Improving initial infectivity of the Turnip mosaic virus (TuMV) infectious clone by an mini binary vector via agro-infiltration

Botanical Studies
Yen-Yu LinShih-Shun Lin

Abstract

The in vivo infectious clone of Turnip mosaic virus (TuMV), p35S-TuMV, was used on plant pathology research for many years. To activate p35S-TuMV, the plasmid was mechanically introduced to the local lesion host Chenopodium quinoa. However, low infectivity occurred when the TuMV from C. quinoa was transferred to the systemic host Nicotiana benthamiana. To increase the efficiency of initial infectivity on N. benthamiana, the expression of the TuMV infectious clone by a binary vector that directly activates viral RNA through agro-infiltration is considered to be a good alternative. The size of the binary vector by agro-infiltration is usually large and its backbone has numerous restriction sites that increase difficulty for construction. In this study, we attempted to construct a mini binary vector (pBD003) with less restriction sites. The full-length cDNA of TuMV genome, with or without green fluorescence protein, was inserted in pBD003 to generate pBD-TuMV constructs, which were then individually introduced to N. benthamiana plants by agro-infiltration. Symptom development and ELISA positivity with TuMV antiserum indicated that the pBD-TuMV constructs are infectious. Moreover, the initial infectivity of a mild strain TuMV-GK, w...Continue Reading

References

May 1, 1989·Virology·L A HeatonT J Morris
Feb 1, 1994·Virology·J C Boyer, A L Haenni
Nov 15, 2000·Proceedings of the National Academy of Sciences of the United States of America·C LlaveJ C Carrington
May 25, 2002·The Plant Journal : for Cell and Molecular Biology·Yule LiuS P Dinesh-Kumar
Nov 17, 2007·Cell Host & Microbe·Takeshi KobayashiTerence S Dermody

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Methods Mentioned

BETA
in vitro transcription
enzyme-linked immunosorbent assay
PCR
ELISA

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