Multiple Coat Protein Mutations Abolish Recognition of Pepino mosaic potexvirus (PepMV) by the Potato Rx Resistance Gene in Transgenic Tomatoes

Despite the fact that Pepino mosaic virus (PepMV) and Potato virus X (PVX) share less than 40% identity in their coat proteins (CP), the known PVX elicitor of Rx, transgenic tomato (cv. Microtom) plants expressing a functional potato Rx resistance gene showed resistance toward PepMV. However, in a low percentage of plants, PepMV accumulation was observed and back inoculation experiments demonstrated that these plants contained resistance-breaking PepMV variants. Sequencing of the CP gene of these variants showed the accumulation of mutations in the amino acid 41 to 125 region the CP, whereas no mutations were observed in the nonevolved isolates. Agroinfiltration-mediated transient expression of the mutant CP demonstrated that they had a greatly attenuated or abolished ability to induce a hypersensitive reaction in Rx-expressing Nicotiana benthamiana leaves. The transient expression of truncated forms of the PepMV CP allowed the identification of a minimal elicitor domain (amino acids 30 to 136). These results demonstrate that the Rx-based sensing system is able to recognize the PepMV CP but, contrary to the situation with PVX, for which only two closely spaced resistance-breaking mutations are known, many mutations over a significant stretch of the PepMV CP allow escape from recognition by Rx.

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Efficient transient expression of human GM-CSF protein in Nicotiana benthamiana using potato virus X vector

Applied Microbiology and Biotechnology ◽

10.1007/s00253-005-0305-2 ◽

2006 ◽

Vol 72 (4) ◽

pp. 756-762 ◽

Cited By ~ 12

Author(s):

Fengyong Zhou ◽

Ming-Li Wang ◽

Henrik H. Albert ◽

Paul H. Moore ◽

Yun J. Zhu

Keyword(s):

Transient Expression ◽

Potato Virus ◽

Nicotiana Benthamiana ◽

Potato Virus X ◽

Gm Csf

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Long-Term Potato Virus X (PVX)-Based Transient Expression of Recombinant GFP Protein in Nicotiana benthamiana Culture In Vitro

Plants ◽

10.3390/plants10102187 ◽

2021 ◽

Vol 10 (10) ◽

pp. 2187

Author(s):

Yana Sindarovska ◽

Mykola Kuchuk

Keyword(s):

Transient Expression ◽

Nicotiana Benthamiana ◽

Molecular Farming ◽

Transient Gene Expression ◽

Leaf Explants ◽

Potato Virus X ◽

Culture In Vitro ◽

High Efficient

Plant molecular farming has a great potential to produce valuable proteins. Transient expression technology provides high yields of recombinant proteins in greenhouse-grown plants, but every plant must be artificially agroinfiltrated, and open greenhouse systems are less controlled. Here, we propose to propagate agrobacteria-free plants with high-efficient long-term self-replicated transient gene expression in a well-controlled closed in vitro system. Nicotiana benthamiana plant tissue culture in vitro, with transient expression of recombinant GFP, was obtained through shoot induction from leaf explants infected by a PVX-based vector. The transient expression occurs in new tissues and regenerants due to the natural systemic distribution of viral RNA carrying the target gene. Gene silencing was delayed in plants grown in vitro, and GFP was detected in plants for five to six months. Agrobacteria-free, GFP-expressing plants can be micropropagated in vitro (avoiding an agroinfiltration step), “rejuvenated” through regeneration (maintaining culture for years), or transferred in soil. The mean GFP in the regenerants was 18% of the total soluble proteins (TSP) (0.52 mg/g of fresh leaf weight (FW). The highest value reached 47% TSP (2 mg/g FW). This study proposes a new method for recombinant protein production combining the advantages of transient expression technology and closed cultural systems.

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Transient expression of Arabidopsis thaliana ascorbate peroxidase 3 in Nicotiana benthamiana plants infected with recombinant potato virus X

Plant Cell Reports ◽

10.1007/s00299-002-0570-9 ◽

2003 ◽

Vol 21 (7) ◽

pp. 699-704 ◽

Cited By ~ 5

Author(s):

C. Escobar ◽

L. E. Hernández ◽

A. Jiménez ◽

G. Creissen ◽

M. T. Ruiz ◽

...

Keyword(s):

Arabidopsis Thaliana ◽

Transient Expression ◽

Ascorbate Peroxidase ◽

Potato Virus ◽

Nicotiana Benthamiana ◽

Potato Virus X

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Harnessing a Transient Gene Expression System in Nicotiana benthamiana to Explore Plant Agrochemical Transporters

Plants ◽

10.3390/plants10030524 ◽

2021 ◽

Vol 10 (3) ◽

pp. 524

Author(s):

Bingqi Wu ◽

Zhiting Chen ◽

Xiaohui Xu ◽

Ronghua Chen ◽

Siwei Wang ◽

...

Keyword(s):

Gene Expression ◽

Transient Expression ◽

Nicotiana Benthamiana ◽

Heterologous Gene Expression ◽

Expression System ◽

Functional Characterization ◽

Transient Gene Expression ◽

High Mobility ◽

Gene Expression System

Functional characterization of plant agrichemical transporters provided an opportunity to discover molecules that have a high mobility in plants and have the potential to increase the amount of pesticides reaching damage sites. Agrobacterium-mediated transient expression in tobacco is simple and fast, and its protein expression efficiency is high; this system is generally used to mediate heterologous gene expression. In this article, transient expression of tobacco nicotine uptake permease (NtNUP1) and rice polyamine uptake transporter 1 (OsPUT1) in Nicotiana benthamiana was performed to investigate whether this system is useful as a platform for studying the interactions between plant transporters and pesticides. The results showed that NtNUP1 increases nicotine uptake in N. benthamiana foliar discs and protoplasts, indicating that this transient gene expression system is feasible for studying gene function. Moreover, yeast expression of OsPUT1 apparently increases methomyl uptake. Overall, this method of constructing a transient gene expression system is useful for improving the efficiency of analyzing the functions of plant heterologous transporter-encoding genes and revealed that this system can be further used to study the functions of transporters and pesticides, especially their interactions.

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Comparative transient expression analyses on two conserved effectors of Colletotrichum orbiculare reveal their distinct cell death‐inducing activities between Nicotiana benthamiana and melon

Molecular Plant Pathology ◽

10.1111/mpp.13078 ◽

2021 ◽

Author(s):

Jinlian Chen ◽

Yoshihiro Inoue ◽

Naoyoshi Kumakura ◽

Kazuyuki Mise ◽

Ken Shirasu ◽

...

Keyword(s):

Cell Death ◽

Transient Expression ◽

Nicotiana Benthamiana ◽

Colletotrichum Orbiculare ◽

Distinct Cell

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First Report of a Resistance-Breaking Strain of Tomato spotted wilt virus Infecting Tomatoes With the Sw-5 Tospovirus-Resistance Gene in California

Plant Disease ◽

10.1094/pdis-09-16-1371-pdn ◽

2017 ◽

Vol 101 (4) ◽

pp. 637-637 ◽

Cited By ~ 12

Author(s):

O. Batuman ◽

T. A. Turini ◽

P. V. Oliveira ◽

M. R. Rojas ◽

M. Macedo ◽

...

Keyword(s):

Resistance Gene ◽

Tomato Spotted Wilt Virus ◽

Breaking Strain ◽

First Report ◽

Tomato Spotted Wilt ◽

Resistance Breaking ◽

Wilt Virus

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The coat protein of tobamovirus acts as elicitor of both L 2 and L 4 gene-mediated resistance in Capsicum

Journal of General Virology ◽

10.1099/vir.0.80017-0 ◽

2004 ◽

Vol 85 (7) ◽

pp. 2077-2085 ◽

Cited By ~ 36

Author(s):

P. Gilardi ◽

I. García-Luque ◽

M. T. Serra

Keyword(s):

Hypersensitive Response ◽

Host Response ◽

Expression System ◽

Potato Virus X ◽

Mottle Virus ◽

Coat Proteins ◽

Pepper Mild Mottle Virus ◽

Defence Genes ◽

Glucuronidase Reporter ◽

Capsicum Chacoense

In Capsicum, the resistance conferred by the L 2 gene is effective against all of the pepper-infecting tobamoviruses except Pepper mild mottle virus (PMMoV), whereas that conferred by the L 4 gene is effective against them all. These resistances are expressed by a hypersensitive response, manifested through the formation of necrotic local lesions (NLLs) at the primary site of infection. The Capsicum L 2 gene confers resistance to Paprika mild mottle virus (PaMMV), while the L 4 gene is effective against both PaMMV and PMMoV. The PaMMV and PMMoV coat proteins (CPs) were expressed in Capsicum frutescens (L 2 L 2) and Capsicum chacoense (L 4 L 4) plants using the heterologous Potato virus X (PVX)-based expression system. In C. frutescens (L 2 L 2) plants, the chimeric PVX virus containing the PaMMV CP was localized in the inoculated leaves and produced NLLs, whereas the chimeric PVX containing the PMMoV CP infected the plants systemically. Thus, the data indicated that the PaMMV CP is the only tobamovirus factor required for the induction of the host response mediated by the Capsicum L 2 resistance gene. In C. chacoense (L 4 L 4) plants, both chimeric viruses were localized to the inoculated leaves and produced NLLs, indicating that either PaMMV or PMMoV CPs are required to elicit the L 4 gene-mediated host response. In addition, transient expression of PaMMV CP into C. frutescens (L 2 L 2) leaves and PMMoV CP into C. chacoense (L 4 L 4) leaves by biolistic co-bombardment with a β-glucuronidase reporter gene led to the induction of cell death and the expression of host defence genes in both hosts. Thus, the tobamovirus CP is the elicitor of the Capsicum L 2 and L 4 gene-mediated hypersensitive response.

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