Expression of Single-Chain Variable Fragments Against Structural and Non-Structural Proteins of Plum Pox Virus in Transgenic Nicotiana Benthamiana Plants to Interfere With Viral Infection

Gil, Maite; Esteban, Olga; Martínez, Mariela; Gorris, M. T.; Peña, Leandro; Cambra, M.; Garcia, J

doi:10.17660/actahortic.2004.657.54

Cited by 4 publications

(3 citation statements)

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“…Both fusions were localized exclusively when transiently expressed in N. benthamiana leaves by agroinfiltration and particle bombardment. The targeting of scFvs to the nucleus and their ability to function therein has been demonstrated [36], and a nuclear-targeted scFv against plum pox virus coat protein has been shown to reduce virus disease symptoms [37], but to our knowledge, we provide here the first report of the nuclear targeting of a scFv fusion protein.…”

Section: Discussionmentioning

confidence: 90%

In vivo expression and binding activity of scFv-RWAV, which recognizes the coat protein of tomato leaf curl New Delhi virus (family Geminiviridae)

Zakri

Ziegler²,

Commandeur

et al. 2012

Arch Virol

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Section: Discussionmentioning

confidence: 90%

In vivo expression and binding activity of scFv-RWAV, which recognizes the coat protein of tomato leaf curl New Delhi virus (family Geminiviridae)

Zakri

Ziegler²,

Commandeur

et al. 2012

Arch Virol

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“…In particular, this strategy was conceived to interfere with PPV NIb or CP functions by expressing in plants single-chain variable fragment (scFv) antibodies specific to these proteins. 124 At 30 dpi in the best performing scFv/NIb and scFv/CP lines plants positive for PPV-D/NAT were only 17% and 21%, respectively. Although this line of research was not further developed, this methodological approach was shown feasible to confer resistance to multiple viruses; 125 hence, scFvs direct against conserved domain of non-structural viral proteins could be another tool for efficiently interfering with viral infection.…”

Section: Transgenic Strategies To Control Ppv Infectionmentioning

confidence: 93%

Genetically engineered resistance to Plum pox virus infection in herbaceous and stone fruit hosts

Ilardi

Nicola-Negri²

2011

GM Crops

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Plum pox virus (PPV), a Potyvirus, is the causal agent of sharka, the most detrimental viral disease affecting stone fruit trees. This review focuses on research carried out to obtain PPV- resistant transgenic plants and on how biotechnological strategies evolved in light of the scientific advances made during the last several years. Successful RNA silencing strategies that confer high level of resistance to strains of PPV have been developed and tested under laboratory and greenhouse conditions. Moreover, field tests showed that transgene-mediated RNA silencing was effective in protecting plum plants against aphid-mediated PPV infection. The new emerging biotechnological approaches for conferring PPV resistance are discussed.

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“…(2003) developed a monoclonal antibody (2A) against the RNA replicase NIb of PPV. Different recombinant versions in the form of single‐chain variable fragments (scFv) against structural and nonstructural proteins of PPV (scFv4C, specific for PPV‐D NAT isolates; scFv2A, specific for NIb protein) were generated (Esteban et al ., 2003; Gil et al ., 2004). Different cytosolic, nuclear and cytosolic facing endoplasmic reticulum membrane targeted versions were expressed in Nicotiana benthamiana .…”

Section: Introductionmentioning

confidence: 99%

Detection and characterization of Plum pox virus: serological methods

Cambra¹,

Boscia²,

Myrta

et al. 2006

EPPO Bulletin

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Tremendous progress has been made in the research and development of Plum pox virus (PPV) serological reagents and methods in recent years. Two facts have revolutionised the serological detection and characterization of the virus: the development of the ELISA method in 1977, and the later emergence of specific monoclonal antibody technology. The availability of commercial kits has popularised PPV diagnosis, now making diagnosis possible at large scale for quarantine purposes, eradication programmes and control of the disease in nurseries. The use of the universal monoclonal antibody 5B-IVIA, used in DASI-ELISA, is the most accurate system for routine PPV detection. Likewise, the use of typing monoclonal antibodies gives exact characterization of the main PPV types described: 4DG5 for PPV-D, AL for PPV-M, EA24 for PPV-EA, and TUV and AC for PPV-C. There is, in general, an excellent correlation between serological data obtained with PPV specific monoclonal antibodies and data obtained by molecular PCR based methods. ELISA using a single or a mixture of monoclonal antibodies will remain the preferred method for universal detection and routine screening of PPV for years to come. Today, other serological methods and reagents are also recommended in the EPPO Diagnostic Protocol, increasing the number of reliable tests available for PPV detection. These developments have helped to control sharka disease in recent years. International co-operation in this field has been crucial to the improvement and validation of serological tools for PPV detection and characterization.

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Expression of Single-Chain Variable Fragments Against Structural and Non-Structural Proteins of Plum Pox Virus in Transgenic Nicotiana Benthamiana Plants to Interfere With Viral Infection

Cited by 4 publications

References 0 publications

In vivo expression and binding activity of scFv-RWAV, which recognizes the coat protein of tomato leaf curl New Delhi virus (family Geminiviridae)

In vivo expression and binding activity of scFv-RWAV, which recognizes the coat protein of tomato leaf curl New Delhi virus (family Geminiviridae)

Genetically engineered resistance to Plum pox virus infection in herbaceous and stone fruit hosts

Detection and characterization of Plum pox virus: serological methods

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