Plant Molecular Biology

, Volume 65, Issue 1–2, pp 163–175 | Cite as

Geminivirus-induced gene silencing of the tobacco retinoblastoma-related gene results in cell death and altered development

  • Chad V. Jordan
  • Wei Shen
  • Linda K. Hanley-Bowdoin
  • Dominique (Niki) Robertson


The retinoblastoma-related protein (RBR) is required for cell cycle control and differentiation and is expressed throughout the life of plants and animals. In this study, the tomato golden mosaic virus (TGMV) geminivirus vector was used to silence NbRBR1 in Nicotiana benthamiana by microprojectile bombardment into fully developed leaves. Similar to previous results using agroinoculation of a tobacco rattle virus silencing vector [Park et al. (Plant J 42:153, 2005)], developmental defects caused by disruptions in cell size and number were seen in new growth. Leaf midvein cross-sections showed tissue-specific differences in size, cell number, and cell morphology. While cortical cell numbers decreased, size increased to maintain overall shape. In contrast, xylem parenchyma cells increased approximately three fold but remained small. Normally straight flowers often curved up to 360° without a significant change in size. However, the most striking phenotype was cell death in mature cells after a delay of 3–4 weeks. Trypan blue staining confirmed cell death and demonstrated that cell death was absent in similarly treated leaves of wild type TGMV-inoculated plants. Quantitative RT-PCR confirmed that the mature TGMV:RBR-inoculated leaves still maintained reduced accumulation of RBR transcript at 4 weeks compared to controls. The results suggest that either inappropriate activation of the cell cycle is lethal in plants or that RBR has other functions, unrelated to the cell cycle. The results also demonstrate that continual transcription of RBR is necessary for cell survival.


DNA VIGS Retinoblastoma-related protein Cell death Geminivirus Hyperplasia Cell differentiation 



Sulfur gene


Mutant TGMV



We thank Petra Epple of the Jeff Dangl/Sarah Grant Lab in the Biology Department at the University of North Carolina at Chapel Hill for assistance with trypan blue staining. This work was supported by the North Carolina Agriculture Research Service (DR) and by NSF IBN-0235251 (LHB).


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Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Chad V. Jordan
    • 1
  • Wei Shen
    • 2
  • Linda K. Hanley-Bowdoin
    • 2
  • Dominique (Niki) Robertson
    • 1
  1. 1.Department of Plant BiologyNorth Carolina State UniversityRaleighUSA
  2. 2.Department of Molecular and Structural BiochemistryNorth Carolina State UniversityRaleighUSA

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