PT  - JOURNAL ARTICLE
AU  - Zhenxing Wang
AU  - Nicolas Butel
AU  - Juan Santos-González
AU  - Filipe Borges
AU  - Jun Yi
AU  - Robert A Martienssen
AU  - German Martinez
AU  - Claudia Köhler
TI  - Functional role of Polymerase IV during pollen development in <em>Capsella</em>
AID  - 10.1101/863522
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 863522
4099  - http://biorxiv.org/content/early/2019/12/03/863522.short
4100  - http://biorxiv.org/content/early/2019/12/03/863522.full
AB  - In Arabidopsis thaliana, the DNA-dependent RNA polymerase IV (Pol IV) is required for the formation of transposable element (TE)-derived small RNA (sRNA) transcripts. These transcripts are processed by DICER-LIKE 3 into 24-nt small interfering RNAs (siRNAs) that guide RNA-dependent DNA methylation. In the pollen grain, Pol IV is also required for the accumulation of 21/22-nt epigenetically-activated siRNAs (easiRNAs) that likely silence TEs by post-transcriptional mechanisms. Despite this proposed functional role, loss of Pol IV function in Arabidopsis does not cause a discernable pollen defect. Here, we show that loss of NRPD1, encoding the largest subunit of Pol IV in the Brassicaceae Capsella rubella, causes post-meiotic arrest of pollen development at the microspore stage. As in Arabidopsis, all TE-derived siRNAs were depleted in Capsella nrpd1 microspores. In wild-type background, we found that the same TEs produced 21/22-nt and 24-nt siRNAs, leading us to propose that Pol IV is generating the direct precursors for 21-24-nt siRNAs, which are targeted by different DICERs. Arrest of Capsella nrpd1 microspores was accompanied by deregulation of genes targeted by Pol IV-dependent siRNAs. The distance of TEs to genes was much closer in Capsella rubella compared to Arabidopsis thaliana, providing a possible explanation for the essential role of Pol IV for pollen development in Capsella. Our study in Capsella uncovers a functional requirement of Pol IV in microspores, emphasizing the relevance of investigating different plant models.One-sentence summary Loss of Polymerase IV function in Capsella rubella causes microspore arrest, revealing an important functional role of Polymerase IV during pollen development.The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantcell.org) is: Claudia Kohler (Claudia.Kohler{at}slu.se)