Plant Cell Advance Online Publication
Published on October 7, 2005; 10.1105/tpc.105.036608
Received July 29, 2005
Returned for revision August 23, 2005
Accepted September 15, 2005
Ectopic DICER-LIKE1 Expression in P1/HC-Pro Arabidopsis Rescues Phenotypic Anomalies but Not Defects in MicroRNA and Silencing Pathways
Sizolwenkosi Mlotshwa 1, Stephen E. Schauer 2, Trenton H. Smith 1, Allison C. Mallory 1, J.M. Herr Jr. 1, Braden Roth 1, Delwin S. Merchant 3, Animesh Ray 4, Lewis H. Bowman 1*, and Vicki B. Vance 1
1 Department of Biological Sciences, University of South Carolina, Columbia, South Carolina 29208
2 Department of Biological Sciences, University of South Carolina, Columbia, South Carolina 29208; Department of Biology, University of Rochester, Rochester, New York 14618
3 Department of Biology, University of Rochester, Rochester, New York 14618
4 Department of Biology, University of Rochester, Rochester, New York 14618; Keck Graduate Institute, Claremont, California 91711
* To whom correspondence should be addressed. E-mail: bowman{at}biol.sc.edu.
Expression of the viral silencing suppressor P1/HC-Pro in plants causes severe developmental anomalies accompanied by defects in both short interfering RNA (siRNA) and microRNA (miRNA) pathways. P1/HC-Pro transgenic lines fail to accumulate the siRNAs that mediate RNA silencing and are impaired in both miRNA processing and function, accumulating abnormally high levels of miRNA/miRNA* processing intermediates as well as miRNA target messages. Both miRNA and RNA silencing pathways require participation of DICER-LIKE (DCL) ribonuclease III-like enzymes. Here, we investigate the effects of overexpressing DCL1, one of four Dicers in Arabidopsis thaliana, on P1/HC-Pro-induced defects in development and small RNA metabolism. Expression of a DCL1 cDNA transgene (35S:DCL1) produced a mild gain-of-function phenotype and largely rescued dcl1 mutant phenotypes. The 35S:DCL1 plants were competent for virus-induced RNA silencing but were impaired in transgene-induced RNA silencing and in the accumulation of some miRNAs. Ectopic DCL1 largely alleviated developmental anomalies in P1/HC-Pro plants but did not correct the P1/HC-Pro-associated defects in small RNA pathways. The ability of P1/HC-Pro plants to suppress RNA silencing and the levels of miRNAs, miRNA*s, and miRNA target messages in these plants were essentially unaffected by ectopic DCL1. These data suggest that P1/HC-Pro defects in development do not result from general impairments in small RNA pathways and raise the possibility that DCL1 participates in processes in addition to miRNA biogenesis.
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