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Transgenic resistance to Citrus tristeza virus in grapefruit

  • Genetic Transformation and Hybridization
  • Published:
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Abstract

Grapefruit (Citrus paradisi) transgenic plants transformed with a variety of constructs derived from the Citrus tristeza virus (CTV) genome were tested for their resistance to the virus. Most transgenic lines were susceptible (27 lines), a few were partially resistant (6 lines) and only one line, transformed with the 3′ end of CTV was resistant. Transgene expression levels and siRNA accumulation were determined to identify whether the resistance observed was RNA-mediated. The responses were varied. At least one resistant plant from a partially resistant line showed no steady-state transgene mRNA, siRNA accumulation and no viral RNA, implicating posttranscriptional gene silencing (PTGS) as the mechanism of resistance. The most resistant line showed no transgene mRNA accumulation and promoter methylation of cytosines in all contexts, the hallmark of RNA-directed DNA methylation and transcriptional gene silencing (TGS). The variety of responses, even among clonally propagated plants, is unexplained but is not unique to citrus. The genetics of CTV, host response or other factors may be responsible for this variability.

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Abbreviations

CP:

Capsid protein

CPm:

Minor capsid protein

CTV:

Citrus tristeza virus

CU:

Cucumber mosaic virus (CMV) 5′ untranslated region

DASI-ELISA:

Double antibody sandwich indirect-enzyme linked immunosorbent assay

GUS:

β-Glucoronidase

NptII:

Neomycin phosphotransferase II

PTGS:

Post-transcriptional gene silencing

RdDM:

RNA-mediated DNA methylation

RdRp:

RNA directed RNA polymerase

RNAi:

RNA interference

SiRNA:

Small interfering RNA

TGS:

Transcriptional gene silencing

UTR:

Untranslated region

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Acknowledgments

This research was supported in part by grants from the Florida Citrus Production Research Advisory Council and the USDA-CSREES Special Grants Program—Citrus tristeza Research.

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Authors and Affiliations

  1. Horticultural Sciences Department, University of Florida, PO BOX 110690, Gainesville, FL, 32611, USA

    Vicente J. Febres & Gloria A. Moore

  2. USDA/ARS. National Clonal Germplasm Repository for Citrus and Dates, Riverside, CA, 92507, USA

    Richard F. Lee

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  1. Vicente J. Febres
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  2. Richard F. Lee
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  3. Gloria A. Moore
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Correspondence to Vicente J. Febres.

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Communicated by S. Merkle.

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Febres, V.J., Lee, R.F. & Moore, G.A. Transgenic resistance to Citrus tristeza virus in grapefruit. Plant Cell Rep 27, 93–104 (2008). https://doi.org/10.1007/s00299-007-0445-1

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  • DOI: https://doi.org/10.1007/s00299-007-0445-1

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