Spontaneous short-range silencing of a GFP transgene in Nicotiana benthamiana is possibly mediated by small quantities of siRNA that do not trigger systemic silencing

The Plant Journal : for Cell and Molecular Biology
Kriton KalantidisMartin Tabler

Abstract

A green fluorescent protein (GFP) transgene under the control of the 35S cauliflower mosaic virus (CaMV) promoter was introduced by Agrobacterium-mediated transformation into Nicotiana benthamiana to generate fourteen transgenic lines. Homozygous lines that contained one or two copies of the transgene showed great variation of GFP expression under ultraviolet (UV) light, which allowed classification into three types of transgenic plants. Plants from more than half of the transgenic lines underwent systemic RNA silencing and produced short interfering RNA (siRNA) as young seedlings, while plants of the remaining lines developed, in a spontaneous manner, defined GFP-silenced zones on their leaves, mostly in the form of circular spots that expanded to about 4-7 mm in size. In some of the latter lines, the GFP-silenced spots remained stable, but no systemic silencing occurred. Here we characterize this phenomenon, which we term spontaneous short-range silencing (SSRS). Biochemical analysis of silenced spot tissue did not reveal detectable levels of siRNA. However, agro-infiltration with the suppressor proteins P19 of cymbidium ring spot virus (CymRSV), HC-Pro of tobacco etch virus (TEV), and crosses to a P19 transgenic line, nevert...Continue Reading

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Citations

May 28, 2011·Planta·Athanasios DalakourasMichael Wassenegger
Dec 8, 2011·Plant Molecular Biology·Megumi KasaiAkira Kanazawa
Jun 29, 2010·Transgenic Research·Athanasios DalakourasKriton Kalantidis
Oct 14, 2010·BMC Plant Biology·Christos KotakisKriton Kalantidis
Apr 2, 2014·Metabolic Engineering·Isabel Mendoza-PoudereuxJuan Segura
Feb 13, 2015·Viruses·Athanasios DalakourasMichael Wassenegger
Dec 13, 2007·Biology of the Cell·Kriton KalantidisJutta Maria Helm
Jan 30, 2009·Plant Biotechnology Journal·Changjun HuangXueping Zhou
Jun 15, 2006·The Plant Journal : for Cell and Molecular Biology·Barthélémy TournierKriton Kalantidis
Aug 25, 2015·Journal of Integrative Plant Biology·Glykeria MermigkaKriton Kalantidis
Sep 16, 2014·Molecular Plant Pathology·Basavaprabhu L Patil, Claude M Fauquet
Jul 28, 2011·Plant Signaling & Behavior·Christos KotakisKriton Kalantidis
Aug 18, 2016·The Plant Journal : for Cell and Molecular Biology·Glykeria MermigkaKriton Kalantidis
Jul 9, 2020·Science Advances·Gozde S DemirerMarkita P Landry
Mar 20, 2021·Frontiers in Plant Science·Neeti Sanan-MishraSunil K Mukherjee
May 14, 2021·Molecular Plant-microbe Interactions : MPMI·Giulia TarquiniGiuseppe Firrao

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