Temperature-dependent survival of turnip crinkle virus-infected Arabidopsis plants relies on an RNA silencing-based defense that requires DCL2, AGO2, and HEN1
Zhang Xiuchun1,2, Zhang Xiaofeng1,3, Singh Jasleen1, Li Dawei3, Qu Feng1*
1Department of Plant Pathology, The Ohio State University, Wooster, Ohio, USA;
2Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture, Haikou Hainan, P.R. China;
3 State Key Laboratory of Agro-Biotechnology, College of Biological Sciences, China Agricultural University, Beijing, P.R. China.
*Corresponding Author：Tel: (001)330-263-3835 E-mail: firstname.lastname@example.org
While RNA silencing is a potent antiviral defense in plants, well-adapted plant viruses are known to encode suppressors of RNA silencing (VSR) that can neutralize the effectiveness of RNA silencing. As a result, most plant genes involved in antiviral silencing were identified by using debilitated viruses lacking silencing suppression capabilities. Therefore, it remains to be resolved whether RNA silencing plays a significant part in defending plants against wild-type viruses. We report here that, at a higher plant growth temperature (26°C) that permits rigorous replication of Turnip crinkle virus (TCV) in Arabidopsis, plants containing loss-of-function mutations within the Dicer-like 2 (DCL2), Argonaute 2 (AGO2), and HEN1 RNA methyltransferase genes died of TCV infection, whereas the wild-type Col-0 plants survived to produce viable seeds. To account for the critical role of DCL2 in ensuring the survival of wild-type plants, we established that higher temperature upregulates the activity of DCL2 to produce viral 22-nucleotide (nt) small interfering RNAs (vsRNAs). We further demonstrated that DCL2-produced 22-nt vsRNAs were fully capable of silencing target genes, but that this activity was suppressed by the TCV VSR. Finally, we provide additional evidence supporting the notion that TCV VSR suppresses RNA silencing through directly interacting with AGO2. Together, these results have revealed a specialized RNA silencing pathway involving DCL2, AGO2, and HEN1 that provides the host plants with a competitive edge against adapted viruses under environmental conditions that facilitates robust virus reproduction.
Key Words: Temperature-Dependent; Turnip Crinkle Virus; RNA Silencing; DCL2; AGO2; HEN1