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| Status |
Public on Nov 01, 2013 |
| Title |
Global and local perturbation of the tomato microRNA pathway by a trans-activated DICER-LIKE 1 mutant |
| Organism |
Solanum lycopersicum |
| Experiment type |
Non-coding RNA profiling by high throughput sequencing
|
| Summary |
DICER-like 1 (DCL1) is a major player in micro RNA (miRNA) biogenesis and accordingly, its few known loss-of-function mutants are either lethal or display arrested seedling development. Consequently, generation of dcl1 mutants by reverse genetics and functional analysis of DCL1 in late-developing organs are challenging. Here, these challenges were resolved through the unique use of trans-activated RNA interference. Global, as well as organ-specific tomato DCL1 (SlDCL1) silencing was induced by crossing the generated responder line (OP:SlDCL1IR) with the appropriate driver line. Constitutive trans-activation knocked down SlDCL1 levels by ~95%, resulting in severe abnormalities including post-germination growth arrest accompanied by decreased miRNA and 21-nucleotide small RNA levels, but prominently elevated levels of 22- nucleotide small RNAs. The elevation in the 22- nucleotide small RNAs was correlated with specific upregulation of SlDCL2b and SlDCL2d, which are likely involved in their biogenesis. Leaf- and flower-specific OP:SlDCL1IR trans-activation inhibited blade outgrowth, induced premature bud senescence and produced pale petals, respectively, emphasizing the importance of SlDCL1-dependent small RNAs in these processes. Together, our results establish OP:SlDCL1IR as an efficient tool for analyzing processes regulated by SlDCL1-mediated gene regulation in tomato.
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| Overall design |
Examination of small RNA populations in control (35S:LhG4) and SlDCL1 silenced seedlings (35S>>SlDCL1IR) 2 biological replicates each
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| Contributor(s) |
Arazi T, Kravchik M, Sunkar R |
| Citation(s) |
24376253 |
| Submission date |
Oct 31, 2013 |
| Last update date |
May 15, 2019 |
| Contact name |
Tzahi Arazi |
| E-mail(s) |
tarazi@agri.gov.il
|
| Phone |
972-3-9683498
|
| Organization name |
ARO - Volcani Center
|
| Department |
Institute of Plant Sciences
|
| Lab |
Tzahi Arazi
|
| Street address |
Institute of Plant Sciences, ARO the Volcani Center
|
| City |
Bet Dagan |
| State/province |
ISRAEL |
| ZIP/Postal code |
50250 |
| Country |
Israel |
| |
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| Platforms (1) |
| GPL16345 |
Illumina HiSeq 2000 (Solanum lycopersicum) |
|
| Samples (4)
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|
| Relations |
| BioProject |
PRJNA225935 |
| SRA |
SRP032450 |
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