transfected into BHK-21 cells, infectious LV was produced and secreted. The virus was rescued by passage to porcine alveolar lung macrophages or CL2621 cells. When infectious transcripts were transfected to porcine alveolar lung macrophages or CL2621 cells, no infectious virus was produced due to the poor transfection efficiency of these cells. The growth properties of the viruses produced by BHK-21 cells transfected with infectious transcripts of LV cDNA resembled the growth properties of the parental virus from which the cDNA was derived. Two nucleotide changes leading to a unique PacI restriction site directly downstream of the ORF7 gene were introduced in the genome-length cDNA clone. The virus recovered from this mutated cDNA clone retained the PacI site, which confirmed the de novo generation of infectious LV from cloned cDNA. These results indicate that the infectious clone of LV enables us to mutagenize the viral genome at specific sites and that it will therefore be useful for detailed molecular characterization of the virus, as well as for the development of a safe and effective live vaccine for use in pigs.

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