The developmental outcomes of P0-mediated ARGONAUTE destabilization in tomato.
The plant protein ARGONAUTE1 (AGO1) functions in multiple RNA-silencing pathways, including those of microRNAs, key regulators of growth and development. Genetic analysis of ago1 mutants with informative defects has provided valuable insights into AGO1's biological functions. Tomato encodes two AGO1 homologs (SlAGO1s), but mutants have not been described to date. To analyze SlAGO1s' involvement in development, we confirmed that both undergo decay in the presence of the Polerovirus silencing suppressor P0 and produce a transgenic responder line (OP:P0HA) that, upon transactivation, expresses P0 C-terminally fused to a hemagglutinin (HA) tag (P0HA) and destabilizes SlAGO1s at the site of expression. By crossing OP:P0HA with a battery of driver lines, constitutive as well as organ- and stage-specific SlAGO1 downregulation was induced in the F1 progeny. Activated plants exhibited various developmental phenotypes that partially overlapped with those of Arabidopsis ago1 mutants. Plants that constitutively expressed P0HA had reduced SlAGO1 levels and increased accumulation of miRNA targets, indicating compromised SlAGO1-mediated silencing. Consistent with this, they exhibited pleiotropic morphological defects and their growth was arrested post-germination. Transactivation of P0HA in young leaf and floral organ primordia dramatically modified corresponding organ morphology, including the radialization of leaflets, petals and anthers, suggesting that SlAGO1s' activities are required for normal lateral organ development and polarity. Overall, our results suggest that the OP:P0HA responder line can serve as a valuable tool to suppress SlAGO1 silencing pathways in tomato. The suppression of additional SlAGOs by P0HA and its contribution to the observed phenotypes awaits investigation.
Institute of Plant Sciences, Agricultural Research Organization, Volcani Center, PO BOX 6, 50250 Bet Dagan, Israel.
Anat
A
Hendelman
Michael
M
Kravchik
Ran
R
Stav
Moriyah
M
Zik
Nitsan
N
Lugassi
Tzahi
T
Arazi
D060565:Argonaute Proteins / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D015153:Blotting, Western; D035264:Flowers / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D015870:Gene Expression; D052679:Luteoviridae / Q000235:genetics / Q000378:metabolism; D018551:Solanum lycopersicum / Q000235:genetics* / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D010641:Phenotype; D010802:Phylogeny; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism
D060565:Argonaute Proteins; D035683:MicroRNAs; D010940:Plant Proteins; D012333:RNA, Messenger; D012097:Repressor Proteins; D014764:Viral Proteins